Sample records for nested sampling algorithm

  1. New prior sampling methods for nested sampling - Development and testing

    NASA Astrophysics Data System (ADS)

    Stokes, Barrie; Tuyl, Frank; Hudson, Irene

    2017-06-01

    Nested Sampling is a powerful algorithm for fitting models to data in the Bayesian setting, introduced by Skilling [1]. The nested sampling algorithm proceeds by carrying out a series of compressive steps, involving successively nested iso-likelihood boundaries, starting with the full prior distribution of the problem parameters. The "central problem" of nested sampling is to draw at each step a sample from the prior distribution whose likelihood is greater than the current likelihood threshold, i.e., a sample falling inside the current likelihood-restricted region. For both flat and informative priors this ultimately requires uniform sampling restricted to the likelihood-restricted region. We present two new methods of carrying out this sampling step, and illustrate their use with the lighthouse problem [2], a bivariate likelihood used by Gregory [3] and a trivariate Gaussian mixture likelihood. All the algorithm development and testing reported here has been done with Mathematica® [4].

  2. Construction of nested maximin designs based on successive local enumeration and modified novel global harmony search algorithm

    NASA Astrophysics Data System (ADS)

    Yi, Jin; Li, Xinyu; Xiao, Mi; Xu, Junnan; Zhang, Lin

    2017-01-01

    Engineering design often involves different types of simulation, which results in expensive computational costs. Variable fidelity approximation-based design optimization approaches can realize effective simulation and efficiency optimization of the design space using approximation models with different levels of fidelity and have been widely used in different fields. As the foundations of variable fidelity approximation models, the selection of sample points of variable-fidelity approximation, called nested designs, is essential. In this article a novel nested maximin Latin hypercube design is constructed based on successive local enumeration and a modified novel global harmony search algorithm. In the proposed nested designs, successive local enumeration is employed to select sample points for a low-fidelity model, whereas the modified novel global harmony search algorithm is employed to select sample points for a high-fidelity model. A comparative study with multiple criteria and an engineering application are employed to verify the efficiency of the proposed nested designs approach.

  3. MultiNest: Efficient and Robust Bayesian Inference

    NASA Astrophysics Data System (ADS)

    Feroz, F.; Hobson, M. P.; Bridges, M.

    2011-09-01

    We present further development and the first public release of our multimodal nested sampling algorithm, called MultiNest. This Bayesian inference tool calculates the evidence, with an associated error estimate, and produces posterior samples from distributions that may contain multiple modes and pronounced (curving) degeneracies in high dimensions. The developments presented here lead to further substantial improvements in sampling efficiency and robustness, as compared to the original algorithm presented in Feroz & Hobson (2008), which itself significantly outperformed existing MCMC techniques in a wide range of astrophysical inference problems. The accuracy and economy of the MultiNest algorithm is demonstrated by application to two toy problems and to a cosmological inference problem focusing on the extension of the vanilla LambdaCDM model to include spatial curvature and a varying equation of state for dark energy. The MultiNest software is fully parallelized using MPI and includes an interface to CosmoMC. It will also be released as part of the SuperBayeS package, for the analysis of supersymmetric theories of particle physics, at this http URL.

  4. Exploring the Energy Landscapes of Protein Folding Simulations with Bayesian Computation

    PubMed Central

    Burkoff, Nikolas S.; Várnai, Csilla; Wells, Stephen A.; Wild, David L.

    2012-01-01

    Nested sampling is a Bayesian sampling technique developed to explore probability distributions localized in an exponentially small area of the parameter space. The algorithm provides both posterior samples and an estimate of the evidence (marginal likelihood) of the model. The nested sampling algorithm also provides an efficient way to calculate free energies and the expectation value of thermodynamic observables at any temperature, through a simple post processing of the output. Previous applications of the algorithm have yielded large efficiency gains over other sampling techniques, including parallel tempering. In this article, we describe a parallel implementation of the nested sampling algorithm and its application to the problem of protein folding in a Gō-like force field of empirical potentials that were designed to stabilize secondary structure elements in room-temperature simulations. We demonstrate the method by conducting folding simulations on a number of small proteins that are commonly used for testing protein-folding procedures. A topological analysis of the posterior samples is performed to produce energy landscape charts, which give a high-level description of the potential energy surface for the protein folding simulations. These charts provide qualitative insights into both the folding process and the nature of the model and force field used. PMID:22385859

  5. Exploring the energy landscapes of protein folding simulations with Bayesian computation.

    PubMed

    Burkoff, Nikolas S; Várnai, Csilla; Wells, Stephen A; Wild, David L

    2012-02-22

    Nested sampling is a Bayesian sampling technique developed to explore probability distributions localized in an exponentially small area of the parameter space. The algorithm provides both posterior samples and an estimate of the evidence (marginal likelihood) of the model. The nested sampling algorithm also provides an efficient way to calculate free energies and the expectation value of thermodynamic observables at any temperature, through a simple post processing of the output. Previous applications of the algorithm have yielded large efficiency gains over other sampling techniques, including parallel tempering. In this article, we describe a parallel implementation of the nested sampling algorithm and its application to the problem of protein folding in a Gō-like force field of empirical potentials that were designed to stabilize secondary structure elements in room-temperature simulations. We demonstrate the method by conducting folding simulations on a number of small proteins that are commonly used for testing protein-folding procedures. A topological analysis of the posterior samples is performed to produce energy landscape charts, which give a high-level description of the potential energy surface for the protein folding simulations. These charts provide qualitative insights into both the folding process and the nature of the model and force field used. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. Constant-pressure nested sampling with atomistic dynamics

    NASA Astrophysics Data System (ADS)

    Baldock, Robert J. N.; Bernstein, Noam; Salerno, K. Michael; Pártay, Lívia B.; Csányi, Gábor

    2017-10-01

    The nested sampling algorithm has been shown to be a general method for calculating the pressure-temperature-composition phase diagrams of materials. While the previous implementation used single-particle Monte Carlo moves, these are inefficient for condensed systems with general interactions where single-particle moves cannot be evaluated faster than the energy of the whole system. Here we enhance the method by using all-particle moves: either Galilean Monte Carlo or the total enthalpy Hamiltonian Monte Carlo algorithm, introduced in this paper. We show that these algorithms enable the determination of phase transition temperatures with equivalent accuracy to the previous method at 1 /N of the cost for an N -particle system with general interactions, or at equal cost when single-particle moves can be done in 1 /N of the cost of a full N -particle energy evaluation. We demonstrate this speed-up for the freezing and condensation transitions of the Lennard-Jones system and show the utility of the algorithms by calculating the order-disorder phase transition of a binary Lennard-Jones model alloy, the eutectic of copper-gold, the density anomaly of water, and the condensation and solidification of bead-spring polymers. The nested sampling method with all three algorithms is implemented in the pymatnest software.

  7. An Improved Nested Sampling Algorithm for Model Selection and Assessment

    NASA Astrophysics Data System (ADS)

    Zeng, X.; Ye, M.; Wu, J.; WANG, D.

    2017-12-01

    Multimodel strategy is a general approach for treating model structure uncertainty in recent researches. The unknown groundwater system is represented by several plausible conceptual models. Each alternative conceptual model is attached with a weight which represents the possibility of this model. In Bayesian framework, the posterior model weight is computed as the product of model prior weight and marginal likelihood (or termed as model evidence). As a result, estimating marginal likelihoods is crucial for reliable model selection and assessment in multimodel analysis. Nested sampling estimator (NSE) is a new proposed algorithm for marginal likelihood estimation. The implementation of NSE comprises searching the parameters' space from low likelihood area to high likelihood area gradually, and this evolution is finished iteratively via local sampling procedure. Thus, the efficiency of NSE is dominated by the strength of local sampling procedure. Currently, Metropolis-Hasting (M-H) algorithm and its variants are often used for local sampling in NSE. However, M-H is not an efficient sampling algorithm for high-dimensional or complex likelihood function. For improving the performance of NSE, it could be feasible to integrate more efficient and elaborated sampling algorithm - DREAMzs into the local sampling. In addition, in order to overcome the computation burden problem of large quantity of repeating model executions in marginal likelihood estimation, an adaptive sparse grid stochastic collocation method is used to build the surrogates for original groundwater model.

  8. Entropy-Based Search Algorithm for Experimental Design

    NASA Astrophysics Data System (ADS)

    Malakar, N. K.; Knuth, K. H.

    2011-03-01

    The scientific method relies on the iterated processes of inference and inquiry. The inference phase consists of selecting the most probable models based on the available data; whereas the inquiry phase consists of using what is known about the models to select the most relevant experiment. Optimizing inquiry involves searching the parameterized space of experiments to select the experiment that promises, on average, to be maximally informative. In the case where it is important to learn about each of the model parameters, the relevance of an experiment is quantified by Shannon entropy of the distribution of experimental outcomes predicted by a probable set of models. If the set of potential experiments is described by many parameters, we must search this high-dimensional entropy space. Brute force search methods will be slow and computationally expensive. We present an entropy-based search algorithm, called nested entropy sampling, to select the most informative experiment for efficient experimental design. This algorithm is inspired by Skilling's nested sampling algorithm used in inference and borrows the concept of a rising threshold while a set of experiment samples are maintained. We demonstrate that this algorithm not only selects highly relevant experiments, but also is more efficient than brute force search. Such entropic search techniques promise to greatly benefit autonomous experimental design.

  9. Improving and Evaluating Nested Sampling Algorithm for Marginal Likelihood Estimation

    NASA Astrophysics Data System (ADS)

    Ye, M.; Zeng, X.; Wu, J.; Wang, D.; Liu, J.

    2016-12-01

    With the growing impacts of climate change and human activities on the cycle of water resources, an increasing number of researches focus on the quantification of modeling uncertainty. Bayesian model averaging (BMA) provides a popular framework for quantifying conceptual model and parameter uncertainty. The ensemble prediction is generated by combining each plausible model's prediction, and each model is attached with a model weight which is determined by model's prior weight and marginal likelihood. Thus, the estimation of model's marginal likelihood is crucial for reliable and accurate BMA prediction. Nested sampling estimator (NSE) is a new proposed method for marginal likelihood estimation. The process of NSE is accomplished by searching the parameters' space from low likelihood area to high likelihood area gradually, and this evolution is finished iteratively via local sampling procedure. Thus, the efficiency of NSE is dominated by the strength of local sampling procedure. Currently, Metropolis-Hasting (M-H) algorithm is often used for local sampling. However, M-H is not an efficient sampling algorithm for high-dimensional or complicated parameter space. For improving the efficiency of NSE, it could be ideal to incorporate the robust and efficient sampling algorithm - DREAMzs into the local sampling of NSE. The comparison results demonstrated that the improved NSE could improve the efficiency of marginal likelihood estimation significantly. However, both improved and original NSEs suffer from heavy instability. In addition, the heavy computation cost of huge number of model executions is overcome by using an adaptive sparse grid surrogates.

  10. Comparison of statistical sampling methods with ScannerBit, the GAMBIT scanning module

    NASA Astrophysics Data System (ADS)

    Martinez, Gregory D.; McKay, James; Farmer, Ben; Scott, Pat; Roebber, Elinore; Putze, Antje; Conrad, Jan

    2017-11-01

    We introduce ScannerBit, the statistics and sampling module of the public, open-source global fitting framework GAMBIT. ScannerBit provides a standardised interface to different sampling algorithms, enabling the use and comparison of multiple computational methods for inferring profile likelihoods, Bayesian posteriors, and other statistical quantities. The current version offers random, grid, raster, nested sampling, differential evolution, Markov Chain Monte Carlo (MCMC) and ensemble Monte Carlo samplers. We also announce the release of a new standalone differential evolution sampler, Diver, and describe its design, usage and interface to ScannerBit. We subject Diver and three other samplers (the nested sampler MultiNest, the MCMC GreAT, and the native ScannerBit implementation of the ensemble Monte Carlo algorithm T-Walk) to a battery of statistical tests. For this we use a realistic physical likelihood function, based on the scalar singlet model of dark matter. We examine the performance of each sampler as a function of its adjustable settings, and the dimensionality of the sampling problem. We evaluate performance on four metrics: optimality of the best fit found, completeness in exploring the best-fit region, number of likelihood evaluations, and total runtime. For Bayesian posterior estimation at high resolution, T-Walk provides the most accurate and timely mapping of the full parameter space. For profile likelihood analysis in less than about ten dimensions, we find that Diver and MultiNest score similarly in terms of best fit and speed, outperforming GreAT and T-Walk; in ten or more dimensions, Diver substantially outperforms the other three samplers on all metrics.

  11. Maximum likelihood estimation for Cox's regression model under nested case-control sampling.

    PubMed

    Scheike, Thomas H; Juul, Anders

    2004-04-01

    Nested case-control sampling is designed to reduce the costs of large cohort studies. It is important to estimate the parameters of interest as efficiently as possible. We present a new maximum likelihood estimator (MLE) for nested case-control sampling in the context of Cox's proportional hazards model. The MLE is computed by the EM-algorithm, which is easy to implement in the proportional hazards setting. Standard errors are estimated by a numerical profile likelihood approach based on EM aided differentiation. The work was motivated by a nested case-control study that hypothesized that insulin-like growth factor I was associated with ischemic heart disease. The study was based on a population of 3784 Danes and 231 cases of ischemic heart disease where controls were matched on age and gender. We illustrate the use of the MLE for these data and show how the maximum likelihood framework can be used to obtain information additional to the relative risk estimates of covariates.

  12. Exploiting molecular dynamics in Nested Sampling simulations of small peptides

    NASA Astrophysics Data System (ADS)

    Burkoff, Nikolas S.; Baldock, Robert J. N.; Várnai, Csilla; Wild, David L.; Csányi, Gábor

    2016-04-01

    Nested Sampling (NS) is a parameter space sampling algorithm which can be used for sampling the equilibrium thermodynamics of atomistic systems. NS has previously been used to explore the potential energy surface of a coarse-grained protein model and has significantly outperformed parallel tempering when calculating heat capacity curves of Lennard-Jones clusters. The original NS algorithm uses Monte Carlo (MC) moves; however, a variant, Galilean NS, has recently been introduced which allows NS to be incorporated into a molecular dynamics framework, so NS can be used for systems which lack efficient prescribed MC moves. In this work we demonstrate the applicability of Galilean NS to atomistic systems. We present an implementation of Galilean NS using the Amber molecular dynamics package and demonstrate its viability by sampling alanine dipeptide, both in vacuo and implicit solvent. Unlike previous studies of this system, we present the heat capacity curves of alanine dipeptide, whose calculation provides a stringent test for sampling algorithms. We also compare our results with those calculated using replica exchange molecular dynamics (REMD) and find good agreement. We show the computational effort required for accurate heat capacity estimation for small peptides. We also calculate the alanine dipeptide Ramachandran free energy surface for a range of temperatures and use it to compare the results using the latest Amber force field with previous theoretical and experimental results.

  13. Comparison of sampling techniques for Bayesian parameter estimation

    NASA Astrophysics Data System (ADS)

    Allison, Rupert; Dunkley, Joanna

    2014-02-01

    The posterior probability distribution for a set of model parameters encodes all that the data have to tell us in the context of a given model; it is the fundamental quantity for Bayesian parameter estimation. In order to infer the posterior probability distribution we have to decide how to explore parameter space. Here we compare three prescriptions for how parameter space is navigated, discussing their relative merits. We consider Metropolis-Hasting sampling, nested sampling and affine-invariant ensemble Markov chain Monte Carlo (MCMC) sampling. We focus on their performance on toy-model Gaussian likelihoods and on a real-world cosmological data set. We outline the sampling algorithms themselves and elaborate on performance diagnostics such as convergence time, scope for parallelization, dimensional scaling, requisite tunings and suitability for non-Gaussian distributions. We find that nested sampling delivers high-fidelity estimates for posterior statistics at low computational cost, and should be adopted in favour of Metropolis-Hastings in many cases. Affine-invariant MCMC is competitive when computing clusters can be utilized for massive parallelization. Affine-invariant MCMC and existing extensions to nested sampling naturally probe multimodal and curving distributions.

  14. Balancing a U-Shaped Assembly Line by Applying Nested Partitions Method

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bhagwat, Nikhil V.

    2005-01-01

    In this study, we applied the Nested Partitions method to a U-line balancing problem and conducted experiments to evaluate the application. From the results, it is quite evident that the Nested Partitions method provided near optimal solutions (optimal in some cases). Besides, the execution time is quite short as compared to the Branch and Bound algorithm. However, for larger data sets, the algorithm took significantly longer times for execution. One of the reasons could be the way in which the random samples are generated. In the present study, a random sample is a solution in itself which requires assignment ofmore » tasks to various stations. The time taken to assign tasks to stations is directly proportional to the number of tasks. Thus, if the number of tasks increases, the time taken to generate random samples for the different regions also increases. The performance index for the Nested Partitions method in the present study was the number of stations in the random solutions (samples) generated. The total idle time for the samples can be used as another performance index. ULINO method is known to have used a combination of bounds to come up with good solutions. This approach of combining different performance indices can be used to evaluate the random samples and obtain even better solutions. Here, we used deterministic time values for the tasks. In industries where majority of tasks are performed manually, the stochastic version of the problem could be of vital importance. Experimenting with different objective functions (No. of stations was used in this study) could be of some significance to some industries where in the cost associated with creation of a new station is not the same. For such industries, the results obtained by using the present approach will not be of much value. Labor costs, task incompletion costs or a combination of those can be effectively used as alternate objective functions.« less

  15. Nested Conjugate Gradient Algorithm with Nested Preconditioning for Non-linear Image Restoration.

    PubMed

    Skariah, Deepak G; Arigovindan, Muthuvel

    2017-06-19

    We develop a novel optimization algorithm, which we call Nested Non-Linear Conjugate Gradient algorithm (NNCG), for image restoration based on quadratic data fitting and smooth non-quadratic regularization. The algorithm is constructed as a nesting of two conjugate gradient (CG) iterations. The outer iteration is constructed as a preconditioned non-linear CG algorithm; the preconditioning is performed by the inner CG iteration that is linear. The inner CG iteration, which performs preconditioning for outer CG iteration, itself is accelerated by an another FFT based non-iterative preconditioner. We prove that the method converges to a stationary point for both convex and non-convex regularization functionals. We demonstrate experimentally that proposed method outperforms the well-known majorization-minimization method used for convex regularization, and a non-convex inertial-proximal method for non-convex regularization functional.

  16. MontePython 3: Parameter inference code for cosmology

    NASA Astrophysics Data System (ADS)

    Brinckmann, Thejs; Lesgourgues, Julien; Audren, Benjamin; Benabed, Karim; Prunet, Simon

    2018-05-01

    MontePython 3 provides numerous ways to explore parameter space using Monte Carlo Markov Chain (MCMC) sampling, including Metropolis-Hastings, Nested Sampling, Cosmo Hammer, and a Fisher sampling method. This improved version of the Monte Python (ascl:1307.002) parameter inference code for cosmology offers new ingredients that improve the performance of Metropolis-Hastings sampling, speeding up convergence and offering significant time improvement in difficult runs. Additional likelihoods and plotting options are available, as are post-processing algorithms such as Importance Sampling and Adding Derived Parameter.

  17. Hybrid Nested Partitions and Math Programming Framework for Large-scale Combinatorial Optimization

    DTIC Science & Technology

    2010-03-31

    optimization problems: 1) exact algorithms and 2) metaheuristic algorithms . This project will integrate concepts from these two technologies to develop...optimal solutions within an acceptable amount of computation time, and 2) metaheuristic algorithms such as genetic algorithms , tabu search, and the...integer programming decomposition approaches, such as Dantzig Wolfe decomposition and Lagrangian relaxation, and metaheuristics such as the Nested

  18. Optimal reservoir operation policies using novel nested algorithms

    NASA Astrophysics Data System (ADS)

    Delipetrev, Blagoj; Jonoski, Andreja; Solomatine, Dimitri

    2015-04-01

    Historically, the two most widely practiced methods for optimal reservoir operation have been dynamic programming (DP) and stochastic dynamic programming (SDP). These two methods suffer from the so called "dual curse" which prevents them to be used in reasonably complex water systems. The first one is the "curse of dimensionality" that denotes an exponential growth of the computational complexity with the state - decision space dimension. The second one is the "curse of modelling" that requires an explicit model of each component of the water system to anticipate the effect of each system's transition. We address the problem of optimal reservoir operation concerning multiple objectives that are related to 1) reservoir releases to satisfy several downstream users competing for water with dynamically varying demands, 2) deviations from the target minimum and maximum reservoir water levels and 3) hydropower production that is a combination of the reservoir water level and the reservoir releases. Addressing such a problem with classical methods (DP and SDP) requires a reasonably high level of discretization of the reservoir storage volume, which in combination with the required releases discretization for meeting the demands of downstream users leads to computationally expensive formulations and causes the curse of dimensionality. We present a novel approach, named "nested" that is implemented in DP, SDP and reinforcement learning (RL) and correspondingly three new algorithms are developed named nested DP (nDP), nested SDP (nSDP) and nested RL (nRL). The nested algorithms are composed from two algorithms: 1) DP, SDP or RL and 2) nested optimization algorithm. Depending on the way we formulate the objective function related to deficits in the allocation problem in the nested optimization, two methods are implemented: 1) Simplex for linear allocation problems, and 2) quadratic Knapsack method in the case of nonlinear problems. The novel idea is to include the nested optimization algorithm into the state transition that lowers the starting problem dimension and alleviates the curse of dimensionality. The algorithms can solve multi-objective optimization problems, without significantly increasing the complexity and the computational expenses. The algorithms can handle dense and irregular variable discretization, and are coded in Java as prototype applications. The three algorithms were tested at the multipurpose reservoir Knezevo of the Zletovica hydro-system located in the Republic of Macedonia, with eight objectives, including urban water supply, agriculture, ensuring ecological flow, and generation of hydropower. Because the Zletovica hydro-system is relatively complex, the novel algorithms were pushed to their limits, demonstrating their capabilities and limitations. The nSDP and nRL derived/learned the optimal reservoir policy using 45 (1951-1995) years historical data. The nSDP and nRL optimal reservoir policy was tested on 10 (1995-2005) years historical data, and compared with nDP optimal reservoir operation in the same period. The nested algorithms and optimal reservoir operation results are analysed and explained.

  19. Reconstruction of calmodulin single-molecule FRET states, dye interactions, and CaMKII peptide binding by MultiNest and classic maximum entropy

    NASA Astrophysics Data System (ADS)

    DeVore, Matthew S.; Gull, Stephen F.; Johnson, Carey K.

    2013-08-01

    We analyzed single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data.

  20. Reconstruction of Calmodulin Single-Molecule FRET States, Dye-Interactions, and CaMKII Peptide Binding by MultiNest and Classic Maximum Entropy

    PubMed Central

    DeVore, Matthew S.; Gull, Stephen F.; Johnson, Carey K.

    2013-01-01

    We analyze single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data. PMID:24223465

  1. Reconstruction of Calmodulin Single-Molecule FRET States, Dye-Interactions, and CaMKII Peptide Binding by MultiNest and Classic Maximum Entropy.

    PubMed

    Devore, Matthew S; Gull, Stephen F; Johnson, Carey K

    2013-08-30

    We analyze single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca 2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data.

  2. Darwinian Model Building

    NASA Astrophysics Data System (ADS)

    Kester, Do; Bontekoe, Romke

    2011-03-01

    We present a way to generate heuristic mathematical models based on the Darwinian principles of variation and selection in a pool of individuals over many generations. Each individual has a genotype (the hereditary properties) and a phenotype (the expression of these properties in the environment). Variation is achieved by cross-over and mutation operations on the genotype which consists in the present case of a single chromosome. The genotypes `live' in the environment of the data. Nested Sampling is used to optimize the free parameters of the models given the data, thus giving rise to the phenotypes. Selection is based on the phenotypes. The evidences which naturally follow from the Nested Sampling Algorithm are used in a second level of Nested Sampling to find increasingly better models. The data in this paper originate from the Leiden Cytology and Pathology Laboratory (LCPL), which screens pap smears for cervical cancer. We have data for 1750 women who on average underwent 5 tests each. The data on individual women are treated as a small time series. We will try to estimate the next value of the prime cancer indicator from previous tests of the same woman.

  3. Dreaming of Atmospheres

    NASA Astrophysics Data System (ADS)

    Waldmann, Ingo

    2016-10-01

    Radiative transfer retrievals have become the standard in modelling of exoplanetary transmission and emission spectra. Analysing currently available observations of exoplanetary atmospheres often invoke large and correlated parameter spaces that can be difficult to map or constrain.To address these issues, we have developed the Tau-REx (tau-retrieval of exoplanets) retrieval and the RobERt spectral recognition algorithms. Tau-REx is a bayesian atmospheric retrieval framework using Nested Sampling and cluster computing to fully map these large correlated parameter spaces. Nonetheless, data volumes can become prohibitively large and we must often select a subset of potential molecular/atomic absorbers in an atmosphere.In the era of open-source, automated and self-sufficient retrieval algorithms, such manual input should be avoided. User dependent input could, in worst case scenarios, lead to incomplete models and biases in the retrieval. The RobERt algorithm is build to address these issues. RobERt is a deep belief neural (DBN) networks trained to accurately recognise molecular signatures for a wide range of planets, atmospheric thermal profiles and compositions. Using these deep neural networks, we work towards retrieval algorithms that themselves understand the nature of the observed spectra, are able to learn from current and past data and make sensible qualitative preselections of atmospheric opacities to be used for the quantitative stage of the retrieval process.In this talk I will discuss how neural networks and Bayesian Nested Sampling can be used to solve highly degenerate spectral retrieval problems and what 'dreaming' neural networks can tell us about atmospheric characteristics.

  4. Limiting Index Sort: A New Non-Dominated Sorting Algorithm and its Comparison to the State-of-the-Art

    DTIC Science & Technology

    2010-05-01

    Skyline Algorithms 2.2.1 Block-Nested Loops A simple way to find the skyline is to use the block-nested loops ( BNL ) algorithm [3], which is the algorithm...by an NDS member are discarded. After every individual has been compared with the NDS, the NDS is the dataset’s skyline. In the best case for BNL ...SFS) algorithm [4] is a variation on BNL that first introduces the idea of initially ordering the individuals by a monotonically increasing scoring

  5. Darwinian Model Building

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kester, Do; Bontekoe, Romke

    2011-03-14

    We present a way to generate heuristic mathematical models based on the Darwinian principles of variation and selection in a pool of individuals over many generations. Each individual has a genotype (the hereditary properties) and a phenotype (the expression of these properties in the environment). Variation is achieved by cross-over and mutation operations on the genotype which consists in the present case of a single chromosome. The genotypes 'live' in the environment of the data. Nested Sampling is used to optimize the free parameters of the models given the data, thus giving rise to the phenotypes. Selection is based onmore » the phenotypes.The evidences which naturally follow from the Nested Sampling Algorithm are used in a second level of Nested Sampling to find increasingly better models.The data in this paper originate from the Leiden Cytology and Pathology Laboratory (LCPL), which screens pap smears for cervical cancer. We have data for 1750 women who on average underwent 5 tests each. The data on individual women are treated as a small time series. We will try to estimate the next value of the prime cancer indicator from previous tests of the same woman.« less

  6. Acceleration of image-based resolution modelling reconstruction using an expectation maximization nested algorithm.

    PubMed

    Angelis, G I; Reader, A J; Markiewicz, P J; Kotasidis, F A; Lionheart, W R; Matthews, J C

    2013-08-07

    Recent studies have demonstrated the benefits of a resolution model within iterative reconstruction algorithms in an attempt to account for effects that degrade the spatial resolution of the reconstructed images. However, these algorithms suffer from slower convergence rates, compared to algorithms where no resolution model is used, due to the additional need to solve an image deconvolution problem. In this paper, a recently proposed algorithm, which decouples the tomographic and image deconvolution problems within an image-based expectation maximization (EM) framework, was evaluated. This separation is convenient, because more computational effort can be placed on the image deconvolution problem and therefore accelerate convergence. Since the computational cost of solving the image deconvolution problem is relatively small, multiple image-based EM iterations do not significantly increase the overall reconstruction time. The proposed algorithm was evaluated using 2D simulations, as well as measured 3D data acquired on the high-resolution research tomograph. Results showed that bias reduction can be accelerated by interleaving multiple iterations of the image-based EM algorithm solving the resolution model problem, with a single EM iteration solving the tomographic problem. Significant improvements were observed particularly for voxels that were located on the boundaries between regions of high contrast within the object being imaged and for small regions of interest, where resolution recovery is usually more challenging. Minor differences were observed using the proposed nested algorithm, compared to the single iteration normally performed, when an optimal number of iterations are performed for each algorithm. However, using the proposed nested approach convergence is significantly accelerated enabling reconstruction using far fewer tomographic iterations (up to 70% fewer iterations for small regions). Nevertheless, the optimal number of nested image-based EM iterations is hard to be defined and it should be selected according to the given application.

  7. Managing Algorithmic Skeleton Nesting Requirements in Realistic Image Processing Applications: The Case of the SKiPPER-II Parallel Programming Environment's Operating Model

    NASA Astrophysics Data System (ADS)

    Coudarcher, Rémi; Duculty, Florent; Serot, Jocelyn; Jurie, Frédéric; Derutin, Jean-Pierre; Dhome, Michel

    2005-12-01

    SKiPPER is a SKeleton-based Parallel Programming EnviRonment being developed since 1996 and running at LASMEA Laboratory, the Blaise-Pascal University, France. The main goal of the project was to demonstrate the applicability of skeleton-based parallel programming techniques to the fast prototyping of reactive vision applications. This paper deals with the special features embedded in the latest version of the project: algorithmic skeleton nesting capabilities and a fully dynamic operating model. Throughout the case study of a complete and realistic image processing application, in which we have pointed out the requirement for skeleton nesting, we are presenting the operating model of this feature. The work described here is one of the few reported experiments showing the application of skeleton nesting facilities for the parallelisation of a realistic application, especially in the area of image processing. The image processing application we have chosen is a 3D face-tracking algorithm from appearance.

  8. Hybridization of Strength Pareto Multiobjective Optimization with Modified Cuckoo Search Algorithm for Rectangular Array

    NASA Astrophysics Data System (ADS)

    Abdul Rani, Khairul Najmy; Abdulmalek, Mohamedfareq; A. Rahim, Hasliza; Siew Chin, Neoh; Abd Wahab, Alawiyah

    2017-04-01

    This research proposes the various versions of modified cuckoo search (MCS) metaheuristic algorithm deploying the strength Pareto evolutionary algorithm (SPEA) multiobjective (MO) optimization technique in rectangular array geometry synthesis. Precisely, the MCS algorithm is proposed by incorporating the Roulette wheel selection operator to choose the initial host nests (individuals) that give better results, adaptive inertia weight to control the positions exploration of the potential best host nests (solutions), and dynamic discovery rate to manage the fraction probability of finding the best host nests in 3-dimensional search space. In addition, the MCS algorithm is hybridized with the particle swarm optimization (PSO) and hill climbing (HC) stochastic techniques along with the standard strength Pareto evolutionary algorithm (SPEA) forming the MCSPSOSPEA and MCSHCSPEA, respectively. All the proposed MCS-based algorithms are examined to perform MO optimization on Zitzler-Deb-Thiele’s (ZDT’s) test functions. Pareto optimum trade-offs are done to generate a set of three non-dominated solutions, which are locations, excitation amplitudes, and excitation phases of array elements, respectively. Overall, simulations demonstrates that the proposed MCSPSOSPEA outperforms other compatible competitors, in gaining a high antenna directivity, small half-power beamwidth (HPBW), low average side lobe level (SLL) suppression, and/or significant predefined nulls mitigation, simultaneously.

  9. Better cancer biomarker discovery through better study design.

    PubMed

    Rundle, Andrew; Ahsan, Habibul; Vineis, Paolo

    2012-12-01

    High-throughput laboratory technologies coupled with sophisticated bioinformatics algorithms have tremendous potential for discovering novel biomarkers, or profiles of biomarkers, that could serve as predictors of disease risk, response to treatment or prognosis. We discuss methodological issues in wedding high-throughput approaches for biomarker discovery with the case-control study designs typically used in biomarker discovery studies, especially focusing on nested case-control designs. We review principles for nested case-control study design in relation to biomarker discovery studies and describe how the efficiency of biomarker discovery can be effected by study design choices. We develop a simulated prostate cancer cohort data set and a series of biomarker discovery case-control studies nested within the cohort to illustrate how study design choices can influence biomarker discovery process. Common elements of nested case-control design, incidence density sampling and matching of controls to cases are not typically factored correctly into biomarker discovery analyses, inducing bias in the discovery process. We illustrate how incidence density sampling and matching of controls to cases reduce the apparent specificity of truly valid biomarkers 'discovered' in a nested case-control study. We also propose and demonstrate a new case-control matching protocol, we call 'antimatching', that improves the efficiency of biomarker discovery studies. For a valid, but as yet undiscovered, biomarker(s) disjunctions between correctly designed epidemiologic studies and the practice of biomarker discovery reduce the likelihood that true biomarker(s) will be discovered and increases the false-positive discovery rate. © 2012 The Authors. European Journal of Clinical Investigation © 2012 Stichting European Society for Clinical Investigation Journal Foundation.

  10. Monitoring bald eagles using lists of nests: Response to Watts and Duerr

    USGS Publications Warehouse

    Sauer, John R.; Otto, Mark C.; Kendall, William L.; Zimmerman, Guthrie S.

    2011-01-01

    The post-delisting monitoring plan for bald eagles (Haliaeetus leucocephalus) roposed use of a dual-frame sample design, in which sampling of known nest sites in combination with additional area-based sampling is used to estimate total number of nesting bald eagle pairs. Watts and Duerr (2010) used data from repeated observations of bald eagle nests in Virginia, USA to estimate a nest turnover rate and used this rate to simulate decline in number of occupied nests in list nests over time. Results of Watts and Duerr suggest that, given the rates of loss of nests from the list of known nest sites in Virginia, the list information will be of little value to sampling unless lists are constantly updated. Those authors criticize the plan for not placing sufficient emphasis on updating and maintaining lists of bald eagle nests. Watts and Duerr's metric of turnover rate does not distinguish detectability or temporary nonuse of nests from permanent loss of nests and likely overestimates turnover rate. We describe a multi-state capture–recapture model that allows appropriate estimation of rates of loss of nests, and we use the model to estimate rates of loss from a sample of nests from Maine, USA. The post-delisting monitoring plan addresses the need to maintain and update the lists of nests, and we show that dual frame sampling is an effective approach for sampling nesting bald eagle populations.

  11. Temnothorax crasecundus sp. n. – a cryptic Eurocaucasian ant species (Hymenoptera, Formicidae) discovered by Nest Centroid Clustering

    PubMed Central

    Seifert, Bernhard; Csösz, Sandor

    2015-01-01

    Abstract The paper integrates two independent studies of numeric morphology-based alpha-taxonomy of the cryptic ant species Temnothorax crassispinus (Karavajev, 1926) and Temnothorax crasecundus sp. n. conducted by different investigators, using different equipment, considering different character combinations and evaluating different samples. Samples investigated included 603 individual workers from 203 nests – thereof 104 nest samples measured by Seifert and 99 by Csösz. The material originated from Europe, Asia Minor and Caucasia. There was a very strong interspecific overlap in any of the 29 shape characters recorded and subjective expert determination failed in many cases. Primary classification hypotheses were formed by the exploratory data analysis Nest Centroid (NC) clustering and corrected to final species hypotheses by an iterative linear discriminant analysis algorithm. The evaluation of Seifert’s and Csösz’s data sets arrived at fully congruent conclusions. NC-Ward and NC-K-means clustering disagreed from the final species hypothesis in only 1.9 and 1.9% of the samples in Seifert’s data set and by 1.1 and 2.1% in Csösz’s data set which is a strong argument for heterospecificity. The type series of Temnothorax crassispinus and Temnothorax crasecundus sp. n. were allocated to different clusters with p = 0.9851 and p = 0.9912 respectively. The type series of the junior synonym Temnothorax slavonicus (Seifert, 1995) was allocated to the Temnothorax crassispinus cluster with p = 0.9927. Temnothorax crasecundus sp. n. and Temnothorax crassispinus are parapatric species with a long contact zone stretching from the Peloponnisos peninsula across Bulgaria northeast to the southern Ukraine. There is no indication for occurrence of interspecifically mixed nests or intraspecific polymorphism. However, a significant reduction of interspecific morphological distance at sites with syntopic occurrence of both species indicates local hybridization. The results are discussed within the context of the Pragmatic Species Concept of Seifert (2014). The taxonomic description and a differential diagnosis of Temnothorax crasecundus sp. n. are given. PMID:25685016

  12. Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP).

    PubMed

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Plutzer, Judit; Noack, Michael J; Mahmoudi, Mohammad Reza; Karanis, Panagiotis

    2016-06-01

    Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. MULTINEST: an efficient and robust Bayesian inference tool for cosmology and particle physics

    NASA Astrophysics Data System (ADS)

    Feroz, F.; Hobson, M. P.; Bridges, M.

    2009-10-01

    We present further development and the first public release of our multimodal nested sampling algorithm, called MULTINEST. This Bayesian inference tool calculates the evidence, with an associated error estimate, and produces posterior samples from distributions that may contain multiple modes and pronounced (curving) degeneracies in high dimensions. The developments presented here lead to further substantial improvements in sampling efficiency and robustness, as compared to the original algorithm presented in Feroz & Hobson, which itself significantly outperformed existing Markov chain Monte Carlo techniques in a wide range of astrophysical inference problems. The accuracy and economy of the MULTINEST algorithm are demonstrated by application to two toy problems and to a cosmological inference problem focusing on the extension of the vanilla Λ cold dark matter model to include spatial curvature and a varying equation of state for dark energy. The MULTINEST software, which is fully parallelized using MPI and includes an interface to COSMOMC, is available at http://www.mrao.cam.ac.uk/software/multinest/. It will also be released as part of the SUPERBAYES package, for the analysis of supersymmetric theories of particle physics, at http://www.superbayes.org.

  14. Probabilistic methods for sensitivity analysis and calibration in the NASA challenge problem

    DOE PAGES

    Safta, Cosmin; Sargsyan, Khachik; Najm, Habib N.; ...

    2015-01-01

    In this study, a series of algorithms are proposed to address the problems in the NASA Langley Research Center Multidisciplinary Uncertainty Quantification Challenge. A Bayesian approach is employed to characterize and calibrate the epistemic parameters based on the available data, whereas a variance-based global sensitivity analysis is used to rank the epistemic and aleatory model parameters. A nested sampling of the aleatory–epistemic space is proposed to propagate uncertainties from model parameters to output quantities of interest.

  15. Probabilistic methods for sensitivity analysis and calibration in the NASA challenge problem

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Safta, Cosmin; Sargsyan, Khachik; Najm, Habib N.

    In this study, a series of algorithms are proposed to address the problems in the NASA Langley Research Center Multidisciplinary Uncertainty Quantification Challenge. A Bayesian approach is employed to characterize and calibrate the epistemic parameters based on the available data, whereas a variance-based global sensitivity analysis is used to rank the epistemic and aleatory model parameters. A nested sampling of the aleatory–epistemic space is proposed to propagate uncertainties from model parameters to output quantities of interest.

  16. Kinematic modelling of disc galaxies using graphics processing units

    NASA Astrophysics Data System (ADS)

    Bekiaris, G.; Glazebrook, K.; Fluke, C. J.; Abraham, R.

    2016-01-01

    With large-scale integral field spectroscopy (IFS) surveys of thousands of galaxies currently under-way or planned, the astronomical community is in need of methods, techniques and tools that will allow the analysis of huge amounts of data. We focus on the kinematic modelling of disc galaxies and investigate the potential use of massively parallel architectures, such as the graphics processing unit (GPU), as an accelerator for the computationally expensive model-fitting procedure. We review the algorithms involved in model-fitting and evaluate their suitability for GPU implementation. We employ different optimization techniques, including the Levenberg-Marquardt and nested sampling algorithms, but also a naive brute-force approach based on nested grids. We find that the GPU can accelerate the model-fitting procedure up to a factor of ˜100 when compared to a single-threaded CPU, and up to a factor of ˜10 when compared to a multithreaded dual CPU configuration. Our method's accuracy, precision and robustness are assessed by successfully recovering the kinematic properties of simulated data, and also by verifying the kinematic modelling results of galaxies from the GHASP and DYNAMO surveys as found in the literature. The resulting GBKFIT code is available for download from: http://supercomputing.swin.edu.au/gbkfit.

  17. Plasma steroid profiles in nesting loggerhead turtles (Caretta caretta) in Queensland, Australia: relationship to nesting episode and season.

    PubMed

    Whittier, J M; Corrie, F; Limpus, C

    1997-04-01

    Plasma levels of four hormones-progesterone (P), testosterone (T), estradiol 17-beta (E2), and corticosterone (B)-were measured in samples taken from nesting female loggerhead turtles (Caretta caretta) by using specific radioimmunoassays. Samples were taken in an early, middle, or late period during the summer nesting season from females at first, second, third, or > fourth nesting episodes, defined as successive within-season nesting events, at Mon Repos Beach, Queensland, Australia. Data on individual patterns of nesting, collected over the past 20 years by the Queensland Turtle Research Project, and the seasonal nesting data, were analyzed with respect to influences on hormonal profiles. Circulating levels of E2 were mostly undetectable, suggesting either that this estrogen is not produced at this time of nesting, or that, perhaps, another estrogen may be present that is not detected by the specific radioimmunoassay. P, T, and B profiles in the nesting females were associated with the individual turtles' progression through successive nesting episodes, with a marked decline in all three hormones by the last (> 4) nesting episode of the season. Nesting episode accounted for significant changes that were related to season, in that nesting episode and season were significantly correlated. These patterns were observed in the population, when singly sampled at each time period or nesting episode, and in individual females sampled repeatedly over time. Moreover, T and B were highly and significantly correlated in females at all nesting episodes and time periods, and in the singly and repeatedly sampled females. The magnitude of change in T and B over time was also highly and significantly correlated in repeatedly sampled females. Together these results suggest the hypothesis that T and B interact over the period of successive nesting and may be involved in reproductive functions such as the mobilization of reserves for egg production in C. caretta.

  18. Human papillomavirus detection and typing using a nested-PCR-RFLP assay.

    PubMed

    Coser, Janaina; Boeira, Thaís da Rocha; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

    2011-01-01

    It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

  19. Variability in nest survival rates and implications to nesting studies

    USGS Publications Warehouse

    Klett, A.T.; Johnson, D.H.

    1982-01-01

    We used four reasonably large samples (83-213) of Mallard (Anas platyrhynchos) and Blue-winged Teal (A. discors) nests on an interstate highway right-of-way in southcentral North Dakota to evaluate potential biases in hatch-rate estimates. Twelve consecutive, weekly searches for nests were conducted with a cable-chain drag in 1976 and 1977. Nests were revisited at weekly intervals. Four methods were used to estimate hatch rates for the four data sets: the Traditional Method, the Mayfield Method, and two modifications of the Mayfield Method that are sometimes appropriate when daily mortality rates of nests are not constant. Hatch rates and the average age of nests at discovery declined as the interval between searches decreased, suggesting that mortality rates were not constant in our samples. An analysis of variance indicated that daily mortality rates varied with the age of nests in all four samples. Mortality was generally highest during the early laying period, moderately high during the late laying period, and lowest during incubation. We speculate that this relationship of mortality to nest age might be due to the presence of hens at nests or to differences in the vulnerability of nest sites to predation. A modification of the Mayfield Method that accounts for age-related variation in nest mortality was most appropriate for our samples. We suggest methods for conducting nesting studies and estimating nest success for species possessing similar nesting habits.

  20. User's Manual MCnest - Markov Chain Nest Productivity Model Version 2.0

    EPA Science Inventory

    The Markov chain nest productivity model, or MCnest, is a set of algorithms for integrating the results of avian toxicity tests with reproductive life-history data to project the relative magnitude of chemical effects on avian reproduction. The mathematical foundation of MCnest i...

  1. Evaluation of ecological risk to populations of a threatened plant from an invasive biocontrol insect

    USGS Publications Warehouse

    Louda, S.M.; Rand, T.A.; Arnett, A.E.; McClay, A.S.; McEachern, A.K.

    2005-01-01

    This study was conducted to determine the influence of residues of organochlorine pollutants on reproductive success in the brown pelican. A sample egg was taken from each of 93 marked nests in the nesting colony in South Carolina. Periodic visits were made to determine the fate of marked nests. Each sample egg was analysed for residues of organochlorine pollutants. Nest success in the brown pelican was related to residues of DDE and dieldrin in sample eggs. DDE seemed primarily responsible for nest failure; however, deleterious effects of this pollutant on nest success were not satisfactorily separated from those induced by dieldrin. Significant intercorrelation of all five organochlorine residues identified in the eggs complicated the relationship of residues to nest success. Maximum DDE residues in an egg from a successful nest were 2.37 ?g/g and 8.48 ?g/g in an egg from an unsuccessful nest. Comparable maximum residues for dieldrin in sample eggs were 0.54 ?g/g (successful) and 0.99 ?g/g (unsuccessful). Nineteen of the 26 eggs from unsuccessful nests contained DDE residues that exceeded 2.37 ?g/g, and 14 of these 26 eggs contained dieldrin residues that exceeded O.54 ?g/g. Residues of DDD, DDT or PCBs in sample eggs were not significantly related to nest success. Reproductive success in the brown pelican colony was subnormal in the years of study--1971 and 1972--but reproductive success was normal in those nests in which the sample egg contained either <2,5 ?g/g of DDE or 0.54 ?g/g of dieldrin.

  2. Clutch sizes and nests of tailed frogs from the Olympic Peninsula, Washington

    USGS Publications Warehouse

    Bury, R. Bruce; Loafman, P.; Rofkar, D.; Mike, K.

    2001-01-01

    In the summers 1995-1998, we sampled 168 streams (1,714 in of randomly selected 1-m bands) to determine distribution and abundance of stream amphibians in Olympic National Park, Washington. We found six nests (two in one stream) of the tailed frog, compared to only two nests with clutch sizes reported earlier for coastal regions. This represents only one nest per 286 in searched and one nest per 34 streams sampled. Tailed frogs occurred only in 94 (60%) of the streams and, for these waters, we found one nest per 171 in searched or one nest per 20 streams sampled. The numbers of eggs for four masses ((x) over bar = 48.3, range 40-55) were low but one single strand in a fifth nest had 96 eggs. One nest with 185 eggs likely represented communal egg deposition. Current evidence indicates a geographic trend with yearly clutches of relatively few eggs in coastal tailed frogs compared to biennial nesting with larger clutches for inland populations in the Rocky Mountains.

  3. Multilevel modelling: Beyond the basic applications.

    PubMed

    Wright, Daniel B; London, Kamala

    2009-05-01

    Over the last 30 years statistical algorithms have been developed to analyse datasets that have a hierarchical/multilevel structure. Particularly within developmental and educational psychology these techniques have become common where the sample has an obvious hierarchical structure, like pupils nested within a classroom. We describe two areas beyond the basic applications of multilevel modelling that are important to psychology: modelling the covariance structure in longitudinal designs and using generalized linear multilevel modelling as an alternative to methods from signal detection theory (SDT). Detailed code for all analyses is described using packages for the freeware R.

  4. An Enhanced Memetic Algorithm for Single-Objective Bilevel Optimization Problems.

    PubMed

    Islam, Md Monjurul; Singh, Hemant Kumar; Ray, Tapabrata; Sinha, Ankur

    2017-01-01

    Bilevel optimization, as the name reflects, deals with optimization at two interconnected hierarchical levels. The aim is to identify the optimum of an upper-level  leader problem, subject to the optimality of a lower-level follower problem. Several problems from the domain of engineering, logistics, economics, and transportation have an inherent nested structure which requires them to be modeled as bilevel optimization problems. Increasing size and complexity of such problems has prompted active theoretical and practical interest in the design of efficient algorithms for bilevel optimization. Given the nested nature of bilevel problems, the computational effort (number of function evaluations) required to solve them is often quite high. In this article, we explore the use of a Memetic Algorithm (MA) to solve bilevel optimization problems. While MAs have been quite successful in solving single-level optimization problems, there have been relatively few studies exploring their potential for solving bilevel optimization problems. MAs essentially attempt to combine advantages of global and local search strategies to identify optimum solutions with low computational cost (function evaluations). The approach introduced in this article is a nested Bilevel Memetic Algorithm (BLMA). At both upper and lower levels, either a global or a local search method is used during different phases of the search. The performance of BLMA is presented on twenty-five standard test problems and two real-life applications. The results are compared with other established algorithms to demonstrate the efficacy of the proposed approach.

  5. Whole-body PET parametric imaging employing direct 4D nested reconstruction and a generalized non-linear Patlak model

    NASA Astrophysics Data System (ADS)

    Karakatsanis, Nicolas A.; Rahmim, Arman

    2014-03-01

    Graphical analysis is employed in the research setting to provide quantitative estimation of PET tracer kinetics from dynamic images at a single bed. Recently, we proposed a multi-bed dynamic acquisition framework enabling clinically feasible whole-body parametric PET imaging by employing post-reconstruction parameter estimation. In addition, by incorporating linear Patlak modeling within the system matrix, we enabled direct 4D reconstruction in order to effectively circumvent noise amplification in dynamic whole-body imaging. However, direct 4D Patlak reconstruction exhibits a relatively slow convergence due to the presence of non-sparse spatial correlations in temporal kinetic analysis. In addition, the standard Patlak model does not account for reversible uptake, thus underestimating the influx rate Ki. We have developed a novel whole-body PET parametric reconstruction framework in the STIR platform, a widely employed open-source reconstruction toolkit, a) enabling accelerated convergence of direct 4D multi-bed reconstruction, by employing a nested algorithm to decouple the temporal parameter estimation from the spatial image update process, and b) enhancing the quantitative performance particularly in regions with reversible uptake, by pursuing a non-linear generalized Patlak 4D nested reconstruction algorithm. A set of published kinetic parameters and the XCAT phantom were employed for the simulation of dynamic multi-bed acquisitions. Quantitative analysis on the Ki images demonstrated considerable acceleration in the convergence of the nested 4D whole-body Patlak algorithm. In addition, our simulated and patient whole-body data in the postreconstruction domain indicated the quantitative benefits of our extended generalized Patlak 4D nested reconstruction for tumor diagnosis and treatment response monitoring.

  6. Different bees, different needs: how nest-site requirements have shaped the decision-making processes in homeless honeybees (Apis spp.).

    PubMed

    Beekman, Madeleine; Oldroyd, Benjamin P

    2018-05-19

    During reproductive swarming, a honeybee swarm needs to decide on a new nest site and then move to the chosen site collectively. Most studies of swarming and nest-site selection are based on one species, Apis mellifera Natural colonies of A. mellifera live in tree cavities. The quality of the cavity is critical to the survival of a swarm. Other honeybee species nest in the open, and have less strict nest-site requirements, such as the open-nesting dwarf honeybee Apis florea Apis florea builds a nest comprised of a single comb suspended from a twig. For a cavity-nesting species, there is only a limited number of potential nest sites that can be located by a swarm, because suitable sites are scarce. By contrast, for an open-nesting species, there is an abundance of equally suitable twigs. While the decision-making process of cavity-nesting bees is geared towards selecting the best site possible, open-nesting species need to coordinate collective movement towards areas with potential nest sites. Here, we argue that the nest-site selection processes of A. florea and A. mellifera have been shaped by each species' specific nest-site requirements. Both species use the same behavioural algorithm, tuned to allow each species to solve their species-specific problem.This article is part of the theme issue 'Collective movement ecology'. © 2018 The Author(s).

  7. Methodologic considerations in the design and analysis of nested case-control studies: association between cytokines and postoperative delirium.

    PubMed

    Ngo, Long H; Inouye, Sharon K; Jones, Richard N; Travison, Thomas G; Libermann, Towia A; Dillon, Simon T; Kuchel, George A; Vasunilashorn, Sarinnapha M; Alsop, David C; Marcantonio, Edward R

    2017-06-06

    The nested case-control study (NCC) design within a prospective cohort study is used when outcome data are available for all subjects, but the exposure of interest has not been collected, and is difficult or prohibitively expensive to obtain for all subjects. A NCC analysis with good matching procedures yields estimates that are as efficient and unbiased as estimates from the full cohort study. We present methodological considerations in a matched NCC design and analysis, which include the choice of match algorithms, analysis methods to evaluate the association of exposures of interest with outcomes, and consideration of overmatching. Matched, NCC design within a longitudinal observational prospective cohort study in the setting of two academic hospitals. Study participants are patients aged over 70 years who underwent scheduled major non-cardiac surgery. The primary outcome was postoperative delirium from in-hospital interviews and medical record review. The main exposure was IL-6 concentration (pg/ml) from blood sampled at three time points before delirium occurred. We used nonparametric signed ranked test to test for the median of the paired differences. We used conditional logistic regression to model the risk of IL-6 on delirium incidence. Simulation was used to generate a sample of cohort data on which unconditional multivariable logistic regression was used, and the results were compared to those of the conditional logistic regression. Partial R-square was used to assess the level of overmatching. We found that the optimal match algorithm yielded more matched pairs than the greedy algorithm. The choice of analytic strategy-whether to consider measured cytokine levels as the predictor or outcome-- yielded inferences that have different clinical interpretations but similar levels of statistical significance. Estimation results from NCC design using conditional logistic regression, and from simulated cohort design using unconditional logistic regression, were similar. We found minimal evidence for overmatching. Using a matched NCC approach introduces methodological challenges into the study design and data analysis. Nonetheless, with careful selection of the match algorithm, match factors, and analysis methods, this design is cost effective and, for our study, yields estimates that are similar to those from a prospective cohort study design.

  8. Chemical analyses of ground-water samples from the Rio Grande Valley in the vicinity of Albuquerque, New Mexico, October 1993 through January 1994

    USGS Publications Warehouse

    Wilkins, D.W.; Schlottmann, J.L.; Ferree, D.M.

    1996-01-01

    A study was conducted to investigate general ground-water- quality conditions and contaminant locations in the Rio Grande Valley in the vicinity of Albuquerque, New Mexico. Water samples from 36 observation wells in 12 well nests were analyzed. The well nests are located along three roads near the Rio Grande--two well nests near Paseo del Norte, five well nests near Monta?o Road, and five well nests near Rio Bravo Boulevard. The water samples were collected from October 19, 1993, through January 18, 1994. Water-quality types by major-ion composition were calcium bicarbonate (found in most samples), sodium sulfate, calcium sulfate, and calcium sulfate chloride. Nutrients were detected in all but one sample. Ammonia was detected in 34 samples, nitrite in 4 samples, and nitrate in 17 samples. Orthophosphate was detected in 31 samples. Organic carbon was detected in all samples collected. The trace elements arsenic and barium were detected in all samples and zinc in 31 samples. Fourteen samples contained detectable copper. Cadmium was detected in one sample, chromium in two samples, lead in four samples, and selenium in two samples. Mercury and silver were not detected.

  9. Comparison of loop-mediated isothermal amplification (LAMP) and nested-PCR assay targeting the RE and B1 gene for detection of Toxoplasma gondii in blood samples of children with leukaemia.

    PubMed

    Fallahi, Shirzad; Seyyed Tabaei, Seyyed Javad; Pournia, Yadollah; Zebardast, Nozhat; Kazemi, Bahram

    2014-07-01

    Toxoplasmosis diagnosis constitutes an important measure for disease prevention and control. In this paper, a newly described DNA amplification technique, loop-mediated isothermal amplification (LAMP), and nested-PCR targeting the repeated element (RE) and B1 gene, were compared to each other for the detection of Toxoplasma gondii DNA in blood samples of children with leukaemia. One hundred ten blood samples from these patients were analyzed by LAMP and nested-PCR. Out of 50 seropositive samples (IgM+, IgG+), positive results were obtained with 92% and 86% on RE, B1-LAMP and 82% and 68% on RE, B1-nested PCR analyses, respectively. Of the 50 seronegative samples, three, two and one samples were detected positive by RE-LAMP, B1-LAMP and RE-nested PCR assays, respectively, while none were detected positive by B1-nested PCR. None of the 10 IgM-, IgG+ samples was detected positive after testing LAMP and nested-PCR assays in duplicate. This is the first report of a study in which the LAMP method was applied with high sensitivity and efficacy for the diagnosis of T. gonii in blood samples of children with leukaemia. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Establishment of a nested-ASP-PCR method to determine the clarithromycin resistance of Helicobacter pylori.

    PubMed

    Luo, Xiao-Feng; Jiao, Jian-Hua; Zhang, Wen-Yue; Pu, Han-Ming; Qu, Bao-Jin; Yang, Bing-Ya; Hou, Min; Ji, Min-Jun

    2016-07-07

    To investigate clarithromycin resistance positions 2142, 2143 and 2144 of the 23SrRNA gene in Helicobacter pylori (H. pylori) by nested-allele specific primer-polymerase chain reaction (nested-ASP-PCR). The gastric tissue and saliva samples from 99 patients with positive results of the rapid urease test (RUT) were collected. The nested-ASP-PCR method was carried out with the external primers and inner allele-specific primers corresponding to the reference strain and clinical strains. Thirty gastric tissue and saliva samples were tested to determine the sensitivity of nested-ASP-PCR and ASP-PCR methods. Then, clarithromycin resistance was detected for 99 clinical samples by using different methods, including nested-ASP-PCR, bacterial culture and disk diffusion. The nested-ASP-PCR method was successfully established to test the resistance mutation points 2142, 2143 and 2144 of the 23SrRNA gene of H. pylori. Among 30 samples of gastric tissue and saliva, the H. pylori detection rate of nested-ASP-PCR was 90% and 83.33%, while the detection rate of ASP-PCR was just 63% and 56.67%. Especially in the saliva samples, nested-ASP-PCR showed much higher sensitivity in H. pylori detection and resistance mutation rates than ASP-PCR. In the 99 RUT-positive gastric tissue and saliva samples, the H. pylori-positive detection rate by nested-ASP-PCR was 87 (87.88%) and 67 (67.68%), in which there were 30 wild-type and 57 mutated strains in gastric tissue and 22 wild-type and 45 mutated strains in saliva. Genotype analysis showed that three-points mixed mutations were quite common, but different resistant strains were present in gastric mucosa and saliva. Compared to the high sensitivity shown by nested-ASP-PCR, the positive detection of bacterial culture with gastric tissue samples was 50 cases, in which only 26 drug-resistant strains were found through analyzing minimum inhibitory zone of clarithromycin. The nested-ASP-PCR assay showed higher detection sensitivity than ASP-PCR and drug sensitivity testing, which could be performed to evaluate clarithromycin resistance of H. pylori.

  11. Establishment of a nested-ASP-PCR method to determine the clarithromycin resistance of Helicobacter pylori

    PubMed Central

    Luo, Xiao-Feng; Jiao, Jian-Hua; Zhang, Wen-Yue; Pu, Han-Ming; Qu, Bao-Jin; Yang, Bing-Ya; Hou, Min; Ji, Min-Jun

    2016-01-01

    AIM: To investigate clarithromycin resistance positions 2142, 2143 and 2144 of the 23SrRNA gene in Helicobacter pylori (H. pylori) by nested-allele specific primer-polymerase chain reaction (nested-ASP-PCR). METHODS: The gastric tissue and saliva samples from 99 patients with positive results of the rapid urease test (RUT) were collected. The nested-ASP-PCR method was carried out with the external primers and inner allele-specific primers corresponding to the reference strain and clinical strains. Thirty gastric tissue and saliva samples were tested to determine the sensitivity of nested-ASP-PCR and ASP-PCR methods. Then, clarithromycin resistance was detected for 99 clinical samples by using different methods, including nested-ASP-PCR, bacterial culture and disk diffusion. RESULTS: The nested-ASP-PCR method was successfully established to test the resistance mutation points 2142, 2143 and 2144 of the 23SrRNA gene of H. pylori. Among 30 samples of gastric tissue and saliva, the H. pylori detection rate of nested-ASP-PCR was 90% and 83.33%, while the detection rate of ASP-PCR was just 63% and 56.67%. Especially in the saliva samples, nested-ASP-PCR showed much higher sensitivity in H. pylori detection and resistance mutation rates than ASP-PCR. In the 99 RUT-positive gastric tissue and saliva samples, the H. pylori-positive detection rate by nested-ASP-PCR was 87 (87.88%) and 67 (67.68%), in which there were 30 wild-type and 57 mutated strains in gastric tissue and 22 wild-type and 45 mutated strains in saliva. Genotype analysis showed that three-points mixed mutations were quite common, but different resistant strains were present in gastric mucosa and saliva. Compared to the high sensitivity shown by nested-ASP-PCR, the positive detection of bacterial culture with gastric tissue samples was 50 cases, in which only 26 drug-resistant strains were found through analyzing minimum inhibitory zone of clarithromycin. CONCLUSION: The nested-ASP-PCR assay showed higher detection sensitivity than ASP-PCR and drug sensitivity testing, which could be performed to evaluate clarithromycin resistance of H. pylori. PMID:27433095

  12. Automatic blocking of nested loops

    NASA Technical Reports Server (NTRS)

    Schreiber, Robert; Dongarra, Jack J.

    1990-01-01

    Blocked algorithms have much better properties of data locality and therefore can be much more efficient than ordinary algorithms when a memory hierarchy is involved. On the other hand, they are very difficult to write and to tune for particular machines. The reorganization is considered of nested loops through the use of known program transformations in order to create blocked algorithms automatically. The program transformations used are strip mining, loop interchange, and a variant of loop skewing in which invertible linear transformations (with integer coordinates) of the loop indices are allowed. Some problems are solved concerning the optimal application of these transformations. It is shown, in a very general setting, how to choose a nearly optimal set of transformed indices. It is then shown, in one particular but rather frequently occurring situation, how to choose an optimal set of block sizes.

  13. Improved parallel data partitioning by nested dissection with applications to information retrieval.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wolf, Michael M.; Chevalier, Cedric; Boman, Erik Gunnar

    The computational work in many information retrieval and analysis algorithms is based on sparse linear algebra. Sparse matrix-vector multiplication is a common kernel in many of these computations. Thus, an important related combinatorial problem in parallel computing is how to distribute the matrix and the vectors among processors so as to minimize the communication cost. We focus on minimizing the total communication volume while keeping the computation balanced across processes. In [1], the first two authors presented a new 2D partitioning method, the nested dissection partitioning algorithm. In this paper, we improve on that algorithm and show that it ismore » a good option for data partitioning in information retrieval. We also show partitioning time can be substantially reduced by using the SCOTCH software, and quality improves in some cases, too.« less

  14. Nest materials as a source of genetic data for avian ecological studies

    USGS Publications Warehouse

    Pearce, J.M.; Fields, R.L.; Scribner, K.T.

    1997-01-01

    We examined the utility of feathers and egg shell membranes, deposited in the nests of Spectacled Eiders (Somateria fischeri), as a source of DNA for genetic studies at both the population and individual level. The potential for feather DNA contamination as a result of female behavioral interactions (e.g. nest parasitism), reuse of nest sites from previous years, or other unknown occurrences was acknowledged and specifically tested. DNA was successfully extracted from both feathers and egg shell membranes and waterfowl microsatellite loci were used to construct individual genotypes. We found no difference in the genotypes obtained from nest feathers or blood of the incubating female. Detection of nest feather contamination was possible with as little as one feather when samples from multiple females were intentionally mixed. Triplicate DNA extractions from 33 nests provided a means of detecting contamination in 3 nests. Egg membranes proved a viable source of offspring DNA and can contribute valuable data to investigations of parentage when assayed jointly with maternal feather DNA. Nest materials provide an efficient, non-invasive method of genetic sampling that can be readily incorporated into field research. However, the natural history traits and mating strategies of a species must be considered during sample collection to identify the possible sources of nest materials (e.g., paternal, maternal, parasite, etc.). Specific experiments should also be designed to test sampling assumptions.

  15. Assessing relative abundance and reproductive success of shrubsteppe raptors

    USGS Publications Warehouse

    Lehman, Robert N.; Carpenter, L.B.; Steenhof, Karen; Kochert, Michael N.

    1998-01-01

    From 1991-1994, we quantified relative abundance and reproductive success of the Ferruginous Hawk (Buteo regalis), Northern Harrier (Circus cyaneus), Burrowing Owl (Speotytoc unicularia), and Short-eared Owl (Asio flammeus) on the shrubsteppe plateaus (benchlands) in and near the Snake River Birds of Prey National Conservation Area in southwestern Idaho. To assess relative abundance, we searched randomly selected plots using four sampling methods: point counts, line transects, and quadrats of two sizes. On a persampling-effort basis, transects were slightly more effective than point counts and quadrats for locating raptor nests (3.4 pairs detected/100 h of effort vs. 2.2-3.1 pairs). Random sampling using quadrats failed to detect a Short-eared Owl population increase from 1993 to 1994. To evaluate nesting success, we tried to determine reproductive outcome for all nesting attempts located during random, historical, and incidental nest searches. We compared nesting success estimates based on all nesting attempts, on attempts found during incubation, and the Mayfield model. Most pairs used to evaluate success were pairs found incidentally. Visits to historical nesting areas yielded the highest number of pairs per sampling effort (14.6/100 h), but reoccupancy rates for most species decreased through time. Estimates based on all attempts had the highest sample sizes but probably overestimated success for all species except the Ferruginous Hawk. Estimates of success based on nesting attempts found during incubation had the lowest sample sizes. All three methods yielded biased nesting snccess estimates for the Northern Harrier and Short-eared Owl. The estimate based on pairs found during incubation probably provided the least biased estimate for the Burrowing Owl. Assessments of nesting success were hindered by difficulties in confirming egg laying and nesting success for all species except the Ferruginous hawk.

  16. A tunable algorithm for collective decision-making.

    PubMed

    Pratt, Stephen C; Sumpter, David J T

    2006-10-24

    Complex biological systems are increasingly understood in terms of the algorithms that guide the behavior of system components and the information pathways that link them. Much attention has been given to robust algorithms, or those that allow a system to maintain its functions in the face of internal or external perturbations. At the same time, environmental variation imposes a complementary need for algorithm versatility, or the ability to alter system function adaptively as external circumstances change. An important goal of systems biology is thus the identification of biological algorithms that can meet multiple challenges rather than being narrowly specified to particular problems. Here we show that emigrating colonies of the ant Temnothorax curvispinosus tune the parameters of a single decision algorithm to respond adaptively to two distinct problems: rapid abandonment of their old nest in a crisis and deliberative selection of the best available new home when their old nest is still intact. The algorithm uses a stepwise commitment scheme and a quorum rule to integrate information gathered by numerous individual ants visiting several candidate homes. By varying the rates at which they search for and accept these candidates, the ants yield a colony-level response that adaptively emphasizes either speed or accuracy. We propose such general but tunable algorithms as a design feature of complex systems, each algorithm providing elegant solutions to a wide range of problems.

  17. Towards a framework for testing general relativity with extreme-mass-ratio-inspiral observations

    NASA Astrophysics Data System (ADS)

    Chua, A. J. K.; Hee, S.; Handley, W. J.; Higson, E.; Moore, C. J.; Gair, J. R.; Hobson, M. P.; Lasenby, A. N.

    2018-07-01

    Extreme-mass-ratio-inspiral observations from future space-based gravitational-wave detectors such as LISA will enable strong-field tests of general relativity with unprecedented precision, but at prohibitive computational cost if existing statistical techniques are used. In one such test that is currently employed for LIGO black hole binary mergers, generic deviations from relativity are represented by N deformation parameters in a generalized waveform model; the Bayesian evidence for each of its 2N combinatorial submodels is then combined into a posterior odds ratio for modified gravity over relativity in a null-hypothesis test. We adapt and apply this test to a generalized model for extreme-mass-ratio inspirals constructed on deformed black hole spacetimes, and focus our investigation on how computational efficiency can be increased through an evidence-free method of model selection. This method is akin to the algorithm known as product-space Markov chain Monte Carlo, but uses nested sampling and improved error estimates from a rethreading technique. We perform benchmarking and robustness checks for the method, and find order-of-magnitude computational gains over regular nested sampling in the case of synthetic data generated from the null model.

  18. Towards a framework for testing general relativity with extreme-mass-ratio-inspiral observations

    NASA Astrophysics Data System (ADS)

    Chua, A. J. K.; Hee, S.; Handley, W. J.; Higson, E.; Moore, C. J.; Gair, J. R.; Hobson, M. P.; Lasenby, A. N.

    2018-04-01

    Extreme-mass-ratio-inspiral observations from future space-based gravitational-wave detectors such as LISA will enable strong-field tests of general relativity with unprecedented precision, but at prohibitive computational cost if existing statistical techniques are used. In one such test that is currently employed for LIGO black-hole binary mergers, generic deviations from relativity are represented by N deformation parameters in a generalised waveform model; the Bayesian evidence for each of its 2N combinatorial submodels is then combined into a posterior odds ratio for modified gravity over relativity in a null-hypothesis test. We adapt and apply this test to a generalised model for extreme-mass-ratio inspirals constructed on deformed black-hole spacetimes, and focus our investigation on how computational efficiency can be increased through an evidence-free method of model selection. This method is akin to the algorithm known as product-space Markov chain Monte Carlo, but uses nested sampling and improved error estimates from a rethreading technique. We perform benchmarking and robustness checks for the method, and find order-of-magnitude computational gains over regular nested sampling in the case of synthetic data generated from the null model.

  19. Introducing a novel highly prognostic grading scheme based on tumour budding and cell nest size for squamous cell carcinoma of the uterine cervix.

    PubMed

    Jesinghaus, Moritz; Strehl, Johanna; Boxberg, Melanie; Brühl, Frido; Wenzel, Adrian; Konukiewitz, Björn; Schlitter, Anna M; Steiger, Katja; Warth, Arne; Schnelzer, Andreas; Kiechle, Marion; Beckmann, Matthias W; Noske, Aurelia; Hartmann, Arndt; Mehlhorn, Grit; Koch, Martin C; Weichert, Wilko

    2018-04-01

    A novel histopathological grading system based on tumour budding and cell nest size has recently been shown to outperform conventional (WHO-based) grading algorithms in several tumour entities such as lung, oral, and oesophageal squamous cell carcinoma (SCC) in terms of prognostic patient stratification. Here, we tested the prognostic value of this innovative grading approach in two completely independent cohorts of SCC of the uterine cervix. To improve morphology-based grading, we investigated tumour budding activity and cell nest size as well as several other histomorphological factors (e.g., keratinization, nuclear size, mitotic activity) in a test cohort (n = 125) and an independent validation cohort (n = 122) of cervical SCC. All parameters were correlated with clinicopathological factors and patient outcome. Small cell nest size and high tumour budding activity were strongly associated with a dismal patient prognosis (p < 0.001 for overall survival [OS], disease-specific survival, and disease-free survival; test cohort) in both cohorts of cervical SCC. A novel grading algorithm combining these two parameters proved to be a highly effective, stage-independent prognosticator in both cohorts (OS: p < 0.001, test cohort; p = 0.001, validation cohort). In the test cohort, multivariate statistical analysis of the novel grade revealed that the hazard ratio (HR) for OS was 2.3 for G2 and 5.1 for G3 tumours compared to G1 neoplasms (p = 0.010). In the validation cohort, HR for OS was 3.0 for G2 and 7.2 for G3 tumours (p = 0.012). In conclusion, our novel grading algorithm incorporating cell nest size and tumour budding allows strongly prognostic histopathological grading of cervical SCC superior to WHO-based grading. Therefore, our data can be regarded as a cross-organ validation of previous results demonstrated for oesophageal, lung, and oral SCC. We suggest this grading algorithm as an additional morphology-based parameter for the routine diagnostic assessment of this tumour entity. © 2018 The Authors The Journal of Pathology: Clinical Research published by The Pathological Society of Great Britain and Ireland and John Wiley & Sons Ltd.

  20. A semi-nested PCR assay for molecular detection of Paracoccidioides brasiliensis in tissue samples.

    PubMed

    Koishi, Andrea Cristine; Vituri, Débora Fonseca; Dionízio Filho, Pedro Sebastião Raimundo; Sasaki, Alexandre Augusto; Felipe, Maria Sueli Soares; Venancio, Emerson José

    2010-01-01

    Paracoccidioidomycosis is a systemic infection caused by Paracoccidioides brasiliensis. In this study, a semi-nested PCR for paracoccidioidomycosis diagnosis was developed. The primers ITS1 and ITS4 were used in the first reaction, while the primers MJ03 and ITS1 primer were used in the second reaction. The semi-nested PCR was used to investigate biopsies of five patients with oral lesions that resembled paracoccidioidomycosis. The semi-nested PCR was positive for four samples and negative for a sample from a patient later diagnosed with leishmaniasis. The new semi-nested PCR describe is useful for paracoccidioidomycosis diagnosis.

  1. Clinical and epidemiological use of nested PCR targeting the repetitive element IS1111 associated with the transposase gene from Coxiella burnetii.

    PubMed

    Mares-Guia, Maria Angélica M M; Guterres, Alexandro; Rozental, Tatiana; Ferreira, Michelle Dos Santos; Lemos, Elba R S

    Q fever is a worldwide zoonosis caused by Coxiella burnetii-a small obligate intracellular Gram-negative bacterium found in a variety of animals. It is transmitted to humans by inhalation of contaminated aerosols from urine, feces, milk, amniotic fluid, placenta, abortion products, wool, and rarely by ingestion of raw milk from infected animals. Nested PCR can improve the sensitivity and specificity of testing while offering a suitable amplicon size for sequencing. Serial dilutions were performed tenfold to test the limit of detection, and the result was 10× detection of C. burnetti DNA with internal nested PCR primers relative to trans-PCR. Different biological samples were tested and identified only in nested PCR. This demonstrates the efficiency and effectiveness of the primers. Of the 19 samples, which amplify the partial sequence of C. burnetii, 12 were positive by conventional PCR and nested PCR. Seven samples-five spleen tissue samples from rodents and two tick samples-were only positive in nested PCR. With these new internal primers for trans-PCR, we demonstrate that our nested PCR assay for C. burnetii can achieve better results than conventional PCR. Published by Elsevier Editora Ltda.

  2. Comparison of Nested Polymerase Chain Reaction and Real-Time Polymerase Chain Reaction with Parasitological Methods for Detection of Strongyloides stercoralis in Human Fecal Samples

    PubMed Central

    Sharifdini, Meysam; Mirhendi, Hossein; Ashrafi, Keyhan; Hosseini, Mostafa; Mohebali, Mehdi; Khodadadi, Hossein; Kia, Eshrat Beigom

    2015-01-01

    This study was performed to evaluate nested polymerase chain reaction (PCR) and real-time PCR methods for detection of Strongyloides stercoralis in fecal samples compared with parasitological methods. A total of 466 stool samples were examined by conventional parasitological methods (formalin ether concentration [FEC] and agar plate culture [APC]). DNA was extracted using an in-house method, and mitochondrial cytochrome c oxidase subunit 1 and 18S ribosomal genes were amplified by nested PCR and real-time PCR, respectively. Among 466 samples, 12.7% and 18.2% were found infected with S. stercoralis by FEC and APC, respectively. DNA of S. stercoralis was detected in 18.9% and 25.1% of samples by real-time PCR and nested PCR, respectively. Considering parasitological methods as the diagnostic gold standard, the sensitivity and specificity of nested PCR were 100% and 91.6%, respectively, and that of real-time PCR were 84.7% and 95.8%, respectively. However, considering sequence analyzes of the selected nested PCR products, the specificity of nested PCR is increased. In general, molecular methods were superior to parasitological methods. They were more sensitive and more reliable in detection of S. stercoralis in comparison with parasitological methods. Between the two molecular methods, the sensitivity of nested PCR was higher than real-time PCR. PMID:26350449

  3. Ground-nesting by the chimpanzees of the Nimba Mountains, Guinea: environmentally or socially determined?

    PubMed

    Koops, Kathelijne; Humle, Tatyana; Sterck, Elisabeth H M; Matsuzawa, Tetsuro

    2007-04-01

    The chimpanzees (Pan troglodytes verus) of the Nimba Mountains, Guinea, West Africa, commonly make both elaborate ("night") and simple ("day") nests on the ground. In this study we investigated which factors might influence ground-nesting in this population, and tested two ecological hypotheses: 1) climatic conditions, such as high wind speeds at high altitudes, may deter chimpanzees from nesting in trees; and 2) a lack of appropriate arboreal nesting opportunities may drive the chimpanzees to nest on the ground. In addition to testing these two hypotheses, we explored whether ground-nesting is a sex-linked behavior. Data were collected monthly between August 2003 and May 2004 along transects and ad libitum. To identify the sex of ground-nesting individuals, we used DNA extracted from hair samples. The results showed that the occurrence and distribution of ground nests were not affected by climatic conditions or a lack of appropriate nest trees. Support was found for the notion that ground-nesting is a sex-linked behavior, as males were responsible for building all of the elaborate ground nests and most of the simple ground nests sampled. Elaborate ground nests occurred mostly in nest groups associated with tree nests, whereas simple ground nests usually occurred without tree nests in their vicinity. These results suggest that ground-nesting may be socially, rather than ecologically, determined.

  4. Nested Machine Learning Facilitates Increased Sequence Content for Large-Scale Automated High Resolution Melt Genotyping

    PubMed Central

    Fraley, Stephanie I.; Athamanolap, Pornpat; Masek, Billie J.; Hardick, Justin; Carroll, Karen C.; Hsieh, Yu-Hsiang; Rothman, Richard E.; Gaydos, Charlotte A.; Wang, Tza-Huei; Yang, Samuel

    2016-01-01

    High Resolution Melt (HRM) is a versatile and rapid post-PCR DNA analysis technique primarily used to differentiate sequence variants among only a few short amplicons. We recently developed a one-vs-one support vector machine algorithm (OVO SVM) that enables the use of HRM for identifying numerous short amplicon sequences automatically and reliably. Herein, we set out to maximize the discriminating power of HRM + SVM for a single genetic locus by testing longer amplicons harboring significantly more sequence information. Using universal primers that amplify the hypervariable bacterial 16 S rRNA gene as a model system, we found that long amplicons yield more complex HRM curve shapes. We developed a novel nested OVO SVM approach to take advantage of this feature and achieved 100% accuracy in the identification of 37 clinically relevant bacteria in Leave-One-Out-Cross-Validation. A subset of organisms were independently tested. Those from pure culture were identified with high accuracy, while those tested directly from clinical blood bottles displayed more technical variability and reduced accuracy. Our findings demonstrate that long sequences can be accurately and automatically profiled by HRM with a novel nested SVM approach and suggest that clinical sample testing is feasible with further optimization. PMID:26778280

  5. Preconditioned Alternating Projection Algorithms for Maximum a Posteriori ECT Reconstruction

    PubMed Central

    Krol, Andrzej; Li, Si; Shen, Lixin; Xu, Yuesheng

    2012-01-01

    We propose a preconditioned alternating projection algorithm (PAPA) for solving the maximum a posteriori (MAP) emission computed tomography (ECT) reconstruction problem. Specifically, we formulate the reconstruction problem as a constrained convex optimization problem with the total variation (TV) regularization. We then characterize the solution of the constrained convex optimization problem and show that it satisfies a system of fixed-point equations defined in terms of two proximity operators raised from the convex functions that define the TV-norm and the constrain involved in the problem. The characterization (of the solution) via the proximity operators that define two projection operators naturally leads to an alternating projection algorithm for finding the solution. For efficient numerical computation, we introduce to the alternating projection algorithm a preconditioning matrix (the EM-preconditioner) for the dense system matrix involved in the optimization problem. We prove theoretically convergence of the preconditioned alternating projection algorithm. In numerical experiments, performance of our algorithms, with an appropriately selected preconditioning matrix, is compared with performance of the conventional MAP expectation-maximization (MAP-EM) algorithm with TV regularizer (EM-TV) and that of the recently developed nested EM-TV algorithm for ECT reconstruction. Based on the numerical experiments performed in this work, we observe that the alternating projection algorithm with the EM-preconditioner outperforms significantly the EM-TV in all aspects including the convergence speed, the noise in the reconstructed images and the image quality. It also outperforms the nested EM-TV in the convergence speed while providing comparable image quality. PMID:23271835

  6. Detection of Mycobacterium tuberculosis in extrapulmonary biopsy samples using PCR targeting IS6110, rpoB, and nested-rpoB PCR Cloning

    PubMed Central

    Meghdadi, Hossein; Khosravi, Azar D.; Ghadiri, Ata A.; Sina, Amir H.; Alami, Ameneh

    2015-01-01

    Present study was aimed to examine the diagnostic utility of polymerase chain reaction (PCR) and nested PCR techniques for the detection of Mycobacterium tuberculosis (MTB) DNA in samples from patients with extra pulmonary tuberculosis (EPTB). In total 80 formalin-fixed, paraffin-embedded (FFPE) samples comprising 70 samples with definite diagnosis of EPTB and 10 samples from known non- EPTB on the basis of histopathology examination, were included in the study. PCR amplification targeting IS6110, rpoB gene and nested PCR targeting the rpoB gene were performed on the extracted DNAs from 80 FFPE samples. The strong positive samples were directly sequenced. For negative samples and those with weak band in nested-rpoB PCR, TA cloning was performed by cloning the products into the plasmid vector with subsequent sequencing. The 95% confidence intervals (CI) for the estimates of sensitivity and specificity were calculated for each method. Fourteen (20%), 34 (48.6%), and 60 (85.7%) of the 70 positive samples confirmed by histopathology, were positive by rpoB-PCR, IS6110-PCR, and nested-rpoB PCR, respectively. By performing TA cloning on samples that yielded weak (n = 8) or negative results (n = 10) in the PCR methods, we were able to improve their quality for later sequencing. All samples with weak band and 7 out of 10 negative samples, showed strong positive results after cloning. So nested-rpoB PCR cloning revealed positivity in 67 out of 70 confirmed samples (95.7%). The sensitivity of these combination methods was calculated as 95.7% in comparison with histopathology examination. The CI for sensitivity of the PCR methods were calculated as 11.39–31.27% for rpoB-PCR, 36.44–60.83% for IS6110- PCR, 75.29–92.93% for nested-rpoB PCR, and 87.98–99.11% for nested-rpoB PCR cloning. The 10 true EPTB negative samples by histopathology, were negative by all tested methods including cloning and were used to calculate the specificity of the applied methods. The CI for 100% specificity of each PCR method were calculated as 69.15–100%. Our results indicated that nested-rpoB PCR combined with TA cloning and sequencing is a preferred method for the detection of MTB DNA in EPTB samples with high sensitivity and specificity which confirm the histopathology results. PMID:26191059

  7. Detection of Mycobacterium tuberculosis in extrapulmonary biopsy samples using PCR targeting IS6110, rpoB, and nested-rpoB PCR Cloning.

    PubMed

    Meghdadi, Hossein; Khosravi, Azar D; Ghadiri, Ata A; Sina, Amir H; Alami, Ameneh

    2015-01-01

    Present study was aimed to examine the diagnostic utility of polymerase chain reaction (PCR) and nested PCR techniques for the detection of Mycobacterium tuberculosis (MTB) DNA in samples from patients with extra pulmonary tuberculosis (EPTB). In total 80 formalin-fixed, paraffin-embedded (FFPE) samples comprising 70 samples with definite diagnosis of EPTB and 10 samples from known non- EPTB on the basis of histopathology examination, were included in the study. PCR amplification targeting IS6110, rpoB gene and nested PCR targeting the rpoB gene were performed on the extracted DNAs from 80 FFPE samples. The strong positive samples were directly sequenced. For negative samples and those with weak band in nested-rpoB PCR, TA cloning was performed by cloning the products into the plasmid vector with subsequent sequencing. The 95% confidence intervals (CI) for the estimates of sensitivity and specificity were calculated for each method. Fourteen (20%), 34 (48.6%), and 60 (85.7%) of the 70 positive samples confirmed by histopathology, were positive by rpoB-PCR, IS6110-PCR, and nested-rpoB PCR, respectively. By performing TA cloning on samples that yielded weak (n = 8) or negative results (n = 10) in the PCR methods, we were able to improve their quality for later sequencing. All samples with weak band and 7 out of 10 negative samples, showed strong positive results after cloning. So nested-rpoB PCR cloning revealed positivity in 67 out of 70 confirmed samples (95.7%). The sensitivity of these combination methods was calculated as 95.7% in comparison with histopathology examination. The CI for sensitivity of the PCR methods were calculated as 11.39-31.27% for rpoB-PCR, 36.44-60.83% for IS6110- PCR, 75.29-92.93% for nested-rpoB PCR, and 87.98-99.11% for nested-rpoB PCR cloning. The 10 true EPTB negative samples by histopathology, were negative by all tested methods including cloning and were used to calculate the specificity of the applied methods. The CI for 100% specificity of each PCR method were calculated as 69.15-100%. Our results indicated that nested-rpoB PCR combined with TA cloning and sequencing is a preferred method for the detection of MTB DNA in EPTB samples with high sensitivity and specificity which confirm the histopathology results.

  8. Algorithm for repairing the damaged images of grain structures obtained from the cellular automata and measurement of grain size

    NASA Astrophysics Data System (ADS)

    Ramírez-López, A.; Romero-Romo, M. A.; Muñoz-Negron, D.; López-Ramírez, S.; Escarela-Pérez, R.; Duran-Valencia, C.

    2012-10-01

    Computational models are developed to create grain structures using mathematical algorithms based on the chaos theory such as cellular automaton, geometrical models, fractals, and stochastic methods. Because of the chaotic nature of grain structures, some of the most popular routines are based on the Monte Carlo method, statistical distributions, and random walk methods, which can be easily programmed and included in nested loops. Nevertheless, grain structures are not well defined as the results of computational errors and numerical inconsistencies on mathematical methods. Due to the finite definition of numbers or the numerical restrictions during the simulation of solidification, damaged images appear on the screen. These images must be repaired to obtain a good measurement of grain geometrical properties. Some mathematical algorithms were developed to repair, measure, and characterize grain structures obtained from cellular automata in the present work. An appropriate measurement of grain size and the corrected identification of interfaces and length are very important topics in materials science because they are the representation and validation of mathematical models with real samples. As a result, the developed algorithms are tested and proved to be appropriate and efficient to eliminate the errors and characterize the grain structures.

  9. Stabilities of ant nests and their adjacent soils

    NASA Astrophysics Data System (ADS)

    Echezona, B. C.; Igwe, C. A.

    2012-10-01

    Nests habour ants and termites and protect them from harsh environmental conditions. The structural stabilities of nests were studied to ascertain their relative vulnerability to environmental stresses. Arboreal-ant nests were pried from different trees, while epigeous-termite nests were excavated from soil surface within the sample area. Soils without any visible sign of ant or termite activity were also sampled 6 m away from the nests as control. Laboratory analysis result showed that irrespective of the tree hosts, the aggregate stabilities of the ant nests were lower than those of the ground termite, with nests formed on Cola nitida significantly showing lower aggregate stability (19.7%) than other antnest structures. Clay dispersion ratio, moisture content, water stable aggregate class <0.25mm and sand mass were each negatively correlated with aggregate stability, while water stable aggregate class1.00-0.50 mm gave a positive correlation. Nest structures were dominated more by water stable aggregate class >2.00 mm but path analysis demonstrated that water stable aggregate class <0.25 mm contributed most to the higher aggregate stability of the termite nest than the other nest. Nest aggregates had greater structural stability compared to the control soil. The higher structural stability of termite nests over other nest and soil was considered a better adaptive mechanism against body desiccation.

  10. Usefulness of in-house PCR methods for hepatitis B virus DNA detection.

    PubMed

    Portilho, Moyra Machado; Baptista, Marcia Leite; da Silva, Messias; de Sousa, Paulo Sérgio Fonseca; Lewis-Ximenez, Lia Laura; Lampe, Elisabeth; Villar, Livia Melo

    2015-10-01

    The aim of the present study was to evaluate the performance of three in-house PCR techniques for HBV DNA detection and compare it with commercial quantitative methods to evaluate the usefulness of in-house methods for HBV diagnosis. Three panels of HBsAg reactive sera samples were evaluated: (i) 50 samples were examined using three methods for in-house qualitative PCR and the Cobas Amplicor HBV Monitor Assay; (ii) 87 samples were assayed using in-house semi-nested PCR and the Cobas TaqMan HBV test; (iii) 11 serial samples obtained from 2 HBV-infected individuals were assayed using the Cobas Amplicor HBV test and semi-nested PCR. In panel I, HBV DNA was detected in 44 samples using the Cobas Amplicor HBV test, 42 samples using semi-nested PCR (90% concordance with Cobas Amplicor), 22 samples using PCR for the core gene (63.6% concordance) and 29 samples using single-round PCR for the pre-S/S gene (75% concordance). In panel II, HBV DNA was quantified in 78 of the 87 HBsAg reactive samples using Cobas TaqMan but 52 samples using semi-nested PCR (67.8% concordance). HBV DNA was detected in serial samples until the 17th and 26th week after first donation using in-house semi-nested PCR and the Cobas Amplicor HBV test, respectively. In-house semi-nested PCR presented adequate concordance with commercial methods as an alternative method for HBV molecular diagnosis in low-resource settings. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Detection of Mycoplasma hyopneumoniae by Air Sampling with a Nested PCR Assay

    PubMed Central

    Stärk, Katharina D. C.; Nicolet, Jacques; Frey, Joachim

    1998-01-01

    This article describes the first successful detection of airborne Mycoplasma hyopneumoniae under experimental and field conditions with a new nested PCR assay. Air was sampled with polyethersulfone membranes (pore size, 0.2 μm) mounted in filter holders. Filters were processed by dissolution and direct extraction of DNA for PCR analysis. For the PCR, two nested pairs of oligonucleotide primers were designed by using an M. hyopneumoniae-specific DNA sequence of a repeated gene segment. A nested PCR assay was developed and used to analyze samples collected in eight pig houses where respiratory problems had been common. Air was also sampled from a mycoplasma-free herd. The nested PCR was highly specific and 104 times as sensitive as a one-step PCR. Under field conditions, the sampling system was able to detect airborne M. hyopneumoniae on 80% of farms where acute respiratory disease was present. No airborne M. hyopneumoniae was detected on infected farms without acute cases. The chance of successful detection was increased if air was sampled at several locations within a room and at a lower air humidity. PMID:9464391

  12. Thermal regimes of Mexican spotted owl nest stands

    Treesearch

    Joseph L. Ganey

    2004-01-01

    To evaluate the hypothesis that spotted owls (Strix occidentalis) select habitats with cool microclimates to avoid high daytime temperatures, I sampled thermal regimes in nest areas used by Mexican spotted owls (S. o. lucida) in northern Arizona. I sampled air temperature at 30-min intervals in 30 pairs of nest and random sites...

  13. Development of Fast Algorithms Using Recursion, Nesting and Iterations for Computational Electromagnetics

    NASA Technical Reports Server (NTRS)

    Chew, W. C.; Song, J. M.; Lu, C. C.; Weedon, W. H.

    1995-01-01

    In the first phase of our work, we have concentrated on laying the foundation to develop fast algorithms, including the use of recursive structure like the recursive aggregate interaction matrix algorithm (RAIMA), the nested equivalence principle algorithm (NEPAL), the ray-propagation fast multipole algorithm (RPFMA), and the multi-level fast multipole algorithm (MLFMA). We have also investigated the use of curvilinear patches to build a basic method of moments code where these acceleration techniques can be used later. In the second phase, which is mainly reported on here, we have concentrated on implementing three-dimensional NEPAL on a massively parallel machine, the Connection Machine CM-5, and have been able to obtain some 3D scattering results. In order to understand the parallelization of codes on the Connection Machine, we have also studied the parallelization of 3D finite-difference time-domain (FDTD) code with PML material absorbing boundary condition (ABC). We found that simple algorithms like the FDTD with material ABC can be parallelized very well allowing us to solve within a minute a problem of over a million nodes. In addition, we have studied the use of the fast multipole method and the ray-propagation fast multipole algorithm to expedite matrix-vector multiplication in a conjugate-gradient solution to integral equations of scattering. We find that these methods are faster than LU decomposition for one incident angle, but are slower than LU decomposition when many incident angles are needed as in the monostatic RCS calculations.

  14. Preconditioned alternating projection algorithms for maximum a posteriori ECT reconstruction

    NASA Astrophysics Data System (ADS)

    Krol, Andrzej; Li, Si; Shen, Lixin; Xu, Yuesheng

    2012-11-01

    We propose a preconditioned alternating projection algorithm (PAPA) for solving the maximum a posteriori (MAP) emission computed tomography (ECT) reconstruction problem. Specifically, we formulate the reconstruction problem as a constrained convex optimization problem with the total variation (TV) regularization. We then characterize the solution of the constrained convex optimization problem and show that it satisfies a system of fixed-point equations defined in terms of two proximity operators raised from the convex functions that define the TV-norm and the constraint involved in the problem. The characterization (of the solution) via the proximity operators that define two projection operators naturally leads to an alternating projection algorithm for finding the solution. For efficient numerical computation, we introduce to the alternating projection algorithm a preconditioning matrix (the EM-preconditioner) for the dense system matrix involved in the optimization problem. We prove theoretically convergence of the PAPA. In numerical experiments, performance of our algorithms, with an appropriately selected preconditioning matrix, is compared with performance of the conventional MAP expectation-maximization (MAP-EM) algorithm with TV regularizer (EM-TV) and that of the recently developed nested EM-TV algorithm for ECT reconstruction. Based on the numerical experiments performed in this work, we observe that the alternating projection algorithm with the EM-preconditioner outperforms significantly the EM-TV in all aspects including the convergence speed, the noise in the reconstructed images and the image quality. It also outperforms the nested EM-TV in the convergence speed while providing comparable image quality.

  15. The Influences of Soil Characteristics on Nest-Site Selection in Painted Turtles (Chrysemys picta)

    NASA Astrophysics Data System (ADS)

    Page, R.

    2017-12-01

    A variety of animals dig nests and lay their eggs in soil, leaving them to incubate and hatch without assistance from the parents. Nesting habitat is important for these organisms many of which exhibit temperature dependent sex determination (TSD) whereby the incubation temperature determines the sex of each hatchling. However, suitable nesting habitat may be limited due to anthropogenic activities and global temperature increases. Soil thermal properties are critical to these organisms and are positively correlated with water retention and soil carbon; carbon-rich soils result in higher incubation temperatures. We investigated nest-site selection in painted turtles (Chrysemys picta) inhabiting an anthropogenic pond in south central Pennsylvania. We surveyed for turtle nests and documented location, depth, width, temperature, canopy coverage, clutch size, and hatch success for a total of 31 turtle nests. To address the influence of soil carbon and particle size on nest selection, we analyzed samples collected from: 1) actual nests that were depredated, 2) false nests, incomplete nests aborted during digging prior to nest completion, and 3) randomized locations. Soil samples were separated into coarse, medium, and fine grain size fractions through a stack of sieves. Samples were combusted in a total carbon analyzer to measure weight percent organic carbon. We found that anthropogenic activity at this site has created homogenous, sandy, compacted soils at the uppermost layer that may limit females' access to appropriate nesting habitat. Turtle nesting activity was limited to a linear region north of the pond and was constrained by an impassable rail line. Relative to other studies, turtle nests were notably shallow (5.8±0.9 cm) and placed close to the pond. Compared to false nests and random locations, turtle-selected sites averaged greater coarse grains (35% compared to 20.24 and 20.57%) and less fine grains (47% compared to 59 and 59, respectively). Despite remarkably high soil carbon along the rail line (47.08%) turtles nested here with slightly higher hatch success. We suggest that the turtles are limited to sandy, compact soils with low heat capacities and may compensate for this by also nesting adjacent to the rail line where high soil carbon could increase incubation temperatures.

  16. Development of a Highly Sensitive Nested-PCR Procedure Using a Single Closed Tube for Detection of Erwinia amylovora in Asymptomatic Plant Material

    PubMed Central

    Llop, Pablo; Bonaterra, Anna; Peñalver, Javier; López, María M.

    2000-01-01

    A novel method, which involves a nested PCR in a single closed tube, was developed for the sensitive detection of Erwinia amylovora in plant material. The external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific DNA fragment from plasmid pEA29. The procedure involved two consecutive PCRs, the first of which was performed at a higher annealing temperature that allowed amplification only by the external primer pair. Using pure cultures of E. amylovora, the sensitivity of the nested PCR in one tube was similar to that of a standard nested PCR in two tubes. The specificity and sensitivity were greater than those of standard PCR procedures that used a single primer pair. The presence of inhibitors in plant material, very common in E. amylovora hosts, is overcome with this system in combination with a simple DNA extraction protocol because it eliminates many of the inhibitory compounds. In addition, it needs a very small sample volume (1 μl of DNA extracted). With 83 samples of naturally infected material, this method achieved better results than any other PCR technique: standard PCR detected 55% of positive samples, two-tube nested PCR detected 71% of positive samples, and nested PCR in a single closed tube detected 78% of positive samples. When analyzing asymptomatic plant material, the number of positive samples detected by the developed nested PCR was also the highest, compared with the PCR protocols indicated previously (17, 20, and 25% of 251 samples analyzed, respectively). This method is proposed for the detection of endophytic and epiphytic populations of E. amylovora in epidemiological studies and for routine use in quarantine surveys, due to its high sensitivity, specificity, speed, and simplicity. PMID:10788384

  17. Genetic Relatedness of Low Solitary Nests of Apis dorsata from Marang, Terengganu, Malaysia

    PubMed Central

    Ali, Abdul Manaf; Tan, Soon Guan; Adam, Nur Azura; Lau, Wei Hong

    2012-01-01

    Knowledge on the population of genetic structure and ecological behaviour of Apis dorsata from Peninsular Malaysia is needed for effective management and conservation of this species since unsustainable whole solitary low nest cutting for product harvesting is the current common practice here. The analysis of 15 single locus DNA microsatellite markers on samples from 20 solitary nests of A. dorsata showed that while these markers were polymorphic, high intracolonial relatedness existed. Furthermore, in general, slightly negative values of intercolony relatedness (R) among the nests of A. dorsata were found. However, positive values of mean intercolony relatedness were observed between 54 pairs of nests out of 190 possible combinations. The R values among nest pairs 3–4 and 3–5 was higher than 0.50 showing that their queens were half siblings, whereas nest pair 19–20 showed relatedness of 0.95 indicating that the same queen was sampled. The results that we obtained could not conclusively support the hypothesis of this study that the honey hunters in Marang district of Malaysia repeatedly harvest the same nest located at a different site and at a different time during the same honey harvesting season. However, our finding of an appreciable level of intercolonial relatedness between several pairs of nests in this pioneer study indicated that a comprehensive study with a larger sample size of solitary nests found throughout the region would be necessary to provide concrete proof for this novel idea. PMID:22911733

  18. Assessment of a Pan-Dermatophyte Nested-PCR Compared with Conventional Methods for Direct Detection and Identification of Dermatophytosis Agents in Animals.

    PubMed

    Piri, Fahimeh; Zarei Mahmoudabadi, Ali; Ronagh, Ali; Ahmadi, Bahram; Makimura, Koichi; Rezaei-Matehkolaei, Ali

    2018-06-26

    Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  19. Methane and carbon dioxide flux in the profile of wood ant (Formica aquilonia) nests and the surrounding forest floor during a laboratory incubation.

    PubMed

    Jílková, Veronika; Picek, Tomáš; Šestauberová, Martina; Krištůfek, Václav; Cajthaml, Tomáš; Frouz, Jan

    2016-10-01

    We compared methane (CH4) and carbon dioxide (CO2) fluxes in samples collected from the aboveground parts of wood ant nests and in the organic and mineral layer of the surrounding forest floor. Gas fluxes were measured during a laboratory incubation, and microbial properties (abundance of fungi, bacteria and methanotrophic bacteria) and nutrient contents (total and available carbon and nitrogen) were also determined. Both CO2 and CH4 were produced from ant nest samples, indicating that the aboveground parts of wood ant nests act as sources of both gases; in comparison, the forest floor produced about four times less CO2 and consumed rather than produced CH4 Fluxes of CH4 and CO2 were positively correlated with contents of available carbon and nitrogen. The methanotrophic community was represented by type II methanotrophic bacteria, but their abundance did not explain CH4 flux. Fungal abundance was greater in ant nest samples than in forest floor samples, but bacterial abundance was similar in both kinds of samples, suggesting that the organic materials in the nests may have been too recalcitrant for bacteria to decompose. The results indicate that the aboveground parts of wood ant nests are hot spots of CO2 and CH4 production in the forest floor. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Design of a Synthetic Aperture Array to Support Experiments in Active Control of Scattering

    DTIC Science & Technology

    1990-06-01

    becomes necessary to validate the theory and test the control system algorithms . While experiments in open water would be most like the anticipated...mathematical development of the beamforming algorithms used as well as an estimate of their applicability to the specifics of beamforming in a reverberant...Chebyshev array have been proposed. The method used in ARRAY, a nested product algorithm , proposed by Bresler [21] is recommended by Pozar [19] and

  1. Comparison of nested PCR and qPCR for the detection and quantitation of BoHV6 DNA.

    PubMed

    Kubiś, Piotr; Materniak, Magdalena; Kuźmak, Jacek

    2013-12-01

    Nested PCR and qPCR (quantitative PCR) tests based on glycoprotein B (gB) gene were designed for detecting Bovine herpesvirus 6 (BoHV6) in bovine whole blood samples and wild ruminant blood clots (deer and roe-deer). This virus, commonly known as BLHV (bovine lymphotropic herpesvirus) belongs to the Herpesviridae family, subfamily Gammaherpesvirinae and Macavirus genus. DNA isolated from 92 dairy cow blood samples and 69 wild ruminant clots were examined for the presence of BoHV6 using nested PCR and qPCR tests. Viral DNA was detected by using nested PCR in 59 out of 92 bovine blood samples (64.1%), and by qPCR in 68 out of 92 bovine blood samples (73.9%), but none out of 69 DNA samples isolated from wild ruminant blood clots, was positive in both assays. The specificity of nested PCR and qPCR was confirmed by using BoHV1, BoHV4, BoHV6, BFV, BIV, and BLV DNA. The sensitivity of nested PCR and qPCR was determined using a serially 10-fold diluted vector pCR2.1HgB (2 × 10(0)-2 × 10(6)copies/reaction). In this testing, qPCR was more sensitive than the nested PCR, detecting two copies of BoHV6 whilst the limit of detection for nested PCR was 20 copies. In all qPCR assays, the coefficients of determination (R(2)) ranged between 0.990 and 0.999, and the calculated amplification efficiencies (Eff%) within the range of 89.7-106.9. The intra- and inter-assay CV (coefficient of variation) values did not exceed 4%. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Vehicle Routing with Three-dimensional Container Loading Constraints—Comparison of Nested and Joint Algorithms

    NASA Astrophysics Data System (ADS)

    Koloch, Grzegorz; Kaminski, Bogumil

    2010-10-01

    In the paper we examine a modification of the classical Vehicle Routing Problem (VRP) in which shapes of transported cargo are accounted for. This problem, known as a three-dimensional VRP with loading constraints (3D-VRP), is appropriate when transported commodities are not perfectly divisible, but they have fixed and heterogeneous dimensions. In the paper restrictions on allowable cargo positionings are also considered. These restrictions are derived from business practice and they extended the baseline 3D-VRP formulation as considered by Koloch and Kaminski (2010). In particular, we investigate how additional restrictions influence relative performance of two proposed optimization algorithms: the nested and the joint one. Performance of both methods is compared on artificial problems and on a big-scale real life case study.

  3. Collecting a sample of loggerhead sea turtle hatchlings before a natural emergence does not reduce nest productivity

    USGS Publications Warehouse

    Salmon, Michael; Carthy, Raymond R.; Lohmann, Catherine M. F.; Lohmann, Kenneth J.; Wyneken, Jeanette

    2012-01-01

     In numerous studies involving hatchling sea turtles, researchers have collected small numbers of hatchlings from nests a few hours before the turtles would otherwise have emerged naturally. This procedure makes it possible to do experiments in which the behavioral or physiological responses of numerous hatchlings must be tested in a limited period of time, and also allows hatchlings to be released back into the sea in time to migrate offshore before dawn. In principle, however, the procedure might inadvertently reduce nest productivity (the number of hatchlings that successfully leave the nest), if digging into a nest prior to emergence somehow reduces the ability of the remaining turtles to emerge. We compared nest productivity in 67 experimental loggerhead nests, from which we removed 10 hatchlings before a natural emergence, to 95 control nests left undisturbed before a natural emergence. The 2 groups showed no statistical differences in productivity. We conclude that taking a few hatchlings from a loggerhead nest shortly before a natural emergence has no negative impact on hatchling production if sampling is done with care at locations where there are few nest predators, and at sites where an emergence can be predicted because nest deposition dates are known.

  4. Linking species richness curves from non-contiguous sampling to contiguous-nested SAR: An empirical study

    NASA Astrophysics Data System (ADS)

    Lazarina, Maria; Kallimanis, Athanasios S.; Pantis, John D.; Sgardelis, Stefanos P.

    2014-11-01

    The species-area relationship (SAR) is one of the few generalizations in ecology. However, many different relationships are denoted as SARs. Here, we empirically evaluated the differences between SARs derived from nested-contiguous and non-contiguous sampling designs, using plants, birds and butterflies datasets from Great Britain, Greece, Massachusetts, New York and San Diego. The shape of SAR depends on the sampling scheme, but there is little empirical documentation on the magnitude of the deviation between different types of SARs and the factors affecting it. We implemented a strictly nested sampling design to construct nested-contiguous SAR (SACR), and systematic nested but non-contiguous, and random designs to construct non-contiguous species richness curves (SASRs for systematic and SACs for random designs) per dataset. The SACR lay below any SASR and most of the SACs. The deviation between them was related to the exponent f of the power law relationship between sampled area and extent. The lower the exponent f, the higher was the deviation between the curves. We linked SACR to SASR and SAC through the concept of "effective" area (Ae), i.e. the nested-contiguous area containing equal number of species with the accumulated sampled area (AS) of a non-contiguous sampling. The relationship between effective and sampled area was modeled as log(Ae) = klog(AS). A Generalized Linear Model was used to estimate the values of k from sampling design and dataset properties. The parameter k increased with the average distance between samples and with beta diversity, while k decreased with f. For both systematic and random sampling, the model performed well in predicting effective area in both the training set and in the test set which was totally independent from the training one. Through effective area, we can link different types of species richness curves based on sampling design properties, sampling effort, spatial scale and beta diversity patterns.

  5. Solving LR Conflicts Through Context Aware Scanning

    NASA Astrophysics Data System (ADS)

    Leon, C. Rodriguez; Forte, L. Garcia

    2011-09-01

    This paper presents a new algorithm to compute the exact list of tokens expected by any LR syntax analyzer at any point of the scanning process. The lexer can, at any time, compute the exact list of valid tokens to return only tokens in this set. In the case than more than one matching token is in the valid set, the lexer can resort to a nested LR parser to disambiguate. Allowing nested LR parsing requires some slight modifications when building the LR parsing tables. We also show how LR parsers can parse conflictive and inherently ambiguous languages using a combination of nested parsing and context aware scanning. These expanded lexical analyzers can be generated from high level specifications.

  6. Marine turtles are not fussy nesters: a novel test of small-scale nest site selection using structure from motion beach terrain information

    PubMed Central

    Leon, Javier X.; Gilby, Ben L.; Olds, Andrew D.; Schlacher, Thomas A.

    2017-01-01

    Background Nest selection is widely regarded as a key process determining the fitness of individuals and viability of animal populations. For marine turtles that nest on beaches, this is particularly pivotal as the nesting environment can significantly control reproductive success.The aim of this study was to identify the environmental attributes of beaches (i.e., morphology, vegetation, urbanisation) that may be associated with successful oviposition in green and loggerhead turtle nests. Methods We quantified the proximity of turtle nests (and surrounding beach locations) to urban areas, measured their exposure to artificial light, and used ultra-high resolution (cm-scale) digital surface models derived from Structure-from-Motion (SfM) algorithms, to characterise geomorphic and vegetation features of beaches on the Sunshine Coast, eastern Australia. Results At small spatial scales (i.e., <100 m), we found no evidence that turtles selected nest sites based on a particular suite of environmental attributes (i.e., the attributes of nest sites were not consistently different from those of surrounding beach locations). Nest sites were, however, typically characterised by occurring close to vegetation, on parts of the shore where the beach- and dune-face was concave and not highly rugged, and in areas with moderate exposure to artificial light. Conclusion This study used a novel empirical approach to identify the attributes of turtle nest sites from a broader ‘envelope’ of environmental nest traits, and is the first step towards optimizing conservation actions to mitigate, at the local scale, present and emerging human impacts on turtle nesting beaches. PMID:28070454

  7. Validation and clinical application of a nested PCR for paracoccidioidomycosis diagnosis in clinical samples from Colombian patients.

    PubMed

    Gaviria, Marcela; Rivera, Vanessa; Muñoz-Cadavid, Cesar; Cano, Luz Elena; Naranjo, Tonny Williams

    2015-01-01

    Paracoccidioidomycosis is a systemic and endemic mycosis, restricted to tropical and subtropical areas of Latin America. The infection is caused by the thermal dimorphic fungus Paracoccidioides brasiliensis and Paracoccidioides lutzii. The diagnosis of paracoccidioidomycosis is usually performed by microscopic examination, culture and immunodiagnostic tests to respiratory specimens, body fluids and/or biopsies; however these methods require laboratory personnel with experience and several days to produce a result. In the present study, we have validated and evaluated a nested PCR assay targeting the gene encoding the Paracoccidioides gp43 membrane protein in 191 clinical samples: 115 samples from patients with proven infections other than paracoccidioidomycosis, 51 samples as negative controls, and 25 samples from patients diagnosed with paracoccidioidomycosis. Additionally, the specificity of the nested PCR assay was also evaluated using purified DNA isolated from cultures of different microorganisms (n=35) previously identified by culture and/or sequencing. The results showed that in our hands, this nested PCR assay for gp43 protein showed specificity and sensitivity rates of 100%. The optimized nested PCR conditions in our laboratory allowed detection down to 1fg of P. brasiliensis DNA. Copyright © 2015 Elsevier Editora Ltda. All rights reserved.

  8. Detection of Toxoplasma gondii oocysts in soils in northwestern China using a new semi-nested PCR assay.

    PubMed

    Wang, Meng; Meng, Peng; Ye, Qiang; Pu, Yuan-Hua; Yang, Xiao-Yu; Luo, Jian-Xun; Zhang, Nian-Zhang; Zhang, De-Lin

    2014-09-28

    Toxoplasma gondii is a zoonotic pathogen that can infect a range of animals and humans. Ingestion of T. gondii oocysts in soil is a significant transmission route for humans and animals acquiring toxoplasmosis. In the present study, we developed a new semi-nested PCR method to determine T. gondii oocysts distribution in soils in northwestern China. The one tube semi-nested PCR assay was developed to detect the oocysts of T. gondii in soil, targeting the repetitive 529 bp fragment of T. gondii genomic DNA. Then a total of 268 soil samples, including 148 samples from Gansu Province and 120 samples from Qinghai Province, northwestern China, were examined by the semi-nested PCR method. One third of the positive samples were sequenced. The sensitivity of the semi-nested PCR assay was 10(2)  T. gondii oocysts in 5 g soil sample. Investigation of soil samples from northwestern China showed that 34 out of 268 soil samples (12.69%) were T. gondii positive. Sequences of the partial 529 bp fragments varied from 0-1.2% among the sequenced samples. The prevalence of T. gondii oocysts in soil from cities (24/163) was slightly higher than that in soils from pasturing areas (10/105) (P = 0.21). Among the different regions in cities, the prevalence of T. gondii oocysts in soils from parks was 14.15%, whereas that in soils from schools was 19.05%. The present study firstly reported the prevalence of T. gondii oocysts in soils in northwest China using a novel semi-nested PCR assay, which provided baseline data for the effective prevention and control of toxoplasmosis in this region.

  9. Molecular diagnosis of strongyloidiasis in a population of an endemic area through nested-PCR.

    PubMed

    Sharifdini, Meysam; Keyhani, Amir; Eshraghian, Mohammad Reza; Beigom Kia, Eshrat

    2018-01-01

    This study is aimed to diagnose and analyze strongyloidiasis in a population of an endemic area of Iran using nested-PCR, coupled with parasitological methods. Screening of strongyloidiasis infected people using reliable diagnostic techniques are essential to decrease the mortality and morbidity associated with this infection. Molecular methods have been proved to be highly sensitive and specific for detection of Strongyloides stercoralis in stool samples. A total of 155 fresh single stool samples were randomly collected from residents of north and northwest of Khouzestan Province, Iran. All samples were examined by parasitological methods including formalin-ether concentration and nutrient agar plate culture, and molecular method of nested-PCR. Infections with S. stercoralis were analyzed according to demographic criteria. Based on the results of nested-PCR method 15 cases (9.7%) were strongyloidiasis positive. Nested-PCR was more sensitive than parasitological techniques on single stool sampling. Elderly was the most important population index for higher infectivity with S. stercoralis . In endemic areas of S. stercoralis , old age should be considered as one of the most important risk factors of infection, especially among the immunosuppressed individuals.

  10. A Cladistic Analysis of Phenotypic Associations with Haplotypes Inferred from Restriction Endonuclease Mapping. IV. Nested Analyses with Cladogram Uncertainty and Recombination

    PubMed Central

    Templeton, A. R.; Sing, C. F.

    1993-01-01

    We previously developed an analytical strategy based on cladistic theory to identify subsets of haplotypes that are associated with significant phenotypic deviations. Our initial approach was limited to segments of DNA in which little recombination occurs. In such cases, a cladogram can be constructed from the restriction site data to estimate the evolutionary steps that interrelate the observed haplotypes to one another. The cladogram is then used to define a nested statistical design for identifying mutational steps associated with significant phenotypic deviations. The central assumption behind this strategy is that a mutation responsible for a particular phenotypic effect is embedded within the evolutionary history that is represented by the cladogram. The power of this approach depends on the accuracy of the cladogram in portraying the evolutionary history of the DNA region. This accuracy can be diminished both by recombination and by uncertainty in the estimated cladogram topology. In a previous paper, we presented an algorithm for estimating the set of likely cladograms and recombination events. In this paper we present an algorithm for defining a nested statistical design under cladogram uncertainty and recombination. Given the nested design, phenotypic associations can be examined using either a nested analysis of variance (for haploids or homozygous strains) or permutation testing (for outcrossed, diploid gene regions). In this paper we also extend this analytical strategy to include categorical phenotypes in addition to quantitative phenotypes. Some worked examples are presented using Drosophila data sets. These examples illustrate that having some recombination may actually enhance the biological inferences that may derived from a cladistic analysis. In particular, recombination can be used to assign a physical localization to a given subregion for mutations responsible for significant phenotypic effects. PMID:8100789

  11. Nested PCR Biases in Interpreting Microbial Community Structure in 16S rRNA Gene Sequence Datasets

    PubMed Central

    Yu, Guoqin; Fadrosh, Doug; Goedert, James J.; Ravel, Jacques; Goldstein, Alisa M.

    2015-01-01

    Background Sequencing of the PCR-amplified 16S rRNA gene has become a common approach to microbial community investigations in the fields of human health and environmental sciences. This approach, however, is difficult when the amount of DNA is too low to be amplified by standard PCR. Nested PCR can be employed as it can amplify samples with DNA concentration several-fold lower than standard PCR. However, potential biases with nested PCRs that could affect measurement of community structure have received little attention. Results In this study, we used 17 DNAs extracted from vaginal swabs and 12 DNAs extracted from stool samples to study the influence of nested PCR amplification of the 16S rRNA gene on the estimation of microbial community structure using Illumina MiSeq sequencing. Nested and standard PCR methods were compared on alpha- and beta-diversity metrics and relative abundances of bacterial genera. The effects of number of cycles in the first round of PCR (10 vs. 20) and microbial diversity (relatively low in vagina vs. high in stool) were also investigated. Vaginal swab samples showed no significant difference in alpha diversity or community structure between nested PCR and standard PCR (one round of 40 cycles). Stool samples showed significant differences in alpha diversity (except Shannon’s index) and relative abundance of 13 genera between nested PCR with 20 cycles in the first round and standard PCR (P<0.01), but not between nested PCR with 10 cycles in the first round and standard PCR. Operational taxonomic units (OTUs) that had low relative abundance (sum of relative abundance <0.167) accounted for most of the distortion (>27% of total OTUs in stool). Conclusions Nested PCR introduced bias in estimated diversity and community structure. The bias was more significant for communities with relatively higher diversity and when more cycles were applied in the first round of PCR. We conclude that nested PCR could be used when standard PCR does not work. However, rare taxa detected by nested PCR should be validated by other technologies. PMID:26196512

  12. Nested PCR Biases in Interpreting Microbial Community Structure in 16S rRNA Gene Sequence Datasets.

    PubMed

    Yu, Guoqin; Fadrosh, Doug; Goedert, James J; Ravel, Jacques; Goldstein, Alisa M

    2015-01-01

    Sequencing of the PCR-amplified 16S rRNA gene has become a common approach to microbial community investigations in the fields of human health and environmental sciences. This approach, however, is difficult when the amount of DNA is too low to be amplified by standard PCR. Nested PCR can be employed as it can amplify samples with DNA concentration several-fold lower than standard PCR. However, potential biases with nested PCRs that could affect measurement of community structure have received little attention. In this study, we used 17 DNAs extracted from vaginal swabs and 12 DNAs extracted from stool samples to study the influence of nested PCR amplification of the 16S rRNA gene on the estimation of microbial community structure using Illumina MiSeq sequencing. Nested and standard PCR methods were compared on alpha- and beta-diversity metrics and relative abundances of bacterial genera. The effects of number of cycles in the first round of PCR (10 vs. 20) and microbial diversity (relatively low in vagina vs. high in stool) were also investigated. Vaginal swab samples showed no significant difference in alpha diversity or community structure between nested PCR and standard PCR (one round of 40 cycles). Stool samples showed significant differences in alpha diversity (except Shannon's index) and relative abundance of 13 genera between nested PCR with 20 cycles in the first round and standard PCR (P<0.01), but not between nested PCR with 10 cycles in the first round and standard PCR. Operational taxonomic units (OTUs) that had low relative abundance (sum of relative abundance <0.167) accounted for most of the distortion (>27% of total OTUs in stool). Nested PCR introduced bias in estimated diversity and community structure. The bias was more significant for communities with relatively higher diversity and when more cycles were applied in the first round of PCR. We conclude that nested PCR could be used when standard PCR does not work. However, rare taxa detected by nested PCR should be validated by other technologies.

  13. Evaluation of efficiency of nested multiplex allele-specific PCR assay for detection of multidrug resistant tuberculosis directly from sputum samples.

    PubMed

    Mistri, S K; Sultana, M; Kamal, S M M; Alam, M M; Irin, F; Nessa, J; Ahsan, C R; Yasmin, M

    2016-05-01

    For an effective control of tuberculosis, rapid detection of multidrug resistant tuberculosis (MDR-TB) is necessary. Therefore, we developed a modified nested multiplex allele-specific polymerase chain reaction (MAS-PCR) method that enables rapid MDR-TB detection directly from sputum samples. The efficacy of this method was evaluated using 79 sputum samples collected from suspected tuberculosis patients. The performance of nested MAS-PCR method was compared with other MDR-TB detection methods like drug susceptibility testing (DST) and DNA sequencing. As rifampicin (RIF) resistance conforms to MDR-TB in greater than 90% cases, only the presence of RIF-associated mutations in rpoB gene was determined by DNA sequencing and nested MAS-PCR to detect MDR-TB. The concordance between nested MAS-PCR and DNA sequencing results was found to be 96·3%. When compared with DST, the sensitivity and specificity of nested MAS-PCR for RIF-resistance detection were determined to be 92·9 and 100% respectively. For developing- and high-TB burden countries, molecular-based tests have been recommended by the World Health Organization for rapid detection of MDR-TB. The results of this study indicate that, nested MAS-PCR assay might be a practical and relatively cost effective molecular method for rapid detection of MDR-TB from suspected sputum samples in developing countries with resource poor settings. © 2016 The Society for Applied Microbiology.

  14. Illustration Watermarking for Digital Images: An Investigation of Hierarchical Signal Inheritances for Nested Object-based Embedding

    DTIC Science & Technology

    2007-02-23

    approach for signal-level watermark inheritance. 15. SUBJECT TERMS EOARD, Steganography , Image Fusion, Data Mining, Image ...in watermarking algorithms , a program interface and protocol has been de - veloped, which allows control of the embedding and retrieval processes by the...watermarks in an image . Watermarking algorithm (DLL) Watermarking editor (Delphi) - User marks all objects: ci - class information oi - object instance

  15. Evaluation of hsp65 Nested PCR-Restriction Analysis (PRA) for Diagnosing Tuberculosis in a High Burden Country

    PubMed Central

    Macente, Sara; Fujimura Leite, Clarice Queico; Santos, Adolfo Carlos Barreto; Siqueira, Vera Lúcia Dias; Machado, Luzia Neri Cosmo; Marcondes, Nadir Rodrigues; Hirata, Mario Hiroyuki; Hirata, Rosário Dominguez Crespo

    2013-01-01

    Current study evaluated the hsp65 Nested PCR Restriction Fragment Length Polymorphism Analysis (hsp65 Nested PCR-PRA) to detect and identify Mycobacterium tuberculosis complex directly in clinical samples for a rapid and specific diagnosis of tuberculosis (TB). hsp65 Nested PCR-PRA was applied directly to 218 clinical samples obtained from 127 patients suspected of TB or another mycobacterial infection from July 2009 to July 2010. The hsp65 Nested PCR-PRA showed 100% sensitivity and 95.0 and 93.1% specificity in comparison with culture and microscopy (acid fast bacillus smear), respectively. hsp65 Nested PCR-PRA was shown to be a fast and reliable assay for diagnosing TB, which may contribute towards a fast diagnosis that could help the selection of appropriate chemotherapeutic and early epidemiological management of the cases which are of paramount importance in a high TB burden country. PMID:24260739

  16. [Sensitivity and specificity of nested PCR pyrosequencing in hepatitis B virus drug resistance gene testing].

    PubMed

    Sun, Shumei; Zhou, Hao; Zhou, Bin; Hu, Ziyou; Hou, Jinlin; Sun, Jian

    2012-05-01

    To evaluate the sensitivity and specificity of nested PCR combined with pyrosequencing in the detection of HBV drug-resistance gene. RtM204I (ATT) mutant and rtM204 (ATG) nonmutant plasmids mixed at different ratios were detected for mutations using nested-PCR combined with pyrosequencing, and the results were compared with those by conventional PCR pyrosequencing to analyze the linearity and consistency of the two methods. Clinical specimens with different viral loads were examined for drug-resistant mutations using nested PCR pyrosequencing and nested PCR combined with dideoxy sequencing (Sanger) for comparison of the detection sensitivity and specificity. The fitting curves demonstrated good linearity of both conventional PCR pyrosequencing and nested PCR pyrosequencing (R(2)>0.99, P<0.05). Nested PCR showed a better consistency with the predicted value than conventional PCR, and was superior to conventional PCR for detection of samples containing 90% mutant plasmid. In the detection of clinical specimens, Sanger sequencing had a significantly lower sensitivity than nested PCR pyrosequencing (92% vs 100%, P<0.01). The detection sensitivity of Sanger sequencing varied with the viral loads, especially in samples with low viral copies (HBV DNA ≤3log10 copies/ml), where the sensitivity was 78%, significantly lower than that of pyrosequencing (100%, P<0.01). Neither of the two methods yielded positive results for the negative control samples, suggesting their good specificity. Compared with nested PCR and Sanger sequencing method, nested PCR pyrosequencing has a higher sensitivity especially in clinical specimens with low viral copies, which can be important for early detection of HBV mutant strains and hence more effective clinical management.

  17. Soil attributes drive nest-site selection by the campo miner Geositta poeciloptera.

    PubMed

    Meireles, Ricardo Camargos de; Teixeira, João Paulo Gusmão; Solar, Ricardo; Vasconcelos, Bruno Nery F; Fernandes, Raphael B A; Lopes, Leonardo Esteves

    2018-01-01

    Substrate type is a key-factor in nest-site selection and nest architecture of burrowing birds. However, little is known about which factors drive nest-site selection for these species, especially in the tropics. We studied the influence of soil attributes on nest-site selection by the campo miner Geositta poeciloptera, an open grassland bird that builds its nests within soil cavities. For all nests found, we measured the depth of the nest cavity and the resistance of the soil to penetration, and identified the soil horizon in which the nest was located. In soil banks with nests, we collected soil samples for granulometric analysis around each nest cavity, while in soil banks without nests we collected these samples at random points. From 43 nests found, 86% were located in the deeper soil horizons (C-horizon), and only 14% in the shallower horizons (B-horizon). Granulometric analysis showed that the C-horizons possessed a high similar granulometric composition, with high silt and low clay contents. These characteristics are associated with a low degree of structural development of the soil, which makes it easier to excavate. Contrarily, soil resistance to penetration does not seem to be an important criterion for nest site selection, although nests in more resistant the soils tend to have shallower nest cavities. Among the soil banks analyzed, 40% of those without cavities possessed a larger proportion of B-horizon relative to the C-horizon, and their texture was more clayey. On the other hand, almost all soil banks containing nest cavities had a larger C-horizon and a silty texture, indicating that soil attributes drive nest-site selection by G. poeciloptera. Thus, we conclude that the patchy distribution of G. poeciloptera can attributed to the infrequent natural exposure of the C-horizon in the tropical region, where well developed, deep and permeable soils are more common.

  18. Soil attributes drive nest-site selection by the campo miner Geositta poeciloptera

    PubMed Central

    Teixeira, João Paulo Gusmão; Solar, Ricardo; Vasconcelos, Bruno Nery F.; Fernandes, Raphael B. A.; Lopes, Leonardo Esteves

    2018-01-01

    Substrate type is a key-factor in nest-site selection and nest architecture of burrowing birds. However, little is known about which factors drive nest-site selection for these species, especially in the tropics. We studied the influence of soil attributes on nest-site selection by the campo miner Geositta poeciloptera, an open grassland bird that builds its nests within soil cavities. For all nests found, we measured the depth of the nest cavity and the resistance of the soil to penetration, and identified the soil horizon in which the nest was located. In soil banks with nests, we collected soil samples for granulometric analysis around each nest cavity, while in soil banks without nests we collected these samples at random points. From 43 nests found, 86% were located in the deeper soil horizons (C-horizon), and only 14% in the shallower horizons (B-horizon). Granulometric analysis showed that the C-horizons possessed a high similar granulometric composition, with high silt and low clay contents. These characteristics are associated with a low degree of structural development of the soil, which makes it easier to excavate. Contrarily, soil resistance to penetration does not seem to be an important criterion for nest site selection, although nests in more resistant the soils tend to have shallower nest cavities. Among the soil banks analyzed, 40% of those without cavities possessed a larger proportion of B-horizon relative to the C-horizon, and their texture was more clayey. On the other hand, almost all soil banks containing nest cavities had a larger C-horizon and a silty texture, indicating that soil attributes drive nest-site selection by G. poeciloptera. Thus, we conclude that the patchy distribution of G. poeciloptera can attributed to the infrequent natural exposure of the C-horizon in the tropical region, where well developed, deep and permeable soils are more common. PMID:29381768

  19. Comparison of the diagnostic performance of microscopic examination with nested polymerase chain reaction for optimum malaria diagnosis in Upper Myanmar.

    PubMed

    Kang, Jung-Mi; Cho, Pyo-Yun; Moe, Mya; Lee, Jinyoung; Jun, Hojong; Lee, Hyeong-Woo; Ahn, Seong Kyu; Kim, Tae Im; Pak, Jhang Ho; Myint, Moe Kyaw; Lin, Khin; Kim, Tong-Soo; Na, Byoung-Kuk

    2017-03-16

    Accurate diagnosis of Plasmodium infection is crucial for prompt malaria treatment and surveillance. Microscopic examination has been widely applied as the gold standard for malaria diagnosis in most part of malaria endemic areas, but its diagnostic value has been questioned, particularly in submicroscopic malaria. In this study, the diagnostic performance of microscopic examination and nested polymerase chain reaction (PCR) was evaluated to establish optimal malaria diagnosis method in Myanmar. A total of 1125 blood samples collected from residents in the villages and towns located in Naung Cho, Pyin Oo Lwin, Tha Beik Kyin townships and Mandalay of Upper Myanmar were screened by microscopic examination and species-specific nested PCR method. Among the 1125 blood samples, 261 samples were confirmed to be infected with malaria by microscopic examination. Evaluation of the 1125 samples by species-specific nested PCR analysis revealed that the agreement between microscopic examination and nested PCR was 87.3% (261/299). Nested PCR successfully detected 38 Plasmodium falciparum or Plasmodium vivax infections, which were missed in microscopic examination. Microscopic examinations also either misdiagnosed the infected Plasmodium species, or did not detect mixed infections with different Plasmodium species in 31 cases. The nested PCR method is more reliable than conventional microscopic examination for the diagnosis of malaria infections, and this is particularly true in cases of mixed infections and submicroscopic infections. Given the observed higher sensitivity and specificity of nested PCR, the molecular method holds enormous promise in malaria diagnosis and species differentiation, and can be applied as an effective monitoring tool for malaria surveillance, control and elimination in Myanmar.

  20. Characteristics of the Least Bell's Vireo Nest Sites Along the Santa Ynez River, Santa Barbara County

    Treesearch

    Thomas E. Olson; M. Violet Gray

    1989-01-01

    Due primarily to alteration of riparian vegetation and nest parasitism by brown-headed cowbirds (Molothrus ater), the least Bell's vireo (Vireo bellii pusillus) has under-gone a tremendous decline in range and numbers since the 1920's. In 1987, we sampled vegetation at 32 nest sites to characterize nesting habitat of...

  1. Optimized nested polymerase chain reaction for antemortem detection of Mycobacteria in Amazon parrots (Amazona aestiva) and orange-winged Amazons (Amazona amazonica).

    PubMed

    Baquião, Arianne Costa; Luna, Janaina Oliveira; Medina, Aziz Orro; Sanfilippo, Luiz Francisco; de Faria, Maria Jacinta; dos Santos, Manuel Armando Azevedo

    2014-03-01

    The objectives of this study were to optimize nested polymerase chain reaction (PCR) for Mycobacterium avium complex and Mycobacterium tuberculosis complex and apply them on samples from parrots. Results were negative for the presence of these Mycobacterium in the samples, and nested PCR was specific, faster, and more sensitive than other tests, thereby justifying its use in antemortem diagnosis.

  2. Estimating nest detection probabilities for white-winged dove nest transects in Tamaulipas, Mexico

    USGS Publications Warehouse

    Nichols, J.D.; Tomlinson, R.E.; Waggerman, G.

    1986-01-01

    Nest transects in nesting colonies provide one source of information on White-winged Dove (Zenaida asiatica asiatica) population status and reproduction. Nests are counted along transects using standardized field methods each year in Texas and northeastern Mexico by personnel associated with Mexico's Office of Flora and Fauna, the Texas Parks and Wildlife Department, and the U.S. Fish and Wildlife Service. Nest counts on transects are combined with information on the size of nesting colonies to estimate total numbers of nests in sampled colonies. Historically, these estimates have been based on the actual nest counts on transects and thus have required the assumption that all nests lying within transect boundaries are detected (seen) with a probability of one. Our objectives were to test the hypothesis that nest detection probability is one and, if rejected, to estimate this probability.

  3. Calculating three loop ladder and V-topologies for massive operator matrix elements by computer algebra

    NASA Astrophysics Data System (ADS)

    Ablinger, J.; Behring, A.; Blümlein, J.; De Freitas, A.; von Manteuffel, A.; Schneider, C.

    2016-05-01

    Three loop ladder and V-topology diagrams contributing to the massive operator matrix element AQg are calculated. The corresponding objects can all be expressed in terms of nested sums and recurrences depending on the Mellin variable N and the dimensional parameter ε. Given these representations, the desired Laurent series expansions in ε can be obtained with the help of our computer algebra toolbox. Here we rely on generalized hypergeometric functions and Mellin-Barnes representations, on difference ring algorithms for symbolic summation, on an optimized version of the multivariate Almkvist-Zeilberger algorithm for symbolic integration, and on new methods to calculate Laurent series solutions of coupled systems of differential equations. The solutions can be computed for general coefficient matrices directly for any basis also performing the expansion in the dimensional parameter in case it is expressible in terms of indefinite nested product-sum expressions. This structural result is based on new results of our difference ring theory. In the cases discussed we deal with iterative sum- and integral-solutions over general alphabets. The final results are expressed in terms of special sums, forming quasi-shuffle algebras, such as nested harmonic sums, generalized harmonic sums, and nested binomially weighted (cyclotomic) sums. Analytic continuations to complex values of N are possible through the recursion relations obeyed by these quantities and their analytic asymptotic expansions. The latter lead to a host of new constants beyond the multiple zeta values, the infinite generalized harmonic and cyclotomic sums in the case of V-topologies.

  4. Nest survival modelling using a multi-species approach in forests managed for timber and biofuel feedstock

    USGS Publications Warehouse

    Loman, Zachary G.; Monroe, Adrian; Riffell, Samuel K.; Miller, Darren A.; Vilella, Francisco; Wheat, Bradley R.; Rush, Scott A.; Martin, James A.

    2018-01-01

    Switchgrass (Panicum virgatum) intercropping is a novel forest management practice for biomass production intended to generate cellulosic feedstocks within intensively managed loblolly pine‐dominated landscapes. These pine plantations are important for early‐successional bird species, as short rotation times continually maintain early‐successional habitat. We tested the efficacy of using community models compared to individual surrogate species models in understanding influences on nest survival. We analysed nest data to test for differences in habitat use for 14 bird species in plots managed for switchgrass intercropping and controls within loblolly pine (Pinus taeda) plantations in Mississippi, USA.We adapted hierarchical models using hyper‐parameters to incorporate information from both common and rare species to understand community‐level nest survival. This approach incorporates rare species that are often discarded due to low sample sizes, but can inform community‐level demographic parameter estimates. We illustrate use of this approach in generating both species‐level and community‐wide estimates of daily survival rates for songbird nests. We were able to include rare species with low sample size (minimum n = 5) to inform a hyper‐prior, allowing us to estimate effects of covariates on daily survival at the community level, then compare this with a single‐species approach using surrogate species. Using single‐species models, we were unable to generate estimates below a sample size of 21 nests per species.Community model species‐level survival and parameter estimates were similar to those generated by five single‐species models, with improved precision in community model parameters.Covariates of nest placement indicated that switchgrass at the nest site (<4 m) reduced daily nest survival, although intercropping at the forest stand level increased daily nest survival.Synthesis and applications. Community models represent a viable method for estimating community nest survival rates and effects of covariates while incorporating limited data for rarely detected species. Intercropping switchgrass in loblolly pine plantations slightly increased daily nest survival at the research plot scale (0.1 km2), although at a local scale (50 m2) switchgrass negatively influenced nest survival. A likely explanation is intercropping shifted community composition, favouring species with greater disturbance tolerance.

  5. Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR.

    PubMed

    Kim, Y H; Cho, K W; Youn, H Y; Yoo, H S; Han, H R

    2001-04-01

    A one step reverse transcription PCR (RT-PCR) combined nested PCR was set up to increase efficiency in the diagnosis of canine distemper virus (CDV) infection after developement of nested PCR. Two PCR primer sets were designed based on the sequence of nucleocapsid gene of CDV Onderstepoort strain. One-step RT-PCR with the outer primer pair was revealed to detect 10(2) PFU/ml. The sensitivity was increased hundredfold using the one-step RT-PCR combined with the nested PCR. Specificity of the PCR was also confirmed using other related canine virus and peripheral blood mononuclear cells (PBMC) and body secretes of healthy dogs. Of the 51 blood samples from dogs clinically suspected of CD, 45 samples were revealed as positive by one-step RT-PCR combined with nested PCR. However, only 15 samples were identified as positive with a single one step RT-PCR. Therefore approximately 60% increase in the efficiency of the diagnosis was observed by the combined method. These results suggested that one step RT-PCR combined with nested PCR could be a sensitive, specific, and practical method for diagnosis of CDV infection.

  6. Changes in plasma levels of adrenaline, noradrenaline, glucose, lactate and CO2 in the green turtle, Chelonia mydas, during peak period of nesting.

    PubMed

    Alkindi, A Y A; Al-Habsi, A A; Mahmoud, I Y

    2008-02-01

    Plasma concentrations of stress hormones [adrenaline (ADR), noradrenaline (NR)], lactate, glucose and CO2 were monitored during peak nesting period (May-October) at different phases of nesting in the green turtle, Chelonia mydas. These include, emergence from sea, excavating body and nest chambers, oviposition, covering and camouflaging the nest and then returning to sea. Turtles that completed all phases of nesting including oviposition before returning to sea were considered "successful" turtles, while those that completed all phases but failed to lay their eggs were "unsuccessful". Blood samples were taken from the cervical sinus within 5min of capture to avoid stress due to handling. The turtles were usually sampled for blood between 20:00 and 1:00h of nesting time to ensure uniformity in the sampling. Plasma ADR and NR values were highly significant (P<0.001) in successful turtles over emergence, excavating and unsuccessful turtles. Plasma glucose levels remained stable throughout the nesting phases while lactate levels were significantly higher in successful turtles over the other phases (P<0.05) which signifies anaerobic metabolism during nesting. Plasma CO2 values were negatively correlated with ADR and NR (r=-0.258, P=0.03; r=-0.304, P=0.010), respectively. Hematocrit was significantly higher in successful phase (P<0.05) compared to other phases, and this may signify a higher degree of stress in successful turtles. Body temperature were significantly lower (P<0.005) in the excavating phase compared to the other three phases. Overall, body temperatures were lower than sand temperatures around the nest, which may indicate a behavioral thermoregulation used by the turtles during nesting. This information will be of value to the ongoing conservation program at Ras Al-Hadd Reserve in the Sultanate of Oman.

  7. Genealogical Properties of Subsamples in Highly Fecund Populations

    NASA Astrophysics Data System (ADS)

    Eldon, Bjarki; Freund, Fabian

    2018-03-01

    We consider some genealogical properties of nested samples. The complete sample is assumed to have been drawn from a natural population characterised by high fecundity and sweepstakes reproduction (abbreviated HFSR). The random gene genealogies of the samples are—due to our assumption of HFSR—modelled by coalescent processes which admit multiple mergers of ancestral lineages looking back in time. Among the genealogical properties we consider are the probability that the most recent common ancestor is shared between the complete sample and the subsample nested within the complete sample; we also compare the lengths of `internal' branches of nested genealogies between different coalescent processes. The results indicate how `informative' a subsample is about the properties of the larger complete sample, how much information is gained by increasing the sample size, and how the `informativeness' of the subsample varies between different coalescent processes.

  8. Sensitivity of nested-PCR for plasmodium detection in pooled whole blood samples and its usefulness to blood donor screening in endemic areas.

    PubMed

    de Freitas, Daniel Roberto Coradi; Gomes, Luciano Teixeira; Fontes, Cor Jesus F; Tauil, Pedro Luiz; Pang, Lorrin W; Duarte, Elisabeth Carmen

    2014-04-01

    Transfusion-transmitted malaria is a severe disease with high fatality rate. Most Brazilian blood banks in the Amazon region perform malaria screening using microscopic examination (thick smears). Since low parasite concentrations are expected in asymptomatic blood donors a high sensitivity test should be used for donor screening. This study determined the sensitivity of a nested-PCR for plasmodium detection in pooled samples. We performed a one-stage criterion validation study with 21 positive samples pooled with samples from ten negative volunteer until three different concentrations were reached (0.33; 0.25; 0.20 parasites/μL - p/μL). Nested PCR was performed as described by Snounou et al. (1993). Sensitivities (and confidence intervals) were determined by stratum of final parasite concentration on the pooled samples. All samples with parasitemia values of 0.33 and 0.25 p/μL had 100% sensitivity (95%CI=86.3-100). One negative result was obtained from a sample with 0.20 p/μL sensitivity=95.2% (95%CI=76.2-99.9). Compared to parasitemia detectable under ideal conditions of thick smear, this nested-PCR in pooled sample was able to detect 40 times more parasites per microliter. Nested-PCR in pooled samples should be considered as a high sensitive alternative to thick smear for donor screening in blood banks at endemic regions. Local authorities need to assess cost:benefit advantages of this method compared to alternatives. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Comprehensive Comparison between Empty Nest and Non-Empty Nest Elderly: A Cross-Sectional Study among Rural Populations in Northeast China

    PubMed Central

    Chang, Ye; Guo, Xiaofan; Guo, Liang; Li, Zhao; Yang, Hongmei; Yu, Shasha; Sun, Guozhe; Sun, Yingxian

    2016-01-01

    This study aimed to comprehensively compare the general characteristics, lifestyles, serum parameters, ultrasonic cardiogram (UCG) parameters, depression, quality of life, and various comorbidities between empty nest and non-empty nest elderly among rural populations in northeast China. This analysis was based on our previous study which was conducted from January 2012 to August 2013, using a multistage, stratified, random cluster sampling scheme. The final analyzed sample consisted of 3208 participants aged no less than 60 years, which was further classified into three groups: non-empty nest group, empty nest group (living as a couple), and empty nest group (living alone). More than half of the participants were empty nest elderly (60.5%). There were no significant statistical differences for serum parameters, UCG parameters, lifestyles, dietary pattern, and scores of Patient Health Questionnaire-9 (PHQ-9) and World Health Organization Quality of Life questionnaire, abbreviated version (WHOQOL-BREF) among the three groups. Empty nest elderly showed no more risk for comorbidities such as general obesity, abdominal obesity, hyperuricemia, hyperhomocysteinemia, diabetes, dyslipidemia, left atrial enlargement (LAE), and stroke. Our study indicated that empty nest elderly showed no more risk for depression, low quality of life and comorbidities such as general obesity, abdominal obesity, hyperuricemia, hyperhomocysteinemia, diabetes, dyslipidemia, LAE, and stroke among rural populations in northeast China. PMID:27618905

  10. Comprehensive Comparison between Empty Nest and Non-Empty Nest Elderly: A Cross-Sectional Study among Rural Populations in Northeast China.

    PubMed

    Chang, Ye; Guo, Xiaofan; Guo, Liang; Li, Zhao; Yang, Hongmei; Yu, Shasha; Sun, Guozhe; Sun, Yingxian

    2016-08-27

    This study aimed to comprehensively compare the general characteristics, lifestyles, serum parameters, ultrasonic cardiogram (UCG) parameters, depression, quality of life, and various comorbidities between empty nest and non-empty nest elderly among rural populations in northeast China. This analysis was based on our previous study which was conducted from January 2012 to August 2013, using a multistage, stratified, random cluster sampling scheme. The final analyzed sample consisted of 3208 participants aged no less than 60 years, which was further classified into three groups: non-empty nest group, empty nest group (living as a couple), and empty nest group (living alone). More than half of the participants were empty nest elderly (60.5%). There were no significant statistical differences for serum parameters, UCG parameters, lifestyles, dietary pattern, and scores of Patient Health Questionnaire-9 (PHQ-9) and World Health Organization Quality of Life questionnaire, abbreviated version (WHOQOL-BREF) among the three groups. Empty nest elderly showed no more risk for comorbidities such as general obesity, abdominal obesity, hyperuricemia, hyperhomocysteinemia, diabetes, dyslipidemia, left atrial enlargement (LAE), and stroke. Our study indicated that empty nest elderly showed no more risk for depression, low quality of life and comorbidities such as general obesity, abdominal obesity, hyperuricemia, hyperhomocysteinemia, diabetes, dyslipidemia, LAE, and stroke among rural populations in northeast China.

  11. Evaluation of a nested-PCR for mycobacterium tuberculosis detection in blood and urine samples.

    PubMed

    da Cruz, Heidi Lacerda Alves; de Albuquerque Montenegro, Rosana; de Araújo Lima, Juliana Falcão; da Rocha Poroca, Diogo; da Costa Lima, Juliana Figueirêdo; Maria Lapa Montenegro, Lílian; Crovella, Sergio; Charifker Schindler, Haiana

    2011-01-01

    The polymerase chain reaction (PCR) and its variations, such as the nested-PCR, have been described as promising techniques for rapid diagnosis of tuberculosis (TB). With the aim of evaluating the usefulness of a nested-PCR method on samples of blood and urine of patients suspected of tuberculosis we analyzed 192 clinical samples, using as a molecular target the insertion element IS6110 specific of M. tuberculosis genome. Nested-PCR method showed higher sensitivity in patients with extrapulmonary tuberculosis (47.8% and 52% in blood and urine) when compared to patients with the pulmonary form of the disease (sensitivity of 29% and 26.9% in blood and urine), regardless of the type of biological sample used. The nested-PCR is a rapid technique that, even if not showing a good sensitivity, should be considered as a helpful tool especially in the extrapulmonary cases or in cases where confirmatory diagnosis is quite difficult to be achieved by routine methods. The performance of PCR-based techniques should be considered and tested in future works on other types of biological specimens besides sputum, like blood and urine, readily obtainable in most cases. The improving of M. tuberculosis nested-PCR detection in TB affected patients will give the possibility of an earlier detection of bacilli thus interrupting the transmission chain of the disease.

  12. An O(n(5)) algorithm for MFE prediction of kissing hairpins and 4-chains in nucleic acids.

    PubMed

    Chen, Ho-Lin; Condon, Anne; Jabbari, Hosna

    2009-06-01

    Efficient methods for prediction of minimum free energy (MFE) nucleic secondary structures are widely used, both to better understand structure and function of biological RNAs and to design novel nano-structures. Here, we present a new algorithm for MFE secondary structure prediction, which significantly expands the class of structures that can be handled in O(n(5)) time. Our algorithm can handle H-type pseudoknotted structures, kissing hairpins, and chains of four overlapping stems, as well as nested substructures of these types.

  13. Development and evaluation of a nested-PCR assay for Senecavirus A diagnosis.

    PubMed

    Feronato, Cesar; Leme, Raquel A; Diniz, Jaqueline A; Agnol, Alais Maria Dall; Alfieri, Alice F; Alfieri, Amauri A

    2018-02-01

    Senecavirus A (SVA) has been associated with vesicular disease in weaned and adult pigs and with high mortality of newborn piglets. This study aimed to establish a nested-PCR assay for the routine diagnosis of SVA infection. Tissue samples (n = 177) were collected from 37 piglets of 18 pig farms located in four different Brazilian states. For the nested-PCR, a primer set was defined to amplify an internal VP1 fragment of 316 bp of SVA genome. Of the 37 piglets, 15 (40.5%) and 23 (62.2%) were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. The SVA RNA was detected in 61/177 (34.5%) samples with the RT-PCR, while the nested-PCR assay showed 84/177 (47.5%) samples with the virus (p < 0.05). According to the herds, 11 (61.1%) and 16 (88.9%) of the 18 pig herds were positive for the SVA in the RT-PCR and nested-PCR assays, respectively. Nucleotide sequencing analysis revealed similarities of 98.7-100% among SVA Brazilian strains and of 86.6-98% with SVA strains from other countries. The nested-PCR assay in this study was suitable to recover the SVA RNA in biological specimens, piglets, and/or herds that were considered as negative in the RT-PCR assay, and is proposed for the routine investigation of the SVA infection in piglets, especially when other techniques are not available or when a great number of samples has to be examined.

  14. Cross-validation pitfalls when selecting and assessing regression and classification models.

    PubMed

    Krstajic, Damjan; Buturovic, Ljubomir J; Leahy, David E; Thomas, Simon

    2014-03-29

    We address the problem of selecting and assessing classification and regression models using cross-validation. Current state-of-the-art methods can yield models with high variance, rendering them unsuitable for a number of practical applications including QSAR. In this paper we describe and evaluate best practices which improve reliability and increase confidence in selected models. A key operational component of the proposed methods is cloud computing which enables routine use of previously infeasible approaches. We describe in detail an algorithm for repeated grid-search V-fold cross-validation for parameter tuning in classification and regression, and we define a repeated nested cross-validation algorithm for model assessment. As regards variable selection and parameter tuning we define two algorithms (repeated grid-search cross-validation and double cross-validation), and provide arguments for using the repeated grid-search in the general case. We show results of our algorithms on seven QSAR datasets. The variation of the prediction performance, which is the result of choosing different splits of the dataset in V-fold cross-validation, needs to be taken into account when selecting and assessing classification and regression models. We demonstrate the importance of repeating cross-validation when selecting an optimal model, as well as the importance of repeating nested cross-validation when assessing a prediction error.

  15. Nest marking behavior and chemical composition of olfactory cues involved in nest recognition in Megachile rotundata.

    PubMed

    Guédot, Christelle; Buckner, James S; Hagen, Marcia M; Bosch, Jordi; Kemp, William P; Pitts-Singer, Theresa L

    2013-08-01

    In-nest observations of the solitary bee, Megachile rotundata (F.), revealed that nesting females apply olfactory cues to nests for nest recognition. On their way in and out of the nest, females drag the abdomen along the entire length of the nest, and sometimes deposit fluid droplets from the tip of the abdomen. The removal of bee-marked sections of the nest resulted in hesitation and searching behavior by females, indicating the loss of olfactory cues used for nest recognition. Chemical analysis of female cuticles and the deposits inside marked nesting tubes revealed the presence of hydrocarbons, wax esters, fatty aldehydes, and fatty alcohol acetate esters. Chemical compositions were similar across tube samples, but proportionally different from cuticular extracts. These findings reveal the importance of lipids as chemical signals for nest recognition and suggest that the nest-marking cues are derived from a source in addition to, or other than, the female cuticle.

  16. Cost-efficient designs for three-arm trials with treatment delivered by health professionals: Sample sizes for a combination of nested and crossed designs

    PubMed Central

    Moerbeek, Mirjam

    2018-01-01

    Background This article studies the design of trials that compare three treatment conditions that are delivered by two types of health professionals. The one type of health professional delivers one treatment, and the other type delivers two treatments, hence, this design is a combination of a nested and crossed design. As each health professional treats multiple patients, the data have a nested structure. This nested structure has thus far been ignored in the design of such trials, which may result in an underestimate of the required sample size. In the design stage, the sample sizes should be determined such that a desired power is achieved for each of the three pairwise comparisons, while keeping costs or sample size at a minimum. Methods The statistical model that relates outcome to treatment condition and explicitly takes the nested data structure into account is presented. Mathematical expressions that relate sample size to power are derived for each of the three pairwise comparisons on the basis of this model. The cost-efficient design achieves sufficient power for each pairwise comparison at lowest costs. Alternatively, one may minimize the total number of patients. The sample sizes are found numerically and an Internet application is available for this purpose. The design is also compared to a nested design in which each health professional delivers just one treatment. Results Mathematical expressions show that this design is more efficient than the nested design. For each pairwise comparison, power increases with the number of health professionals and the number of patients per health professional. The methodology of finding a cost-efficient design is illustrated using a trial that compares treatments for social phobia. The optimal sample sizes reflect the costs for training and supervising psychologists and psychiatrists, and the patient-level costs in the three treatment conditions. Conclusion This article provides the methodology for designing trials that compare three treatment conditions while taking the nesting of patients within health professionals into account. As such, it helps to avoid underpowered trials. To use the methodology, a priori estimates of the total outcome variances and intraclass correlation coefficients must be obtained from experts’ opinions or findings in the literature. PMID:29316807

  17. Incidence of pulmonary aspergillosis and correlation of conventional diagnostic methods with nested PCR and real-time PCR assay using BAL fluid in intensive care unit patients.

    PubMed

    Zarrinfar, Hossein; Makimura, Koichi; Satoh, Kazuo; Khodadadi, Hossein; Mirhendi, Hossein

    2013-05-01

    Although the incidence of invasive aspergillosis in the intensive care unit (ICU) is scarce, it has emerged as major problems in critically ill patients. In this study, the incidence of pulmonary aspergillosis (PA) in ICU patients has evaluated and direct microscopy and culture has compared with nested polymerase chain reaction (PCR) and real-time PCR for detection of Aspergillus fumigatus and A. flavus in bronchoalveolar lavage (BAL) samples of the patients. Thirty BAL samples obtained from ICU patients during a 16-month period were subjected to direct examinations on 20% potassium hydroxide (KOH) and culture on two culture media. Nested PCR targeting internal transcribed spacer ribosomal DNA and TaqMan real-time PCR assay targeting β-tubulin gene were used for the detection of A. fumigatus and A. flavus. Of 30 patients, 60% were men and 40% were women. The diagnosis of invasive PA was probable in 1 (3%), possible in 11 (37%), and not IPA in 18 (60%). Nine samples were positive in nested PCR including seven samples by A. flavus and two by A. fumigatus specific primers. The lowest amount of DNA that TaqMan real-time PCR could detect was ≥40 copy numbers. Only one of the samples had a positive result of A. flavus real-time PCR with Ct value of 37.5. Although a significant number of specimens were positive in nested PCR, results of this study showed that establishment of a correlation between the conventional methods with nested PCR and real-time PCR needs more data confirmed by a prospective study with a larger sample group. © 2013 Wiley Periodicals, Inc.

  18. Cuckoo Search Algorithm Based on Repeat-Cycle Asymptotic Self-Learning and Self-Evolving Disturbance for Function Optimization

    PubMed Central

    Wang, Jie-sheng; Li, Shu-xia; Song, Jiang-di

    2015-01-01

    In order to improve convergence velocity and optimization accuracy of the cuckoo search (CS) algorithm for solving the function optimization problems, a new improved cuckoo search algorithm based on the repeat-cycle asymptotic self-learning and self-evolving disturbance (RC-SSCS) is proposed. A disturbance operation is added into the algorithm by constructing a disturbance factor to make a more careful and thorough search near the bird's nests location. In order to select a reasonable repeat-cycled disturbance number, a further study on the choice of disturbance times is made. Finally, six typical test functions are adopted to carry out simulation experiments, meanwhile, compare algorithms of this paper with two typical swarm intelligence algorithms particle swarm optimization (PSO) algorithm and artificial bee colony (ABC) algorithm. The results show that the improved cuckoo search algorithm has better convergence velocity and optimization accuracy. PMID:26366164

  19. A Minimally Invasive Method for Sampling Nest and Roost Cavities for Fungi: a Novel Approach to Identify the Fungi Associated with Cavity-Nesting Birds

    Treesearch

    Michelle A. Jusino; Daniel Lindner; John K. Cianchetti; Adam T. Grisé; Nicholas J. Brazee; Jeffrey R. Walters

    2014-01-01

    Relationships among cavity-nesting birds, trees, and wood decay fungi pose interesting management challenges and research questions in many systems. Ornithologists need to understand the relationships between cavity-nesting birds and fungi in order to understand the habitat requirements of these birds. Typically, researchers rely on fruiting body surveys to identify...

  20. [Effect of landscape change on the structure of the sting-less bee community (Hymenoptera: Apidae) in Meta, Colombia].

    PubMed

    Nates-Parra, Guiomar; Palacios, Eliana; Parra-H, Alejandro

    2008-09-01

    Stingless bees represent one of the most diversified components of the natural Apoidea fauna of pollinators in the tropics. They use diverse kinds of substrates and inhabit varied habitats. Some species are typical for some natural either artificial place. The landscape alteration were this group of bees nests, has and important impact on the natural composition of its community structure, fact which is reflected in the nest density. We analyzed the structure composition of the stingless bees' community in three environments in the Colombian Ilanos piedmont, an important region that represents the transition between Andean ecosystems and a savannah that is seriously threatened by cattle practices. We made systematic samples in secondary forest, agro-ecosystems and urban areas, recording the presence of 204 nests from 11 genera (24 species). The nest density per landscape was heterogeneous and never higher than 16 nests/Ha. We observed two nesting patterns and an effect of sampling criterion on the measured biodiversity.

  1. Factors influencing nesting success of king eiders on northern Alaska's Coastal Plain

    USGS Publications Warehouse

    Bentzen, R.L.; Powell, A.N.; Suydam, R.S.

    2008-01-01

    King eider (Somateria spectabilis) populations have declined markedly in recent decades for unknown reasons. Nest survival is one component of recruitment, and a female's chance of reproductive success increases with her ability to choose an appropriate nesting strategy. We estimated variation in daily nest survival of king eiders at 2 sites, Teshekpuk and Kuparuk, Alaska, USA, 2002-2005. We evaluated both a priori and exploratory competing models of nest survival that considered importance of nest concealment, seclusion, and incubation constancy as strategies to avoid 2 primary egg predators, avian (Larus spp., Stercorarius spp., and Corvus corax) and fox (Alopex lagopus). We used generalized nonlinear techniques to examine factors affecting nest survival rates and information-theoretic approaches to select among competing models. Estimated nest survival, accounting for a nest visitation effect, varied considerably across sites and years (0.21-0.57); however, given our small sample size, much of this variation maybe attributable to sampling variation (??process = 0.007, 95% CI: 0.003-0.070). Nest survival was higher at Kuparuk than Teshekpuk in all years; however, due to the correlative nature of our data, we cannot determine the underlying causes with any certainty. We found mixed support for the concealed breeding strategy, females derived no benefit from nesting in areas with more willow (Salix spp.; measure of concealment) except that the observer effect diminished as willow cover increased. We suggest these patterns are due to conflicting predation pressures. Nest survival was not higher on islands (measure of seclusion) or with increased incubation constancy but was higher post-fox removal, indicating that predator control on breeding grounds could be a viable management option. Nest survival was negatively affected by our nest visitations, most likely by exposing the nest to avian scavengers. We recommend precautions be taken to limit the effects of nest visits in future studies and to consider them as a possible negative bias in estimated nest survival. Future models of the impacts of development within the breeding grounds of king eider should consider the influence of humans in the vicinity of nests.

  2. The effects of ant nests on soil fertility and plant performance: a meta-analysis.

    PubMed

    Farji-Brener, Alejandro G; Werenkraut, Victoria

    2017-07-01

    Ants are recognized as one of the major sources of soil disturbance world-wide. However, this view is largely based on isolated studies and qualitative reviews. Here, for the first time, we quantitatively determined whether ant nests affect soil fertility and plant performance, and identified the possible sources of variation of these effects. Using Bayesian mixed-models meta-analysis, we tested the hypotheses that ant effects on soil fertility and plant performance depend on the substrate sampled, ant feeding type, latitude, habitat and the plant response variable measured. Ant nests showed higher nutrient and cation content than adjacent non-nest soil samples, but similar pH. Nutrient content was higher in ant refuse materials than in nest soils. The fertilizer effect of ant nests was also higher in dry habitats than in grasslands or savannas. Cation content was higher in nests of plant-feeding ants than in nests of omnivorous species, and lower in nests from agro-ecosystems than in nests from any other habitat. Plants showed higher green/root biomass and fitness on ant nests soils than in adjacent, non-nest sites; but plant density and diversity were unaffected by the presence of ant nests. Root growth was particularly higher in refuse materials than in ant nest soils, in leaf-cutting ant nests and in deserts habitats. Our results confirm the major role of ant nests in influencing soil fertility and vegetation patterns and provide information about the factors that mediate these effects. First, ant nests improve soil fertility mainly through the accumulation of refuse materials. Thus, different refuse dump locations (external or in underground nest chambers) could benefit different vegetation life-forms. Second, ant nests could increase plant diversity at larger spatial scales only if the identity of favoured plants changes along environmental gradients (i.e. enhancing β-diversity). Third, ant species that feed on plants play a relevant role fertilizing soils, which may balance their known influence as primary consumers. Fourth, the effects of ant nests as fertility islands are larger in arid lands, possibly because fertility is intrinsically lower in these habitats. Overall, this study provide novel and quantitative evidence confirming that ant nests are key soil modifiers, emphasizing their role as ecological engineers. © 2017 The Authors. Journal of Animal Ecology © 2017 British Ecological Society.

  3. Probability of detection of nests and implications for survey design

    USGS Publications Warehouse

    Smith, P.A.; Bart, J.; Lanctot, Richard B.; McCaffery, B.J.; Brown, S.

    2009-01-01

    Surveys based on double sampling include a correction for the probability of detection by assuming complete enumeration of birds in an intensively surveyed subsample of plots. To evaluate this assumption, we calculated the probability of detecting active shorebird nests by using information from observers who searched the same plots independently. Our results demonstrate that this probability varies substantially by species and stage of the nesting cycle but less by site or density of nests. Among the species we studied, the estimated single-visit probability of nest detection during the incubation period varied from 0.21 for the White-rumped Sandpiper (Calidris fuscicollis), the most difficult species to detect, to 0.64 for the Western Sandpiper (Calidris mauri), the most easily detected species, with a mean across species of 0.46. We used these detection probabilities to predict the fraction of persistent nests found over repeated nest searches. For a species with the mean value for detectability, the detection rate exceeded 0.85 after four visits. This level of nest detection was exceeded in only three visits for the Western Sandpiper, but six to nine visits were required for the White-rumped Sandpiper, depending on the type of survey employed. Our results suggest that the double-sampling method's requirement of nearly complete counts of birds in the intensively surveyed plots is likely to be met for birds with nests that survive over several visits of nest searching. Individuals with nests that fail quickly or individuals that do not breed can be detected with high probability only if territorial behavior is used to identify likely nesting pairs. ?? The Cooper Ornithological Society, 2009.

  4. Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples.

    PubMed

    Foord, A J; Heine, H G; Pritchard, L I; Lunt, R A; Newberry, K M; Rootes, C L; Boyle, D B

    2006-07-01

    To evaluate and implement molecular diagnostic tests for the detection of lyssaviruses in Australia. A published hemi-nested reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of all lyssavirus genotypes was modified to a fully nested RT-PCR format and compared with the original assay. TaqMan assays for the detection of Australian bat lyssavirus (ABLV) were compared with both the nested and hemi-nested RT-PCR assays. The sequences of RT-PCR products were determined to assess sequence variations of the target region (nucleocapsid gene) in samples of ABLV originating from different regions. The nested RT-PCR assay was highly analytically specific, and at least as analytically sensitive as the hemi-nested assay. The TaqMan assays were highly analytically specific and more analytically sensitive than either RT-PCR assay, with a detection level of approximately 10 genome equivalents per microl. Sequence of the first 544 nucleotides of the nucleocapsid protein coding sequence was obtained from all samples of ABLV received at Australian Animal Health Laboratory during the study period. The nested RT-PCR provided a means for molecular diagnosis of all tested genotypes of lyssavirus including classical rabies virus and Australian bat lyssavirus. The published TaqMan assay proved to be superior to the RT-PCR assays for the detection of ABLV in terms of analytical sensitivity. The TaqMan assay would also be faster and cross contamination is less likely. Nucleotide sequence analyses of samples of ABLV from a wide geographical range in Australia demonstrated the conserved nature of this region of the genome and therefore the suitability of this region for molecular diagnosis.

  5. Rapid direct identification of Cryptococcus neoformans from pigeon droppings by nested PCR using CNLAC1 gene.

    PubMed

    Chae, H S; Park, G N; Kim, S H; Jo, H J; Kim, J T; Jeoung, H Y; An, D J; Kim, N H; Shin, B W; Kang, Y I; Chang, K S

    2012-08-01

    Isolation and identification of Cryptococcus neoformans and pathogenic yeast-like fungi from pigeon droppings has been taken for a long time and requires various nutrients for its growth. In this study, we attempted to establish a rapid direct identification method of Cr. neoformans from pigeon dropping samples by nested-PCR using internal transcribed spacer (ITS) CAP64 and CNLAC1 genes, polysaccharide capsule gene and laccase-associated gene to produce melanin pigment, respectively, which are common genes of yeasts. The ITS and CAP64 genes were amplified in all pathogenic yeasts, but CNLAC1 was amplified only in Cr. neoformans. The ITS gene was useful for yeast genotyping depending on nucleotide sequence. Homology of CAP64 genes among the yeasts were very high. The specificity of PCR using CNLAC1 was demonstrated in Cr. neoformans environmental strains but not in other yeast-like fungi. The CNLAC1 gene was detected in 5 serotypes of Cr. neoformans. The nested-PCR amplified up to 10(-11) μg of the genomic DNA and showed high sensitivity. All pigeon droppings among 31 Cr. neoformans-positive samples were positive and all pigeon droppings among 348 Cr. neoformans-negative samples were negative by the direct nested-PCR. In addition, after primary enrichment of pigeon droppings in Sabouraud dextrose broth, all Cr. neoformans-negative samples were negative by the nested-PCR, which showed high specificity. The nested-PCR showed high sensitivity without culture of pigeon droppings. Nested-PCR using CNLAC1 provides a rapid and reliable molecular diagnostic method to overcome weak points such as long culture time of many conventional methods.

  6. Detection of Mycobacterium bovis in Bovine and Bubaline Tissues Using Nested-PCR for TbD1

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A. R.; Jorge, Kláudia S. G.; Ramos, Carlos Alberto N.; Filho, Antonio Francisco S.; Vidal, Carlos Eugênio S.; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F.; Júnior, Antônio A. F.; Silva, Marcio R.; Neto, José Diomedes B.; Cerqueira, Valíria D.; Zumárraga, Martín J.; Araújo, Flábio R.

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:24618787

  7. Detection of Mycobacterium bovis in bovine and bubaline tissues using nested-PCR for TbD1.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Kláudia S G; Ramos, Carlos Alberto N; Filho, Antonio Francisco S; Vidal, Carlos Eugênio S; Roxo, Eliana; Nishibe, Christiane; Almeida, Nalvo F; Júnior, Antônio A F; Silva, Marcio R; Neto, José Diomedes B; Cerqueira, Valíria D; Zumárraga, Martín J; Araújo, Flábio R

    2014-01-01

    In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  8. Organochlorine contaminant exposure and reproductive success of Black-Crowned Night Herons (Nycticorax nycticorax) nesting in Baltimore Harbor, Maryland

    USGS Publications Warehouse

    Rattner, B.A.; McGowan, P.C.; Hatfield, J.S.; Hong, C.-S.; Chu, S.G.

    2001-01-01

    The declining size of the Baltimore Harbor black-crowned night heron (Nycticorax nycticorax) colony has been hypothesized to be linked to PCB exposure. In 1998, a 'sample egg' was collected from 65 black-crowned night heron nests (each containing > three eggs) for contaminant analysis, and the remaining eggs in these 65 nests, plus four two-egg nests, were monitored for hatching and fledging success. Eggs were also collected from 12 nests at Holland Island, a reference site in southern Chesapeake Bay. Samples were analyzed for 26 organochlorine pesticides and metabolites, and 145 PCB congeners. Pesticide and metabolite concentrations, including p,p'-DDE, were well below thresholds associated with adverse reproductive effects at both sites. Average concentration of total PCBs, 12 Ah receptor-active PCB congeners, and toxic equivalents in eggs from Baltimore Harbor were greater (up to 35- fold) than that observed in Holland Island samples. Overall nest success at the Baltimore Harbor heronry was estimated by the Mayfield method to be 0.74, and the mean number of young fledged/hen was 2.05, which is within published productivity estimates for maintaining a stable black-crowned night heron population. Using logistic regression, no significant relationships were found between organochlorine contaminant concentrations in sample eggs and hatching, fledging, or overall reproductive success. Processes other than poor reproduction (e.g., low post- fledging survival, emigration, habitat degradation) may be responsible for the declining size of the Baltimore Harbor colony.

  9. Toxic industrial deposit remediation by ant activity

    NASA Astrophysics Data System (ADS)

    Jilkova, Veronika; Frouz, Jan

    2016-04-01

    Toxic industrial deposits are often contaminated by heavy metals and the substrates have low pH values. In such systems, soil development is thus slowed down by high toxicity and acidic conditions which are unfavourable to soil fauna. Ants (Hymenoptera, Formicidae) are considered tolerant to heavy metal pollution and are known to increase organic matter content and microbial activity in their nests. Here, we focused on soil remediation caused by three ant species (Formica sanguinea, Lasius niger, and Tetramorium sp.) in an ore-washery sedimentation basin near Chvaletice (Czech Republic). Soil samples were taken from the centre of ant nests and from the nest surroundings (>3 m from nests). Samples were then analyzed for microbial activity and biomass and contents of organic matter and nutrients. As a result, ant species that most influenced soil properties was F. sanguinea as there were higher microbial activity and total nitrogen and ammonia contents in ant nests than in the surrounding soil. We expected such a result because F. sanguinea builds conspicuous large nests and is a carnivorous species that brings substantial amounts of nitrogen in insect prey to their nests. Effects of the other two ant species might be lower because of smaller nests and different feeding habits as they rely mainly on honeydew from aphids or on plant seeds that do not contain much nutrients.

  10. [Application of Nested PCR in the Diagnosis of Imported Plasmodium Ovale Infection].

    PubMed

    Huang, Bing-cheng; Xu, Chao; Li, Jin; Xiao, Ting; Yin, Kun; Liu, Gong-zhen; Wang, Wei-yan; Zhao, Gui-hua; Wei, Yan-bin; Wang, Yong-bin; Zhao, Chang-lei; Wei, Qing-kuan

    2015-02-01

    To identity Plasmodium ovale infection by 18S rRNA gene nested PCR. Whole blood and filter paper blood samples of malaria patients in Shandong Province were collected during 2012-2013. The parasites were observed under a microscope with Giemsa staining. The genome DNA of blood samples were extracted as PCR templates. Genus- and species-specific primers were designed according to the Plasmodium 18S rRNA gene sequences. Plasmodium ovale-positive specimens were identified by nested PCR as well as verified by sequencing. There were 7 imported cases of P. ovale infection in the province during 2012-2013. Nested PCR results showed that the P. ovale specific band (800 bp) was amplified in all the 7 specimens. Blast results indicated that the PCR products were consistent with the Plasmodium ovale reference sequence in GenBank. Seven imported cases of ovale malaria in Shandong Province in 2012-2013 are confirmed by nested PCR.

  11. Conducting Three-Level Longitudinal Analyses

    ERIC Educational Resources Information Center

    Peugh, James L.; Heck, Ronald H.

    2017-01-01

    Researchers in the field of early adolescence interested in quantifying the environmental influences on a response variable of interest over time would use cluster sampling (i.e., obtaining repeated measures from students nested within classrooms and/or schools) to obtain the needed sample size. The resulting longitudinal data would be nested at…

  12. Nest Mosquito Trap quantifies contact rates between nesting birds and mosquitoes.

    PubMed

    Caillouët, Kevin A; Riggan, Anna E; Rider, Mark; Bulluck, Lesley P

    2012-06-01

    Accurate estimates of host-vector contact rates are required for precise determination of arbovirus transmission intensity. We designed and tested a novel mosquito collection device, the Nest Mosquito Trap (NMT), to collect mosquitoes as they attempt to feed on unrestrained nesting birds in artificial nest boxes. In the laboratory, the NMT collected nearly one-third of the mosquitoes introduced to the nest boxes. We then used these laboratory data to estimate our capture efficiency of field-collected bird-seeking mosquitoes collected over 66 trap nights. We estimated that 7.5 mosquitoes per trap night attempted to feed on nesting birds in artificial nest boxes. Presence of the NMT did not have a negative effect on avian nest success when compared to occupied nest boxes that were not sampled with the trap. Future studies using the NMT may elucidate the role of nestlings in arbovirus transmission and further refine estimates of nesting bird and vector contact rates. © 2012 The Society for Vector Ecology.

  13. Nest Mosquito Trap quantifies contact rates between nesting birds and mosquitoes

    PubMed Central

    Caillouët, Kevin A.; Riggan, Anna E.; Rider, Mark; Bulluck, Lesley P.

    2012-01-01

    Accurate estimates of host-vector contact rates are required for precise determination of arbovirus transmission intensity. We designed and tested a novel mosquito collection device, the Nest Mosquito Trap (NMT), to collect mosquitoes as they attempt to feed on unrestrained nesting birds in artificial nest boxes. In the laboratory, the NMT collected nearly one-third of the mosquitoes introduced to the nest boxes. We then used these laboratory data to estimate our capture efficiency of field-collected bird-seeking mosquitoes collected over 66 trap nights. We estimated that 7.5 mosquitoes per trap night attempted to feed on nesting birds in artificial nest boxes. Presence of the NMT did not have a negative effect on avian nest success when compared to occupied nest boxes that were not sampled with the trap. Future studies using the NMT may elucidate the role of nestlings in arbovirus transmission and further refine estimates of nesting bird and vector contact rates. PMID:22548555

  14. Use of radio-telemetry to reduce bias in nest searching

    USGS Publications Warehouse

    Powell, L.A.; Lang, J.D.; Krementz, D.G.; Conroy, M.J.

    2005-01-01

    We used traditional searching, as well as radio-telemetry, to find 125 Wood Thrush (Hylocichla mustelina) nests during 1994?1996 at the Piedmont National Wildlife Refuge in Georgia, USA. We compared daily nest survival rates for 66 nests of radio-marked birds with 59 nests of birds found through systematic searching. By using radio-telemetry, we found Wood Thrush nests in higher elevation pine habitats, in addition to the more usual hardwood forests with moist soils. We found nests of radio-marked birds farther from streams than nests found by systematic searching. Thirty-two percent of radio-marked birds' nests were found at the tops of slopes, compared to 15% of the nests found by traditional searching. In addition, radio-marked birds generally moved up-slope for re-nesting attempts. Although the distribution of nests found with telemetry and searching varied, daily nest survival did not vary between the two groups. Radio-telemetry provided new information about Wood Thrush nesting habitats. We believe radio-telemetry can be a valuable addition to traditional searching techniques; it has the potential to provide a sample of nests free from a priori habitat biases.

  15. Leaf Cutter Ant (Atta cephalotes) Soil Modification and In Situ CO2 Gas Dynamics in a Neotropical Wet Forest

    NASA Astrophysics Data System (ADS)

    Fernandez Bou, A. S.; Carrasquillo Quintana, O.; Dierick, D.; Harmon, T. C.; Johnson, S.; Schwendenmann, L.; Zelikova, T. J.

    2016-12-01

    The goal of this work is to advance our understanding of soil carbon cycling in highly productive neotropical wet forests. More specifically, we are investigating the influence of leaf cutter ants (LCA) on soil CO2 gas dynamics in primary and secondary forest soils at La Selva Biological Station, Costa Rica. LCA are the dominant herbivore in tropical Americas, responsible for as much as 50% of the total herbivory. Their presence is increasing and their range is expanding because of forest fragmentation and other human impacts. We installed gas sampling wells in LCA (Atta cephalotes) nest and control sites (non-nests in the same soil and forest settings). The experimental design encompassed land cover (primary and secondary forest) and soil type (residual and alluvial). We collected gas samples monthly over an 18-month period. Several of the LCA nests were abandoned during this period. Nevertheless, we continued to sample these sites for LCA legacy effects. In several of the sites, we also installed sensors to continuously monitor soil moisture content, temperature, and CO2 levels. Within the 18-month period we conducted a 2-month field campaign to collect soil and nest vent CO2 efflux data from 3 of the nest-control pairs. Integrating the various data sets, we observed that for most of the sites nest and control soils behaved similarly during the tropical dry season. However, during the wet season gas well CO2 concentrations increased in the control sites while levels in the nests remained at dry season levels. This outcome suggests that ants modify soil gas transport properties (e.g., tortuosity). In situ time series and efflux sampling campaign data corroborated these findings. Abandoned nest CO2 levels were similar to those of the active nests, supporting the notion of a legacy effect from LCA manipulations. For this work, the period of abandonment was relatively short (several months to 1 year maximum), which appears to be insufficient for estimating the duration effect. Overall, these results demonstrate that LCA exert a significant effect on carbon cycling in rain forest soils.

  16. A Constraint-Based Planner for Data Production

    NASA Technical Reports Server (NTRS)

    Pang, Wanlin; Golden, Keith

    2005-01-01

    This paper presents a graph-based backtracking algorithm designed to support constrain-tbased planning in data production domains. This algorithm performs backtracking at two nested levels: the outer- backtracking following the structure of the planning graph to select planner subgoals and actions to achieve them and the inner-backtracking inside a subproblem associated with a selected action to find action parameter values. We show this algorithm works well in a planner applied to automating data production in an ecological forecasting system. We also discuss how the idea of multi-level backtracking may improve efficiency of solving semi-structured constraint problems.

  17. Highly controlled nest homeostasis of honey bees helps deactivate phenolics in nectar

    NASA Astrophysics Data System (ADS)

    Liu, Fanglin; He, Jianzhong; Fu, Wenjun

    2005-06-01

    Honey bees have a highly developed nest homeostasis, for example, maintaining low CO2 levels and stable nest temperatures at 35°C.We investigate the role of nest homeostasis in deactivating phenolic compounds present in the nectar of Aloe littoralis. We show that the phenolic content in nectar was reduced (from 0.65% to 0.49%) after nectar was incubated in a nest of Apis cerana, and that it was reduced still more (from 0.65% to 0.37%) if nectar was mixed with hypopharyngeal gland proteins (HGP) of worker bees before being placed inside a nest. HGP had little effect on samples outside a nest, indicating that nest conditions are necessary for HGP to deactivate phenolics in nectar. Consequently, the highly controlled nest homeostasis of honey bees facilitates direct deactivation of phenolics in nectar, and plays a role in the action of HGP as well.

  18. Description of Nesting Habitat for Least Bell's Vireo in San Diego County

    Treesearch

    Bonnie J. Hendricks; John P. Rieger

    1989-01-01

    Least Bell's Vireo (Vireo bellii pusillus) nesting sites on three rivers in coastal southern California were characterized to provide data for a habitat restoration plan for this endangered species. In addition, riparian areas outside vireo territories were sampled to compare with nesting habitat. The parameters measured were percent cover,...

  19. Double-Crested Cormorant ( Phalacrocorax auritus) Nesting Effects on Understory Composition and Diversity on Island Ecosystems in Lake Erie

    NASA Astrophysics Data System (ADS)

    McGrath, Darby M.; Murphy, Stephen D.

    2012-08-01

    The context for this study is the management concerns over the severity and extent of the impact of cormorants on island flora in the recent past on Lake Erie islands. Accordingly, this study sought to quantify the nesting colonies' influence on coarse woody litter and how nest densities and litter depth may influence the herbaceous layer, the seed bank composition and viability across the extent of three Lake Erie islands. The data for this study were collected from 2004 to 2008 on East Sister Island and Middle Island using two main strategies. First, herbaceous layer surveys, cormorant nest counts, soil seed bank cores, and litter depth measurements were executed using a plotless-point quarter method to test island-wide impacts from nesting activities (data were also collected on a third island, West Sister Island as a reference for the other two islands). Secondly, a sub-sample of the entire plot set was examined in particularly high nesting density areas for two islands (Middle Island and East Sister Island). Kruskal-Wallis tests indicated that there are subtle changes in the herbaceous diversity (total, native and exotic) and seed bank composition across the islands. The sub sample set of the plots demonstrated that Phalacrocorax auritus nest density does influence litter depth, herbaceous species abundance and diversity. Cormorant nesting pressures are restricted to areas of high nesting pressures and competition. However, there remains a risk to the interior herbaceous layer of the island if the effects of nesting pressures at the edges advance inward from this perimeter.

  20. LP and NLP decomposition without a master problem

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fuller, D.; Lan, B.

    We describe a new algorithm for decomposition of linear programs and a class of convex nonlinear programs, together with theoretical properties and some test results. Its most striking feature is the absence of a master problem; the subproblems pass primal and dual proposals directly to one another. The algorithm is defined for multi-stage LPs or NLPs, in which the constraints link the current stage`s variables to earlier stages` variables. This problem class is general enough to include many problem structures that do not immediately suggest stages, such as block diagonal problems. The basic algorithmis derived for two-stage problems and extendedmore » to more than two stages through nested decomposition. The main theoretical result assures convergence, to within any preset tolerance of the optimal value, in a finite number of iterations. This asymptotic convergence result contrasts with the results of limited tests on LPs, in which the optimal solution is apparently found exactly, i.e., to machine accuracy, in a small number of iterations. The tests further suggest that for LPs, the new algorithm is faster than the simplex method applied to the whole problem, as long as the stages are linked loosely; that the speedup over the simpex method improves as the number of stages increases; and that the algorithm is more reliable than nested Dantzig-Wolfe or Benders` methods in its improvement over the simplex method.« less

  1. Maternal transfer and sublethal immune system effects of brevetoxin exposure in nesting loggerhead sea turtles (Caretta caretta) from western Florida.

    PubMed

    Perrault, Justin R; Bauman, Katherine D; Greenan, Taylor M; Blum, Patricia C; Henry, Michael S; Walsh, Catherine J

    2016-11-01

    Blooms of Karenia brevis (also called red tides) occur almost annually in the Gulf of Mexico. The health effects of the neurotoxins (i.e., brevetoxins) produced by this toxic dinoflagellate on marine turtles are poorly understood. Florida's Gulf Coast represents an important foraging and nesting area for a number of marine turtle species. Most studies investigating brevetoxin exposure in marine turtles thus far focus on dead and/or stranded individuals and rarely examine the effects in apparently "healthy" free-ranging individuals. From May-July 2014, one year after the last red tide bloom, we collected blood from nesting loggerhead sea turtles (Caretta caretta) on Casey Key, Florida USA. These organisms show both strong nesting and foraging site fidelity. The plasma was analyzed for brevetoxin concentrations in addition to a number of health and immune-related parameters in an effort to establish sublethal effects of this toxin. Lastly, from July-September 2014, we collected unhatched eggs and liver and yolk sacs from dead-in-nest hatchlings from nests laid by the sampled females and tested these samples for brevetoxin concentrations to determine maternal transfer and effects on reproductive success. Using a competitive enzyme-linked immunosorbent assay (ELISA), all plasma samples from nesting females tested positive for brevetoxin (reported as ng brevetoxin-3[PbTx-3] equivalents [eq]/mL) exposure (2.1-26.7ng PbTx-3eq/mL). Additionally, 100% of livers (1.4-13.3ng PbTx-3eq/mL) and yolk sacs (1.7-6.6ng PbTx-3eq/mL) from dead-in-nest hatchlings and 70% of eggs (<1.0-24.4ng PbTx-3eq/mL) tested positive for brevetoxin exposure with the ELISA. We found that plasma brevetoxin concentrations determined by an ELISA in nesting females positively correlated with gamma-globulins, indicating a potential for immunomodulation as a result of brevetoxin exposure. While the sample sizes were small, we also found that plasma brevetoxin concentrations determined by an ELISA in nesting females significantly correlated with liver brevetoxin concentrations of dead-in-nest hatchlings and that brevetoxins could be related to a decreased reproductive success in this species. This study suggests that brevetoxins can still elicit negative effects on marine life long after a bloom has dissipated. These results improve our understanding of maternal transfer and sublethal effects of brevetoxin exposure in marine turtles. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Abundance of enterovirus C in RD-L20B cell culture-negative stool samples from acute flaccid paralysis cases in Nigeria is geographically defined.

    PubMed

    Donbraye, Emmanuel; Olasunkanmi, Oluwatayo Israel; Opabode, Babatunde Ayoola; Ishola, Temitayo Rachael; Faleye, Temitope Oluwasegun Cephas; Adewumi, Olubusuyi Moses; Adeniji, Johnson Adekunle

    2018-06-01

    We recently showed that enteroviruses (EVs) andenterovirus species C (EV-C) in particular were abundant in faecal samples from children who had been diagnosed with acute flaccid paralysis (AFP) in Nigeria but declared to be EV-free by the RD-L20B cell culture-based algorithm. In this study, we investigated whether this observed preponderance of EVs (and EV-Cs) in such samples varies by geographical region. One hundred and eight samples (i.e. 54 paired stool suspensions from 54 AFP cases) that had previously been confirmed to be negative for EVs by the WHO-recommended RD-L20B cell culture-based algorithm were analysed. The 108 samples were made into 54 pools (27 each from North-West and South-South Nigeria). All were subjected to RNA extraction, cDNA synthesis and the WHO-recommended semi-nested PCR assay and its modifications. All of the amplicons were sequenced, and the enteroviruses identified, using the enterovirus genotyping tool and phylogenetic analysis. EVs were detected in 16 (29.63 %) of the 54 samples that were screened and successfully identified in 14 (25.93 %). Of these, 10 were from North-West and 4 were from South-South Nigeria. One (7.14 %), 2 (14.29 %) and 11 (78.57 %) of the strains detected were EV-A, EV-B and EV-C, respectively. The 10 strains from North-West Nigeria included 7 EV types, namely CV-A10, E29, CV-A13, CV-A17, CV-A19, CV-A24 and EV-C99. The four EV types recovered from South-South Nigeria were E31, CV-A1, EV-C99 and EV-C116. The results of this study showed that the presence of EVs and consequently EV-Cs in AFP samples declared to be EV-free by the RD-L20B cell culture-based algorithm varies by geographical region in Nigeria.

  3. Bayesian model comparison and parameter inference in systems biology using nested sampling.

    PubMed

    Pullen, Nick; Morris, Richard J

    2014-01-01

    Inferring parameters for models of biological processes is a current challenge in systems biology, as is the related problem of comparing competing models that explain the data. In this work we apply Skilling's nested sampling to address both of these problems. Nested sampling is a Bayesian method for exploring parameter space that transforms a multi-dimensional integral to a 1D integration over likelihood space. This approach focuses on the computation of the marginal likelihood or evidence. The ratio of evidences of different models leads to the Bayes factor, which can be used for model comparison. We demonstrate how nested sampling can be used to reverse-engineer a system's behaviour whilst accounting for the uncertainty in the results. The effect of missing initial conditions of the variables as well as unknown parameters is investigated. We show how the evidence and the model ranking can change as a function of the available data. Furthermore, the addition of data from extra variables of the system can deliver more information for model comparison than increasing the data from one variable, thus providing a basis for experimental design.

  4. Development of a nested polymerase chain reaction for amplification of a sequence of the p57 gene of Renibacterium salmoninarum that provides a highly sensitive method for detection of the bacterium in salmonid kidney

    USGS Publications Warehouse

    Chase, D.M.; Pascho, R.J.

    1998-01-01

    Nucleic acid-based assays have shown promise for diagnosing Renibacterium salmoninarum in tissues and body fluids of salmonids. DeVelopment of a nested polymerase chain reaction (PCR) method to detect a 320 bp DNA segment of the gene encoding the p57 protein of R. salmoninarum is described. Whereas a conventional PCR for a 383 bp segment of the p57 gene reliably detected 1000 R. salmoninarum cells per reaction in kidney tissue, the nested PCR detected as few as 10 R. salmoninarum per reaction in kidney tissue. Two DNA extraction methods for the nested PCR were compared and the correlation between replicate samples was generally higher in samples extracted by the QIAamp system compared with those extracted by the phenol/chloroform method. The specificity of the nested PCR was confirmed by testing DNA extracts of common bacterial fish pathogens and a panel of bacterial species reported to cause false-positive reactions in the enzyme-linked immunosorbent assay (ELISA) and the fluorescent antibody test (FAT) for R. salmoninarum. Kidney samples from 74 naturally infected chinook Salmon were examined by the nested PCR, the ELISA, and the FAT, and the detected prevalences of R. salmoninarum were 61, 47, and 43%, respectively.

  5. Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods.

    PubMed

    Camargo, Vinícius da Silva; Santana, Bruna Nicoleti; Ferrari, Elis Domingos; Nakamura, Alex Akira; Nagata, Walter Bertequini; Nardi, Ana Rita Moraes; Meireles, Marcelo Vasconcelos

    2018-01-01

    This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.

  6. Nested PCR Assay for Detection of Leishmania donovani in Slit Aspirates from Post-Kala-Azar Dermal Leishmaniasis Lesions

    PubMed Central

    Sreenivas, Gannavaram; Ansari, N. A.; Kataria, Joginder; Salotra, Poonam

    2004-01-01

    A nested PCR assay to detect parasite DNA in slit aspirates from skin lesions of patients with post-kala-azar dermal lesihmaniasis (PKDL) is described. PCR results were positive in 27 of 29 (93%) samples by nested PCR assay, while only 20 of 29 (69%) were positive in a primary PCR assay. The nested PCR assay allowed reliable diagnosis of PKDL in a noninvasive manner. PMID:15071047

  7. Determination of an effective sampling regime to detect salmonella enteritidis in the environment of poultry units.

    PubMed

    Davies, R H; Wray, C

    1996-05-01

    A study of the dissemination of Salmonella enteritidis in the poultry breeder industry in the UK showed that the choice of sites for sampling the environment of occupied houses and empty houses which had been disinfected after depopulation had a significant influence on the outcome. Increased isolation rates could be achieved by sampling nest box floors and dust in open slave feed hoppers in occupied poultry houses. Nest box floors were the most sensitive sites for detection of residual environmental contamination in poultry houses where enrofloxacin treatment had been used. Floor sweepings, nest box floors, slave feed hoppers, hydrated wall fabric junctions and high beams and pipes were the most sensitive sample sites in cleansed and disinfected poultry houses. The use of universal disinfectant neutralisers gave good results in laboratory trials but appeared to reduce the isolation rate from field samples.

  8. Baseline plasma corticosterone, haematological and biochemical results in nesting and rehabilitating loggerhead sea turtles (Caretta caretta).

    PubMed

    Flower, Jennifer E; Norton, Terry M; Andrews, Kimberly M; Nelson, Steven E; Parker, Clare E; Romero, L Michael; Mitchell, Mark A

    2015-01-01

    The evaluation of hormonal responses to stress in reptiles relies on acquisition of baseline corticosterone concentrations; however, the stress associated with the restraint needed to collect the blood samples can affect the results. The purpose of this study was to determine a time limit for the collection of blood samples to evaluate baseline corticosterone, haematological and biochemical results in nesting (n = 11) and rehabilitating (n = 16) loggerhead sea turtles (Caretta caretta). Blood samples were collected from the dorsal cervical sinus of each turtle immediately after touching the animal (t 0; 0-3 min) and 3 (t 3; 3-6 min), 6 (t 6; 6-9 min; nesting turtles only), 10 (t 10; 10-13 min) and 30 min (t 30; rehabilitating turtles only) after the initial hands-on time. Consistent between the rehabilitating and nesting turtles, there was a subtle yet significant increase in white blood cell counts over time. Despite the fact that white blood cell counts increased during the sampling period, there was no direct correlation between white blood cell count and corticosterone in the sampled turtles. In the nesting turtles, significant elevations in corticosterone were noted between t 0 and t 3 (P = 0.014) and between t 0 and t 6 (P = 0.022). Values at t 10 were not significantly different from those at t 0 (P = 0.102); however, there was a trend for the corticosterone values to continue to increase. These results suggest that sampling of nesting loggerhead sea turtles within 3 min of handling will provide baseline corticosterone concentrations in their natural environment. Significant elevations in corticosterone were also noted in the rehabilitating loggerhead sea turtles between t 0 and t 10 (P = 0.02) and between t 0 and t 30 of sampling (P = 0.0001). These results suggest that sampling of loggerhead sea turtles within 6 min of handling should provide baseline corticosterone concentrations in a rehabilitation setting. The delay in the corticosterone response noted in the rehabilitating turtles may be associated with the daily contact (visual or direct) they have with their human caretakers.

  9. The effect of canopy closure on chimpanzee nest abundance in Lagoas de Cufada National Park, Guinea-Bissau.

    PubMed

    Sousa, Joana; Casanova, Catarina; Barata, André V; Sousa, Cláudia

    2014-04-01

    The present study aimed to gather baseline information about chimpanzee nesting and density in Lagoas de Cufada Natural Park (LCNP), in Guinea-Bissau. Old and narrow trails were followed to estimate chimpanzee density through marked-nest counts and to test the effect of canopy closure (woodland savannah, forest with a sparse canopy, and forest with a dense canopy) on nest distribution. Chimpanzee abundance was estimated at 0.79 nest builders/km(2), the lowest among the areas of Guinea-Bissau with currently studied chimpanzee populations. Our data suggest that sub-humid forest with a dense canopy accounts for significantly higher chimpanzee nest abundance (1.50 nests/km of trail) than sub-humid forest with a sparse canopy (0.49 nests/km of trail) or woodland savannah (0.30 nests/km of trail). Dense-canopy forests play an important role in chimpanzee nesting in the patchy and highly humanized landscape of LCNP. The tree species most frequently used for nesting are Dialium guineense (46%) and Elaeis guineensis (28%). E. guineensis contain nests built higher in the canopy, while D. guineense contain nests built at lower heights. Nests observed during baseline sampling and replications suggest seasonal variations in the tree species used for nest building.

  10. An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection.

    PubMed

    Pitz, Amanda de Faveri; Koishi, Andrea Cristine; Tavares, Eliandro Reis; Andrade, Fábio Goulart de; Loth, Eduardo Alexandre; Gandra, Rinaldo Ferreira; Venancio, Emerson José

    2013-01-01

    Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

  11. High-tech or field techs: Radio-telemetry is a cost-effective method for reducing bias in songbird nest searching

    USGS Publications Warehouse

    Peterson, Sean M.; Streby, Henry M.; Lehman, Justin A.; Kramer, Gunnar R.; Fish, Alexander C.; Andersen, David E.

    2015-01-01

    We compared the efficacy of standard nest-searching methods with finding nests via radio-tagged birds to assess how search technique influenced our determination of nest-site characteristics and nest success for Golden-winged Warblers (Vermivora chrysoptera). We also evaluated the cost-effectiveness of using radio-tagged birds to find nests. Using standard nest-searching techniques for 3 populations, we found 111 nests in locations with habitat characteristics similar to those described in previous studies: edges between forest and relatively open areas of early successional vegetation or shrubby wetlands, with 43% within 5 m of forest edge. The 83 nests found using telemetry were about half as likely (23%) to be within 5 m of forest edge. We spent little time searching >25 m into forest because published reports state that Golden-winged Warblers do not nest there. However, 14 nests found using telemetry (18%) were >25 m into forest. We modeled nest success using nest-searching method, nest age, and distance to forest edge as explanatory variables. Nest-searching method explained nest success better than nest age alone; we estimated that nests found using telemetry were 10% more likely to fledge young than nests found using standard nest-searching methods. Although radio-telemetry was more expensive than standard nest searching, the cost-effectiveness of both methods differed depending on searcher experience, amount of equipment owned, and bird population density. Our results demonstrate that telemetry can be an effective method for reducing bias in Golden-winged Warbler nest samples, can be cost competitive with standard nest-searching methods in some situations, and is likely to be a useful approach for finding nests of other forest-nesting songbirds.

  12. Survey of pathogens in threatened wild red-tailed Amazon parrot (Amazona brasiliensis) nestlings in Rasa Island, Brazil.

    PubMed

    Vaz, Frederico Fontanelli; Serafini, Patrícia Pereira; Locatelli-Dittrich, Rosangela; Meurer, Rafael; Durigon, Edison Luiz; de Araújo, Jansen; Thomazelli, Luciano Matsumiya; Ometto, Tatiana; Sipinski, Elenise Angelotti Bastos; Sezerban, Rafael Meirelles; Abbud, Maria Cecília; Raso, Tânia Freitas

    The red-tailed Amazon parrot (Amazona brasiliensis) is a threatened species of psittacine bird that inhabit coastal regions of Brazil. In view of the threat of this species, the aim of this study was to perform a health evaluation in wild nestlings in Rasa Island, determining the prevalence of enterobacteria and infectious agents according to type of nest. Blood samples were collected from 64 birds and evaluated for antibodies of Chlamydia psittaci by commercial dot-blot ELISA. Cloacal and oropharyngeal swabs samples were collected from 23 birds from artificial wooden nests, 15 birds from PVC nests and 2 birds from natural nests for microbiological analysis. Swab samples were collected from 58 parrots for C. psittaci detection by PCR and from 50 nestlings for Avian Influenza, Newcastle Disease and West Nile viruses' detection analysis by real-time RT-PCR. Ten bacterial genera and 17 species were identified, and the most prevalent were Escherichia coli and Klebsiella oxytoca. There was no influence of the type of nest in the nestlings' microbiota. All samples tested by ELISA and PCR were negative. There is currently insufficient information available about the health of A. brasiliensis and data of this study provide a reference point for future evaluations and aid in conservation plans. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  13. Organochlorine and heavy metal residues in bald eagle eggs

    USGS Publications Warehouse

    Krantz, W.C.; Mulhern, B.M.; Bagley, George E.; Sprunt, A.; Ligas, F.J.; Robertson, W.B.

    1970-01-01

    Bald eagle eggs collected in 1968 from nests in Wisconsin, Maine, and Florida all contained residues of DDE, DDD, dieldrin, heptachlor epoxide, and polychlorinated biphenyls. Many also contained traces of DDT. Eggs from five nonproductive nests sampled in Maine contained much higher residues than did eggs collected from either productive or nonproductive nests in Wisconsin and Florida.

  14. Molecular forensics in avian conservation: a DNA-based approach for identifying mammalian predators of ground-nesting birds and eggs.

    PubMed

    Hopken, Matthew W; Orning, Elizabeth K; Young, Julie K; Piaggio, Antoinette J

    2016-01-07

    The greater sage-grouse (Centrocercus urophasianus) is a ground-nesting bird from the Northern Rocky Mountains and a species at risk of extinction in in multiple U.S. states and Canada. Herein we report results from a proof of concept that mitochondrial and nuclear DNAs from mammalian predator saliva could be non-invasively collected from depredated greater sage-grouse eggshells and carcasses and used for predator species identification. Molecular forensic approaches have been applied to identify predators from depredated remains as one strategy to better understand predator-prey dynamics and guide management strategies. This can aid conservation efforts by correctly identifying predators most likely to impact threatened and endangered species. DNA isolated from non-invasive samples around nesting sites (e.g. fecal or hair samples) is one method that can increase the success and accuracy of predator species identification when compared to relying on nest remains alone. Predator saliva DNA was collected from depredated eggshells and carcasses using swabs. We sequenced two partial fragments of two mitochondrial genes and obtained microsatellite genotypes using canid specific primers for species and individual identification, respectively. Using this multilocus approach we were able to identify predators, at least down to family, from 11 out of 14 nests (79%) and three out of seven carcasses (47%). Predators detected most frequently were canids (86%), while other taxa included rodents, a striped skunk, and cattle. We attempted to match the genotypes of individual coyotes obtained from eggshells and carcasses with those obtained from fecal samples and coyotes collected in the areas, but no genotype matches were found. Predation is a main cause of nest failure in ground-nesting birds and can impact reproduction and recruitment. To inform predator management for ground-nesting bird conservation, accurate identification of predator species is necessary. Considering predation can have a high impact on recruitment, predation events are very difficult to observe, and predator species are difficult to identify visually from nest remains, molecular approaches that reduce the need to observe or handle animals offer an additional tool to better understand predator-prey dynamics at nesting sites.

  15. Swarm Intelligence for Urban Dynamics Modelling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghnemat, Rawan; Bertelle, Cyrille; Duchamp, Gerard H. E.

    2009-04-16

    In this paper, we propose swarm intelligence algorithms to deal with dynamical and spatial organization emergence. The goal is to model and simulate the developement of spatial centers using multi-criteria. We combine a decentralized approach based on emergent clustering mixed with spatial constraints or attractions. We propose an extension of the ant nest building algorithm with multi-center and adaptive process. Typically, this model is suitable to analyse and simulate urban dynamics like gentrification or the dynamics of the cultural equipment in urban area.

  16. Swarm Intelligence for Urban Dynamics Modelling

    NASA Astrophysics Data System (ADS)

    Ghnemat, Rawan; Bertelle, Cyrille; Duchamp, Gérard H. E.

    2009-04-01

    In this paper, we propose swarm intelligence algorithms to deal with dynamical and spatial organization emergence. The goal is to model and simulate the developement of spatial centers using multi-criteria. We combine a decentralized approach based on emergent clustering mixed with spatial constraints or attractions. We propose an extension of the ant nest building algorithm with multi-center and adaptive process. Typically, this model is suitable to analyse and simulate urban dynamics like gentrification or the dynamics of the cultural equipment in urban area.

  17. Monitoring and improving the sensitivity of dengue nested RT-PCR used in longitudinal surveillance in Thailand.

    PubMed

    Klungthong, Chonticha; Manasatienkij, Wudtichai; Phonpakobsin, Thipwipha; Chinnawirotpisan, Piyawan; Rodpradit, Prinyada; Hussem, Kittinun; Thaisomboonsuk, Butsaya; Ong-ajchaowlerd, Prapapun; Nisalak, Ananda; Kalayanarooj, Siripen; Buddhari, Darunee; Gibbons, Robert V; Jarman, Richard G; Yoon, In-Kyu; Fernandez, Stefan

    2015-02-01

    AFRIMS longitudinal dengue surveillance in Thailand depends on the nested RT-PCR and the dengue IgM/IgG ELISA. To examine and improve the sensitivity of the nested RT-PCR using a panel of archived samples collected during dengue surveillance. A retrospective analysis of 16,454 dengue IgM/IgG ELISA positive cases collected between 2000 and 2013 was done to investigate the sensitivity of the nested RT-PCR. From these cases, 318 acute serum specimens or extracted RNA, previously found to be negative by the nested RT-PCR, were tested using TaqMan real-time RT-PCR (TaqMan rRT-PCR). To improve the sensitivity of nested RT-PCR, we designed a new primer based on nucleotide sequences from contemporary strains found to be positive by the TaqMan rRT-PCR. Sensitivity of the new nested PCR was calculated using a panel of 87 samples collected during 2011-2013. The percentage of dengue IgM/IgG ELISA positive cases that were negative by the nested RT-PCR varied from 17% to 42% for all serotypes depending on the year. Using TaqMan rRT-PCR, dengue RNA was detected in 194 (61%) of the 318 acute sera or extracted RNA previously found to be negative by the nested RT-PCR. The newly designed DENV-1 specific primer increased the sensitivity of DENV-1 detection by the nested RT-PCR from 48% to 88%, and of all 4 serotypes from 73% to 87%. These findings demonstrate the impact of genetic diversity and signal erosion on the sensitivity of PCR-based methods. Published by Elsevier B.V.

  18. Estimating stage-specific daily survival probabilities of nests when nest age is unknown

    USGS Publications Warehouse

    Stanley, T.R.

    2004-01-01

    Estimation of daily survival probabilities of nests is common in studies of avian populations. Since the introduction of Mayfield's (1961, 1975) estimator, numerous models have been developed to relax Mayfield's assumptions and account for biologically important sources of variation. Stanley (2000) presented a model for estimating stage-specific (e.g. incubation stage, nestling stage) daily survival probabilities of nests that conditions on “nest type” and requires that nests be aged when they are found. Because aging nests typically requires handling the eggs, there may be situations where nests can not or should not be aged and the Stanley (2000) model will be inapplicable. Here, I present a model for estimating stage-specific daily survival probabilities that conditions on nest stage for active nests, thereby obviating the need to age nests when they are found. Specifically, I derive the maximum likelihood function for the model, evaluate the model's performance using Monte Carlo simulations, and provide software for estimating parameters (along with an example). For sample sizes as low as 50 nests, bias was small and confidence interval coverage was close to the nominal rate, especially when a reduced-parameter model was used for estimation.

  19. Mourning Dove nesting habitat and nest success in Central Missouri

    USGS Publications Warehouse

    Drobney, R.D.; Schulz, J.H.; Sheriff, S.L.; Fuemmeler, W.J.

    1998-01-01

    Previous Mourning Dove (Zenaida macroura) nesting studies conducted in areas containing a mixture of edge and continuous habitats have focused on edge habitats. Consequently, little is known about the potential contribution of continuous habitats to dove production. In this study we evaluated the relative importance of these two extensive habitat types by monitoring the habitat use and nest success of 59 radio-marked doves during 1990-1991 in central Missouri. Of 83 nests initiated by our marked sample, most (81.9%) were located in edge habitats. Although continuous habitats were selected less as nest sites, the proportion of successful nests did not differ significantly from that in edge habitats. Our data indicate that continuous habitats should not be considered marginal nesting habitat. If the intensity of use and nest success that we observed are representative regionally or nationally, continuous habitats could contribute substantially to annual Mourning Dove production because of the high availability of these habitats throughout much of the Mourning Dove breeding range.

  20. Optimization of internet content filtering-Combined with KNN and OCAT algorithms

    NASA Astrophysics Data System (ADS)

    Guo, Tianze; Wu, Lingjing; Liu, Jiaming

    2018-04-01

    The face of the status quo that rampant illegal content in the Internet, the result of traditional way to filter information, keyword recognition and manual screening, is getting worse. Based on this, this paper uses OCAT algorithm nested by KNN classification algorithm to construct a corpus training library that can dynamically learn and update, which can be improved on the filter corpus for constantly updated illegal content of the network, including text and pictures, and thus can better filter and investigate illegal content and its source. After that, the research direction will focus on the simplified updating of recognition and comparison algorithms and the optimization of the corpus learning ability in order to improve the efficiency of filtering, save time and resources.

  1. Nestedness in assemblages of helminth parasites of bats: a function of geography, environment, or host nestedness?

    PubMed

    Warburton, Elizabeth M; Van Der Mescht, Luther; Khokhlova, Irina S; Krasnov, Boris R; Vonhof, Maarten J

    2018-05-01

    Nested subsets occur in ecological communities when species-poor communities are subsets of larger, species-rich communities. Understanding this pattern can help elucidate species colonization abilities, extinction risks, and general structuring of biological communities. Here, we evaluate nestedness in a poorly studied host-parasite system, bats and their helminths, across the Japanese archipelago and within its different bioclimatic regions. We hypothesized that (1) if helminth communities are nested across geographic sites at the level of the archipelago, then broad-scale processes, like colonization-extinction dynamics, mainly structure parasite assemblages; (2) if helminth communities are nested across geographic sites at the level of the bioclimatic region, then fine-scale environmental variation plays a significant role in species nestedness; (3) if helminth community nestedness mirrors host species nestedness, then communities are nested because the habitats they occupy are nested; and (4) if nestedness does not occur or if it is not correlated with any geographical or host data, then passive sampling could be responsible for the patterns of parasite assemblage in our sample. We found that helminth communities were nested across host species throughout the archipelago but, when considering each bioclimatic region, helminths in only one region were significantly more nested than the null model. Helminth communities were also nested across sites within all four bioclimatic regions. These results suggest that helminths form nested subsets across the archipelago due to broad-scale processes that reflect the overall lineages of their mammalian hosts; however, at the regional scale, environmental processes related to nestedness of their habitats drive parasite community nestedness.

  2. Nested effects models for learning signaling networks from perturbation data.

    PubMed

    Fröhlich, Holger; Tresch, Achim; Beissbarth, Tim

    2009-04-01

    Targeted gene perturbations have become a major tool to gain insight into complex cellular processes. In combination with the measurement of downstream effects via DNA microarrays, this approach can be used to gain insight into signaling pathways. Nested Effects Models were first introduced by Markowetz et al. as a probabilistic method to reverse engineer signaling cascades based on the nested structure of downstream perturbation effects. The basic framework was substantially extended later on by Fröhlich et al., Markowetz et al., and Tresch and Markowetz. In this paper, we present a review of the complete methodology with a detailed comparison of so far proposed algorithms on a qualitative and quantitative level. As an application, we present results on estimating the signaling network between 13 genes in the ER-alpha pathway of human MCF-7 breast cancer cells. Comparison with the literature shows a substantial overlap.

  3. [Optimized application of nested PCR method for detection of malaria].

    PubMed

    Yao-Guang, Z; Li, J; Zhen-Yu, W; Li, C

    2017-04-28

    Objective To optimize the application of the nested PCR method for the detection of malaria according to the working practice, so as to improve the efficiency of malaria detection. Methods Premixing solution of PCR, internal primers for further amplification and new designed primers that aimed at two Plasmodium ovale subspecies were employed to optimize the reaction system, reaction condition and specific primers of P . ovale on basis of routine nested PCR. Then the specificity and the sensitivity of the optimized method were analyzed. The positive blood samples and examination samples of malaria were detected by the routine nested PCR and the optimized method simultaneously, and the detection results were compared and analyzed. Results The optimized method showed good specificity, and its sensitivity could reach the pg to fg level. The two methods were used to detect the same positive malarial blood samples simultaneously, the results indicated that the PCR products of the two methods had no significant difference, but the non-specific amplification reduced obviously and the detection rates of P . ovale subspecies improved, as well as the total specificity also increased through the use of the optimized method. The actual detection results of 111 cases of malarial blood samples showed that the sensitivity and specificity of the routine nested PCR were 94.57% and 86.96%, respectively, and those of the optimized method were both 93.48%, and there was no statistically significant difference between the two methods in the sensitivity ( P > 0.05), but there was a statistically significant difference between the two methods in the specificity ( P < 0.05). Conclusion The optimized PCR can improve the specificity without reducing the sensitivity on the basis of the routine nested PCR, it also can save the cost and increase the efficiency of malaria detection as less experiment links.

  4. Effects of temporally persistent ant nests of soil protozoan communities and the abundance of morphological types of amoeba

    USDA-ARS?s Scientific Manuscript database

    We compared soil protozoan communities near ant nests with soil protozoans in reference soils 5m from the edge of any mounds. We sampled three species of Chihuahuan Desert ants that construct nests that persist for more than a decade: a seed harvester, Pogonomymex rugosus, a liquid feeding honey-po...

  5. Detection of small number of Giardia in biological materials prepared from stray dogs.

    PubMed

    Esmailikia, Leila; Ebrahimzade, Elahe; Shayan, Parviz; Amininia, Narges

    2017-12-20

    Giardia lamblia is an intestinal protozoa with intermittent and low shedding especially in dogs, and the detection of Giardia is accompanied with problems such as sampling and diagnostic method. The objective of this study was to detection of Giardia in biological materials with low number of parasite using parasitological and molecular methods, and also to determine whether the examined stray dogs harbor known zoonotic genotype of Giardia. For this aim 85 fecal and duodenal samples were studied from which 1 was positive by Trichrome staining of stool, 4 were positive by staining of duodenal samples. The nested PCR analysis with primers derived from 18 SrRNA showed that the specific PCR product could be amplified in 4 stool and 4 duodenal samples. All positive samples in staining analysis were also positive in nested PCR. No amplification could be observed by nested PCR with primers derived from β giardin gene due to the single copy of gene. Interestingly, the extracted DNA from old fixed stained Giardia positive smears could be also amplified with primers derived from 18SrRNA gene. The sequence analysis of nested PCR products showed that they belong to the genotype D. In conclusion, it is to denote that the Trichrome or Giemsa methods were not suitable for the detection of small number of this parasite in stool and the nested PCR with primers derived from 18S rRNA gene can replace the traditional methods successfully. For detection of Giardia in stool, primers derived from β giardin will not be recommended.

  6. Nested Cohort - R software package

    Cancer.gov

    NestedCohort is an R software package for fitting Kaplan-Meier and Cox Models to estimate standardized survival and attributable risks for studies where covariates of interest are observed on only a sample of the cohort.

  7. Sampling considerations for demographic and habitat studies of Northern Goshawks

    Treesearch

    Richard T. Reynolds; J. David Wiens; Suzanne M. Joy; Susan R. Salafsky

    2005-01-01

    We used mark-recapture methods to monitor Northern Goshawks (Accipiter gentilis) and their nests over 12 yr in an increasing sample of breeding territories (37 in 1991 to 121 in 2002) in northern Arizona. As many as 8 yr of repeated nest searching were required to identify the population of breeders, as individuals skipped egg-laying on territories...

  8. Arboreal ant colonies as 'hot-points' of cryptic diversity for myrmecophiles: the weaver ant Camponotus sp. aff. textor and its interaction network with its associates.

    PubMed

    Pérez-Lachaud, Gabriela; Lachaud, Jean-Paul

    2014-01-01

    Systematic surveys of macrofaunal diversity within ant colonies are lacking, particularly for ants nesting in microhabitats that are difficult to sample. Species associated with ants are generally small and rarely collected organisms, which makes them more likely to be unnoticed. We assumed that this tendency is greater for arthropod communities in microhabitats with low accessibility, such as those found in the nests of arboreal ants that may constitute a source of cryptic biodiversity. We investigated the invertebrate diversity associated with an undescribed, but already threatened, Neotropical Camponotus weaver ant. As most of the common sampling methods used in studies of ant diversity are not suited for evaluating myrmecophile diversity within ant nests, we evaluated the macrofauna within ant nests through exhaustive colony sampling of three nests and examination of more than 80,000 individuals. We identified invertebrates from three classes belonging to 18 taxa, some of which were new to science, and recorded the first instance of the co-occurrence of two brood parasitoid wasp families attacking the same ant host colony. This diversity of ant associates corresponded to a highly complex interaction network. Agonistic interactions prevailed, but the prevalence of myrmecophiles was remarkably low. Our data support the hypothesis of the evolution of low virulence in a variety of symbionts associated with large insect societies. Because most myrmecophiles found in this work are rare, strictly specific, and exhibit highly specialized biology, the risk of extinction for these hitherto unknown invertebrates and their natural enemies is high. The cryptic, far unappreciated diversity within arboreal ant nests in areas at high risk of habitat loss qualifies these nests as 'hot-points' of biodiversity that urgently require special attention as a component of conservation and management programs.

  9. Human herpesvirus infections of the central nervous system: laboratory diagnosis based on DNA detection by nested PCR in plasma and cerebrospinal fluid samples.

    PubMed

    Rimério, Carla Aparecida Tavares; De Oliveira, Renato Souza; de Almeida Bonatelli, Murilo Queiroz; Nucci, Anamarli; Costa, Sandra Cecília Botelho; Bonon, Sandra Helena Alves

    2015-04-01

    Infections of the central nervous systems (CNS) present a diagnostic problem for which an accurate laboratory diagnosis is essential. Invasive practices, such as cerebral biopsy, have been replaced by obtaining a polymerase chain reaction (PCR) diagnosis using cerebral spinal fluid (CSF) as a reference method. Tests on DNA extracted from plasma are noninvasive, thus avoiding all of the collateral effects and patient risks associated with CSF collection. This study aimed to determine whether plasma can replace CSF in nested PCR analysis for the detection of CNS human herpesvirus (HHV) diseases by analysing the proportion of patients whose CSF nested PCR results were positive for CNS HHV who also had the same organism identified by plasma nested PCR. In this study, CSF DNA was used as the "gold standard," and nested PCR was performed on both types of samples. Fifty-two patients with symptoms of nervous system infection were submitted to CSF and blood collection. For the eight HHV, one positive DNA result-in plasma and/or CSF nested PCR-was considered an active HHV infection, whereas the occurrence of two or more HHVs in the same sample was considered a coinfection. HHV infections were positively detected in 27/52 (51.9%) of the CSF and in 32/52 (61.5%) of the plasma, difference not significant, thus nested PCR can be performed on plasma instead of CSF. In conclusion, this findings suggest that plasma as a useful material for the diagnosis of cases where there is any difficulty to perform a CSF puncture. © 2015 Wiley Periodicals, Inc.

  10. PCR-based Diagnosis of Toxoplasma Parasite in Ocular Infections Having Clinical Indications of Toxoplasmosis.

    PubMed

    Farhadi, Atieh; Haniloo, Ali; Fazaeli, Asghar; Moradian, Siamak; Farhadi, Mehdi

    2017-01-01

    The diagnosis of ocular toxoplasmosis is mainly based on clinical features. However, ocular fluid testing by PCR may be very helpful for approval or rejection of this etiology. In this study, we utilized a nested-PCR technique, targeting the B1 partial sequence to analyze the aqueous and vitreous samples for evaluating the presence of the Toxoplasma DNA. Fifty aqueous or vitreous humor samples were obtained from patients with clinical features of ocular toxoplasmosis admitted to ophthalmology hospitals and clinics in Iran, within 2014. The samples were subsequently subjected to DNA extraction and purification. For nested amplification of the Toxoplasma B1 gene, two primer pairs were used. The outer and inner primers are expected to produce a 193 bp and a 96 bp fragments, respectively. The first-round PCR resulted in the detection of T. gondii in 58% of samples by amplification of the expected 193bp DNA fragment. The nested-PCR using the inner primers, detected 15 additional samples from those with negative amplicons in the first round PCR (overall positivity of 88%). In addition, vitreous samples showed relatively more positive cases than aqueous humor in detection of the infection. The nested-PCR protocol using the B1 gene, with the high detection power, could be a useful complimentary method to clinical diagnose of ocular toxoplasmosis.

  11. Detection of Pneumocystis jirovecii by nested PCR in HIV-negative patients with pulmonary disease.

    PubMed

    Santos, Cristina Rodrigues; de Assis, Ângela M; Luz, Edson A; Lyra, Luzia; Toro, Ivan F; Seabra, José Claudio C; Daldin, Dira H; Marcalto, Tathiane U; Galasso, Marcos T; Macedo, Ronaldo F; Schreiber, Angélica Z; Aoki, Francisco H

    Nested PCR can be used to determine the status of Pneumocystis jirovecii infection in other lung diseases. This study sought to detect a target DNA fragment (mitochondrial large subunit rRNA or mtL SUrRNA) of P. jirovecii in patients with lung disease who underwent bronchoscopy with collection of bronchoalveolar lavage (BAL). The results from toluidine blue staining were compared with those obtained using molecular methods that included an "in house" DNA extraction procedure, PCR and nested PCR. Fifty-five BAL samples from patients with atypical chest X-rays were screened for P. jirovecii. None of the samples was positive for P. jirovecii using toluidine blue staining. In contrast, P. jirovecii DNA was detected by nested PCR in BAL samples from 36 of 55 patients (65.5%). The lung diseases in the patients included cancer, pneumonia, tuberculosis, and chronic obstructive pulmonary disease (COPD). Other chronic problems in the patients included hypertension, diabetes, smoking, and alcoholism. Nested PCR showed high sensitivity for detecting P. jirovecii, especially when compared with toluidine blue staining. Using this method, P. jirovecii infection was detected in HIV-negative patients with lung disease. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  12. Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India.

    PubMed

    Dinoop, K P; Parija, Subhash Chandra; Mandal, Jharna; Swaminathan, R P; Narayanan, P

    2016-01-01

    Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated ( P<0.0001). The nested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods.

  13. Rapid detection of human fecal Eubacterium species and related genera by nested PCR method.

    PubMed

    Kageyama, A; Benno, Y

    2001-01-01

    PCR procedures based on 16S rDNA gene sequence specific for seven Eubacterium spp. and Eggerthella lenta that predominate in the human intestinal tract were developed, and used for direct detection of these species in seven human feces samples. Three species of Eggerthella lenta, Eubacterium rectale, and Eubacterium eligens were detected from seven fecal samples. Eubacterium biforme was detected from six samples. It was reported that E. rectale, E. eligens, and E. biforme were difficult to detect by traditional culture method, but the nested PCR method is available for the detection of these species. This result shows that the nested PCR method utilizing a universal primer pair, followed by amplification with species-specific primers, would allow rapid detection of Eubacterium species in human feces.

  14. Can avian reproductive outcomes estimated with MCnest be made more robust using stochastic parameterizations?

    EPA Science Inventory

    The Markov chain nest productivity model, or MCnest, is a set of algorithms for integrating the results of avian toxicity tests with reproductive life-history data to project the relative magnitude of chemical effects on avian reproduction. The mathematical foundation of MCnest i...

  15. Age-specific productivity and nest site characteristics of Cooper's hawks (Accipiter cooperii)

    USGS Publications Warehouse

    Moore, K.R.; Henny, C.J.

    1984-01-01

    Nesting Cooper's Hawks (Accipiter cooperii) were studied in northeastern Oregon. Second-year (SY) males did not breed, but 22 percent of the breeding females were SY's. Mean clutch size (P = 0.012) and mean number of young fledged per pair that laid eggs (P < 0.10) were lower for SY females than for adult (after second year [ASY}) females; however, an equal percentage of the eggs (excluding a collected sample egg) yielded fledged young for each age class. Stepwise discriminant analysis suggested differences in structural characteristics of the nest site habitat for ASY and SY females, i.e., SY female nest sites were associated with younger successional stages than ASY female nest sites. Nests of both age groups were built in trees with high crown volume, but ASY females utilized mistletoe (Arceuthobium sp.) for nest structures more frequently (P < 0.01) than SY females.

  16. Density and Distribution of Xylocopa Nests (Hymenoptera: Apidae) in Caatinga Areas in the Surroundings of Passion Fruit Crops.

    PubMed

    Martins, C F; de Siqueira, K M M; Kiill, L H P; Sá, I I S; Aguiar, C M L

    2014-08-01

    Due to their importance as pollinators of many plant species, this study aimed to know the nest density, spatial distribution, and nesting substrates used by Xylocopa species in the Caatinga, a xerophilous vegetation of Northeastern Brazil. Three areas of Caatinga in the surroundings of passion fruit crops were sampled. The bee species found in these areas were Xylocopa grisescens Lepeletier and Xylocopa frontalis (Olivier). All nests were in Commiphora leptophloeos (Burseraceae) trees (n = 113). Phytosociological analysis showed that this tree species presented the highest absolute density (212.5 individuals/ha) and index of importance value (52.7). The distribution pattern of the C. leptophloeos was aggregated. The nests were located in dead and dried branches with an average diameter of 5.3 ± 2.0 cm (n = 43). The mean number of nests/tree was 3.1 ± 2.8 (n = 113). The less disturbed area showed 6.7 nests/ha and 4.2 nests/tree. In the disturbed areas, 0.9 nests/ha and 2.4 to 2.7 nests/tree were observed. The availability of substrate for nesting in the studied areas and its importance as a limiting factor for nesting are discussed.

  17. Nesting habitat of the Tule Greater White-fronted Goose Anser albifrons elgasi

    USGS Publications Warehouse

    Densmore, R.V.; Ely, Craig R.; Bollinger, K.S.; Kratzer, S.; Udevitz, M.S.; Fehringer, D.J.; Rothe, T.C.

    2006-01-01

    This paper presents the first information on the availability and use of nesting habitat by the rare Tule Greater White-fronted Goose Anser albifrons elgasi. The breeding range was sampled by marking geese with radio transmitters on wintering and moulting areas, and tracking them to nest sites in Alaska. Nesting habitat was described at the scales of ecoregion, wetland ecosystem (National Wetlands Inventory (NWI) maps), vegetation type within wetland (Alaska Vegetation Classification (AVC) maps based on satellite imagery), and nest site. Tule Greater White-fronted Goose nests were located in boreal forest wetlands in the upper Cook Inlet Basin ecoregion. Nesting Tule Greater White-fronted Geese selected NWT Palustrine Seasonally Flooded wetlands and used NWI Palustrine Saturated wetlands in proportion to availability. Within these wetlands, Tule Greater White-fronted Geese used Needleleaf Forest, Low Shrub and Herbaceous (mostly graminoid) AVC classes for nest sites in proportion to availability Most (93%) Tule Greater White-fronted Geese nested > 75 m from open water ponds or lakes, and many nested in wetlands with little or no open water. Tule Greater White-fronted Geese nest only in a small breeding area near the most human-impacted area of the state, and continued development may limit the use of suitable nesting habitat.

  18. Baseline plasma corticosterone, haematological and biochemical results in nesting and rehabilitating loggerhead sea turtles (Caretta caretta)

    PubMed Central

    Flower, Jennifer E.; Norton, Terry M.; Andrews, Kimberly M.; Nelson, Steven E.; Parker, Clare E.; Romero, L. Michael; Mitchell, Mark A.

    2015-01-01

    The evaluation of hormonal responses to stress in reptiles relies on acquisition of baseline corticosterone concentrations; however, the stress associated with the restraint needed to collect the blood samples can affect the results. The purpose of this study was to determine a time limit for the collection of blood samples to evaluate baseline corticosterone, haematological and biochemical results in nesting (n = 11) and rehabilitating (n = 16) loggerhead sea turtles (Caretta caretta). Blood samples were collected from the dorsal cervical sinus of each turtle immediately after touching the animal (t0; 0–3 min) and 3 (t3; 3–6 min), 6 (t6; 6–9 min; nesting turtles only), 10 (t10; 10–13 min) and 30 min (t30; rehabilitating turtles only) after the initial hands-on time. Consistent between the rehabilitating and nesting turtles, there was a subtle yet significant increase in white blood cell counts over time. Despite the fact that white blood cell counts increased during the sampling period, there was no direct correlation between white blood cell count and corticosterone in the sampled turtles. In the nesting turtles, significant elevations in corticosterone were noted between t0 and t3 (P = 0.014) and between t0 and t6 (P = 0.022). Values at t10 were not significantly different from those at t0 (P = 0.102); however, there was a trend for the corticosterone values to continue to increase. These results suggest that sampling of nesting loggerhead sea turtles within 3 min of handling will provide baseline corticosterone concentrations in their natural environment. Significant elevations in corticosterone were also noted in the rehabilitating loggerhead sea turtles between t0 and t10 (P = 0.02) and between t0 and t30 of sampling (P = 0.0001). These results suggest that sampling of loggerhead sea turtles within 6 min of handling should provide baseline corticosterone concentrations in a rehabilitation setting. The delay in the corticosterone response noted in the rehabilitating turtles may be associated with the daily contact (visual or direct) they have with their human caretakers. PMID:27293688

  19. Nest site characteristics, nesting movements, and lack of long-term nest site fidelity in Agassiz's desert tortoises at a wind energy facility in southern California

    USGS Publications Warehouse

    Lovich, Jeffrey E.; Agha, Mickey; Yackulic, Charles B.; Meyer-Wilkins, Kathie; Bjurlin, Curtis; Ennen, Joshua R.; Arundel, Terry R.; Austin, Meaghan

    2014-01-01

    Nest site selection has important consequences for maternal and offspring survival and fitness. Females of some species return to the same nesting areas year after year. We studied nest site characteristics, fidelity, and daily pre-nesting movements in a population of Agassiz’s desert tortoises (Gopherus agassizii) at a wind energy facility in southern California during two field seasons separated by over a decade. No females returned to the same exact nest site within or between years but several nested in the same general area. However, distances between first and second clutches within a year (2000) were not significantly different from distances between nests among years (2000 and 2011) for a small sample of females, suggesting some degree of fidelity within their normal activity areas. Environmental attributes of nest sites did not differ significantly among females but did among years due largely to changes in perennial plant structure as a result of multiple fires. Daily pre-nesting distances moved by females decreased consistently from the time shelled eggs were first visible in X-radiographs until oviposition, again suggesting some degree of nest site selection. Tortoises appear to select nest sites that are within their long-term activity areas, inside the climate-moderated confines of one of their self-constructed burrows, and specifically, at a depth in the burrow that minimizes exposure of eggs and embryos to lethal incubation temperatures. Nesting in “climate-controlled” burrows and nest guarding by females relaxes some of the constraints that drive nest site selection in other oviparous species.

  20. Flexible social organization and high incidence of drifting in the sweat bee, Halictus scabiosae.

    PubMed

    Ulrich, Yuko; Perrin, Nicolas; Chapuisat, Michel

    2009-04-01

    The very diverse social systems of sweat bees make them interesting models to study social evolution. Here we focus on the dispersal behaviour and social organization of Halictus scabiosae, a common yet poorly known species of Europe. By combining field observations and genetic data, we show that females have multiple reproductive strategies, which generates a large diversity in the social structure of nests. A detailed microsatellite analysis of 60 nests revealed that 55% of the nests contained the offspring of a single female, whereas the rest had more complex social structures, with three clear cases of multiple females reproducing in the same nest and frequent occurrence of unrelated individuals. Drifting among nests was surprisingly common, as 16% of the 122 nests in the overall sample and 44% of the nests with complex social structure contained females that had genotypes consistent with being full-sisters of females sampled in other nests of the population. Drifters originated from nests with an above-average productivity and were unrelated to their nestmates, suggesting that drifting might be a strategy to avoid competition among related females. The sex-specific comparison of genetic differentiation indicated that dispersal was male-biased, which would reinforce local resource competition among females. The pattern of genetic differentiation among populations was consistent with a dynamic process of patch colonization and extinction, as expected from the unstable, anthropogenic habitat of this species. Overall, our data show that H. scabiosae varies greatly in dispersal behaviour and social organization. The surprisingly high frequency of drifters echoes recent findings in wasps and bees, calling for further investigation of the adaptive basis of drifting in the social insects.

  1. [A study on identification of edible bird's nests by DNA barcodes].

    PubMed

    Chen, Yue-Juan; Liu, Wen-Jian; Chen, Dan-Na; Chieng, Sing-Hock; Jiang, Lin

    2017-12-01

    To provide theoretical basis for the traceability and quality evaluation of edible bird's nests (EBNs), the Cytb sequence was applied to identify the origin of EBNs. A total of 39 experiment samples were collected from Malaysia, Indonesia, Vietnam and Thailand. Genomic DNA was extracted for the PCR reaction. The amplified products were sequenced. 36 sequences were downloaded from Gen Bank including edible nest swiftlet, black nest swiftlet, mascarene swiftlet, pacific swiftlet and germain's swiftlet. MEGA 7.0 was used to analyze the distinction of sequences by the method of calculating the distances in intraspecific and interspecific divergences and constructing NJ and UPMGA phylogenetic tree based on Kimera-2-parameter model. The results showed that 39 samples were from three kinds of EBNs. Interspecific divergences were significantly greater than the intraspecific one. Samples could be successfully distinguished by NJ and UPMGA phylogenetic tree. In conclusion, Cytb sequence could be used to distinguish the origin of EBNs and it is efficient for tracing the origin species of EBNs. Copyright© by the Chinese Pharmaceutical Association.

  2. [Contribution of nested PCR in the diagnosis of imported malaria in southern Algeria].

    PubMed

    Bouiba, L; Gassen, B; Gasmi, M; Hammadi, D; Harrat, Z

    2016-12-01

    The nested PCR was used to estimate its inputs in malaria diagnosis and in the performance of the microscope operators involved in the surveillance of malaria in remote areas of South Algeria. For the period 2010 to 2015, 112 patients (93 febrile and 19 asymptomatic) coming from sub-Saharan Africa were tested for malaria in the hospital of Tamanrasset. One part of the blood taken from fingertip was used for blood smears and the second part was absorbed in filter paper for molecular diagnosis. Overall, the infection was detected by nested PCR in 63 samples versus 53 by direct examination. In addition, 11 mixed infections and 6 positive asymptomatic cases not detected by microscopy were diagnosed by PCR. Moreover, two negative samples in nested PCR were tested positive by direct examination. The molecular tool is more sensitive than the direct examination in detecting infra-microscopic parasitaemia and mixed infections...

  3. Quantification of periodontal pathogens in vascular, blood, and subgingival samples from patients with peripheral arterial disease or abdominal aortic aneurysms.

    PubMed

    Figuero, Elena; Lindahl, Christeel; Marín, María José; Renvert, Stefan; Herrera, David; Ohlsson, Ola; Wetterling, Thomas; Sanz, Mariano

    2014-09-01

    The aim of this investigation is to quantify periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, and Tannerella forsythia) in vascular, blood, and subgingival samples. As a secondary objective, two molecular bacterial identification methods (nested polymerase chain reaction [PCR] and quantitative PCR [qPCR]) are compared. Seventy consecutive patients provided a vascular lesion, a blood sample, and 36 subgingival samples. Bacterial DNA was extracted, and qPCR was used to determine the prevalence and amounts of the target pathogens in each sample. Nested PCR was performed only in the samples from vascular lesions. Periodontal examination was performed in 42 patients. Mann-Whitney U or χ(2) tests were used to compare microbiologic results according to periodontal diagnosis. All targeted periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, T. forsythia, or C. rectus) were detected in subgingival samples, with a prevalence rate of 72.2%, 47.2%, 74.3%, and 82.9%, respectively. In 7.1% and 11.4% of vascular and blood samples, bacterial DNA was detected. One patient was positive for A. actinomycetemcomitans in the three types of samples. No differences were found in the levels of targeted bacteria when comparing patients with and without periodontitis. Prevalence rates obtained with nested PCR were significantly higher than those obtained with qPCR. The presence of A. actinomycetemcomitans was demonstrated in vascular, blood, and subgingival samples in one of 36 patients. These results, although with a very low frequency, may support the hypothesis of a translocation of periodontal pathogens from subgingival microbiota to the bloodstream and then to atheromatous plaques in carotid or other peripheral arteries. Nested PCR is not an adequate method for identifying DNA of periodontal pathogens in low quantities because of the high number of false-negative results.

  4. Nest size is predicted by female identity and the local environment in the blue tit (Cyanistes caeruleus), but is not related to the nest size of the genetic or foster mother

    PubMed Central

    Parker, Timothy H.; Griffith, Simon C.

    2018-01-01

    The potential for animals to respond to changing climates has sparked interest in intraspecific variation in avian nest structure since this may influence nest microclimate and protect eggs and offspring from inclement weather. However, there have been relatively few large-scale attempts to examine variation in nests or the determinates of individual variation in nest structure within populations. Using a set of mostly pre-registered analyses, we studied potential predictors of variation in the size of a large sample (803) of blue tit (Cyanistes caeruleus) nests across three breeding seasons at Wytham Woods, UK. While our pre-registered analyses found that individual females built very similar nests across years, there was no evidence in follow-up (post hoc) analyses that their nest size correlated to that of their genetic mother or, in a cross-fostering experiment, to the nest where they were reared. In further pre-registered analyses, spatial environmental variability explained nest size variability at relatively broad spatial scales, and especially strongly at the scale of individual nest boxes. Our study indicates that nest structure is a characteristic of individuals, but is not strongly heritable, indicating that it will not respond rapidly to selection. Explaining the within-individual and within-location repeatability we observed requires further study. PMID:29765658

  5. Analysis of sugarcane herbicides in marine turtle nesting areas and assessment of risk using in vitro toxicity assays.

    PubMed

    Allan, Hannah L; van de Merwe, Jason P; Finlayson, Kimberly A; O'Brien, Jake W; Mueller, Jochen F; Leusch, Frederic D L

    2017-10-01

    Agricultural processes are associated with many different herbicides that can contaminate surrounding environments. In Queensland, Australia, herbicides applied to agricultural crops may pose a threat to valuable coastal habitats including nesting beaches for threatened loggerhead turtles (Caretta caretta). This study 1) measured concentrations of herbicides in the beach sand of Mon Repos, an important marine turtle nesting beach in Australia that is adjacent to significant sugarcane crops, and 2) investigated the toxicity of these herbicides to marine turtles using a cell-based assay. Samples of sand from turtle nest depth and water from surrounding agricultural drains and wetlands were collected during the wet season when herbicide runoff was expected to be the greatest and turtles were nesting. Samples were extracted using solid phase extraction and extracts were analysed using chemical analysis targeting herbicides, as well as bioanalytical techniques (IPAM-assay and loggerhead turtle skin cell cytotoxicity assay). Twenty herbicides were detected in areas between sugarcane crops and the nesting beach, seven of which were also detected in the sand extracts. Herbicides present in the nearby wetland were also detected in the beach sand, indicating potential contamination of the nesting beach via the river outlet as well as ground water. Although herbicides were detected in nesting sand, bioassays using loggerhead turtle skin cells indicated a low risk of acute toxicity at measured environmental concentrations. Further research should investigate potentially more subtle effects, such as endocrine disruption and mixture effects, to better assess the threat that herbicides pose to this population of marine turtles. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. [Investigation of Coxiella burnetii contamination in commercial milk and PCR method for the detection of C. burnetii in egg].

    PubMed

    Hirai, Akihiko; Kaneko, Seiji; Nakama, Akiko; Ishizaki, Naoto; Odagiri, Megumi; Kai, Akemi; Sadamasu, Kenji; Shinkai, Takayuki; Yano, Kazuyoshi; Morozumi, Satoshi

    2005-06-01

    A total of 244 milk samples collected from supermarkets in Tokyo were examined for contamination with Coxiella burnetii. C. burnetii DNA was detected in 131 (53.7%) of the samples by nested PCR. PCR-positive samples were injected into immunosuppressed A/J strain mice. Of the 22 PCR-positive milk samples tested, none resulted in isolation of C. burnetii from the mice. Heat-treatment was sufficient to inactivate C. burnetii in commercial milk. In addition, a PCR detection method for C. burnetii in chicken egg was developed. Egg yolk was added to an equal volume of 1 mol/L of NaCl phosphate buffer and homogenized for removal of protein and lipid. After centrifugal separation, the supernatant was removed, and template DNA in the precipitate was extracted using SDS, proteinase K and NaI. Using such prepared samples, 3.2 x 10(1) C. burnetii particles in 1 g of egg yolk could be detected by nested PCR. All of 200 chicken egg samples collected from supermarkets in Tokyo were negative for C. burnetii by the nested PCR method.

  7. Nest-site fidelity and dispersal of Gyrfalcons estimated by noninvasive genetic sampling

    USGS Publications Warehouse

    Booms, T.L.; Talbot, S.L.; Sage, G.K.; McCaffery, B.J.; McCracken, K.G.; Schempf, P.F.

    2011-01-01

    We used feathers from adult Gyrfalcons (Falco rusticolus) molted in breeding territories and blood samples from nestlings to document nest-site fidelity and dispersal of breeding adults and juveniles at three areas 100- 350 km apart in Yukon Delta National Wildlife Refuge, Alaska, 2003-2007. We used genotypes from seven polymorphic microsatellite loci that provided a mean probability of identity of 0.91 ??10 -5. Breeding Gyrfalcons were highly faithful to study area and territory; we documented no dispersals of breeding birds among study areas and only one dispersal between territories. But their fidelity to nest sites was low; 22% of birds returned to the same nest site the following year. Distance among alternate nests within a territory averaged 750 m and was similar for both sexes. Mean tenure in a territory was 2.8 years, similar for both sexes, and distributed bimodally with peaks at 1 and 4 years. Mean annual turnover rate at the Ingakslugwat Hills (Volcanoes) study area was 20%. We detected three young that established breeding territories at distances ranging from 0 to 254 km from their natal territory, representing 2.5% apparent recruitment. Gyrfalcons in the Askinuk Mountains study area were slightly but statistically significantly differentiated genetically from those in the Volcanoes and Kilbuck Mountain study areas. These data are the first published on the nest-site fidelity, breeding dispersal, and natal dispersal of the Gyrfalcon in North America and demonstrate the utility of noninvasive genetic sampling to greatly improve our understanding of avian dispersal and its underlying mechanisms. ?? The Cooper Ornithological Society 2011.

  8. A novel joint-processing adaptive nonlinear equalizer using a modular recurrent neural network for chaotic communication systems.

    PubMed

    Zhao, Haiquan; Zeng, Xiangping; Zhang, Jiashu; Liu, Yangguang; Wang, Xiaomin; Li, Tianrui

    2011-01-01

    To eliminate nonlinear channel distortion in chaotic communication systems, a novel joint-processing adaptive nonlinear equalizer based on a pipelined recurrent neural network (JPRNN) is proposed, using a modified real-time recurrent learning (RTRL) algorithm. Furthermore, an adaptive amplitude RTRL algorithm is adopted to overcome the deteriorating effect introduced by the nesting process. Computer simulations illustrate that the proposed equalizer outperforms the pipelined recurrent neural network (PRNN) and recurrent neural network (RNN) equalizers. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Parallel tiled Nussinov RNA folding loop nest generated using both dependence graph transitive closure and loop skewing.

    PubMed

    Palkowski, Marek; Bielecki, Wlodzimierz

    2017-06-02

    RNA secondary structure prediction is a compute intensive task that lies at the core of several search algorithms in bioinformatics. Fortunately, the RNA folding approaches, such as the Nussinov base pair maximization, involve mathematical operations over affine control loops whose iteration space can be represented by the polyhedral model. Polyhedral compilation techniques have proven to be a powerful tool for optimization of dense array codes. However, classical affine loop nest transformations used with these techniques do not optimize effectively codes of dynamic programming of RNA structure predictions. The purpose of this paper is to present a novel approach allowing for generation of a parallel tiled Nussinov RNA loop nest exposing significantly higher performance than that of known related code. This effect is achieved due to improving code locality and calculation parallelization. In order to improve code locality, we apply our previously published technique of automatic loop nest tiling to all the three loops of the Nussinov loop nest. This approach first forms original rectangular 3D tiles and then corrects them to establish their validity by means of applying the transitive closure of a dependence graph. To produce parallel code, we apply the loop skewing technique to a tiled Nussinov loop nest. The technique is implemented as a part of the publicly available polyhedral source-to-source TRACO compiler. Generated code was run on modern Intel multi-core processors and coprocessors. We present the speed-up factor of generated Nussinov RNA parallel code and demonstrate that it is considerably faster than related codes in which only the two outer loops of the Nussinov loop nest are tiled.

  10. Changes in gonadal and adrenal steroid levels in the leatherback sea turtle (Dermochelys coriacea) during the nesting cycle.

    PubMed

    Rostal, D C; Grumbles, J S; Palmer, K S; Lance, V A; Spotila, J R; Paladino, F V

    2001-05-01

    The reproductive endocrinology of nesting leatherback turtles (Dermochelys coriacea) was studied during the 1996-1997 and 1997-1998 nesting seasons at Los Baulas National Park, Playa Grande, Costa Rica. Blood samples were collected from nesting females throughout the season. Females were observed to nest up to 10 times during the nesting season. Plasma steroids were measured by radioimmunoassay and total plasma calcium was measured by flame atomic absorption spectrophotometry. Plasma testosterone and plasma estradiol levels declined throughout the nesting cycle of the female. Testosterone declined from a mean of 10.18 +/- 0.77 ng/ml at the beginning of the nesting cycle to 1.73 +/- 0.34 ng/ml at the end of the nesting cycle. Estradiol declined in a similar manner, ranging from a mean of 190.95 +/- 16.80 pg/ml at the beginning of the nesting cycle to 76.52 +/- 12.66 pg/ml at the end of the nesting cycle. Plasma progesterone and total calcium levels were relatively constant throughout the nesting cycle. Lack of fluctuation of total calcium levels, ranging from a mean high of 97.46 +/- 11.37 microg/ml to a mean low of 64.85 +/- 11.20 microg/ml, further suggests that vitellogenesis is complete prior to the arrival of the female at the nesting beach. Clutch size (both yolked and yolkless eggs) did not vary over the course of the nesting cycle. Copyright 2001 Academic Press.

  11. A Machine-Checked Proof of A State-Space Construction Algorithm

    NASA Technical Reports Server (NTRS)

    Catano, Nestor; Siminiceanu, Radu I.

    2010-01-01

    This paper presents the correctness proof of Saturation, an algorithm for generating state spaces of concurrent systems, implemented in the SMART tool. Unlike the Breadth First Search exploration algorithm, which is easy to understand and formalise, Saturation is a complex algorithm, employing a mutually-recursive pair of procedures that compute a series of non-trivial, nested local fixed points, corresponding to a chaotic fixed point strategy. A pencil-and-paper proof of Saturation exists, but a machine checked proof had never been attempted. The key element of the proof is the characterisation theorem of saturated nodes in decision diagrams, stating that a saturated node represents a set of states encoding a local fixed-point with respect to firing all events affecting only the node s level and levels below. For our purpose, we have employed the Prototype Verification System (PVS) for formalising the Saturation algorithm, its data structures, and for conducting the proofs.

  12. A rapid single-tube protocol for HAV detection by nested real-time PCR.

    PubMed

    Hu, Yuan; Arsov, Ivica

    2014-09-01

    Infections by food-borne viruses such as hepatitis A virus (HAV) and norovirus are significant public health concerns worldwide. Since food-borne viruses are rarely confirmed through direct isolation from contaminated samples, highly sensitive molecular techniques remain the methods of choice for the detection of viral genetic material. Our group has previously developed a specific nested real-time PCR (NRT-PCR) assay for HAV detection that improved overall sensitivity. Furthermore in this study, we have developed a single-tube NRT-PCR approach for HAV detection in food samples that reduces the likelihood of cross contamination between tubes during sample manipulation. HAV RNA was isolated from HAV-spiked food samples and HAV-infected cell cultures. All reactions following HAV RNA isolation, including conventional reverse transcriptase PCR, nested-PCR, and RT-PCR were performed in a single tube. Our results demonstrated that all the samples tested positive by RT-PCR and nested-PCR were also positive by a single-tube NRT-PCR. The detection limits observed for HAV-infected cell cultures and HAV-spiked green onions were 0.1 and 1 PFU, respectively. This novel method retained the specificity and robustness of the original NRT-PCR method, while greatly reducing sample manipulation, turnaround time, and the risk of carry-over contamination. Single-tube NRT-PCR thus represents a promising new tool that can potentially facilitate the detection of HAV in foods thereby improving food safety and public health.

  13. Whole blood Nested PCR and Real-time PCR amplification of Talaromyces marneffei specific DNA for diagnosis.

    PubMed

    Lu, Sha; Li, Xiqing; Calderone, Richard; Zhang, Jing; Ma, Jianchi; Cai, Wenying; Xi, Liyan

    2016-02-01

    Talaromyces marneffei is a dimorphic pathogenic fungus, which is a life-threatening invasive mycosis in the immunocompromised host. Prompt diagnosis of T. marneffei infection remains difficult although there has been progress in attempts to expedite the diagnosis of this infection. We previously demonstrated the value of nested polymerase chain reaction (PCR) to detect T. marneffei in paraffin embedded tissue samples with high sensitivity and specificity. In this study, this assay was used to detect the DNA of T. marneffei in whole blood samples. Real-time PCR assay was also evaluated to identify T. marneffei in the same samples. Twenty out of 30 whole blood samples (67%) collected from 23 patients were found positive by using the nested PCR assay, while 23/30 (77%) samples were found positive by using the real-time PCR assay. In order to express accurately the fungal loads, we used a normalized linearized plasmid as an internal control for real-time PCR. The assay results were correlated as the initial quantity (copies/μl) with fungal burden. These data indicate that combination of nested PCR and real-time PCR assay provides an attractive alternative for identification of T. marneffei DNA in whole blood samples of HIV-infected patients. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. Impact of human cytomegalovirus infection UL55-nested polymerase chain reaction method in hematopoietic stem cell transplant donors and recipients.

    PubMed

    Banan, A A; Yaghobi, R; Ramzi, M; Mehrabani, D

    2009-09-01

    Human cytomegalovirus (HCMV) is one of the most important and critical viral causes of graft rejection among hematopoietic stem cell transplant (HSCT) recipients. Monitoring of this viral infection has a critical role in the management of HSCT clinical complications. In this retrospective cohort, blood (plasma and buffy coat) and urine samples were collected from 110 HSCT patients and 95 donors pretransplantation and weekly for 100 days posttransplantation. An HCMV-optimized UL55-nested polymerase chain reaction (PCR) method was used to detect HCMV infection. Genotyping of the HCMV UL55 gene was performed for all UL55-nested, PCR-positive samples. HSCT donor and recipient laboratory and clinical data were statistically analyzed using SPSS version 15 software. UL55-nested, PCR-positive results were obtained in 3540/4950 (71.5%), 3634/4950 (73.4%), and 3292/4950 (66.5%) of these plasma, buffy coat, and urine samples, respectively. Twenty-five percent of transplant donors were infected with HCMV. An increase in HCMV infection was observed from pre- to post-HSCT conditions. Detection of the gB2 UL55 genotype in most transplant patient samples suggested the need to examine the possible impact of HCMV UL55 genotypes and HCMV infections among stem cell transplant recipients.

  15. A seismic fault recognition method based on ant colony optimization

    NASA Astrophysics Data System (ADS)

    Chen, Lei; Xiao, Chuangbai; Li, Xueliang; Wang, Zhenli; Huo, Shoudong

    2018-05-01

    Fault recognition is an important section in seismic interpretation and there are many methods for this technology, but no one can recognize fault exactly enough. For this problem, we proposed a new fault recognition method based on ant colony optimization which can locate fault precisely and extract fault from the seismic section. Firstly, seismic horizons are extracted by the connected component labeling algorithm; secondly, the fault location are decided according to the horizontal endpoints of each horizon; thirdly, the whole seismic section is divided into several rectangular blocks and the top and bottom endpoints of each rectangular block are considered as the nest and food respectively for the ant colony optimization algorithm. Besides that, the positive section is taken as an actual three dimensional terrain by using the seismic amplitude as a height. After that, the optimal route from nest to food calculated by the ant colony in each block is judged as a fault. Finally, extensive comparative tests were performed on the real seismic data. Availability and advancement of the proposed method were validated by the experimental results.

  16. Use of radio-tagging to map spatial organization and social interactions in insects.

    PubMed

    Moreau, Mathieu; Arrufat, Patrick; Latil, Gérard; Jeanson, Raphaël

    2011-01-01

    Understanding of the organization of animal societies often requires knowledge of the identity of group members and their spatial location. We propose an original experimental design to track automatically the position of individuals using radio frequency identification technology (RFID). Ants equipped with passive transponders were detected by a reader mounted on a mobile arm moving across the nest surface. We developed an algorithm to accurately extract the positions of individuals moving in two dimensions. Our method was validated on synthetic test cases and then used for characterization of the spatial distribution of ants within nests. This approach provides an amenable system for monitoring large populations of individuals over long periods of time.

  17. Phenology largely explains taller grass at successful nests in greater sage-grouse.

    PubMed

    Smith, Joseph T; Tack, Jason D; Doherty, Kevin E; Allred, Brady W; Maestas, Jeremy D; Berkeley, Lorelle I; Dettenmaier, Seth J; Messmer, Terry A; Naugle, David E

    2018-01-01

    Much interest lies in the identification of manageable habitat variables that affect key vital rates for species of concern. For ground-nesting birds, vegetation surrounding the nest may play an important role in mediating nest success by providing concealment from predators. Height of grasses surrounding the nest is thought to be a driver of nest survival in greater sage-grouse ( Centrocercus urophasianus ; sage-grouse), a species that has experienced widespread population declines throughout their range. However, a growing body of the literature has found that widely used field methods can produce misleading inference on the relationship between grass height and nest success. Specifically, it has been demonstrated that measuring concealment following nest fate (failure or hatch) introduces a temporal bias whereby successful nests are measured later in the season, on average, than failed nests. This sampling bias can produce inference suggesting a positive effect of grass height on nest survival, though the relationship arises due to the confounding effect of plant phenology, not an effect on predation risk. To test the generality of this finding for sage-grouse, we reanalyzed existing datasets comprising >800 sage-grouse nests from three independent studies across the range where there was a positive relationship found between grass height and nest survival, including two using methods now known to be biased. Correcting for phenology produced equivocal relationships between grass height and sage-grouse nest survival. Viewed in total, evidence for a ubiquitous biological effect of grass height on sage-grouse nest success across time and space is lacking. In light of these findings, a reevaluation of land management guidelines emphasizing specific grass height targets to promote nest success may be merited.

  18. Field Testing New Plot Designs and Methods for Determining Hydrophytic Vegetation during Wetland Delineations in the United States

    DTIC Science & Technology

    2014-03-01

    Trees and woody vines are sampled in large plots with 9 m (30 ft) radii. Saplings, shrubs , and herbs are sampled in nested smaller plots with 2 m (5 ft... woody vines in 9 m (30 ft) radius plots and saplings, shrubs , and herbaceous species in 2 m (5 ft) radius plots. In herbaceous meadows, only the 2 m (5...suggests stratifying vegetation by growth forms of trees, shrubs , herbs, and vines and sampling plant communities by using nested circular plots

  19. Detection of environmental sources of Histoplasma capsulatum in Chiang Mai, Thailand, by nested PCR.

    PubMed

    Norkaew, Treepradab; Ohno, Hideaki; Sriburee, Pojana; Tanabe, Koichi; Tharavichitkul, Prasit; Takarn, Piyawan; Puengchan, Tanpalang; Bumrungsri, Sara; Miyazaki, Yoshitsugu

    2013-12-01

    Histoplasmosis is a systemic mycosis caused by inhaling spores of Histoplasma capsulatum, a dimorphic fungus. This fungus grows in soil contaminated with bat and avian excreta. Each year, patients with disseminated histoplasmosis have been diagnosed in Chiang Mai, northern Thailand. No published information is currently available on the environmental sources of this fungus in Chiang Mai or anywhere else in Thailand. The aim of this study was to detect H. capsulatum in soil samples contaminated with bat guano and avian droppings by nested PCR. Two hundred and sixty-five samples were collected from the following three sources: soil contaminated with bat guano, 88 samples; soil contaminated with bird droppings, 86 samples; and soil contaminated with chicken droppings, 91 samples. Genomic DNA was directly extracted from each sample, and H. capsulatum was detected by nested PCR using a primer set specific to a gene encoding 100-kDa-like protein (HcI, HcII and HcIII, HcIV). Histoplasma capsulatum was detected in seven of 88 soil samples contaminated with bat guano, one of 21 soil samples contaminated with pigeon droppings and 10 of 91 soil samples contaminated with chicken droppings. The results indicate the possibility of the association of bat guano and chicken droppings with H. capsulatum in this area of Thailand.

  20. Microplastic at nesting grounds used by the northern Gulf of Mexico loggerhead recovery unit.

    PubMed

    Beckwith, Valencia K; Fuentes, Mariana M P B

    2018-06-01

    Microplastics can impact key habitats used by endangered species, such as marine turtles. They impact the environment by transporting toxicants and altering sediment properties affecting temperature and sediment permeability. Our study determined the exposure of the ten most important nesting sites for the Northern Gulf of Mexico Loggerhead Recovery Unit to microplastic. Sand samples were obtained at each nesting site during the 2017 nesting season and analyzed for abundance and characteristics of microplastic. Microplastic was found at all sites, with an average abundance of 61.08 ± 34.61 pieces/m 2 , and 59.9% located at the dunes, where turtles primarily nest. A gradual decrease in microplastics abundance was observed from the most western nesting ground to the east. The results from this study indicate that microplastic accumulation on nesting sites for the Northern Gulf of Mexico may be of great concern, and could negatively affect the incubating environment for marine turtles. Copyright © 2018 Elsevier Ltd. All rights reserved.

  1. COMPARISON OF A GENUS-SPECIFIC CONVENTIONAL PCR AND A SPECIES-SPECIFIC NESTED-PCR FOR MALARIA DIAGNOSIS USING FTA COLLECTED SAMPLES FROM KINGDOM OF SAUDI ARABIA.

    PubMed

    Al-Harthi, Saeed A

    2015-12-01

    Molecular tools are increasingly accepted as the most sensitive and reliable techniques for malaria diagnosis and epidemiological surveys. Also, collection of finger prick blood spots onto filter papers is the most simple and affordable method for samples preservation and posterior molecular analysis, especially in rural endemic regions where malaria remains a major health problem. Two malaria molecular diagnostic tests, a Plasmodium genus-specific conventional PCR and a Plasmodium species-specific Nested PCR, were evaluated using DNA templates prepared from Whatman-FTA cards' dry blood spots using both, Methanol-fixation/Heat-extraction and FTA commercial purification kit. A total of 121 blood samples were collected from six Saudi south-western endemic districts both, as thick and thin films for routine microscopic screening and onto FTA cards for molecular studies. Out of the 121 samples, 75 were P. falciparum positive by at least one technique. No other species of Plasmodium were detected. P. falciparum parasites were identified in 69/75 (92%) samples by microscopic screening in health care centers. P. genus-specific PCR was able to amplify P. falciparum DNA in 41/75 (55%) and 59/75 (79%) samples using Methanol-fixation/Heat-extraction and FTA purification kit, respectively. P. species-specific Nested PCR revealed 68/75 (91%) and 75/75 (100%) positive samples using DNA templates were isolated by Methanol-fixation/Heat- extraction and FTA purification methods, respectively. The species-specific Nested PCR applied to Whatman-FTA preserved and processed blood samples represents the best alternative to classical microscopy for malaria diagnosis, particularly in epidemiological screening.

  2. A new real-time PCR protocol for detection of avian haemosporidians.

    PubMed

    Bell, Jeffrey A; Weckstein, Jason D; Fecchio, Alan; Tkach, Vasyl V

    2015-07-19

    Birds possess the most diverse assemblage of haemosporidian parasites; including three genera, Plasmodium, Haemoproteus, and Leucocytozoon. Currently there are over 200 morphologically identified avian haemosporidian species, although true species richness is unknown due to great genetic diversity and insufficient sampling in highly diverse regions. Studies aimed at surveying haemosporidian diversity involve collecting and screening samples from hundreds to thousands of individuals. Currently, screening relies on microscopy and/or single or nested standard PCR. Although effective, these methods are time and resource consuming, and in the case of microscopy require substantial expertise. Here we report a newly developed real-time PCR protocol designed to quickly and reliably detect all three genera of avian haemosporidians in a single biochemical reaction. Using available DNA sequences from avian haemosporidians we designed primers R330F and R480RL, which flank a 182 base pair fragment of mitochondrial conserved rDNA. These primers were initially tested using real-time PCR on samples from Malawi, Africa, previously screened for avian haemosporidians using traditional nested PCR. Our real time protocol was further tested on 94 samples from the Cerrado biome of Brazil, previously screened using a single PCR assay for haemosporidian parasites. These samples were also amplified using modified nested PCR protocols, allowing for comparisons between the three different screening methods (single PCR, nested PCR, real-time PCR). The real-time PCR protocol successfully identified all three genera of avian haemosporidians from both single and mixed infections previously detected from Malawi. There was no significant difference between the three different screening protocols used for the 94 samples from the Brazilian Cerrado (χ(2) = 0.3429, df = 2, P = 0.842). After proving effective, the real-time protocol was used to screen 2113 Brazilian samples, identifying 693 positive samples. Our real-time PCR assay proved as effective as two widely used molecular screening techniques, single PCR and nested PCR. However, the real-time protocol has the distinct advantage of detecting all three genera in a single reaction, which significantly increases efficiency by greatly decreasing screening time and cost. Our real-time PCR protocol is therefore a valuable tool in the quickly expanding field of avian haemosporidian research.

  3. New insights in permafrost modelling

    NASA Astrophysics Data System (ADS)

    Tubini, Niccolò; Serafin, Francesco; Gruber, Stephan; Casulli, Vincenzo; Rigon, Riccardo

    2017-04-01

    Simulating freezing soil has ignored for long time in mainstream surface hydrology. However, it has indubitably a large influence on soil infiltrability and an even larger influence on the soil energy budget, and, over large spatial scales, a considerable feedback on climate. The topic is difficult because it involves concepts of disequilibrium Thermodynamics and also because, once solved the theoretical problem, integration of the resulting partial differential equations in a robust manner, is not trivial at all. In this abstract, we are presenting a new algorithm to estimate the water and energy budget in freezing soils. The first step is a derivation of a new equation for freezing soil mass budget (called generalized Richards equation) based on the freezing equals drying hypothesis (Miller 1965). The second step is the re-derivation of the energy budget. Finally there is the application of new techniques based on the double nested Newton algorithm (Casulli and Zanolli, 2010) to integrate the coupled equations. Some examples of the freezing dynamics and comparison with the Dall'Amico et al. (2011) algorithm are also shown. References Casulli, V., & Zanolli,P. (2010). A nested newton-type algorithm for finite colume methods solving Richards' equation in mixed form. SIAM J. SCI. Comput., 32(4), 2225-2273. Dall'Amico, M., Endrizzi, S., Gruber, S., & Rigon, R. (2011). A robust and energy-conserving model of freezing variably-saturated soil. The Cryosphere, 5(2), 469-484. http://doi.org/10.5194/tc-5-469-2011 Miller, R.: Phase equilibria and soil freezing, in: Permafrost: Proceedings of the Second International Conference. Washington DC: National Academy of Science-National Research Council, 287, 193-197, 1965.

  4. Detection of oral HPV infection - Comparison of two different specimen collection methods and two HPV detection methods.

    PubMed

    de Souza, Marjorie M A; Hartel, Gunter; Whiteman, David C; Antonsson, Annika

    2018-04-01

    Very little is known about the natural history of oral HPV infection. Several different methods exist to collect oral specimens and detect HPV, but their respective performance characteristics are unknown. We compared two different methods for oral specimen collection (oral saline rinse and commercial saliva kit) from 96 individuals and then analyzed the samples for HPV by two different PCR detection methods (single GP5+/6+ PCR and nested MY09/11 and GP5+/6+ PCR). For the oral rinse samples, the oral HPV prevalence was 10.4% (GP+ PCR; 10% repeatability) vs 11.5% (nested PCR method; 100% repeatability). For the commercial saliva kit samples, the prevalences were 3.1% vs 16.7% with the GP+ PCR vs the nested PCR method (repeatability 100% for both detection methods). Overall the agreement was fair or poor between samples and methods (kappa 0.06-0.36). Standardizing methods of oral sample collection and HPV detection would ensure comparability between future oral HPV studies. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Arboreal Ant Colonies as ‘Hot-Points’ of Cryptic Diversity for Myrmecophiles: The Weaver Ant Camponotus sp. aff. textor and Its Interaction Network with Its Associates

    PubMed Central

    Pérez-Lachaud, Gabriela; Lachaud, Jean-Paul

    2014-01-01

    Introduction Systematic surveys of macrofaunal diversity within ant colonies are lacking, particularly for ants nesting in microhabitats that are difficult to sample. Species associated with ants are generally small and rarely collected organisms, which makes them more likely to be unnoticed. We assumed that this tendency is greater for arthropod communities in microhabitats with low accessibility, such as those found in the nests of arboreal ants that may constitute a source of cryptic biodiversity. Materials and Methods We investigated the invertebrate diversity associated with an undescribed, but already threatened, Neotropical Camponotus weaver ant. As most of the common sampling methods used in studies of ant diversity are not suited for evaluating myrmecophile diversity within ant nests, we evaluated the macrofauna within ant nests through exhaustive colony sampling of three nests and examination of more than 80,000 individuals. Results We identified invertebrates from three classes belonging to 18 taxa, some of which were new to science, and recorded the first instance of the co-occurrence of two brood parasitoid wasp families attacking the same ant host colony. This diversity of ant associates corresponded to a highly complex interaction network. Agonistic interactions prevailed, but the prevalence of myrmecophiles was remarkably low. Conclusions Our data support the hypothesis of the evolution of low virulence in a variety of symbionts associated with large insect societies. Because most myrmecophiles found in this work are rare, strictly specific, and exhibit highly specialized biology, the risk of extinction for these hitherto unknown invertebrates and their natural enemies is high. The cryptic, far unappreciated diversity within arboreal ant nests in areas at high risk of habitat loss qualifies these nests as ‘hot-points’ of biodiversity that urgently require special attention as a component of conservation and management programs. PMID:24941047

  6. Development and Validation of an Improved PCR Method Using the 23S-5S Intergenic Spacer for Detection of Rickettsiae in Dermacentor variabilis Ticks and Tissue Samples from Humans and Laboratory Animals

    PubMed Central

    Kakumanu, Madhavi L.; Ponnusamy, Loganathan; Sutton, Haley T.; Meshnick, Steven R.; Nicholson, William L.

    2016-01-01

    A novel nested PCR assay was developed to detect Rickettsia spp. in ticks and tissue samples from humans and laboratory animals. Primers were designed for the nested run to amplify a variable region of the 23S-5S intergenic spacer (IGS) of Rickettsia spp. The newly designed primers were evaluated using genomic DNA from 11 Rickettsia species belonging to the spotted fever, typhus, and ancestral groups and, in parallel, compared to other Rickettsia-specific PCR targets (ompA, gltA, and the 17-kDa protein gene). The new 23S-5S IGS nested PCR assay amplified all 11 Rickettsia spp., but the assays employing other PCR targets did not. The novel nested assay was sensitive enough to detect one copy of a cloned 23S-5S IGS fragment from “Candidatus Rickettsia amblyommii.” Subsequently, the detection efficiency of the 23S-5S IGS nested assay was compared to those of the other three assays using genomic DNA extracted from 40 adult Dermacentor variabilis ticks. The nested 23S-5S IGS assay detected Rickettsia DNA in 45% of the ticks, while the amplification rates of the other three assays ranged between 5 and 20%. The novel PCR assay was validated using clinical samples from humans and laboratory animals that were known to be infected with pathogenic species of Rickettsia. The nested 23S-5S IGS PCR assay was coupled with reverse line blot hybridization with species-specific probes for high-throughput detection and simultaneous identification of the species of Rickettsia in the ticks. “Candidatus Rickettsia amblyommii,” R. montanensis, R. felis, and R. bellii were frequently identified species, along with some potentially novel Rickettsia strains that were closely related to R. bellii and R. conorii. PMID:26818674

  7. Development and Validation of an Improved PCR Method Using the 23S-5S Intergenic Spacer for Detection of Rickettsiae in Dermacentor variabilis Ticks and Tissue Samples from Humans and Laboratory Animals.

    PubMed

    Kakumanu, Madhavi L; Ponnusamy, Loganathan; Sutton, Haley T; Meshnick, Steven R; Nicholson, William L; Apperson, Charles S

    2016-04-01

    A novel nested PCR assay was developed to detectRickettsiaspp. in ticks and tissue samples from humans and laboratory animals. Primers were designed for the nested run to amplify a variable region of the 23S-5S intergenic spacer (IGS) ofRickettsiaspp. The newly designed primers were evaluated using genomic DNA from 11Rickettsiaspecies belonging to the spotted fever, typhus, and ancestral groups and, in parallel, compared to otherRickettsia-specific PCR targets (ompA,gltA, and the 17-kDa protein gene). The new 23S-5S IGS nested PCR assay amplified all 11Rickettsiaspp., but the assays employing other PCR targets did not. The novel nested assay was sensitive enough to detect one copy of a cloned 23S-5S IGS fragment from "CandidatusRickettsia amblyommii." Subsequently, the detection efficiency of the 23S-5S IGS nested assay was compared to those of the other three assays using genomic DNA extracted from 40 adultDermacentor variabilisticks. The nested 23S-5S IGS assay detectedRickettsiaDNA in 45% of the ticks, while the amplification rates of the other three assays ranged between 5 and 20%. The novel PCR assay was validated using clinical samples from humans and laboratory animals that were known to be infected with pathogenic species ofRickettsia The nested 23S-5S IGS PCR assay was coupled with reverse line blot hybridization with species-specific probes for high-throughput detection and simultaneous identification of the species ofRickettsiain the ticks. "CandidatusRickettsia amblyommii,"R. montanensis,R. felis, andR. belliiwere frequently identified species, along with some potentially novelRickettsiastrains that were closely related toR. belliiandR. conorii. Copyright © 2016 Kakumanu et al.

  8. Comparison of nested-multiplex, Taqman & SYBR Green real-time PCR in diagnosis of amoebic liver abscess in a tertiary health care institute in India

    PubMed Central

    Dinoop, K.P.; Parija, Subhash Chandra; Mandal, Jharna; Swaminathan, R.P.; Narayanan, P.

    2016-01-01

    Background & objectives: Amoebiasis is a common parasitic infection caused by Entamoeba histolytica and amoebic liver abscess (ALA) is the most common extraintestinal manifestation of amoebiasis. The aim of this study was to standardise real-time PCR assays (Taqman and SYBR Green) to detect E. histolytica from liver abscess pus and stool samples and compare its results with nested-multiplex PCR. Methods: Liver abscess pus specimens were subjected to DNA extraction. The extracted DNA samples were subjected to amplification by nested-multiplex PCR, Taqman (18S rRNA) and SYBR Green real-time PCR (16S-like rRNA assays to detect E. histolytica/E. dispar/E. moshkovskii). The amplification products were further confirmed by DNA sequence analysis. Receiver operator characteristic (ROC) curve analysis was done for nested-multiplex and SYBR Green real-time PCR and the area under the curve was calculated for evaluating the accuracy of the tests to dignose ALA. Results: In all, 17, 19 and 25 liver abscess samples were positive for E. histolytica by nested-multiplex PCR, SYBR Green and Taqman real-time PCR assays, respectively. Significant differences in detection of E. histolytica were noted in the real-time PCR assays evaluated (P<0.0001). The nested-multiplex PCR, SYBR Green real-time PCR and Taqman real-time PCR evaluated showed a positivity rate of 34, 38 and 50 per cent, respectively. Based on ROC curve analysis (considering Taqman real-time PCR as the gold standard), it was observed that SYBR Green real-time PCR was better than conventional nested-multiplex PCR for the diagnosis of ALA. Interpretation & conclusions: Taqman real-time PCR targeting the 18S rRNA had the highest positivity rate evaluated in this study. Both nested multiplex and SYBR Green real-time PCR assays utilized were evaluated to give accurate results. Real-time PCR assays can be used as the gold standard in rapid and reliable diagnosis, and appropriate management of amoebiasis, replacing the conventional molecular methods. PMID:26997014

  9. Differential parental care by adult Mountain Plovers, Charadrius montanus

    USGS Publications Warehouse

    Dinsmore, S.J.; Knopf, F.L.

    2005-01-01

    We studied chick survival of the Mountain Plover (Charadrius montanus) in Montana and found that chicks tended by females had higher survival rates than chicks tended by males, and that chick survival generally increased during the nesting season. Differences in chick survival were most pronounced early in the nesting season, and may be related to a larger sample of nests during this period. When compared to information about the nest survival of male- and female-tended plover nests, our chick data suggest a trade-off for adult plovers between the egg and chick phases of reproduction. Because Mountain Plover pairs have clutches at two nests at two different locations and show differential success between the sexes during the egg and chick phases, we offer that the Mountain Plover breeding system favours optimizing annual recruitment in a dynamic ecologic setting driven by annually unpredictable drought, grazing, and predation pressures.

  10. Use of geostatistics to determine the spatial distribution and infestation rate of leaf-cutting ant nests (Hymenoptera: Formicidae) in eucalyptus plantations.

    PubMed

    Lasmar, O; Zanetti, R; dos Santos, A; Fernandes, B V

    2012-08-01

    One of the fundamental steps in pest sampling is the assessment of the population distribution in the field. Several studies have investigated the distribution and appropriate sampling methods for leaf-cutting ants; however, more reliable methods are still required, such as those that use geostatistics. The objective of this study was to determine the spatial distribution and infestation rate of leaf-cutting ant nests in eucalyptus plantations by using geostatistics. The study was carried out in 2008 in two eucalyptus stands in Paraopeba, Minas Gerais, Brazil. All of the nests in the studied area were located and used for the generation of GIS maps, and the spatial pattern of distribution was determined considering the number and size of nests. Each analysis and map was made using the R statistics program and the geoR package. The nest spatial distribution in a savanna area of Minas Gerais was clustered to a certain extent. The models generated allowed the production of kriging maps of areas infested with leaf-cutting ants, where chemical intervention would be necessary, reducing the control costs, impact on humans, and the environment.

  11. Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR).

    PubMed

    Rigotto, C; Sincero, T C M; Simões, C M O; Barardi, C R M

    2005-01-01

    We tested three PCR based methodologies to detect adenoviruses associated with cultivated oysters. Conventional-PCR, nested-PCR, and integrated cell culture-PCR (ICC/PCR) were first optimized using oysters seeded with know amounts of Adenovirus serotype 5 (Ad5). The maximum sensitivity for Ad5 detection was determined for each method, and then used to detect natural adenovirus contamination in oysters from three aquiculture farms in Florianopolis, Santa Catarina State, Brazil, over a period of 6 months. The results showed that the nested-PCR was more sensitive (limit of detection: 1.2 PFU/g of tissue) than conventional-PCR and ICC-PCR (limit of detection for both: 1.2 x 10(2)PFU/g of tissue) for detection of Ad5 in oyster extracts. Nested-PCR was able to detect 90% of Ad5 contamination in harvested oyster samples, while conventional-PCR was unable to detect Ad5 in any of the samples. The present work suggests that detection of human adenoviruses can be used as a tool to monitor the presence of human viruses in marine environments where shellfish grow, and that nested-PCR is the method of choice.

  12. Antennal responses of an oligolectic bee and its cleptoparasite to plant volatiles.

    PubMed

    Dötterl, Stefan

    2008-05-01

    Cleptoparasitic or cuckoo bees lay their eggs in nests of other bees, and the parasitic larvae feed the food that had been provided for the host larvae. Nothing is known about the specific signals used by the cuckoo bees for host nest finding, but previous studies have shown that olfactory cues originating from the host bee alone, or the host bee and the larval provision are essential. Here, I compared by using gas chromatography coupled to electroantennographic detection (GC-EAD) the antennal responses of the oligolectic oil-bee Macropis fulvipes and their cleptoparasite, Epeoloides coecutiens, to dynamic headspace scent samples of Lysimachia punctata, a pollen and oil host of Macropis. Both bee species respond to some scent compounds emitted by L. punctata, and two compounds, which were also found in scent samples collected from a Macropis nest entrance, elicited clear signals in the antennae of both species. These compounds may not only play a role for host plant detection by Macropis, but also for host nest detection by Epeoloides. I hypothesise that oligolectic bees and their cleptoparasites use the same compounds for host plant and host nest detection, respectively.

  13. [Pollen analysis of the post-emergence residue of Centris tarsata Smith (Hymenoptera: Apidae, Centridini) nests].

    PubMed

    Dórea, Marcos da C; Dos Santos, Francisco de A R; Lima, Luciene C de L E; Figueroa, Luís E R

    2009-01-01

    A new treatment protocol was developed to analyze pollen residues found in nests of Centris tarsata Smith harvested from nest-traps. The study area was located in the Canudos Biological Station in the municipality of Canudos (09 masculine56'34'S; 38 masculine59'17'W), in the northeastern micro-region of Bahia State, Brazil. The local vegetation is an open caatinga (deciduous dryland vegetation), the regional climate is semi-arid, the average annual temperature is 24.1 masculineC, and the annual regional rainfall rate is 454 mm. Ten nests of C. tarsata were collected in trap-nests during the first semester of 2004. Pollen analysis from the nests required the development of a new methodology that combined techniques of palynological sediment analysis with the more common pollinic analysis by acetolysis. Microscopic analyses employed optical microscopy techniques. The pollinic spectrum of the samples from C. tarsata indicated the presence of 17 pollen types from seven plant families, which were present in assemblage of five to eleven pollen types, pointed to the plants used by bees to feed on their offspring. The most represented plant families were Leguminosae (49.3%) and Solanaceae (43.2%). The most frequent pollen types in the samples were from Solanum paniculatum (43.8%) and Senna rizzini (32.1%). The protocol developed provides a new tool for diet assessment of Centris and other groups of solitary bees.

  14. A two-stage adaptive stochastic collocation method on nested sparse grids for multiphase flow in randomly heterogeneous porous media

    NASA Astrophysics Data System (ADS)

    Liao, Qinzhuo; Zhang, Dongxiao; Tchelepi, Hamdi

    2017-02-01

    A new computational method is proposed for efficient uncertainty quantification of multiphase flow in porous media with stochastic permeability. For pressure estimation, it combines the dimension-adaptive stochastic collocation method on Smolyak sparse grids and the Kronrod-Patterson-Hermite nested quadrature formulas. For saturation estimation, an additional stage is developed, in which the pressure and velocity samples are first generated by the sparse grid interpolation and then substituted into the transport equation to solve for the saturation samples, to address the low regularity problem of the saturation. Numerical examples are presented for multiphase flow with stochastic permeability fields to demonstrate accuracy and efficiency of the proposed two-stage adaptive stochastic collocation method on nested sparse grids.

  15. Evaluation of nested PCR in diagnosis of fungal rhinosinusitis.

    PubMed

    Badiee, Parisa; Gandomi, Behrooz; Sabz, Gholamabbass; Khodami, Bijan; Choopanizadeh, Maral; Jafarian, Hadis

    2015-02-01

    Given the importance of rapid diagnosis for fungal rhinosinusitis, this study aimed to evaluate the use of nested PCR to identify Aspergillus and Mucor species in clinical samples from patients with suspected fungal rhinosinusitis. Functional endoscopic sinus surgery specimens were collected from 98 patients with rhinosinusitis from 2012 to 2013. All samples were ground and cultured on sabouraud dextrose agar. The isolated fungi were identified based on their macroscopic and microscopic features. Fungal DNA was extracted from the tissue samples and nested PCR was performed with two sets of primers for Mucor and Aspergillus. Direct microscopic showed that 5.1% contained fungal components and 9.2% exhibited growth of fungi in culture. The most common agents isolated were Aspergillus fumigatus (n= 3), Aspergillus flavus (n=2), Penicillium sp (n=3) and Alternaria sp. (n=1). Mucor sp. was identified in the pathology smear from 1 patient. Positive results for fungal rhinosinusitis were obtained for a total of 10.2% by culture or pathology smear. Positive PCR results were obtained in 72 samples for Aspergillus and 31 samples for Mucor. Our results suggest that endoscopic sinus surgery specimens are not suitable for nested PCR, probably because of the accumulation of fungi that contaminate the environmental air. This drawback is a limiting factor for diagnosis with nasal cavity specimens. Therefore, molecular methods and conventional culture techniques are helpful complementary diagnostic methods to detect fungal rhinosinusitis and determine appropriate management for these patients.

  16. Sampling animal sign in heterogeneous environments: how much is enough?

    USGS Publications Warehouse

    Holbrook, Joseph D.; Arkle, Robert S.; Rachlow, Janet L.; Vierling, Kerri T.; Pilliod, David S.

    2015-01-01

    Animal ecologists often use animal sign as a surrogate for direct observation of organisms, especially when species are secretive or difficult to observe. Spatial heterogeneity in arid environments makes it challenging to consistently detect and precisely characterize animal sign, which can bias estimates of animal abundance or habitat use. Piute ground squirrels (Urocitellus mollis) and Owyhee harvester ants (Pogonomyrmex salinus) live in arid environments and are fossorial, which can make them difficult to observe directly. Their relative abundance can be assessed using sign (i.e., burrows and nests). We implemented an over-sampling framework (i.e., recorded an excessive amount of information) with two observers to 1) identify a sampling intensity that balanced precision with our resource constraints, and 2) assess classification and detection of squirrel burrows and ant nests across vegetation conditions. We sampled 20 1-ha plots for ground squirrel burrows and ant nests using six 4 m × 100 m belt transects. Analyses of precision and sampling effort indicated that three belt transects covering 1200 m2 per ha provided sufficient precision, while minimizing effort. Regardless of vegetation conditions, counts by two observers were strongly correlated for ground squirrel burrows (r = 0.99, P < 0.001, df = 18; slope = 0.92) and harvester ant nests (r = 0.99, P < 0.001, df = 18; slope = 1.01) indicating observer consistency and perhaps high detection probability. These findings illustrate an approach for evaluating sampling designs in many ecological contexts.

  17. Efficiency of Nested-PCR in Detecting Asymptomatic Cases toward Malaria Elimination Program in an Endemic Area of Iran.

    PubMed

    Turki, Habibollah; Raeisi, Ahmad; Malekzadeh, Kianoosh; Ghanbarnejad, Amin; Zoghi, Samaneh; Yeryan, Masoud; Abedi Nejad, Masoumeh; Mohseni, Fatemeh; Shekari, Mohammad

    2015-01-01

    The aim of this study was to detect low parasite and asymptomatic malaria infections by means of three malaria diagnostic tests, in a low transmission region of Minab district, Hormozgan Province, southern Iran. Blood samples of 200 healthy volunteers from Bagh-e-Malek area were evaluated using microscopic, rapid diagnostic tests (RDT) and nested-PCR to inspect malaria parasite. The results showed no Plasmodium parasite in subjects by means of microscopy and RDT. However, 3 P. vivax positive samples (1.5%) were discovered by Nested-PCR while microscopy and RDT missed the cases. Microscopy as the gold standard method and RDT correctly identified 98.5% of cases, and molecular analysis is sensitive and reliable, especially in the detection of "asymptomatic" infections for active case surveillance. Regarding the existence of asymptomatic malaria in endemic area of Hormozgan, Iran, nested-PCR could be considered as a sensitive tool to interrupt malaria transmission in the country, beside the microscopic and RDT methods.

  18. Elevated temperatures are associated with stress in rooftop-nesting Common Nighthawk (Chordeiles minor) chicks.

    PubMed

    Newberry, Gretchen N; Swanson, David L

    2018-01-01

    Grasslands and riparian forests in southeastern South Dakota have been greatly reduced since historical times, primarily due to conversion to row-crop agriculture. Common Nighthawk ( Chordeiles minor ) nesting habitat includes grasslands, open woodlands and urban rooftops, but nesting sites in southeastern South Dakota are confined to rooftops, as natural nesting habitat is limited. Nighthawks nesting on exposed rooftop habitats may encounter thermal conditions that increase operative temperatures relative to vegetated land cover types. Mean humidity has increased and mean wind speed and cloud cover have decreased during the nighthawk breeding season from 1948 to 2016 in southeastern South Dakota. These changes might contribute to increasing operative temperatures at exposed rooftop nest sites and this could influence chick condition. We studied nest micro-climate and the plasma stress response for 24 rooftop-nesting nighthawk chicks from 17 nests during 2015 and 2016. High humidity prior to blood collection reduced both baseline and stress-induced plasma corticosterone (CORT). In contrast, high maximum temperatures during the day before sampling increased stress-induced CORT. The magnitude of the chick stress response was significantly negatively related to maximum wind speed for the week prior to CORT measurement. Other weather and micro-climate variables were not significant effectors of CORT metrics. Most chicks had low baseline CORT and were able to mount a stress response, but a subset of chicks ( n = 4) showed elevated baseline CORT and a negative association between the magnitude of stress response and ambient temperature. For this subset, mean ambient temperature for the day before sampling was significantly higher (2.3°C) than for chicks with typical baseline CORT levels. These data suggest that regional climate change trends could affect the ability of nighthawk chicks to mount a stress response, which, in turn, might influence the susceptibility of nighthawk chicks to climate change in the Northern Prairie region.

  19. Contaminants in ospreys from the Pacific Northwest: I. Trends and Patterns in polychlorinated dibenzo-p-dioxins and -dibenzofurans in eggs and plasma

    USGS Publications Warehouse

    Elliott, J.E.; Machmer, M.M.; Henny, Charles J.; Wilson, L.K.; Norstrom, R.J.

    1998-01-01

    Osprey (Pandion haliaetus) eggs were collected from 1991 to 1997 at nests (na??=a??121) upstream and downstream of bleached kraft pulp mills and at reference sites in the Fraser and Columbia River drainage systems of British Columbia, Washington, and Oregon. Blood samples were collected from nestling ospreys during the 1992 breeding season on the Thompson River. Samples were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs) and -dibenzofurans (PCDFs). Mean concentrations of 2,3,7,8-TCDD were significantly higher in eggs collected in 1991 at downstream compared to upstream nests near pulp mills at Kamloops and Castlegar, British Columbia. There were no significant temporal trends in 2,3,7,8-TCDD, -TCDF or other measured compounds at a sample of nests monitored between 1991 and 1994 downstream of the Castlegar pulp mill, despite changes in bleaching technology (CIO2 substitution). However, by 1997 concentrations of 2,3,7,8-TCDD and -TCDF were significantly lower than previous years in nests sampled downstream at both Castlegar and Kamloops. An unusual pattern of higher chlorinated PCDDs and PCDFs was found in many of the osprey eggs collected in this study, and considerable individual variation in the pattern existed among eggs from the same site. For example, eggs from four different nests at one study area (Quesnel) on the Fraser River had concentrations of 1,2,3,4,6,7,8-HpCDD ranging from <1 to 1,100 ng/kg and OCDD from <1 to 7,000 ng/kg wet weight. Higher mean concentrations of HpCDD and OCDD were found in eggs from the Thompson River, a tributary of the Fraser, compared to the Columbia River, and concentrations were generally higher at nests upstream of pulp mills. In plasma samples, 1,2,3,4,6,7,8-HpCDD and OCDD were the main compounds detected, with no significant differences measured between samples upstream versus downstream or earlier versus later in the breeding season. Use of chlorophenolic wood preservatives by lumber processors was considered the main source of higher chlorinated PCDD/Fs throughout the systems, based on patterns of trace PCDFs in eggs and significant correlations between egg concentrations of pentachlorophenol and both HpCDD (ra??=a??0.891, pa??

  20. Contaminants in ospreys from the Pacific Northwest: I. Trends and patterns in polychlorinated dibenzo-p-dioxins and -dibenzofurans in eggs and plasma.

    PubMed

    Elliott, J E; Machmer, M M; Henny, C J; Wilson, L K; Norstrom, R J

    1998-11-01

    Osprey (Pandion haliaetus) eggs were collected from 1991 to 1997 at nests (n = 121) upstream and downstream of bleached kraft pulp mills and at reference sites in the Fraser and Columbia River drainage systems of British Columbia, Washington, and Oregon. Blood samples were collected from nestling ospreys during the 1992 breeding season on the Thompson River. Samples were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs) and -dibenzofurans (PCDFs). Mean concentrations of 2,3,7,8-TCDD were significantly higher in eggs collected in 1991 at downstream compared to upstream nests near pulp mills at Kamloops and Castlegar, British Columbia. There were no significant temporal trends in 2,3,7,8-TCDD, -TCDF or other measured compounds at a sample of nests monitored between 1991 and 1994 downstream of the Castlegar pulp mill, despite changes in bleaching technology (CIO2 substitution). However, by 1997 concentrations of 2, 3,7,8-TCDD and -TCDF were significantly lower than previous years in nests sampled downstream at both Castlegar and Kamloops. An unusual pattern of higher chlorinated PCDDs and PCDFs was found in many of the osprey eggs collected in this study, and considerable individual variation in the pattern existed among eggs from the same site. For example, eggs from four different nests at one study area (Quesnel) on the Fraser River had concentrations of 1,2,3,4,6,7,8-HpCDD ranging from <1 to 1,100 ng/kg and OCDD from <1 to 7,000 ng/kg wet weight. Higher mean concentrations of HpCDD and OCDD were found in eggs from the Thompson River, a tributary of the Fraser, compared to the Columbia River, and concentrations were generally higher at nests upstream of pulp mills. In plasma samples, 1,2,3,4,6,7,8-HpCDD and OCDD were the main compounds detected, with no significant differences measured between samples upstream versus downstream or earlier versus later in the breeding season. Use of chlorophenolic wood preservatives by lumber processors was considered the main source of higher chlorinated PCDD/Fs throughout the systems, based on patterns of trace PCDFs in eggs and significant correlations between egg concentrations of pentachlorophenol and both HpCDD (r = 0.891, p < 0.01) and OCDD (r = 0.870, p < 0.01).

  1. Detection of Histoplasma capsulatum in Organic Fertilizers by Hc100 Nested Polymerase Chain Reaction and Its Correlation with the Physicochemical and Microbiological Characteristics of the Samples.

    PubMed

    Gómez, Luisa F; Torres, Isaura P; Jiménez-A, María Del Pilar; McEwen, Juan Gmo; de Bedout, Catalina; Peláez, Carlos A; Acevedo, José M; Taylor, María L; Arango, Myrtha

    2018-05-01

    Histoplasma capsulatum is the causative agent of histoplasmosis and this fungus inhabits soils rich in phosphorus and nitrogen that are enriched with bird and bat manure. The replacement of organic matter in agroecosystems is necessary in the tropics, and the use of organic fertilizers has increased. Cases and outbreaks due to the presence of the fungus in these components have been reported. The Instituto Colombiano Agropecuario resolution 150 of 2003 contains the parameters set by the Colombian Technical Standard (NTC 5167) on the physicochemical and microbiological features of fertilizers, but it does not regulate the search for H. capsulatum . The aim of this study was to demonstrate H. capsulatum presence in organic fertilizers by nested polymerase chain reaction (PCR). A total of 239 samples were collected: 201 (84.1%) corresponded to organic fertilizers, 30 (12.5%) to bird excrement, and 8 (3.4%) to cave soils. The Hc100 nested PCR had a detection limit of 0.1 pg/µL and a specificity of 100%. A total of 25 (10.5%) samples were positive and validated by sequencing. Seven of the positive samples represented locations where H. capsulatum was previously detected, suggesting the persistence of the fungus. No significant correlations were detected between the physicochemical and microbiological parameters with the presence of H. capsulatum by nested PCR, indicating the fungus existence in organic fertilizers that complied with the NTC 5167. The Hc100 nested PCR targeting H. capsulatum standardized in this work will improve the evaluation of organic fertilizers and ensure the prevention of outbreaks and cases due to manufacturing, marketing, and use of fertilizers contaminated with H. capsulatum .

  2. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR.

    PubMed

    Araújo, Cristina P; Osório, Ana Luiza A R; Jorge, Klaudia S G; Ramos, Carlos A N; Souza Filho, Antonio F; Vidal, Carlos E S; Vargas, Agueda P C; Roxo, Eliana; Rocha, Adalgiza S; Suffys, Philip N; Fonseca, Antônio A; Silva, Marcio R; Barbosa Neto, José D; Cerqueira, Valíria D; Araújo, Flábio R

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

  3. Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

    PubMed Central

    Araújo, Cristina P.; Osório, Ana Luiza A.R.; Jorge, Klaudia S.G.; Ramos, Carlos A.N.; Souza Filho, Antonio F.; Vidal, Carlos E.S.; Vargas, Agueda P.C.; Roxo, Eliana; Rocha, Adalgiza S.; Suffys, Philip N.; Fonseca, Antônio A.; Silva, Marcio R.; Barbosa Neto, José D.; Cerqueira, Valíria D.; Araújo, Flábio R.

    2014-01-01

    Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis. PMID:25242951

  4. Real-time PCR assay in differentiating Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii infections in Orang Asli settlements in Malaysia.

    PubMed

    Lau, Yee Ling; Anthony, Claudia; Fakhrurrazi, Siti Aminah; Ibrahim, Jamaiah; Ithoi, Init; Mahmud, Rohela

    2013-08-28

    Amebiasis caused by Entamoeba histolytica is the third leading cause of death worldwide. This pathogenic amoeba is morphologically indistinguishable from E. dispar and E. moshkovskii, the non-pathogenic species. Polymerase chain reaction is the current method of choice approved by World Health Organization. Real-time PCR is another attractive molecular method for diagnosis of infectious diseases as post-PCR analyses are eliminated and turnaround times are shorter. The present work aimed to compare the results of Entamoeba species identification using the real-time assay against the established nested PCR method. In this study, a total of 334 human faecal samples were collected from different Orang Asli settlements. Faecal samples were processed by direct wet smear and formalin ethyl acetate concentration methods followed by iodine staining and was microscopically examined for Entamoeba species and other intestinal parasites. Microscopically positive samples were then subject to nested PCR and real-time PCR. The overall prevalence of Entamoeba infection was 19.5% (65/334). SK Posh Piah recorded highest Entamoeba prevalence (63.3%) while Kampung Kemensah had the lowest prevalence (3.7%) of Entamoeba. Microscopically positive samples were then tested by real-time PCR and nested PCR for the presence of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii infection. Real-time PCR showed higher Entamoeba detection (86.2%) compared to nested PCR (80%), although the McNemar test value showed no significant difference between the two methods (p = 0.221). This study is the first in Malaysia to report the use of real-time PCR in identifying and differentiating the three Entamoeba infections. It is also proven to be more effective compared to the conventional nested PCR molecular method.

  5. Real-time PCR assay in differentiating Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii infections in Orang Asli settlements in Malaysia

    PubMed Central

    2013-01-01

    Background Amebiasis caused by Entamoeba histolytica is the third leading cause of death worldwide. This pathogenic amoeba is morphologically indistinguishable from E. dispar and E. moshkovskii, the non-pathogenic species. Polymerase chain reaction is the current method of choice approved by World Health Organization. Real-time PCR is another attractive molecular method for diagnosis of infectious diseases as post-PCR analyses are eliminated and turnaround times are shorter. The present work aimed to compare the results of Entamoeba species identification using the real-time assay against the established nested PCR method. Methods In this study, a total of 334 human faecal samples were collected from different Orang Asli settlements. Faecal samples were processed by direct wet smear and formalin ethyl acetate concentration methods followed by iodine staining and was microscopically examined for Entamoeba species and other intestinal parasites. Microscopically positive samples were then subject to nested PCR and real-time PCR. Results The overall prevalence of Entamoeba infection was 19.5% (65/334). SK Posh Piah recorded highest Entamoeba prevalence (63.3%) while Kampung Kemensah had the lowest prevalence (3.7%) of Entamoeba. Microscopically positive samples were then tested by real-time PCR and nested PCR for the presence of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii infection. Real-time PCR showed higher Entamoeba detection (86.2%) compared to nested PCR (80%), although the McNemar test value showed no significant difference between the two methods (p = 0.221). Conclusions This study is the first in Malaysia to report the use of real-time PCR in identifying and differentiating the three Entamoeba infections. It is also proven to be more effective compared to the conventional nested PCR molecular method. PMID:23985047

  6. A simple and rapid method for detection of Trypanosoma evansi in the dromedary camel using a nested polymerase chain reaction

    PubMed Central

    Aradaib, Imadeldin E; Majid, Ali A

    2006-01-01

    A nested polymerase chain reaction (nPCR)-based assay, was developed and evaluated for rapid detection of Trypanosoma evansi in experimentally infected mice and naturally infected camels (Camelus dromedarius). Four oligonucleotide primers (TE1, TE2, TE3 and TE4), selected from nuclear repetitive gene of T. evansi, were designed and used for PCR amplifications. The first amplification, using a pair of outer primers TE1 and TE2, produced a 821-bp primary PCR product from T. evansi DNA. The second amplification, using nested (internal) pair of primers TE3 and TE4, produced a 270-bp PCR product. T. evansi DNAs extracted from blood samples of experimentally infected mice and naturally infected Sudanese breed of dromedary camels were detected by this nested PCR-based assay. The nested primers TE3 and TE4 increased the sensitivity of the PCR assay and as little as 10 fg of T. evansi DNA (equivalent to a single copy of the putative gene of the parasite) was amplified and visualized onto ethidium bromide-stained agarose gels. Amplification products were not detected when the PCR-based assay was applied to DNA from other blood parasites including Thieleria annulata, Babesia bigemina or nucleic acid free samples. Application of this nPCR-based assay to clinical samples resulted in direct detection of T. evansi from a variety of tissue samples collected from experimentally infected mice and blood from naturally infected camels. The described nPCR-based assay provides a valuable tool to study the epidemiology of T. evansi infection in camels and other susceptible animal populations. PMID:16712737

  7. Test plan for evaluating the operational performance of the prototype nested, fixed-depth fluidic sampler

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    REICH, F.R.

    The PHMC will provide Low Activity Wastes (LAW) tank wastes for final treatment by a privatization contractor from two double-shell feed tanks, 241-AP-102 and 241-AP-104. Concerns about the inability of the baseline ''grab'' sampling to provide large volume samples within time constraints has led to the development of a nested, fixed-depth sampling system. This sampling system will provide large volume, representative samples without the environmental, radiation exposure, and sample volume impacts of the current base-line ''grab'' sampling method. A plan has been developed for the cold testing of this nested, fixed-depth sampling system with simulant materials. The sampling system willmore » fill the 500-ml bottles and provide inner packaging to interface with the Hanford Sites cask shipping systems (PAS-1 and/or ''safe-send''). The sampling system will provide a waste stream that will be used for on-line, real-time measurements with an at-tank analysis system. The cold tests evaluate the performance and ability to provide samples that are representative of the tanks' content within a 95 percent confidence interval, to sample while mixing pumps are operating, to provide large sample volumes (1-15 liters) within a short time interval, to sample supernatant wastes with over 25 wt% solids content, to recover from precipitation- and settling-based plugging, and the potential to operate over the 20-year expected time span of the privatization contract.« less

  8. Euschoengastia suzukii (Acari: Trombiculidae): A new species of chigger mite collected from soil samples in the nest burrows of streaked shearwater in Japan.

    PubMed

    Takahashi, Mamoru; Fukaya, Hajime; Takahashi, Hisae

    2005-07-01

    Euschoengastia suzukii Takahashi, Fukaya & Takahashi is described and illustrated. The type material was collected from soil samples in the nest burrows of the sea bird streaked shearwater, Calonectris leucomelas (Temminck), living on Mikurajima Island in Tokyo; Oomorijima Island, Oki, Shimane Prefecture; and Awashima Island, Niigata Prefecture, in Japan, indicating that C. leucomelas is the main parasitic host of E. suzukii new species.

  9. Evaluating nest supplementation as a recovery strategy for the endangered rodents of the Florida Keys

    USGS Publications Warehouse

    Cove, Michael V.; Simons, Theodore R.; Gardner, Beth; Maurer, Andrew S.; O'Connell, Allan F.

    2017-01-01

    The Key Largo woodrat (Neotoma floridana smalli) and Key Largo cotton mouse (Peromyscus gossypinus allapaticola) are federally endangered subspecies endemic to the tropical hardwood hammocks of Key Largo, Florida. Woodrats are considered generalists in habitat and diet, yet a steady decline in natural stick nests and capture rates over the past several decades suggests that they are limited by the availability of nesting habitat due to habitat loss and fragmentation. The more specialized Key Largo cotton mouse appears to rely on old growth hammock, a habitat type that is rare following past land clearing. In 2004, the U.S. Fish and Wildlife Service started building supplemental nest structures to restore habitat quality and connectivity for these endangered rodents, but nest use requires evaluation. We used camera traps and occupancy models to evaluate the factors influencing woodrat and cotton mouse use of the supplemental nests. We detected woodrats at 65 and cotton mice at 175 of 284 sampled nest structures, with co-occurrence at 38 nests. Woodrat nest use followed a gradient from low nest use in the north to high nest use in the south, which might relate to the proximity of free-ranging domestic cat (Felis catus) colonies in residential developments. Cotton mouse nest use, however, was related positively to mature hammock and related negatively to disturbed areas (e.g. scarified lands). The two species occurred independently of each other. Stick-stacking behavior was observed at supplemental nests and, although it was correlated with detection of woodrats, it was not a strong predictor of their occurrence. We suggest that nest supplementation can be an important tool for species recovery as habitat quality continues to improve with succession.

  10. HELIOS–RETRIEVAL: An Open-source, Nested Sampling Atmospheric Retrieval Code; Application to the HR 8799 Exoplanets and Inferred Constraints for Planet Formation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lavie, Baptiste; Mendonça, João M.; Malik, Matej

    We present an open-source retrieval code named HELIOS–RETRIEVAL, designed to obtain chemical abundances and temperature–pressure profiles by inverting the measured spectra of exoplanetary atmospheres. In our forward model, we use an exact solution of the radiative transfer equation, in the pure absorption limit, which allows us to analytically integrate over all of the outgoing rays. Two chemistry models are considered: unconstrained chemistry and equilibrium chemistry (enforced via analytical formulae). The nested sampling algorithm allows us to formally implement Occam’s Razor based on a comparison of the Bayesian evidence between models. We perform a retrieval analysis on the measured spectra ofmore » the four HR 8799 directly imaged exoplanets. Chemical equilibrium is disfavored for HR 8799b and c. We find supersolar C/H and O/H values for the outer HR 8799b and c exoplanets, while the inner HR 8799d and e exoplanets have a range of C/H and O/H values. The C/O values range from being superstellar for HR 8799b to being consistent with stellar for HR 8799c and being substellar for HR 8799d and e. If these retrieved properties are representative of the bulk compositions of the exoplanets, then they are inconsistent with formation via gravitational instability (without late-time accretion) and consistent with a core accretion scenario in which late-time accretion of ices occurred differently for the inner and outer exoplanets. For HR 8799e, we find that spectroscopy in the K band is crucial for constraining C/O and C/H. HELIOS–RETRIEVAL is publicly available as part of the Exoclimes Simulation Platform (http://www.exoclime.org).« less

  11. Observation of quantum criticality with ultracold atoms in optical lattices

    NASA Astrophysics Data System (ADS)

    Zhang, Xibo

    As biological problems are becoming more complex and data growing at a rate much faster than that of computer hardware, new and faster algorithms are required. This dissertation investigates computational problems arising in two of the fields: comparative genomics and epigenomics, and employs a variety of computational techniques to address the problems. One fundamental question in the studies of chromosome evolution is whether the rearrangement breakpoints are happening at random positions or along certain hotspots. We investigate the breakpoint reuse phenomenon, and show the analyses that support the more recently proposed fragile breakage model as opposed to the conventional random breakage models for chromosome evolution. The identification of syntenic regions between chromosomes forms the basis for studies of genome architectures, comparative genomics, and evolutionary genomics. The previous synteny block reconstruction algorithms could not be scaled to a large number of mammalian genomes being sequenced; neither did they address the issue of generating non-overlapping synteny blocks suitable for analyzing rearrangements and evolutionary history of large-scale duplications prevalent in plant genomes. We present a new unified synteny block generation algorithm based on A-Bruijn graph framework that overcomes these shortcomings. In the epigenome sequencing, a sample may contain a mixture of epigenomes and there is a need to resolve the distinct methylation patterns from the mixture. Many sequencing applications, such as haplotype inference for diploid or polyploid genomes, and metagenomic sequencing, share the similar objective: to infer a set of distinct assemblies from reads that are sequenced from a heterogeneous sample and subsequently aligned to a reference genome. We model the problem from both a combinatorial and a statistical angles. First, we describe a theoretical framework. A linear-time algorithm is then given to resolve a minimum number of assemblies that are consistent with all reads, substantially improving on previous algorithms. An efficient algorithm is also described to determine a set of assemblies that is consistent with a maximum subset of the reads, a previously untreated problem. We then prove that allowing nested reads or permitting mismatches between reads and their assemblies renders these problems NP-hard. Second, we describe a mixture model-based approach, and applied the model for the detection of allele-specific methylations.

  12. Approximate Bayesian Computation by Subset Simulation using hierarchical state-space models

    NASA Astrophysics Data System (ADS)

    Vakilzadeh, Majid K.; Huang, Yong; Beck, James L.; Abrahamsson, Thomas

    2017-02-01

    A new multi-level Markov Chain Monte Carlo algorithm for Approximate Bayesian Computation, ABC-SubSim, has recently appeared that exploits the Subset Simulation method for efficient rare-event simulation. ABC-SubSim adaptively creates a nested decreasing sequence of data-approximating regions in the output space that correspond to increasingly closer approximations of the observed output vector in this output space. At each level, multiple samples of the model parameter vector are generated by a component-wise Metropolis algorithm so that the predicted output corresponding to each parameter value falls in the current data-approximating region. Theoretically, if continued to the limit, the sequence of data-approximating regions would converge on to the observed output vector and the approximate posterior distributions, which are conditional on the data-approximation region, would become exact, but this is not practically feasible. In this paper we study the performance of the ABC-SubSim algorithm for Bayesian updating of the parameters of dynamical systems using a general hierarchical state-space model. We note that the ABC methodology gives an approximate posterior distribution that actually corresponds to an exact posterior where a uniformly distributed combined measurement and modeling error is added. We also note that ABC algorithms have a problem with learning the uncertain error variances in a stochastic state-space model and so we treat them as nuisance parameters and analytically integrate them out of the posterior distribution. In addition, the statistical efficiency of the original ABC-SubSim algorithm is improved by developing a novel strategy to regulate the proposal variance for the component-wise Metropolis algorithm at each level. We demonstrate that Self-regulated ABC-SubSim is well suited for Bayesian system identification by first applying it successfully to model updating of a two degree-of-freedom linear structure for three cases: globally, locally and un-identifiable model classes, and then to model updating of a two degree-of-freedom nonlinear structure with Duffing nonlinearities in its interstory force-deflection relationship.

  13. The ART of mating: alternative reproductive tactics and mating success in a nest-guarding fish.

    PubMed

    Mascolino, S; Benvenuto, C; Gubili, C; Sacchi, C; Boufana, B; Mariani, S

    2016-12-01

    Behavioural observations in the field of male Mediterranean damselfish Chromis chromis were combined with molecular analyses, using bi-parentally and maternally inherited markers, to investigate reproductive success patterns of alternative reproductive tactics (ARTs) in terms of number of eggs sired and number of females contributing to each nest. Cuckoldry was observed in every nest sampled, with at least two and up to seven sneaker males per nest. The nesting male, however, always significantly fertilized the greater number of eggs (on average 49%) in each clutch, whereas each sneaker fertilized around 7% of the clutch. The average number of females whose eggs were fertilized by nesting males was 6·76 (range 2-13), while each sneaker on average fertilized the eggs of 1·74 (range 1-8) females. Using this sibship reconstruction, some of the factors involved in the regulation of the dynamic equilibrium of reproductive success were investigated between the two ARTs shown by C. chromis males. Results show that the sneakers' reproductive success was positively linked to egg clutch size; the density of individuals in the nesting area negatively affected the size of egg clutches; the rate of defence behaviours performed by nesting males negatively influenced the number of females contributing to each nest. © 2016 The Fisheries Society of the British Isles.

  14. Robustness-Based Design Optimization Under Data Uncertainty

    NASA Technical Reports Server (NTRS)

    Zaman, Kais; McDonald, Mark; Mahadevan, Sankaran; Green, Lawrence

    2010-01-01

    This paper proposes formulations and algorithms for design optimization under both aleatory (i.e., natural or physical variability) and epistemic uncertainty (i.e., imprecise probabilistic information), from the perspective of system robustness. The proposed formulations deal with epistemic uncertainty arising from both sparse and interval data without any assumption about the probability distributions of the random variables. A decoupled approach is proposed in this paper to un-nest the robustness-based design from the analysis of non-design epistemic variables to achieve computational efficiency. The proposed methods are illustrated for the upper stage design problem of a two-stage-to-orbit (TSTO) vehicle, where the information on the random design inputs are only available as sparse point and/or interval data. As collecting more data reduces uncertainty but increases cost, the effect of sample size on the optimality and robustness of the solution is also studied. A method is developed to determine the optimal sample size for sparse point data that leads to the solutions of the design problem that are least sensitive to variations in the input random variables.

  15. Identifying Kittlitz's Murrelet nesting habitat in North America at the landscape scale

    USGS Publications Warehouse

    Felis, Jonathan J.; Kissling, Michelle L.; Kaler, Robb S.A.; Kenney, Leah A.; Lawonn, Matthew J.

    2016-01-01

    The Kittlitz's Murrelet (Brachyramphus brevirostris) is a small, non-colonial seabird endemic to marine waters of Alaska and eastern Russia that may have experienced significant population decline in recent decades, in part because of low reproductive success and terrestrial threats. Although recent studies have shed new light on Kittlitz's Murrelet nesting habitat in a few discrete areas, the location and extent of suitable nesting habitat throughout most of its range remains unclear. Here, we have compiled all existing nest records and locations to identify landscape-scale parameters (distance to coast, elevation, slope, and land cover) that provide potential nesting habitat in four regions: northern Alaska, Aleutian Islands, Alaska Peninsula Mountains and Kodiak Island, and Pacific Coastal Mountains (including nearshore interior Canada). We produced a final map classifying 12% (70,411 km2) of the lands assessed as potential Kittlitz's Murrelet nesting habitat, with dense but distinct patches in northern Alaska and a more uninterrupted, narrow band extending across the Pacific Coastal Mountains, Alaska Peninsula Mountains, and Aleutian Islands. The extent of habitat-capable parameter values varied regionally, indicating that the Kittlitz's Murrelet may be able to use a variety of habitats for nesting, depending on availability. Future nesting habitat studies could employ spatially random sampling designs to allow for quantitatively robust modeling of nesting habitat and predictive extrapolation to areas where nests have not been located but likely exist.

  16. Development of a species-diagnostic marker and its application for population genetics studies of the stingless bee Trigona collina in Thailand.

    PubMed

    Theeraapisakkun, M; Klinbunga, S; Sittipraneed, S

    2010-05-18

    A molecular maker for authenticating species origin of the stingless bee (Trigona collina) was developed. Initially, amplified fragment length polymorphism analysis was made of 11 stingless bee species using 64 primer combinations. A 316-bp band found only in T. collina was cloned and sequenced. A primer pair (CUTc1-F/R) was designed and tested for species-specificity in 15 stingless bee species (239 nests). The expected 259-bp fragment was consistently amplified in all T. collina individuals (134/134 nests, 100%). Cross-species amplification was observed in T. pagdeni (43/51 nests; 84.3%), but not in other species. SSCP analysis of CUTc1 unambiguously differentiated T. collina from T. pagdeni. CUTc1 generated three genotypes in Thai T. collina (134 nests). An AA (259/259 bp) genotype was found in all stingless bees from the north (21 nests) and northeast (32 nests), and 23/28 nests from the Central region, whereas a BB (253/253 bp) genotype was observed in most samples from peninsular Thailand (42/53 nests). Heterozygotes exhibiting the AB (253/259 bp) genotype were observed in 5 of 28 nests from Prachuap Khiri Khan located slightly above the Kra ecotone and 11 of 53 nests originated further south of the Kra ecotone. Genotype distribution patterns of CUTc1 clearly indicated intraspecific population differentiation of Thai T. collina.

  17. Power lines, roads, and avian nest survival: effects on predator identity and predation intensity

    PubMed Central

    DeGregorio, Brett A; Weatherhead, Patrick J; Sperry, Jinelle H

    2014-01-01

    1 Anthropogenic alteration of landscapes can affect avian nest success by influencing the abundance, distribution, and behavior of predators. Understanding avian nest predation risk necessitates understanding how landscapes affect predator distribution and behavior. 2 From a sample of 463 nests of 17 songbird species, we evaluated how landscape features (distance to forest edge, unpaved roads, and power lines) influenced daily nest survival. We also used video cameras to identify nest predators at 137 nest predation events and evaluated how landscape features influenced predator identity. Finally, we determined the abundance and distribution of several of the principal predators using surveys and radiotelemetry. 3 Distance to power lines was the best predictor of predator identity: predation by brown-headed cowbirds (Molothrus ater), corvids (Corvus sp. and Cyanocitta cristata), racers (Coluber constrictor), and coachwhips (Masticophis flagellum) increased with proximity to power lines, whereas predation by rat snakes (Elaphe obsoleta) and raptors decreased. In some cases, predator density may reliably indicate nest predation risk because racers, corvids, and cowbirds frequently used power line right-of-ways. 4 Of five bird species with enough nests to analyze individually, daily nest survival of only indigo buntings (Passerina cyanea) decreased with proximity to power lines, despite predation by most predators at our site being positively associated with power lines. For all nesting species combined, distance to unpaved road was the model that most influenced daily nest survival. This pattern is likely a consequence of rat snakes, the locally dominant nest predator (28% of predation events), rarely using power lines and associated areas. Instead, rat snakes were frequently associated with road edges, indicating that not all edges are functionally similar. 5 Our results suggest that interactions between predators and landscape features are likely to be specific to both the local predators and landscape. Thus, predicting how anthropogenic changes to landscapes affect nesting birds requires that we know more about how landscape changes affect the behavior of nest predators and which nest predators are locally important. PMID:24967077

  18. Power lines, roads, and avian nest survival: effects on predator identity and predation intensity.

    PubMed

    DeGregorio, Brett A; Weatherhead, Patrick J; Sperry, Jinelle H

    2014-05-01

    1 Anthropogenic alteration of landscapes can affect avian nest success by influencing the abundance, distribution, and behavior of predators. Understanding avian nest predation risk necessitates understanding how landscapes affect predator distribution and behavior. 2 From a sample of 463 nests of 17 songbird species, we evaluated how landscape features (distance to forest edge, unpaved roads, and power lines) influenced daily nest survival. We also used video cameras to identify nest predators at 137 nest predation events and evaluated how landscape features influenced predator identity. Finally, we determined the abundance and distribution of several of the principal predators using surveys and radiotelemetry. 3 Distance to power lines was the best predictor of predator identity: predation by brown-headed cowbirds (Molothrus ater), corvids (Corvus sp. and Cyanocitta cristata), racers (Coluber constrictor), and coachwhips (Masticophis flagellum) increased with proximity to power lines, whereas predation by rat snakes (Elaphe obsoleta) and raptors decreased. In some cases, predator density may reliably indicate nest predation risk because racers, corvids, and cowbirds frequently used power line right-of-ways. 4 Of five bird species with enough nests to analyze individually, daily nest survival of only indigo buntings (Passerina cyanea) decreased with proximity to power lines, despite predation by most predators at our site being positively associated with power lines. For all nesting species combined, distance to unpaved road was the model that most influenced daily nest survival. This pattern is likely a consequence of rat snakes, the locally dominant nest predator (28% of predation events), rarely using power lines and associated areas. Instead, rat snakes were frequently associated with road edges, indicating that not all edges are functionally similar. 5 Our results suggest that interactions between predators and landscape features are likely to be specific to both the local predators and landscape. Thus, predicting how anthropogenic changes to landscapes affect nesting birds requires that we know more about how landscape changes affect the behavior of nest predators and which nest predators are locally important.

  19. HELIOS-R: An Ultrafast, Open-Source Retrieval Code For Exoplanetary Atmosphere Characterization

    NASA Astrophysics Data System (ADS)

    LAVIE, Baptiste

    2015-12-01

    Atmospheric retrieval is a growing, new approach in the theory of exoplanet atmosphere characterization. Unlike self-consistent modeling it allows us to fully explore the parameter space, as well as the degeneracies between the parameters using a Bayesian framework. We present HELIOS-R, a very fast retrieving code written in Python and optimized for GPU computation. Once it is ready, HELIOS-R will be the first open-source atmospheric retrieval code accessible to the exoplanet community. As the new generation of direct imaging instruments (SPHERE, GPI) have started to gather data, the first version of HELIOS-R focuses on emission spectra. We use a 1D two-stream forward model for computing fluxes and couple it to an analytical temperature-pressure profile that is constructed to be in radiative equilibrium. We use our ultra-fast opacity calculator HELIOS-K (also open-source) to compute the opacities of CO2, H2O, CO and CH4 from the HITEMP database. We test both opacity sampling (which is typically used by other workers) and the method of k-distributions. Using this setup, we compute a grid of synthetic spectra and temperature-pressure profiles, which is then explored using a nested sampling algorithm. By focusing on model selection (Occam’s razor) through the explicit computation of the Bayesian evidence, nested sampling allows us to deal with current sparse data as well as upcoming high-resolution observations. Once the best model is selected, HELIOS-R provides posterior distributions of the parameters. As a test for our code we studied HR8799 system and compared our results with the previous analysis of Lee, Heng & Irwin (2013), which used the proprietary NEMESIS retrieval code. HELIOS-R and HELIOS-K are part of the set of open-source community codes we named the Exoclimes Simulation Platform (www.exoclime.org).

  20. Breeding ecology and nesting habitat associations of five marsh bird species in western New York

    USGS Publications Warehouse

    Lor, S.; Malecki, R.A.

    2006-01-01

    Nesting habitats and nest success of five species of marsh birds were studied during 1997 and 1998 at the Iroquois National Wildlife Refuge (NWR) and the adjacent Oak Orchard and Tonawanda State Wildlife Management Areas (WMA) located in western New York. Nest searches located 18 American Bittern (Botaurus lentiginosus), 117 Least Bittern (Ixobrychus exilis), 189 Pied-billed Grebe (Podilymbus podiceps), 23 Sora (Porzana carolina), and 72 Virginia Rail (Rallus limicola) nests. Average nest densities in 1998, our best nest searching year, ranged from 0.01/ha for Soras (N = 8) to 0.28/ha for Pied-billed Grebes (N = 160). Mayfield nest success estimates for Least Bittern were 80% (N = 16) in 1997 and 46% (N = 37) in 1998. Nest success estimates were 72% (N = 55) for Pied-billed Grebe, 43% (N = 6) for Sora, and 38% (N = 20) for Virginia Rail. Nests of all five species were located in ???70% emergent vegetation with a mean water depth of 24-56 cm and an average vegetation height that ranged from 69-133 cm. Logistic regression models were developed for each species using habitat variables at nest and random site locations. Each model was ranked with Akaike's Information Criterion for small sample size (AICc). In general, our best models indicated that increased emergent vegetation and horizontal cover with shallow water depths improved the odds of encountering marsh bird nests in the wetlands of western New York. We suggest that managing wetlands as a complex, at different stages of succession, would best benefit marsh bird species.

  1. Optimal path planning for a mobile robot using cuckoo search algorithm

    NASA Astrophysics Data System (ADS)

    Mohanty, Prases K.; Parhi, Dayal R.

    2016-03-01

    The shortest/optimal path planning is essential for efficient operation of autonomous vehicles. In this article, a new nature-inspired meta-heuristic algorithm has been applied for mobile robot path planning in an unknown or partially known environment populated by a variety of static obstacles. This meta-heuristic algorithm is based on the levy flight behaviour and brood parasitic behaviour of cuckoos. A new objective function has been formulated between the robots and the target and obstacles, which satisfied the conditions of obstacle avoidance and target-seeking behaviour of robots present in the terrain. Depending upon the objective function value of each nest (cuckoo) in the swarm, the robot avoids obstacles and proceeds towards the target. The smooth optimal trajectory is framed with this algorithm when the robot reaches its goal. Some simulation and experimental results are presented at the end of the paper to show the effectiveness of the proposed navigational controller.

  2. A two-stage adaptive stochastic collocation method on nested sparse grids for multiphase flow in randomly heterogeneous porous media

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liao, Qinzhuo, E-mail: liaoqz@pku.edu.cn; Zhang, Dongxiao; Tchelepi, Hamdi

    A new computational method is proposed for efficient uncertainty quantification of multiphase flow in porous media with stochastic permeability. For pressure estimation, it combines the dimension-adaptive stochastic collocation method on Smolyak sparse grids and the Kronrod–Patterson–Hermite nested quadrature formulas. For saturation estimation, an additional stage is developed, in which the pressure and velocity samples are first generated by the sparse grid interpolation and then substituted into the transport equation to solve for the saturation samples, to address the low regularity problem of the saturation. Numerical examples are presented for multiphase flow with stochastic permeability fields to demonstrate accuracy and efficiencymore » of the proposed two-stage adaptive stochastic collocation method on nested sparse grids.« less

  3. A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients

    PubMed Central

    2014-01-01

    Background Nosocomial candidaemia is associated with high mortality rates in critically ill paediatric patients; thus, the early detection and identification of the infectious agent is crucial for successful medical intervention. The PCR-based techniques have significantly increased the detection of Candida species in bloodstream infections. In this study, a multiplex nested PCR approach was developed for candidaemia detection in neonatal and paediatric intensive care patients. Methods DNA samples from the blood of 54 neonates and children hospitalised in intensive care units with suspected candidaemia were evaluated by multiplex nested PCR with specific primers designed to identify seven Candida species, and the results were compared with those obtained from blood cultures. Results The multiplex nested PCR had a detection limit of four Candida genomes/mL of blood for all Candida species. Blood cultures were positive in 14.8% of patients, whereas the multiplex nested PCR was positive in 24.0% of patients, including all culture-positive patients. The results obtained with the molecular technique were available within 24 hours, and the assay was able to identify Candida species with 100% of concordance with blood cultures. Additionally, the multiplex nested PCR detected dual candidaemia in three patients. Conclusions Our proposed PCR method may represent an effective tool for the detection and identification of Candida species in the context of candidaemia diagnosis in children, showing highly sensitive detection and the ability to identify the major species involved in this infection. PMID:25047415

  4. A multiplex nested PCR for the detection and identification of Candida species in blood samples of critically ill paediatric patients.

    PubMed

    Taira, Cleison Ledesma; Okay, Thelma Suely; Delgado, Artur Figueiredo; Ceccon, Maria Esther Jurfest Rivero; de Almeida, Margarete Teresa Gottardo; Del Negro, Gilda Maria Barbaro

    2014-07-21

    Nosocomial candidaemia is associated with high mortality rates in critically ill paediatric patients; thus, the early detection and identification of the infectious agent is crucial for successful medical intervention. The PCR-based techniques have significantly increased the detection of Candida species in bloodstream infections. In this study, a multiplex nested PCR approach was developed for candidaemia detection in neonatal and paediatric intensive care patients. DNA samples from the blood of 54 neonates and children hospitalised in intensive care units with suspected candidaemia were evaluated by multiplex nested PCR with specific primers designed to identify seven Candida species, and the results were compared with those obtained from blood cultures. The multiplex nested PCR had a detection limit of four Candida genomes/mL of blood for all Candida species. Blood cultures were positive in 14.8% of patients, whereas the multiplex nested PCR was positive in 24.0% of patients, including all culture-positive patients. The results obtained with the molecular technique were available within 24 hours, and the assay was able to identify Candida species with 100% of concordance with blood cultures. Additionally, the multiplex nested PCR detected dual candidaemia in three patients. Our proposed PCR method may represent an effective tool for the detection and identification of Candida species in the context of candidaemia diagnosis in children, showing highly sensitive detection and the ability to identify the major species involved in this infection.

  5. Rapid and sensitive detection of mink circovirus by recombinase polymerase amplification.

    PubMed

    Ge, Junwei; Shi, Yunjia; Cui, Xingyang; Gu, Shanshan; Zhao, Lili; Chen, Hongyan

    2018-06-01

    To date, the pathogenic role of mink circovirus (MiCV) remains unclear, and its prevalence and economic importance are unknown. Therefore, a rapid and sensitive molecular diagnosis is necessary for disease management and epidemiological surveillance. However, only PCR methods can identify MiCV infection at present. In this study, we developed a nested PCR and established a novel recombinase polymerase amplification (RPA) assay for MiCV detection. Sensitivity analysis showed that the detection limit of nested PCR and RPA assay was 10 1 copies/reaction, and these methods were more sensitive than conventional PCR, which has a detection limit of 10 5 copies/reaction. The RPA assay had no cross-reactivity with other related viral pathogens, and amplification was completed in less than 20 min with a simple device. Further assessment of clinical samples showed that the two assays were accurate in identifying positive and negative conventional PCR samples. The detection rate of MiCV by the RPA assay in clinical samples was 38.09%, which was 97% consistent with that by the nested PCR. The developed nested PCR is a highly sensitive tool for practical use, and the RPA assay is a simple, sensitive, and potential alternative method for rapid and accurate MiCV diagnosis. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. Prevalence of equine herpesvirus-1 and equine herpesvirus-4 infections in equidae species in Turkey as determined by ELISA and multiplex nested PCR.

    PubMed

    Ataseven, Veysel S; Dağalp, Seval B; Güzel, Murat; Başaran, Zeynep; Tan, Mehmet T; Geraghty, Bob

    2009-04-01

    In this report we examined the presence of specific antibodies against equine herpesvirus type 1 (EHV-1), and equine herpesvirus type 4 (EHV-4) in several equidae, including mules, donkeys, horses. The presence of EHV-1 and EHV-4 in respiratory diseases of equids, and ability of multiplex nested polymerase chain reaction (PCR) screening in simultaneous diagnosis of horses acutely infected by EHV-1 and EHV-4 were also investigated. Sera from 504 horses, mules and donkeys sampled were tested for the presence of EHV-1 and EHV-4 specific antibodies. Blood samples taken from 21 symptomatic horses and nasal swabs taken from 40 symptomatic horses were tested for the presence of EHV-1 and EHV-4 by a multiplex nested PCR. A total of 14.3% (3/21) of buffy coat samples and 32.5% (13/40) nasal swab samples were found to contain EHV-1 DNA, while 19% (4/21) buffy coat samples and 22.5% (9/40) nasal swab samples were found to be positive for EHV-4 DNA. By species, 14.5% of horses, 37.2% of mules and 24.2% of donkeys tested were EHV-1 seropositive. EHV-4 specific antibodies were detected in 237 (81.7%) of 290 horse sera tested. Results from this investigation demonstrate that EHV-1 and EHV-4 are prevalent throughout the equid population, and that donkeys and mules might also represent an important source of infection for other equids. We also showed that the multiplex nested PCR assay might be useful for diagnosis of mixed respiratory infections in horses due to EHV-1 and EHV-4.

  7. Evaluation of the efficiency of nested q-PCR in the detection of Mycobacterium tuberculosis complex directly from tuberculosis-suspected lesions in post-mortem macroscopic inspections of bovine carcasses slaughtered in the state of Mato Grosso, Brazil.

    PubMed

    Carvalho, Ricardo César Tavares; Furlanetto, Leone Vinícius; Maruyama, Fernanda Harumy; Araújo, Cristina Pires de; Barros, Sílvia Letícia Bomfim; Ramos, Carlos Alberto do Nascimento; Dutra, Valéria; Araújo, Flábio Ribeiro de; Paschoalin, Vânia Margaret Flosi; Nakazato, Luciano; Figueiredo, Eduardo Eustáquio de Souza

    2015-08-01

    Bovine tuberculosis (BTB) is a zoonotic disease caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTC). The quick and specific detection of this species is of extreme importance, since BTB may cause economic impacts, in addition to presenting imminent risks to human health. In the present study a nested real-time PCR test (nested q-PCR) was used in post-mortem evaluations to assess cattle carcasses with BTB-suspected lesions. A total of 41,193 cattle slaughtered in slaughterhouses located in the state of Mato Grosso, were examined. Of the examined animals, 198 (0.48%) showed BTB-suspected lesions. M. bovis was isolated in 1.5% (3/198) of the samples. Multiplex-PCR detected MTC in 7% (14/198) of the samples. The nested q-PCR test detected MTC in 28% (56/198) of the BTB-suspected lesions, demonstrating higher efficiency when compared to the multiplex-PCR and conventional microbiology. Nested q-PCR can therefore be used as a complementary test in the national program for control and eradication of bovine tuberculosis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Improved assay to detect Plasmodium falciparum using an uninterrupted, semi-nested PCR and quantitative lateral flow analysis

    PubMed Central

    2013-01-01

    Background A rapid, non-invasive, and inexpensive point-of-care (POC) diagnostic for malaria followed by therapeutic intervention would improve the ability to control infection in endemic areas. Methods A semi-nested PCR amplification protocol is described for quantitative detection of Plasmodium falciparum and is compared to a traditional nested PCR. The approach uses primers that target the P. falciparum dihydrofolate reductase gene. Results This study demonstrates that it is possible to perform an uninterrupted, asymmetric, semi-nested PCR assay with reduced assay time to detect P. falciparum without compromising the sensitivity and specificity of the assay using saliva as a testing matrix. Conclusions The development of this PCR allows nucleic acid amplification without the need to transfer amplicon from the first PCR step to a second reaction tube with nested primers, thus reducing both the chance of contamination and the time for analysis to < two hours. Analysis of the PCR amplicon yield was adapted to lateral flow detection using the quantitative up-converting phosphor (UCP) reporter technology. This approach provides a basis for migration of the assay to a POC microfluidic format. In addition the assay was successfully evaluated with oral samples. Oral fluid collection provides a simple non-invasive method to collect clinical samples. PMID:23433252

  9. Detection and Resolution of Cryptosporidium Species and Species Mixtures by Genus-Specific Nested PCR-Restriction Fragment Length Polymorphism Analysis, Direct Sequencing, and Cloning ▿

    PubMed Central

    Ruecker, Norma J.; Hoffman, Rebecca M.; Chalmers, Rachel M.; Neumann, Norman F.

    2011-01-01

    Molecular methods incorporating nested PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene of Cryptosporidium species were validated to assess performance based on limit of detection (LoD) and for detecting and resolving mixtures of species and genotypes within a single sample. The 95% LoD was determined for seven species (Cryptosporidium hominis, C. parvum, C. felis, C. meleagridis, C. ubiquitum, C. muris, and C. andersoni) and ranged from 7 to 11 plasmid template copies with overlapping 95% confidence limits. The LoD values for genomic DNA from oocysts on microscope slides were 7 and 10 template copies for C. andersoni and C. parvum, respectively. The repetitive nested PCR-RFLP slide protocol had an LoD of 4 oocysts per slide. When templates of two species were mixed in equal ratios in the nested PCR-RFLP reaction mixture, there was no amplification bias toward one species over another. At high ratios of template mixtures (>1:10), there was a reduction or loss of detection of the less abundant species by RFLP analysis, most likely due to heteroduplex formation in the later cycles of the PCR. Replicate nested PCR was successful at resolving many mixtures of Cryptosporidium at template concentrations near or below the LoD. The cloning of nested PCR products resulted in 17% of the cloned sequences being recombinants of the two original templates. Limiting-dilution nested PCR followed by the sequencing of PCR products resulted in no sequence anomalies, suggesting that this method is an effective and accurate way to study the species diversity of Cryptosporidium, particularly for environmental water samples, in which mixtures of parasites are common. PMID:21498746

  10. Origin and Alteration of Organic Matter in Termite Mounds from Different Feeding Guilds of the Amazon Rainforests

    PubMed Central

    Siebers, Nina; Martius, Christopher; Eckhardt, Kai-Uwe; Garcia, Marcos V. B.; Leinweber, Peter; Amelung, Wulf

    2015-01-01

    The impact of termites on nutrient cycling and tropical soil formation depends on their feeding habits and related material transformation. The identification of food sources, however, is difficult, because they are variable and changed by termite activity and nest construction. Here, we related the sources and alteration of organic matter in nests from seven different termite genera and feeding habits in the Terra Firme rainforests to the properties of potential food sources soil, wood, and microepiphytes. Chemical analyses comprised isotopic composition of C and N, cellulosic (CPS), non-cellulosic (NCPS), and N-containing saccharides, and molecular composition screening using pyrolysis-field ionization mass spectrometry (Py-FIMS). The isotopic analysis revealed higher soil δ13C (-27.4‰) and δ15N (6.6‰) values in nests of wood feeding Nasutitermes and Cornitermes than in wood samples (δ13C = -29.1‰, δ15N = 3.4‰), reflecting stable-isotope enrichment with organic matter alterations during or after nest construction. This result was confirmed by elevated NCPS:CPS ratios, indicating a preferential cellulose decomposition in the nests. High portions of muramic acid (MurAc) pointed to the participation of bacteria in the transformation processes. Non-metric multidimensional scaling (NMDS) revealed increasing geophagy in the sequence Termes < Embiratermes < Anoplotermes and increasing xylophagy for Cornitermes < Nasutitermes, and that the nest material of Constrictotermes was similar to the microepiphytes sample, confirming the report that Constrictotermes belongs to the microepiphyte-feeders. We therewith document that nest chemistry of rainforest termites shows variations and evidence of modification by microbial processes, but nevertheless it primarily reflects the trophic niches of the constructors. PMID:25909987

  11. Origin and alteration of organic matter in termite mounds from different feeding guilds of the Amazon rainforests.

    PubMed

    Siebers, Nina; Martius, Christopher; Eckhardt, Kai-Uwe; Garcia, Marcos V B; Leinweber, Peter; Amelung, Wulf

    2015-01-01

    The impact of termites on nutrient cycling and tropical soil formation depends on their feeding habits and related material transformation. The identification of food sources, however, is difficult, because they are variable and changed by termite activity and nest construction. Here, we related the sources and alteration of organic matter in nests from seven different termite genera and feeding habits in the Terra Firme rainforests to the properties of potential food sources soil, wood, and microepiphytes. Chemical analyses comprised isotopic composition of C and N, cellulosic (CPS), non-cellulosic (NCPS), and N-containing saccharides, and molecular composition screening using pyrolysis-field ionization mass spectrometry (Py-FIMS). The isotopic analysis revealed higher soil δ13C (-27.4‰) and δ15N (6.6‰) values in nests of wood feeding Nasutitermes and Cornitermes than in wood samples (δ13C = -29.1‰, δ15N = 3.4‰), reflecting stable-isotope enrichment with organic matter alterations during or after nest construction. This result was confirmed by elevated NCPS:CPS ratios, indicating a preferential cellulose decomposition in the nests. High portions of muramic acid (MurAc) pointed to the participation of bacteria in the transformation processes. Non-metric multidimensional scaling (MDS) revealed increasing geophagy in the sequence Termes < Embiratermes < Anoplotermes and increasing xylophagy for Cornitermes < Nasutitermes., and that the nest material of Constrictotermes was similar to the microepiphytes sample, confirming the report that Constrictotermes belongs to the microepiphyte-feeders. We therewith document that nest chemistry of rainforest termites shows variations and evidence of modification by microbial processes, but nevertheless it primarily reflects the trophic niches of the constructors.

  12. Nesting habitat relationships of sympatric Crested Caracaras, Red-tailed Hawks, and White-tailed Hawks in South Texas

    USGS Publications Warehouse

    Actkinson, M.A.; Kuvlesky, W.P.; Boal, C.W.; Brennan, L.A.; Hernandez, F.

    2007-01-01

    We quantified nesting-site habitats for sympatric White-tailed Hawks (Buteo albicaudatus) (n = 40), Red-tailed Hawks (B. jamaicensis) (n = 39), and Crested Caracaras (Caracara cheriway) (n = 24) in the Coastal Sand Plain of south Texas. White-tailed Hawks and Crested Caracara nest sites occurred in savannas, whereas Red-tailed Hawk nest sites occurred in woodlands on the edge of savannas. White-tailed Hawk nest sites were in shrubs and trees that were shorter (3.5 ?? 1.0 m) and had smaller canopy diameters (5.5 ?? 2.1 m) than those of Red-tailed Hawks (10.1 ?? 2.0 m, 13.7 ?? 5.8 m) and Crested Caracaras (5.6 ?? 1.7 m, 8.5 ?? 3.5 m). Red-tailed Hawk nest sites had higher woody densities (15.7 ?? 9.6 plants) and more woody cover (84 ?? 19%) than those of White-tailed Hawks (5.6 ?? 5.8 plants, 20 ?? 21%) and Crested Caracaras (9.9 ?? 6.7 plants, 55 ?? 34%). Crested Caracara nest sites were in dense, multi-branched shrubs composed of more living material (97 ?? 3%) than those of White-tailed (88 ?? 18%) and Red-tailed hawks (88 ?? 18%). Nest sites of White-tailed Hawks, Red-tailed Hawks, and Crested Caracaras were similar to random samples from the surrounding habitat indicating that preferred nesting habitat was available for each of these species at least within 60 m of active nest sites. Nest tree height, along with woody plant and native grass cover best discriminated nest sites among the three raptor species. There was no overlap at Red-tailed and White-tailed hawk nest sites in vegetation structure, while Crested Caracara nests were in habitat intermediate between the two other species. Partitioning of nesting habitat may be how these raptor species co-exist at the broader landscape scale of our study area in the Coastal Sand Plain of Texas.

  13. A Cell-Centered Multigrid Algorithm for All Grid Sizes

    NASA Technical Reports Server (NTRS)

    Gjesdal, Thor

    1996-01-01

    Multigrid methods are optimal; that is, their rate of convergence is independent of the number of grid points, because they use a nested sequence of coarse grids to represent different scales of the solution. This nesting does, however, usually lead to certain restrictions of the permissible size of the discretised problem. In cases where the modeler is free to specify the whole problem, such constraints are of little importance because they can be taken into consideration from the outset. We consider the situation in which there are other competing constraints on the resolution. These restrictions may stem from the physical problem (e.g., if the discretised operator contains experimental data measured on a fixed grid) or from the need to avoid limitations set by the hardware. In this paper we discuss a modification to the cell-centered multigrid algorithm, so that it can be used br problems with any resolution. We discuss in particular a coarsening strategy and choice of intergrid transfer operators that can handle grids with both an even or odd number of cells. The method is described and applied to linear equations obtained by discretization of two- and three-dimensional second-order elliptic PDEs.

  14. Using full-cohort data in nested case-control and case-cohort studies by multiple imputation.

    PubMed

    Keogh, Ruth H; White, Ian R

    2013-10-15

    In many large prospective cohorts, expensive exposure measurements cannot be obtained for all individuals. Exposure-disease association studies are therefore often based on nested case-control or case-cohort studies in which complete information is obtained only for sampled individuals. However, in the full cohort, there may be a large amount of information on cheaply available covariates and possibly a surrogate of the main exposure(s), which typically goes unused. We view the nested case-control or case-cohort study plus the remainder of the cohort as a full-cohort study with missing data. Hence, we propose using multiple imputation (MI) to utilise information in the full cohort when data from the sub-studies are analysed. We use the fully observed data to fit the imputation models. We consider using approximate imputation models and also using rejection sampling to draw imputed values from the true distribution of the missing values given the observed data. Simulation studies show that using MI to utilise full-cohort information in the analysis of nested case-control and case-cohort studies can result in important gains in efficiency, particularly when a surrogate of the main exposure is available in the full cohort. In simulations, this method outperforms counter-matching in nested case-control studies and a weighted analysis for case-cohort studies, both of which use some full-cohort information. Approximate imputation models perform well except when there are interactions or non-linear terms in the outcome model, where imputation using rejection sampling works well. Copyright © 2013 John Wiley & Sons, Ltd.

  15. Optical systolic solutions of linear algebraic equations

    NASA Technical Reports Server (NTRS)

    Neuman, C. P.; Casasent, D.

    1984-01-01

    The philosophy and data encoding possible in systolic array optical processor (SAOP) were reviewed. The multitude of linear algebraic operations achievable on this architecture is examined. These operations include such linear algebraic algorithms as: matrix-decomposition, direct and indirect solutions, implicit and explicit methods for partial differential equations, eigenvalue and eigenvector calculations, and singular value decomposition. This architecture can be utilized to realize general techniques for solving matrix linear and nonlinear algebraic equations, least mean square error solutions, FIR filters, and nested-loop algorithms for control engineering applications. The data flow and pipelining of operations, design of parallel algorithms and flexible architectures, application of these architectures to computationally intensive physical problems, error source modeling of optical processors, and matching of the computational needs of practical engineering problems to the capabilities of optical processors are emphasized.

  16. Acceleration of the direct reconstruction of linear parametric images using nested algorithms.

    PubMed

    Wang, Guobao; Qi, Jinyi

    2010-03-07

    Parametric imaging using dynamic positron emission tomography (PET) provides important information for biological research and clinical diagnosis. Indirect and direct methods have been developed for reconstructing linear parametric images from dynamic PET data. Indirect methods are relatively simple and easy to implement because the image reconstruction and kinetic modeling are performed in two separate steps. Direct methods estimate parametric images directly from raw PET data and are statistically more efficient. However, the convergence rate of direct algorithms can be slow due to the coupling between the reconstruction and kinetic modeling. Here we present two fast gradient-type algorithms for direct reconstruction of linear parametric images. The new algorithms decouple the reconstruction and linear parametric modeling at each iteration by employing the principle of optimization transfer. Convergence speed is accelerated by running more sub-iterations of linear parametric estimation because the computation cost of the linear parametric modeling is much less than that of the image reconstruction. Computer simulation studies demonstrated that the new algorithms converge much faster than the traditional expectation maximization (EM) and the preconditioned conjugate gradient algorithms for dynamic PET.

  17. Nested-PCR real time as alternative molecular tool for detection of Borrelia burgdorferi compared to the classical serological diagnosis of the blood.

    PubMed

    Sroka-Oleksiak, Agnieszka; Ufir, Krzysztof; Salamon, Dominika; Bulanda, Malgorzata; Gosiewski, Tomasz

    Lyme disease, caused by Borrelia burgdorferi, is a multisystem disease that often makes difficulties to recognize caused by their genetic heterogenity. Currently, the gold standard for the detection of Lyme disease (LD) is serologic diagnostics based mainly on tests: ELISA and Western blot (WB). These methods, however, are subject to consider- able defect, especially in the initial phase of infection due to the occurrence of so-called serological window period and low specificity. For this reason, they might be replaced by molecular methods, for example polymerase chain reaction (PCR), which should be more sensitivity and specificity. In the present study we attempt to optimize the PCR reaction conditions and enhance existing test sensitivity by applying the equivalent of real time PCR - nested PCR for detection B. burgdorferi DNA in the patient's blood. The study involved 94 blood samples of patients with suspected LD. From each sample, 1.5 ml of blood was used for the isolation of bacterial DNA and PCR real time am- plification and its equivalent, in nested version. The remaining part earmarked for serologi- cal testing. Optimization of the reaction conditions made experimentally, using gradient of the temperature and gradient of the magnesium ions concentration for reaction real time in nested-PCR and PCR version. The results show that the nested-PCR real time, has a much higher sensitivity 45 (47.8%) of positive results for the detection of B. burgdorferi compared to the single- variety, without a preceding pre-amplification 2 (2.1%). Serological methods allowed the detection of infection in 41 (43.6%) samples. These results support of the nested PCR method as a better molecular tool for the detection of B. burgdorferi infection than classical PCR real time reaction. The nested-PCR real time method may be considered as a complement to ELISA and WB mainly in the early stages of infection, when in the blood circulating B. burgdorferi cells. By contrast, the results of serological and molecular tests should always be carried out tak- ing into account the patient's clinical status.

  18. Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR

    PubMed Central

    2012-01-01

    Background Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax. Methods A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax, respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. Results The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum-infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. Conclusions The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa. PMID:22682065

  19. Simultaneous detection of Plasmodium vivax and Plasmodium falciparum gametocytes in clinical isolates by multiplex-nested RT-PCR.

    PubMed

    Kuamsab, Napaporn; Putaporntip, Chaturong; Pattanawong, Urassaya; Jongwutiwes, Somchai

    2012-06-10

    Gametocyte carriage is essential for malaria transmission and endemicity of disease; thereby it is a target for malaria control strategies. Malaria-infected individuals may harbour gametocytes below the microscopic detection threshold that can be detected by reverse transcription polymerase chain reaction (RT-PCR) targeting gametocyte-specific mRNA. To date, RT-PCR has mainly been applied to the diagnosis of Plasmodium falciparum gametocytes but very limited for that of Plasmodium vivax. A multiplex-nested RT-PCR targeting Pfs25 and Pvs25 mRNA specific to mature gametocytes of P. falciparum and P. vivax, respectively, was developed. The assay was evaluated using blood samples collected in rainy and dry seasons from febrile patients,in a malaria-endemic area in Thailand. Malaria diagnosis was performed by Giemsa-stained blood smears and 18S rRNA PCR. The multiplex-nested RT-PCR detected Pfs25 mRNA in 75 of 86 (87.2%) P. falciparum-infected individuals and Pvs25 mRNA in 82 of 90 (91.1%) P. vivax malaria patients diagnosed by 18S rRNA PCR. Gametocytes were detected in 38 (eight P. falciparum and 30 P. vivax) of 157 microscopy positive samples, implying that a large number of patients harbour sub-microscopic gametocytaemia. No seasonal differences in gametocyte carriage were observed for both malaria species diagnosed by multiplex-nested RT-PCR. With single-nested RT-PCR targeting Pfs25 or Pvs25 mRNA as standard, the multiplex-nested RT-PCR offered sensitivities of 97.4% and 98.9% and specificities of 100% and 98.8% for diagnosing mature gametocytes of P. falciparum and P. vivax, respectively. The minimum detection limit of the multiplex-nested PCR was 10 copies of templates. The multiplex-nested RT-PCR developed herein is useful for simultaneous assessment of both P. falciparum and P. vivax gametocyte carriage that is prevalent and generally sympatric in several malaria-endemic areas outside Africa.

  20. High-speed scanning: an improved algorithm

    NASA Astrophysics Data System (ADS)

    Nachimuthu, A.; Hoang, Khoi

    1995-10-01

    In using machine vision for assessing an object's surface quality, many images are required to be processed in order to separate the good areas from the defective ones. Examples can be found in the leather hide grading process; in the inspection of garments/canvas on the production line; in the nesting of irregular shapes into a given surface... . The most common method of subtracting the total area from the sum of defective areas does not give an acceptable indication of how much of the `good' area can be used, particularly if the findings are to be used for the nesting of irregular shapes. This paper presents an image scanning technique which enables the estimation of useable areas within an inspected surface in terms of the user's definition, not the supplier's claims. That is, how much useable area the user can use, not the total good area as the supplier estimated. An important application of the developed technique is in the leather industry where the tanner (the supplier) and the footwear manufacturer (the user) are constantly locked in argument due to disputed quality standards of finished leather hide, which disrupts production schedules and wasted costs in re-grading, re- sorting... . The developed basic algorithm for area scanning of a digital image will be presented. The implementation of an improved scanning algorithm will be discussed in detail. The improved features include Boolean OR operations and many other innovative functions which aim at optimizing the scanning process in terms of computing time and the accurate estimation of useable areas.

  1. Use of nested PCR to detect Ceratocystis fagacearum in sapwood of diseased northern oak species

    USDA-ARS?s Scientific Manuscript database

    Early and accurate diagnosis of oak wilt, caused by Ceratocystis fagacearum (Cf), is important when disease control action is planned. When laboratory diagnosis is needed, standard isolation protocols that are used rely on high quality samples and require > 14 days for incubation. Use of a nested P...

  2. Flying the Nest: How the Home Department Shapes Researchers' Career Paths

    ERIC Educational Resources Information Center

    Hottenrott, Hanna; Lawson, Cornelia

    2017-01-01

    This paper studies the importance of the socialization environment--nest--for the career destinations of early career researchers. In a sample of research groups in the fields of science and engineering at universities in Germany, we identify research orientation, output, funding as well as openness to industry and commercialization as relevant…

  3. Parental Representations and Attachment Security in Young Israeli Mothers' Bird's Nest Drawings

    ERIC Educational Resources Information Center

    Goldner, Limor; Golan, Yifat

    2016-01-01

    The Bird's Nest Drawing (BND; Kaiser, 1996) is an art-based technique developed to assess attachment security. In an attempt to expand the BND's validity, the authors explored the possible associations between parental representations and the BND's dimensions and attachment classifications in a sample of 80 young Israeli mothers. Positive…

  4. Isolation and confirmation of viral nervous necrosis (VNN) disease in golden grey mullet (Liza aurata) and leaping mullet (Liza saliens) in the Iranian waters of the Caspian Sea.

    PubMed

    Zorriehzahra, M E J; Ghasemi, M; Ghiasi, M; Karsidani, S Haghighi; Bovo, G; Nazari, A; Adel, M; Arizza, V; Dhama, K

    2016-07-15

    The present study was conducted on 428 moribund mullet fish samples to isolate and identify the causative agent of a mysterious acute mortality which recently occurred in wild mullets in Iranian waters of Caspian Sea, suspected to be due to viral nervous necrosis (VNN) disease. Disease investigation was carried out employing various diagnostic procedures such as virology, bacteriology, parasitology, haematology, histopathology, IFAT, IHC and nested RT-PCR. Brain and eye samples of affected fishes were collected in sterile conditions and then kept at -80°C for cell culture isolation and nested RT-PCR detection of the causative agent. Other tissue samples were also collected and fixed for histopathology, IHC and EM examinations. CPE was observed in cell cultures at 6days after inoculation. Nine samples were found positive with virological assay. Nested RT-PCR, performed on suspected tissues and CPE positive samples, showed that about 21 tissue samples and all the CPE positive samples were positive for VNN virus (VNNV). IFAT was selected as a confirmatory method for detecting the presence of Betanodavirus antigen, cell culture isolation results and nested RT-PCR findings. Moreover, VNNV particles with 25-30nm in diameter were also visualized in the infected brain and retina. In pathogenicity studies, guppy fishes bathed in VNNV-infected tissue culture (10(-4) TCID50) showed clinical signs similar to naturally infected mullet after 15days post infection (dpi), with mortality rates reaching up to 100% at 30dpi. Affected organ samples as examined by cell culture isolation, IFAT, IHC and histopathology, revealed the presence of VNNV in the guppy fishes. In conclusion, it was confirmed that VNNV was the main causative agent for the disease outbreak in mullet fish in the Caspian Sea, and this is such first official report of VNN disease from Iran. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Facile synthesis of bird's nest-like TiO2 microstructure with exposed (001) facets for photocatalytic degradation of methylene blue

    NASA Astrophysics Data System (ADS)

    Zhang, Guozhong; Zhang, Shuqu; Wang, Longlu; Liu, Ran; Zeng, Yunxiong; Xia, Xinnian; Liu, Yutang; Luo, Shenglian

    2017-01-01

    The scrupulous design of hierarchical structure and highly active crystal facets exposure is essential for the creation of photocatalytic system. However, it is still a big challenge for scrupulous design of TiO2 architectures. In this paper, bird's nest-like anatase TiO2 microstructure with exposed highly active (001) surface has been successfully synthesized by a facile one-step solvothermal method. Methylene blue (MB) is chosen as a model pollutant to evaluate photocatalytic activity of as-obtained TiO2 samples. The results show that the photocatalytic activity of the bird's nest-like sample is more excellent than P25 in the degradation of MB due to high specific surface area and highly active (001) crystal facets exposure when tested under simulated solar light. Besides, it can be readily separated from the photocatalytic system by sedimentation after photocatalytic reaction, which is a significant advantage against conventional powder photocatalyst. The bird's nest-like microspheres with novel structure may have potential application in photocatalysis and other fields.

  6. New insights on multiplicity and clustering in Taurus.

    NASA Astrophysics Data System (ADS)

    Joncour, Isabelle; Duchene, Gaspard; Moraux, Estelle; Mundy, Lee

    2018-01-01

    Multiplicity and clustering of young stars are critical clues to constraint star formation process. The Taurus molecular complex is the archetype of a quiescent star forming region that may retain primeval signature of star formation.Using statistical and clustering tools such as nearest neighbor statistics, correlation functions and the density-Based Spatial Clustering of Applications with Noise (DBSCAN) algorithm, this work reveals new spatial substructures in Taurus.We have identified unexpected ultra wide pairs (UWPs) candidates of high order multiplicity in Taurus in the 5-60 kAU separation range (Joncour et al 2017), beyond the separation assessed for wide pairs (Kraus & Hillenbrand 2009).Our work reveals 20 local stellar substructures, the Nested Elementary Structures (NESTs). These NESTs contain nearly half the stars of Taurus and 75% of the Class 0/I objects probing that they are the preferred sites of star formation (Joncour et al, sub.). The NESTs size ranges from few kAU up to 80 kAU making a length scale bridge between wide pairs and loose group (few hundreds kAU, Kirk & Myers, 2011). The NESTs mass ranges from 0.5-10 solar mass. The balance between Class I, II and III in NESTs suggests that they may be ordered as an evolutionary temporal scheme, some of them got infertile, while other shelter stars in infancy.The UWPs and the NESTs may be pristine imprints of their spatial configuration at birth. The UWPs population may result from a cascade fragmentation scenario of the natal molecular core. They could be the older counterparts, to the 0.5 Myr prestellar cores/Class 0 multiple objects observed at radio/millimeter wavelengths (Tobin et al 2010, 2016) and the precursors of the large number of UWPs (10–100 kAU) recently identified in older moving groups (Floriano-Alonso et al, 2015 ; Elliot et al 2016). The NESTs may result from the gravitational collapse of a gas clump that fragments to give a tight collection of stars within few millions years.This project has been partly supported by the StarFormMapper project funded by the European Union's Horizon 2020 Research and Innovation Action (RIA) program under grant agreement number 687528.

  7. Development of a rapid and sensitive one-step reverse transcription-nested polymerase chain reaction in a single tube using the droplet-polymerase chain reaction machine.

    PubMed

    Yamaguchi, Akemi; Matsuda, Kazuyuki; Sueki, Akane; Taira, Chiaki; Uehara, Masayuki; Saito, Yasunori; Honda, Takayuki

    2015-08-25

    Reverse transcription (RT)-nested polymerase chain reaction (PCR) is a time-consuming procedure because it has several handling steps and is associated with the risk of cross-contamination during each step. Therefore, a rapid and sensitive one-step RT-nested PCR was developed that could be performed in a single tube using a droplet-PCR machine. The K562 BCR-ABL mRNA-positive cell line as well as bone marrow aspirates from 5 patients with chronic myelogenous leukemia (CML) and 5 controls without CML were used. We evaluated one-step RT-nested PCR using the droplet-PCR machine. One-step RT-nested PCR performed in a single tube using the droplet-PCR machine enabled the detection of BCR-ABL mRNA within 40min, which was 10(3)-fold superior to conventional RT nested PCR using three steps in separate tubes. The sensitivity of the one-step RT-nested PCR was 0.001%, with sample reactivity comparable to that of the conventional assay. One-step RT-nested PCR was developed using the droplet-PCR machine, which enabled all reactions to be performed in a single tube accurately and rapidly and with high sensitivity. This one-step RT-nested PCR may be applicable to a wide spectrum of genetic tests in clinical laboratories. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Impact of pest control strategies on the arthropodofauna living in bird nests built in nestboxes in pear and apple orchards.

    PubMed

    Roy, Lise; Bouvier, Jean-Charles; Lavigne, Claire; Galès, Mathieu; Buronfosse, Thierry

    2013-08-01

    Pesticide applications have a strong impact on biodiversity in agroecosystems. The present study aimed to assess the impact of pest control strategies on the arthropodofauna of Parus major nests built within nestboxes installed in orchards. Unlike many studied groups, these arthropod communities are not in direct contact with pesticide sprays (on account of their being sheltered by nestboxes) and are also unable to move away from the treated area. In this pilot study, we estimated the prevalence and the taxonomic and ecological diversities of arthropodofauna sampled in the nests and assessed the extent to which the whole and nest-specific arthropodofauna were affected by pest control strategies. Sixteen different insect and arachnid Primary Taxonomic Groups (PTGs, order level or below) were found in nests. The best represented PTGs (≥10% occurrence in years 2007 and 2008) were Psocoptera (Insecta, detritivorous/saprophagous), detritivorous/saprophagous Astigmata (Acari) and hematophagous Mesostigmata (Acari). Pest control strategies had a large impact on the prevalence of arthropods in nests, with higher proportions of nests hosting arthropods in organic orchards than in conventional orchards and with intermediate proportions in nests in Integrated Pest Management orchards. In contrast, pest control strategies had no significant effect on the composition of the arthropod communities when only nests hosting nidicolous arthropods were considered.

  9. Patch size and edge proximity are useful predictors of brood parasitism but not nest survival of grassland birds.

    PubMed

    Benson, Thomas J; Chiavacci, Scott J; Ward, Michael P

    2013-06-01

    Declines of migratory birds have led to increased focus on causative factors for these declines, including the potential adverse effects of habitat fragmentation on reproductive success. Although numerous studies have addressed how proximity to a habitat edge, patch size, or landscape context influence nest survival or brood parasitism, many have failed to find the purported effects. Furthermore, many have sought to generalize patterns across large geographic areas and habitats. Here, we examined evidence for effects of edge proximity, patch size, and landscape context on nest survival and brood parasitism of grassland birds, a group of conservation concern. The only consistent effect was a positive association between edge proximity and brood parasitism. We examined effects of patch size on nest survival (37 studies) and brood parasitism (30 studies) representing 170 and 97 different estimates, respectively, with a total sample size of > 14000 nests spanning eastern North America. Nest survival weakly increased with patch size in the Great Plains, but not in the Midwestern or Eastern United States, and brood parasitism was inversely related to patch size and consistently greater in the Great Plains. The consistency in brood parasitism relative to nest survival patterns is likely due to parasitism being caused by one species, while nest survival is driven by a diverse and variable suite of nest predators. Often, studies assume that predators responsible for nest predation, the main driver of nest success, either are the same or exhibit the same behaviors across large geographic areas. These results suggest that a better mechanistic understanding of nest predation is needed to provide meaningful conservation recommendations for improving grassland bird productivity, and that the use of general recommendations across large geographic areas should only be undertaken when sufficient data are available from all regions.

  10. Diagnostic value of nested-PCR for identification of Malassezia species in dandruff

    NASA Astrophysics Data System (ADS)

    Jusuf, N. K.; Nasution, T. A.; Ullyana, S.

    2018-03-01

    Dandruff or pityriasis simplex is a condition of abnormal occurrence of formation of yellowish white scales from the scalp. Many factors play a role in the pathogenesis of dandruff, i.e.colonization of Malassezia species. Examination of Malassezia species previously done by culture as the gold standard. However, there are various difficulties in doing the culture. Identification method with anested-polymerase chain reaction (nested-PCR) is expected to provide quickly and easily detected. This study aimedto determine the diagnostic value of nested-PCR in the identification of Malassezia species in dandruff. From 21 subjects, scales from the scalp were taken and sent to the laboratory for nested-PCR identification. Statistical analysis of diagnostic test carried out to determine sensitivity, specificity, positive predictive value, and negative predictive value. The results showed nested-PCR detected 10 sample (47.6%) positive for Malassezia species consist of M. sympodialis (23.8%); M. slooffiae (9.5%); M. furfur (4.8%); M. globosa and M. furfur (4.8%); and M. restricta and M. sympodialis (4.8%). Detection of Malassezia species by nested-PCR has 100% in sensitivity whereas the specificity was 55%. Nested-PCR test has high sensitivity. Therefore nested-PCR may be considered for a faster and simpler alternative examination in identification for Malassezia species in dandruff.

  11. Comparison of Antigen Detection and Nested PCR in CSF Samples of HIV Positive and Negative Patients with Suspected Cryptococcal Meningitis in a Tertiary Care Hospital.

    PubMed

    Kumari, Sunita; Verma, Rajesh Kumar; Singh, Dharmendra Prasad; Yadav, Ramakant

    2016-04-01

    The cases of cryptococcal meningitis and other forms of cryptococcosis have increased in recent time and the present scenario of the condition with significant morbidity and mortality is actually posing a serious threat to the community, so an early and prompt diagnosis is necessary to prevent serious complications and thus improving the overall disease outcome. Comparison of diagnostic efficacy of nested Polymerase Chain Reaction (PCR) with Latex Agglutination Test (LAT) in the Cerebro Spinal Fluid (CSF) samples of the cases of meningitis in HIV positive and negative cases. We have compared the diagnostic efficacy of Latex Agglutination Test (LAT) with nested Polymerase Chain Reaction (PCR) in 200 Cerebrospinal Fluid (CSF) samples, including 14 HIV positive also, in the cases of suspected cryptococcal meningitis. Nested PCR was done in all cases reporting positive by LAT and results were then compared with that of India ink and culture on Sabouraud Dextrose Agar (SDA), and the isolates were further identified by urease, nitrate and sugar assimilation tests. Of the 200 cases, including 14 HIV positive, LAT was positive in 46 cases while 154 were negative. Out of these 46 LAT positive cases, nested PCR was positive in 40 cases only, while culture and India ink was positive in 38 and 33 cases respectively. Majority of the cases, 30 (65.2%) were between age group 21-50 years, while 2 (4.3%) in 0-20, and 14 (30.4%) in 51-80 years age group. Although negative staining like India ink and nigrosin are most widely used techniques, but these suffer with subjective error. Rapid method like LAT is available but it always has the scope of false positive and negative results. In such cases nested PCR can help in establishing final diagnosis.

  12. Optimization of the elution buffer and concentration method for detecting hepatitis E virus in swine liver using a nested reverse transcription-polymerase chain reaction and real-time reverse transcription-polymerase chain reaction.

    PubMed

    Son, Na Ry; Seo, Dong Joo; Lee, Min Hwa; Seo, Sheungwoo; Wang, Xiaoyu; Lee, Bog-Hieu; Lee, Jeong-Su; Joo, In-Sun; Hwang, In-Gyun; Choi, Changsun

    2014-09-01

    The aim of this study was to develop an optimal technique for detecting hepatitis E virus (HEV) in swine livers. Here, three elution buffers and two concentration methods were compared with respect to enhancing recovery of HEV from swine liver samples. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested RT-PCR were performed to detect HEV RNA. When phosphate-buffered saline (PBS, pH 7.4) was used to concentrate HEV in swine liver samples using ultrafiltration, real-time RT-PCR detected HEV in 6 of the 26 samples. When threonine buffer was used to concentrate HEV using polyethylene glycol (PEG) precipitation and ultrafiltration, real-time RT-PCR detected HEV in 1 and 3 of the 26 samples, respectively. When glycine buffer was used to concentrate HEV using ultrafiltration and PEG precipitation, real-time RT-PCR detected HEV in 1 and 3 samples of the 26 samples, respectively. When nested RT-PCR was used to detect HEV, all samples tested negative regardless of the type of elution buffer or concentration method used. Therefore, the combination of real-time RT-PCR and ultrafiltration with PBS buffer was the most sensitive and reliable method for detecting HEV in swine livers. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Detection of Giardia intestinalis in water samples collected from natural water reservoirs and wells in northern and north-eastern Poland using LAMP, real-time PCR and nested PCR.

    PubMed

    Lass, Anna; Szostakowska, Beata; Korzeniewski, Krzysztof; Karanis, Panagiotis

    2017-10-01

    Giardia intestinalis is a protozoan parasite, transmitted to humans and animals by the faecal-oral route, mainly through contaminated water and food. Knowledge about the distribution of this parasite in surface water in Poland is fragmentary and incomplete. Accordingly, 36 environmental water samples taken from surface water reservoirs and wells were collected in Pomerania and Warmia-Masuria provinces, Poland. The 50 L samples were filtered and subsequently analysed with three molecular detection methods: loop-mediated isothermal amplification (LAMP), real-time polymerase chain reaction (real-time PCR) and nested PCR. Of the samples examined, Giardia DNA was found in 15 (42%) samples with the use of LAMP; in 12 (33%) of these samples, Giardia DNA from this parasite was also detected using real-time PCR; and in 9 (25%) using nested PCR. Sequencing of selected positive samples confirmed that the PCR products were fragments of the Giardia intestinalis small subunit rRNA gene. Genotyping using multiplex real-time PCR indicated the presence of assemblages A and B, with the latter predominating. The results indicate that surface water in Poland, as well as water taken from surface wells, may be a source of Giardia strains which are potentially pathogenic for humans. It was also demonstrated that LAMP assay is more sensitive than the other two molecular assays.

  14. First record of interspecific breeding of Least Bell's Vireo and White-eyed Vireo

    USGS Publications Warehouse

    Blundell, Melissa A.; Kus, Barbara E.

    2011-01-01

    We provide the first known documentation of a male Least Bell's Vireo (Vireo bellii pusillus) breeding with a female White-eyed Vireo (V. griseus) and the first report of a White-eyed Vireo breeding in California at the San Luis Rey River, Oceanside, San Diego County. We discovered the pair building a nest on 12 May 2010. The female laid four eggs, and the pair successfully raised and fledged four nestlings. We collected DNA samples from each nestling and the female during the nestling stage and banded them with a numbered federal leg band for future identification. We obtained detailed nest measurements after fledging and monitored the territory for further nesting attempts. No additional nesting attempts were detected.

  15. Territory and nest site selection patterns by Grasshopper Sparrows in southeastern Arizona

    USGS Publications Warehouse

    Ruth, Janet M.; Skagen, Susan K.

    2017-01-01

    Grassland bird populations are showing some of the greatest rates of decline of any North American birds, prompting measures to protect and improve important habitat. We assessed how vegetation structure and composition, habitat features often targeted for management, affected territory and nest site selection by Grasshopper Sparrows (Ammodramus savannarum ammolegus) in southeastern Arizona. To identify features important to males establishing territories, we compared vegetation characteristics of known territories and random samples on 2 sites over 5 years. We examined habitat selection patterns of females by comparing characteristics of nest sites with territories over 3 years. Males selected territories in areas of sparser vegetation structure and more tall shrubs (>2 m) than random plots on the site with low shrub densities. Males did not select territories based on the proportion of exotic grasses. Females generally located nest sites in areas with lower small shrub (1–2 m tall) densities than territories overall when possible and preferentially selected native grasses for nest construction. Whether habitat selection was apparent depended upon the range of vegetation structure that was available. We identified an upper threshold above which grass structure seemed to be too high and dense for Grasshopper Sparrows. Our results suggest that some management that reduces vegetative structure may benefit this species in desert grasslands at the nest and territory scale. However, we did not assess initial male habitat selection at a broader landscape scale where their selection patterns may be different and could be influenced by vegetation density and structure outside the range of values sampled in this study.

  16. A nested real-time PCR assay for the quantification of Plasmodium falciparum DNA extracted from dried blood spots.

    PubMed

    Tran, Tuan M; Aghili, Amirali; Li, Shanping; Ongoiba, Aissata; Kayentao, Kassoum; Doumbo, Safiatou; Traore, Boubacar; Crompton, Peter D

    2014-10-04

    As public health efforts seek to eradicate malaria, there has been an emphasis on eliminating low-density parasite reservoirs in asymptomatic carriers. As such, diagnosing submicroscopic Plasmodium infections using PCR-based techniques has become important not only in clinical trials of malaria vaccines and therapeutics, but also in active malaria surveillance campaigns. However, PCR-based quantitative assays that rely on nucleic acid extracted from dried blood spots (DBS) have demonstrated lower sensitivity than assays that use cryopreserved whole blood as source material. The density of Plasmodium falciparum asexual parasites was quantified using genomic DNA extracted from dried blood spots (DBS) and the sensitivity of two approaches was compared: quantitative real-time PCR (qPCR) targeting the P. falciparum 18S ribosomal RNA gene, either with an initial conventional PCR amplification prior to qPCR (nested qPCR), or without an initial amplification (qPCR only). Parasite densities determined by nested qPCR, qPCR only, and light microscopy were compared. Nested qPCR results in 10-fold higher sensitivity (0.5 parasites/μl) when compared to qPCR only (five parasites/ul). Among microscopy-positive samples, parasite densities calculated by nested qPCR correlated strongly with microscopy for both asymptomatic (Pearson's r=0.58, P<0.001) and symptomatic (Pearson's r=0.70, P<0.0001) P. falciparum infections. Nested qPCR improves the sensitivity for the detection of P. falciparum blood-stage infection from clinical DBS samples. This approach may be useful for active malaria surveillance in areas where submicroscopic asymptomatic infections are prevalent.

  17. Diagnosis of theileria equi infections in horses in the Azores using cELISA and nested PCR

    USDA-ARS?s Scientific Manuscript database

    Equine piroplasmosis is a tick-borne disease of equids that is often caused by the parasite Theileria equi. We applied competitive ELISA (cELISA) and nested PCR diagnostic methods to detect this parasite in horses by screening 162 samples from mainland Portugal where the parasite is endemic, and 143...

  18. Genome-wide association study of carbon and nitrogen metabolism in the maize nested association mapping population

    USDA-ARS?s Scientific Manuscript database

    Carbon (C) and nitrogen (N) metabolism are critical to plant growth and development and at the basis of yield and adaptation. We have applied high throughput metabolite analyses to over 12,000 diverse field grown samples from the maize nested association mapping population. This allowed us to identi...

  19. Effects of industrial and investigator disturbance on Arctic-nesting geese

    USGS Publications Warehouse

    Meixell, Brandt W.; Flint, Paul L.

    2017-01-01

    Oil and gas development on the Arctic Coastal Plain (ACP) of Alaska, USA may have effects on Arctic-nesting birds. To estimate effects of industrial activity and investigator disturbance on avian productivity, we monitored nests of greater white-fronted geese (Anser albifrons) with digital cameras and periodic nest visits during 2013–2014 at 2 sites on the ACP. A disturbed site was adjacent to human-made infrastructure and industrial clean-up activities initiated at the onset of the study and a control site was >2 km from sources of industrial disturbance. We assessed variation in estimates of incubation constancy, nest survival, and predator behavior relative to site, year, and distance from industrial activity using nest photographs obtained at 1-minute intervals. We compared analysis of hourly nest survival informed by intensive monitoring with cameras to analysis of daily nest survival informed by traditional nest visit data obtained at intervals of 5–7 days to assess how method and time scale of sampling affect ecological inference. Geese in both sites exhibited high levels of nest attendance and initiated incubation breaks less than once per day. Observer-caused incubation breaks associated with nest visits ( = 37.8 min) were longer than other types of incubation breaks ( = 8.7 min), demonstrating a differential response by nesting geese to direct human encroachment versus indirect vehicular and aircraft traffic. During both years, geese were absent from nests more frequently in the disturbed ( = 0.9 breaks/day) than control ( = 0.6 breaks/day) site, and this break frequency was slightly higher for nests closer to industrial activity. In the year with high rates of depredation, nest survival was positively related to distance from industrial activity and abandoned infrastructure, consistent with predictions of industry-caused effects. This relationship, however, was not evident in the year with reduced predation pressure, likely because of annual variation in arctic fox (Vulpes lagopus) behavior. Analysis of nest survival probability informed by camera data allowed for detection of detailed patterns of variation that were not supported when using only visit data for the same nests. Observer visits were responsible for reductions of 7–35% in nest survival probability, highlighting the importance of minimizing, and controlling for, observer effects in studies of avian productivity. Indirect vehicular and aircraft disturbance posed less risk to nest survival than direct encroachment by observers at nest sites. Therefore, effects of industrial activities on avian productivity in the Arctic can be minimized through practices that limit direct encounters with nests. 

  20. Improvement of DGGE analysis by modifications of PCR protocols for analysis of microbial community members with low abundance.

    PubMed

    Wang, Yong-Feng; Zhang, Fang-Qiu; Gu, Ji-Dong

    2014-06-01

    Denaturing gradient gel electrophoresis (DGGE) is a powerful technique to reveal the community structures and composition of microorganisms in complex natural environments and samples. However, positive and reproducible polymerase chain reaction (PCR) products, which are difficult to acquire for some specific samples due to low abundance of the target microorganisms, significantly impair the effective applications of DGGE. Thus, nested PCR is often introduced to generate positive PCR products from the complex samples, but one problem is also introduced: The total number of thermocycling in nested PCR is usually unacceptably high, which results in skewed community structures by generation of random or mismatched PCR products on the DGGE gel, and this was demonstrated in this study. Furthermore, nested PCR could not resolve the uneven representative issue with PCR products of complex samples with unequal richness of microbial population. In order to solve the two problems in nested PCR, the general protocol was modified and improved in this study. Firstly, a general PCR procedure was used to amplify the target genes with the PCR primers without any guanine cytosine (GC) clamp, and then, the resultant PCR products were purified and diluted to 0.01 μg ml(-1). Subsequently, the diluted PCR products were utilized as templates to amplify again with the same PCR primers with the GC clamp for 17 cycles, and the products were finally subjected to DGGE analysis. We demonstrated that this is a much more reliable approach to obtain a high quality DGGE profile with high reproducibility. Thus, we recommend the adoption of this improved protocol in analyzing microorganisms of low abundance in complex samples when applying the DGGE fingerprinting technique to avoid biased results.

  1. Do weather conditions correlate with findings in failed, provision-filled nest cells of Megachile rotundata (Hymenoptera: Megachilidae) in western North America?

    PubMed

    Pitts-Singer, Theresa L; James, Rosalind R

    2008-06-01

    Cavity-nesting alfalfa leafcutting bees, Megachile rotundata (F.) (Hymenoptera: Megachilidae), are excellent pollinators of alfalfa, Medicago savita L., for seed production. In commercial settings, artificial cavities are placed in field domiciles for nesting and, thereby, bee populations are sustained for future use. For this study, cells from leafcutting bee nests were collected in late summer from commercial seed fields. Over 3 yr (2003-2005), 39 samples in total of approximately equal to 1,000 cells each were taken from several northwestern U.S. states and from Manitoba, Canada. X-radiography of 500 cells from each sample was used to identify "pollen balls" (i.e., cells in which the pollen-nectar provision remained, but the egg or larva, if present, was not detectable on an x-radiograph). Most U.S. samples seemed to have higher proportions of pollen ball cells than Manitoba samples. Pollen ball cells were dissected to determine the moisture condition of the mass provision and true contents of each cell. Most pollen ball cells from Manitoba samples contained fungus, the frequency of which was positively correlated with cool, wet weather. In the United States, most pollen ball cells had moist provisions, and many of them lacked young brood. Correlation analysis revealed that pollen ball cells occurred in greater proportions in fields with more hot days (above 38 degrees C). Broodless pollen ball cells occurred in greater proportions under cool conditions, but dead small larvae (second-third instars) seemed to occur in greater proportions under hot conditions. Pollen ball cells with unhatched eggs and first instars (in the chorion) occurred in lesser proportions under hot conditions.

  2. Diagnostic potential of nested PCR, galactomannan EIA, and beta-D-glucan for invasive aspergillosis in pediatric patients.

    PubMed

    Badiee, Parisa; Alborzi, Abdolvahab; Karimi, Mahammad; Pourabbas, Bahman; Haddadi, Pedram; Mardaneh, Jalal; Moieni, Mahsa

    2012-04-13

    Limited specific data and investigations are available for invasive aspergillosis (IA) in pediatric patients. We evaluated the diagnostic potential of three noninvasive tests including the Platelia Aspergillus EIA kit for using galactomannan antigen, (1,3)-β-D-glucan Detection Reagent Kit, and nested-PCR for Aspergillus DNA in sera. We evaluated the diagnostic potential of three noninvasive tests including EIA for galactomannan antigen  (Platelia Aspergillus), nested  PCR assay for Aspergillus DNA and test for (1→3)-β-D-glucan (Glucatell assay Kit). All pediatric patients treated at the hematology/oncology unit who were at increased risk of developing invasive aspergillosis were enrolled. Clinical samples were examined for Aspergillus infections by mycological methods. Serial blood samples were collected twice weekly and evaluated by noninvasive tests. We analyzed 230 consecutive blood samples from 62 pediatric patients. The incidence rate of invasive aspergillosis in the patients was found to be 27.4%, and the etiologic agents were Aspergillus flavus, Aspergillus fumigatus, and Aspergillus spp.  The sensitivity, specificity, positive and negative predictive values, and likelihood ratios for positive and negative results of galactomannan in patients with proven and probable IA were 90%, 92%, 81.8%, 96%, 11.25, and 0.1; for beta-D-glucan they were 50%, 46%, 26%, 70.6%, 0.9, 0.9; and for nested-PCR they were 80%, 96.2%, 88.9%, 92.6%, 21, and 0.2, respectively. The conventional methods are not able to detect IA, due to the lack of valid and proper sampling. Galactomannan and nested-PCR tests in serum, with enough accuracy and reliability, can serve as noninvasive methods for the detection of IA in pediatric patients. However, the beta-D-glucan test cannot serve as an efficient diagnostic tool in those with hematologic disorders. 

  3. Analysis of Clinical Cohort Data Using Nested Case-control and Case-cohort Sampling Designs. A Powerful and Economical Tool.

    PubMed

    Ohneberg, K; Wolkewitz, M; Beyersmann, J; Palomar-Martinez, M; Olaechea-Astigarraga, P; Alvarez-Lerma, F; Schumacher, M

    2015-01-01

    Sampling from a large cohort in order to derive a subsample that would be sufficient for statistical analysis is a frequently used method for handling large data sets in epidemiological studies with limited resources for exposure measurement. For clinical studies however, when interest is in the influence of a potential risk factor, cohort studies are often the first choice with all individuals entering the analysis. Our aim is to close the gap between epidemiological and clinical studies with respect to design and power considerations. Schoenfeld's formula for the number of events required for a Cox' proportional hazards model is fundamental. Our objective is to compare the power of analyzing the full cohort and the power of a nested case-control and a case-cohort design. We compare formulas for power for sampling designs and cohort studies. In our data example we simultaneously apply a nested case-control design with a varying number of controls matched to each case, a case cohort design with varying subcohort size, a random subsample and a full cohort analysis. For each design we calculate the standard error for estimated regression coefficients and the mean number of distinct persons, for whom covariate information is required. The formula for the power of a nested case-control design and the power of a case-cohort design is directly connected to the power of a cohort study using the well known Schoenfeld formula. The loss in precision of parameter estimates is relatively small compared to the saving in resources. Nested case-control and case-cohort studies, but not random subsamples yield an attractive alternative for analyzing clinical studies in the situation of a low event rate. Power calculations can be conducted straightforwardly to quantify the loss of power compared to the savings in the num-ber of patients using a sampling design instead of analyzing the full cohort.

  4. A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus

    PubMed Central

    2010-01-01

    A multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR) method was developed for the detection and differentiation of wild-type and vaccine strains of canine distemper virus (CDV). A pair of primers (P1 and P4) specific for CDV corresponding to the highly conserved region of the CDV genome were used as a common primer pair in the first-round PCR of the nested PCR. Primers P2 specific for CDV wild-type strains, were used as the forward primer together with the common reverse primer P4 in the second round of nested PCR. Primers P3, P5 specific for CDV wild-type strain or vaccine strain, were used as the forward primer together with the common reverse primer P4+P6 in the second round of nested PCR. A fragment of 177 bp was amplified from vaccine strain genomic RNA, and a fragment of 247 bp from wild-type strain genomic RNA in the RT-nPCR, and two fragments of 247 bp and 177 bp were amplified from the mixed samples of vaccine and wild-type strains. No amplification was achieved for uninfected cells, or cells infected with Newcastle disease virus (NDV), canine parvovirus (CPV), canine coronavirus (CCV), rabies virus (RV), or canine adenovirus (CAV). The RT-nPCR method was used to detect 30 field samples suspected of canine distemper from Heilongjiang and Jilin Provinces, and 51 samples in Shandong province. As a result of 30 samples, were found to be wild-type-like, and 5 to be vaccine-strain-like. The RT-nPCR method can be used to effectively detect and differentiate wild-type CDV-infected dogs from dogs vaccinated with CDV vaccine, and thus can be used in clinical detection and epidemiological surveillance. PMID:20433759

  5. Body Size Correlates with Fertilization Success but not Gonad Size in Grass Goby Territorial Males

    PubMed Central

    Pujolar, Jose Martin; Locatello, Lisa; Zane, Lorenzo; Mazzoldi, Carlotta

    2012-01-01

    In fish species with alternative male mating tactics, sperm competition typically occurs when small males that are unsuccessful in direct contests steal fertilization opportunities from large dominant males. In the grass goby Zosterisessor ophiocephalus, large territorial males defend and court females from nest sites, while small sneaker males obtain matings by sneaking into nests. Parentage assignment of 688 eggs from 8 different nests sampled in the 2003–2004 breeding season revealed a high level of sperm competition. Fertilization success of territorial males was very high but in all nests sneakers also contributed to the progeny. In territorial males, fertilization success correlated positively with male body size. Gonadal investment was explored in a sample of 126 grass gobies collected during the period 1995–1996 in the same area (61 territorial males and 65 sneakers). Correlation between body weight and testis weight was positive and significant for sneaker males, while correlation was virtually equal to zero in territorial males. That body size in territorial males is correlated with fertilization success but not gonad size suggests that males allocate much more energy into growth and relatively little into sperm production once the needed size to become territorial is attained. The increased paternity of larger territorial males might be due to a more effective defense of the nest in comparison with smaller territorial males. PMID:23056415

  6. Body size correlates with fertilization success but not gonad size in grass goby territorial males.

    PubMed

    Pujolar, Jose Martin; Locatello, Lisa; Zane, Lorenzo; Mazzoldi, Carlotta

    2012-01-01

    In fish species with alternative male mating tactics, sperm competition typically occurs when small males that are unsuccessful in direct contests steal fertilization opportunities from large dominant males. In the grass goby Zosterisessor ophiocephalus, large territorial males defend and court females from nest sites, while small sneaker males obtain matings by sneaking into nests. Parentage assignment of 688 eggs from 8 different nests sampled in the 2003-2004 breeding season revealed a high level of sperm competition. Fertilization success of territorial males was very high but in all nests sneakers also contributed to the progeny. In territorial males, fertilization success correlated positively with male body size. Gonadal investment was explored in a sample of 126 grass gobies collected during the period 1995-1996 in the same area (61 territorial males and 65 sneakers). Correlation between body weight and testis weight was positive and significant for sneaker males, while correlation was virtually equal to zero in territorial males. That body size in territorial males is correlated with fertilization success but not gonad size suggests that males allocate much more energy into growth and relatively little into sperm production once the needed size to become territorial is attained. The increased paternity of larger territorial males might be due to a more effective defense of the nest in comparison with smaller territorial males.

  7. Infections of nervous necrosis virus in wild and cage-reared marine fish from South China Sea with unexpected wide host ranges.

    PubMed

    Liu, X D; Huang, J N; Weng, S P; Hu, X Q; Chen, W J; Qin, Z D; Dong, X X; Liu, X L; Zhou, Y; Asim, M; Wang, W M; He, J G; Lin, L

    2015-06-01

    The concerns about the impact of the nervous necrosis virus (NNV) infections in wild fish have been raised. This paper presents the results of quarterly surveys of NNV in wild and cage-reared marine fish from South China Sea. Samples of 892 wild fish belonging to 69 species and 381 cage-reared fish belonging to 11 species were collected and were detected by seminested PCR and nested PCR. In the case of seminested PCR, the positive signal was detected in 3.0% and 3.1% samples of wild and cage-reared fish, respectively. However, by nested RT-PCR, the positive signal was observed in 42.3% and 63.0% samples of wild and cage-reared fish, respectively. If the fish species were considered, the positive signal was detected in 21.7% and 72.7% species of wild and cage-reared fish by seminested PCR assay, respectively. However, by nested RT-PCR, the positive signal was observed in 65.2% and 100% species of wild and cage-reared fish, respectively. The nucleotide sequences of the nested PCR products were determined. Phylogenetic tree showed that all the obtained viral isolates belonged to the red-spotted grouper nervous necrosis virus (RGNNV) genotype. Thirty-five species of the marine fish were the new hosts of NNV. © 2014 John Wiley & Sons Ltd.

  8. Weather radar data correlate to hail-induced mortality in grassland birds

    USGS Publications Warehouse

    Carver, Amber; Ross, Jeremy D.; Augustine, David J.; Skagen, Susan K.; Dwyer, Angela M.; Tomback, Diana F.; Wunder, Michael B.

    2017-01-01

    Small-bodied terrestrial animals such as songbirds (Order Passeriformes) are especially vulnerable to hail-induced mortality; yet, hail events are challenging to predict, and they often occur in locations where populations are not being studied. Focusing on nesting grassland songbirds, we demonstrate a novel approach to estimate hail-induced mortality. We quantify the relationship between the probability of nests destroyed by hail and measured Level-III Next Generation Radar (NEXRAD) data, including atmospheric base reflectivity, maximum estimated size of hail and maximum estimated azimuthal wind shear. On 22 June 2014, a hailstorm in northern Colorado destroyed 102 out of 203 known nests within our research site. Lark bunting (Calamospiza melanocorys) nests comprised most of the sample (n = 186). Destroyed nests were more likely to be found in areas of higher storm intensity, and distributions of NEXRAD variables differed between failed and surviving nests. For 133 ground nests where nest-site vegetation was measured, we examined the ameliorative influence of woody vegetation, nest cover and vegetation density by comparing results for 13 different logistic regression models incorporating the independent and additive effects of weather and vegetation variables. The most parsimonious model used only the interactive effect of hail size and wind shear to predict the probability of nest survival, and the data provided no support for any of the models without this predictor. We conclude that vegetation structure may not mitigate mortality from severe hailstorms and that weather radar products can be used remotely to estimate potential for hail mortality of nesting grassland birds. These insights will improve the efficacy of grassland bird population models under predicted climate change scenarios.

  9. Predation on seabirds by red foxes at Shaiak Island, Alaska

    USGS Publications Warehouse

    Petersen, M.R.

    1982-01-01

    Two Red Foxes (Vulpes fulva) that invaded Shaiak Island before the 1976 nesting season had a marked impact on the nesting success of five of seven species of seabirds breeding on the island that year. Common Eiders (Somateria mollissima), Glaucous-winged Gulls (Larus glaucescens), and Common Murres (Uria aalge), that nest in areas accessible to foxes, did not raise any young to fledging. Double-crested Cormorants (Phalacrocorax auritus) were only slightly more successful; 13 (4.3%) of 300 pairs raised one or more young to fledging. Evidence suggested that 21 (35.6%) of 62 pairs of Tufted Puffins (Lunda cirrhata) lost eggs or chicks to foxes, and foxes killed at least 13 (8.3%) of 156 adult puffins on ten sample plots. Conversely, Black-Legged Kittiwakes (Rissa tridactyla) and Pelagic Cormorants (Phalacrocorax pelagicus), which nested primarily on cliffs inaccessible to foxes, lost very few nests. There was no apparent change in general nest site selections by seabirds the following year, when foxes were no longer present. Any avoidance by birds of areas vulnerable to fox predation would probably be discernible only after several years of continuous predation.

  10. Avian growth and development rates and age-specific mortality: the roles of nest predation and adult mortality.

    PubMed

    Remes, V

    2007-01-01

    Previous studies have shown that avian growth and development covary with juvenile mortality. Juveniles of birds under strong nest predation pressure grow rapidly, have short incubation and nestling periods, and leave the nest at low body mass. Life-history theory predicts that parental investment increases with adult mortality rate. Thus, developmental traits that depend on the parental effort exerted (pre- and postnatal growth rate) should scale positively with adult mortality, in contrast to those that do not have a direct relationship with parental investment (timing of developmental events, e.g. nest leaving). I tested this prediction on a sample of 84 North American songbirds. Nestling growth rate scaled positively and incubation period duration negatively with annual adult mortality rates even when controlled for nest predation and other covariates, including phylogeny. On the contrary, neither the duration of the nestling period nor body mass at fledging showed any relationship. Proximate mechanisms generating the relationship of pre- and postnatal growth rates to adult mortality may include increased feeding, nest attentiveness during incubation and/or allocation of hormones, and deserve further attention.

  11. Ghrelin and leptin modulate the feeding behaviour of the hawksbill turtle Eretmochelys imbricata during nesting season

    PubMed Central

    Goldberg, Daphne Wrobel; Leitão, Santiago Alonso Tobar; Godfrey, Matthew H.; Lopez, Gustave Gilles; Santos, Armando José Barsante; Neves, Fabiana Alves; de Souza, Érica Patrícia Garcia; Moura, Anibal Sanchez; Bastos, Jayme da Cunha; Bastos, Vera Lúcia Freire da Cunha

    2013-01-01

    Female sea turtles have rarely been observed foraging during the nesting season. This suggests that prior to their migration to nesting beaches the females must store sufficient energy and nutrients at their foraging grounds and must be physiologically capable of undergoing months without feeding. Leptin (an appetite-suppressing protein) and ghrelin (a hunger-stimulating peptide) affect body weight by influencing energy intake in all vertebrates. We investigated the levels of these hormones and other physiological and nutritional parameters in nesting hawksbill sea turtles in Rio Grande do Norte State, Brazil, by collecting consecutive blood samples from 41 turtles during the 2010–2011 and 2011–2012 reproductive seasons. We found that levels of serum leptin decreased over the nesting season, which potentially relaxed suppression of food intake and stimulated females to begin foraging either during or after the post-nesting migration. Concurrently, we recorded an increasing trend in ghrelin, which may have stimulated food intake towards the end of the nesting season. Both findings are consistent with the prediction that post-nesting females will begin to forage, either during or immediately after their post-nesting migration. We observed no seasonal trend for other physiological parameters (values of packed cell volume and serum levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyl transferase, low-density lipoprotein, and high-density lipoprotein). The observed downward trends in general serum biochemistry levels were probably due to the physiological challenge of vitellogenesis and nesting in addition to limited energy resources and probable fasting. PMID:27293600

  12. Ghrelin and leptin modulate the feeding behaviour of the hawksbill turtle Eretmochelys imbricata during nesting season.

    PubMed

    Goldberg, Daphne Wrobel; Leitão, Santiago Alonso Tobar; Godfrey, Matthew H; Lopez, Gustave Gilles; Santos, Armando José Barsante; Neves, Fabiana Alves; de Souza, Érica Patrícia Garcia; Moura, Anibal Sanchez; Bastos, Jayme da Cunha; Bastos, Vera Lúcia Freire da Cunha

    2013-01-01

    Female sea turtles have rarely been observed foraging during the nesting season. This suggests that prior to their migration to nesting beaches the females must store sufficient energy and nutrients at their foraging grounds and must be physiologically capable of undergoing months without feeding. Leptin (an appetite-suppressing protein) and ghrelin (a hunger-stimulating peptide) affect body weight by influencing energy intake in all vertebrates. We investigated the levels of these hormones and other physiological and nutritional parameters in nesting hawksbill sea turtles in Rio Grande do Norte State, Brazil, by collecting consecutive blood samples from 41 turtles during the 2010-2011 and 2011-2012 reproductive seasons. We found that levels of serum leptin decreased over the nesting season, which potentially relaxed suppression of food intake and stimulated females to begin foraging either during or after the post-nesting migration. Concurrently, we recorded an increasing trend in ghrelin, which may have stimulated food intake towards the end of the nesting season. Both findings are consistent with the prediction that post-nesting females will begin to forage, either during or immediately after their post-nesting migration. We observed no seasonal trend for other physiological parameters (values of packed cell volume and serum levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyl transferase, low-density lipoprotein, and high-density lipoprotein). The observed downward trends in general serum biochemistry levels were probably due to the physiological challenge of vitellogenesis and nesting in addition to limited energy resources and probable fasting.

  13. Uniform Atmospheric Retrievals of Ultracool Late-T and Early-Y dwarfs

    NASA Astrophysics Data System (ADS)

    Garland, Ryan; Irwin, Patrick

    2017-10-01

    A significant number of ultracool (<600K) extrasolar objects have been discovered in the past decade thanks to wide-field surveys such as WISE. These objects present a perfect testbed for examining the evolution of atmospheric structure as we transition from typically hot extrasolar temperatures to the temperatures found within our Solar System.By examining these types of objects with a uniform retrieval method, we hope to elucidate any trends and (dis)similarities found in atmospheric parameters, such as chemical abundances, temperature-pressure profile, and cloud structure, for a sample of 7 ultracool brown dwarfs as we transition from hotter (~700K) to colder objects (~450K).We perform atmospheric retrievals on two late-T and five early-Y dwarfs. We use the NEMESIS atmospheric retrieval code coupled to a Nested Sampling algorithm, along with a standard uniform model for all of our retrievals. The uniform model assumes the atmosphere is described by a gray radiative-convective temperature profile, (optionally) a gray cloud, and a number of relevant gases. We first verify our methods by comparing it to a benchmark retrieval for Gliese 570D, which is found to be consistent. Furthermore, we present the retrieved gaseous composition, temperature structure, spectroscopic mass and radius, cloud structure and the trends associated with decreasing temperature found in this small sample of objects.

  14. Ecological factors related to the widespread distribution of sylvatic Rhodnius ecuadoriensis populations in southern Ecuador

    PubMed Central

    2012-01-01

    Background Chagas disease transmission risk is a function of the presence of triatomines in domestic habitats. Rhodnius ecuadoriensis is one of the main vectors implicated in transmission of Trypanosoma cruzi in Ecuador. This triatomine species is present in domestic, peridomestic and sylvatic habitats in the country. To determine the distribution of sylvatic populations of R. ecuadoriensis and the factors related to this distribution, triatomine searches were conducted between 2005 and 2009 in southern Ecuador. Methods Manual triatomine searches were conducted by skilled bug collectors in 23 communities. Sylvatic searched sites were selected by a) directed sampling, where microhabitats were selected by the searchers and b) random sampling, where sampling points where randomly generated. Domiciliary triatomine searches were conducted using the one man-hour method. Natural trypanosome infection was determined by microscopic examination and PCR. Generalized linear models were used to test the effect of environmental factors on the presence of sylvatic triatomines. Results In total, 1,923 sylvatic individuals were collected representing a sampling effort of 751 man-hours. Collected sylvatic triatomines were associated with mammal and bird nests. The 1,219 sampled nests presented an infestation index of 11.9%, a crowding of 13 bugs per infested nest, and a colonization of 80% of the nests. Triatomine abundance was significantly higher in squirrel (Sciurus stramineus) nests located above five meters from ground level and close to the houses. In addition, 8.5% of the 820 examined houses in the same localities were infested with triatomines. There was a significant correlation between R. ecuadoriensis infestation rates found in sylvatic and synanthropic environments within communities (p = 0.012). Parasitological analysis revealed that 64.7% and 15.7% of the sylvatic bugs examined (n = 300) were infected with Trypanosoma cruzi and T. rangeli respectively, and 8% of the bugs presented mixed infections. Conclusions The wide distribution of sylvatic R. ecuadoriensis populations may jeopardize the effectiveness of control campaigns conducted to eliminate domestic populations of this species. Also, the high T. cruzi infection rates found in sylvatic R. ecuadoriensis populations in southern Ecuador could constitute a risk for house re-infestation and persistent long-term Chagas disease transmission in the region. PMID:22243930

  15. Ecological factors related to the widespread distribution of sylvatic Rhodnius ecuadoriensis populations in southern Ecuador.

    PubMed

    Grijalva, Mario J; Suarez-Davalos, Victoria; Villacis, Anita G; Ocaña-Mayorga, Sofia; Dangles, Olivier

    2012-01-13

    Chagas disease transmission risk is a function of the presence of triatomines in domestic habitats. Rhodnius ecuadoriensis is one of the main vectors implicated in transmission of Trypanosoma cruzi in Ecuador. This triatomine species is present in domestic, peridomestic and sylvatic habitats in the country. To determine the distribution of sylvatic populations of R. ecuadoriensis and the factors related to this distribution, triatomine searches were conducted between 2005 and 2009 in southern Ecuador. Manual triatomine searches were conducted by skilled bug collectors in 23 communities. Sylvatic searched sites were selected by a) directed sampling, where microhabitats were selected by the searchers and b) random sampling, where sampling points where randomly generated. Domiciliary triatomine searches were conducted using the one man-hour method. Natural trypanosome infection was determined by microscopic examination and PCR. Generalized linear models were used to test the effect of environmental factors on the presence of sylvatic triatomines. In total, 1,923 sylvatic individuals were collected representing a sampling effort of 751 man-hours. Collected sylvatic triatomines were associated with mammal and bird nests. The 1,219 sampled nests presented an infestation index of 11.9%, a crowding of 13 bugs per infested nest, and a colonization of 80% of the nests. Triatomine abundance was significantly higher in squirrel (Sciurus stramineus) nests located above five meters from ground level and close to the houses. In addition, 8.5% of the 820 examined houses in the same localities were infested with triatomines. There was a significant correlation between R. ecuadoriensis infestation rates found in sylvatic and synanthropic environments within communities (p = 0.012). Parasitological analysis revealed that 64.7% and 15.7% of the sylvatic bugs examined (n = 300) were infected with Trypanosoma cruzi and T. rangeli respectively, and 8% of the bugs presented mixed infections. The wide distribution of sylvatic R. ecuadoriensis populations may jeopardize the effectiveness of control campaigns conducted to eliminate domestic populations of this species. Also, the high T. cruzi infection rates found in sylvatic R. ecuadoriensis populations in southern Ecuador could constitute a risk for house re-infestation and persistent long-term Chagas disease transmission in the region.

  16. A Newly Developed Nested PCR Assay for the Detection of Helicobacter pylori in the Oral Cavity.

    PubMed

    Ismail, Hawazen; Morgan, Claire; Griffiths, Paul; Williams, John; Jenkins, Gareth

    2016-01-01

    To develop a new nested polymerase chain reaction (PCR) assay for identifying Helicobacter pylori DNA from dental plaque. H. pylori is one of the most common chronic bacterial pathogens in humans. The accurate detection of this organism is essential for proper patient management and for the eradication of the bacteria following treatment. Forty-nine patients (24 males and 25 females; mean age: 51; range, 19 to 94 y) were investigated for the presence of H. pylori in dental plaque by single-step PCR and nested PCR and in the stomach by single-step PCR, nested PCR, and histologic examination. The newly developed nested PCR assay identified H. pylori DNA in gastric biopsies of 18 patients who were histologically classified as H. pylori-positive and 2 additional biopsies of patients who were H. pylori-negative by histologic examination (20/49; 40.8%). Dental plaque samples collected before and after endoscopy from the 49 patients revealed that single-step PCR did not detect H. pylori but nested PCR was able to detect H. pylori DNA in 40.8% (20/49) patients. Nested PCR gave a higher detection rate (40.8%, 20/49) than that of histology (36.7%, 18/49) and single-step PCR. When nested PCR results were compared with histology results there was no significant difference between the 2 methods. Our newly developed nested PCR assay is at least as sensitive as histology and may be useful for H. pylori detection in patients unfit for endoscopic examination.

  17. Trap-Nesting Bees in Montane Grassland (Campo Rupestre) and Cerrado in Brazil: Collecting Generalist or Specialist Nesters.

    PubMed

    Araújo, P C S; Lourenço, A P; Raw, A

    2016-10-01

    Species richness and seasonal abundance of solitary bees were investigated in rocky, montane grassland (campo rupestre) (1180 m asl) and cerrado sensu stricto (680 m asl) in the Biribiri State Park, Diamantina, state of Minas Gerais, Brazil. Three hundred nineteen nest traps of bamboo canes and black cardboard tubes were monthly inspected at each site during 15 months. A total of eight species of bees built 97 nests. Four species were common to both sites. Tetrapedia aff. curvitarsis Friese and Tetrapedia aff. peckoltii Friese were the most abundant at campo rupestre and cerrado s.s., respectively, followed by Centris analis (Fabricius) in campo rupestre and Centris tarsata Smith in cerrado s.s. The nesting peaks occurred in May in campo rupestre and in February in cerrado s.s. Three cuckoo bees and one bee-fly were collected as natural enemies. The findings suggest that differences between the sites were related more to ecological factors (floral resources, natural nest sites) than to the altitudinal difference. The species richness was similar to that in other habitats with open vegetation. We demonstrate the need to use several types of trap-nest to increase the range of species sampled; some species used only one of the two types traps provided. We also comment on the limitations of trap-nests in cerrado vegetation. Most cerrado species of bees are very selective in their choice for a nesting site, but bees that use trap-nests are more generalists.

  18. Pre- and post-experimental manipulation assessments confirm the increase in number of birds due to the addition of nest boxes.

    PubMed

    Cuatianquiz Lima, Cecilia; Macías Garcia, Constantino

    2016-01-01

    Secondary cavity nesting (SCN) birds breed in holes that they do not excavate themselves. This is possible where there are large trees whose size and age permit the digging of holes by primary excavators and only rarely happens in forest plantations, where we expected a deficit of both breeding holes and SCN species. We assessed whether the availability of tree cavities influenced the number of SCNs in two temperate forest types, and evaluated the change in number of SCNs after adding nest boxes. First, we counted all cavities within each of our 25-m radius sampling points in mature and young forest plots during 2009. We then added nest boxes at standardised locations during 2010 and 2011 and conducted fortnightly bird counts (January-October 2009-2011). In 2011 we added two extra plots of each forest type, where we also conducted bird counts. Prior to adding nest boxes, counts revealed more SCNs in mature than in young forest. Following the addition of nest boxes, the number of SCNs increased significantly in the points with nest boxes in both types of forest. Counts in 2011 confirmed the increase in number of birds due to the addition of nest boxes. Given the likely benefits associated with a richer bird community we propose that, as is routinely done in some countries, forest management programs preserve old tree stumps and add nest boxes to forest plantations in order to increase bird numbers and bird community diversity.

  19. Conservation implications when the nest predators are known

    USGS Publications Warehouse

    Ribic, Christine; Thompson, Frank

    2012-01-01

    Conservation and management of passerines has largely focused on habitat manipulation or restoration because the natural communities on which these birds depend have been destroyed and fragmented. However, productivity is another important aspect of avian conservation, and nest predation can be a large source of nesting mortality for passerines. Recent studies using video surveillance to identify nest predators allow researchers to start evaluating what methods could be used to mitigate nest predation to help passerines of conservation concern. From recent studies, we identified latitudinal and habitat-related patterns in the importance of predator groups that depredate passerine nests. We then reviewed how knowledge of specific nest predators can benefit conservation of bird species of concern. Mammals were the dominant predator group in northern grasslands. Snakes were the dominant predator group in southern habitats. Fire ants were only a nest predator in southern latitudes. Differences in the importance of predator species or groups were likely the result of both their geographic patterns of distribution and habitat preferences. Some direct and indirect predator control measures developed for waterfowl management potentially could be used to benefit passerine productivity. We reviewed three examples-cowbirds, snakes in shrublands, and ground squirrels in grasslands-to illustrate how different predator control strategies may be needed in different situations. Mitigation of passerine nest predation will need to be based on knowledge of predator communities to be effective. This requires large samples of predation events with identified predators; video technology is essential for this task.

  20. Range resolution improvement in passive bistatic radars using nested FM channels and least squares approach

    NASA Astrophysics Data System (ADS)

    Arslan, Musa T.; Tofighi, Mohammad; Sevimli, Rasim A.; ćetin, Ahmet E.

    2015-05-01

    One of the main disadvantages of using commercial broadcasts in a Passive Bistatic Radar (PBR) system is the range resolution. Using multiple broadcast channels to improve the radar performance is offered as a solution to this problem. However, it suffers from detection performance due to the side-lobes that matched filter creates for using multiple channels. In this article, we introduce a deconvolution algorithm to suppress the side-lobes. The two-dimensional matched filter output of a PBR is further analyzed as a deconvolution problem. The deconvolution algorithm is based on making successive projections onto the hyperplanes representing the time delay of a target. Resulting iterative deconvolution algorithm is globally convergent because all constraint sets are closed and convex. Simulation results in an FM based PBR system are presented.

  1. Mining Distance Based Outliers in Near Linear Time with Randomization and a Simple Pruning Rule

    NASA Technical Reports Server (NTRS)

    Bay, Stephen D.; Schwabacher, Mark

    2003-01-01

    Defining outliers by their distance to neighboring examples is a popular approach to finding unusual examples in a data set. Recently, much work has been conducted with the goal of finding fast algorithms for this task. We show that a simple nested loop algorithm that in the worst case is quadratic can give near linear time performance when the data is in random order and a simple pruning rule is used. We test our algorithm on real high-dimensional data sets with millions of examples and show that the near linear scaling holds over several orders of magnitude. Our average case analysis suggests that much of the efficiency is because the time to process non-outliers, which are the majority of examples, does not depend on the size of the data set.

  2. Identification of and Screening for Human Helicobacter cinaedi Infections and Carriers via Nested PCR

    PubMed Central

    Oyama, Kohta; Khan, Shahzada; Okamoto, Tatsuya; Fujii, Shigemoto; Ono, Katsuhiko; Matsunaga, Tetsuro; Yoshitake, Jun; Sawa, Tomohiro; Tomida, Junko; Kawamura, Yoshiaki

    2012-01-01

    Helicobacter cinaedi is the most frequently reported enterohepatic Helicobacter species isolated from humans. Earlier research suggested that certain patients with H. cinaedi infection may remain undiagnosed or incorrectly diagnosed because of difficulties in detecting the bacteria by conventional culture methods. Here, we report a nested PCR assay that rapidly detects the cytolethal distending toxin gene (cdt) of H. cinaedi with high specificity and sensitivity. Specificity of the assay was validated by using different species of Helicobacter and Campylobacter, as well as known H. cinaedi-positive and -negative samples. The sensitivity of detection for the cdt gene in the assay was 102 CFU/ml urine or 102 CFU/105 infected RAW 264.7 cells. In an H. cinaedi-infected mouse model, the cdt gene of H. cinaedi was effectively detected via the assay with urine (6/7), stool (2/3), and blood (2/6) samples. Importantly, it detected H. cinaedi in blood, urine, and stool samples from one patient with a suspected H. cinaedi infection and three patients with known infections. The assay was further used clinically to follow up two H. cinaedi-infected patients after antibiotic treatment. Stool samples from these two patients evaluated by nested PCR after antibiotic therapy showed clearance of bacterial DNA. Finally, analysis of stool specimens from healthy volunteers showed occasional positive reactions (4/30) to H. cinaedi DNA, which suggests intestinal colonization by H. cinaedi in healthy subjects. In conclusion, this nested PCR assay may be useful for the rapid diagnosis, antimicrobial treatment evaluation, and epidemiological study of H. cinaedi infection. PMID:23015666

  3. Prevalence of coliforms, Salmonella, Listeria, and Campylobacter associated with eggs and the environment of conventional cage and free-range egg production.

    PubMed

    Jones, D R; Anderson, K E; Guard, J Y

    2012-05-01

    There is a desire by US consumers for eggs produced by hens in alternative production systems. As the retail shell-egg market offers these products to accommodate consumer demands, additional information is needed to ensure processing methodologies result in safe eggs from all egg sources. A study was conducted to determine if there were differences in the prevalence of coliforms, Salmonella, Listeria, and Campylobacter on and within eggs and in the environment of a sister flock of conventional cage and free-range laying hens. Microbial sampling occurred approximately every 6 wk between 20 and 79 wk of age. A random sampling of typical coliform colonies produced 371 viable isolates for biochemical identification. Twenty-nine genera or species of bacteria were identified. There was a significantly greater (P < 0.0001) prevalence of Campylobacter in the free-range nest box swabs compared with that in the free-range grass and conventional cage swab samples (number of positives: 8 nest box, 1 grass, 0 cage). Seven isolates of Listeria innocua were detected with no significant difference in prevalence between the treatments. Isolates were associated with eggshells (2 free-range floor, 1 cage) and the free-range environment (2 nest box, 2 grass). There were 21 Salmonella isolates detected between all sample locations, with no significant difference in the prevalence of Salmonella detection between the treatments. Additional studies are needed to fully understand the effect of alternative production methods on the prevalence of pathogens and coliforms associated with nest-run eggs and the production environment.

  4. Sound absorption of a new oblique-section acoustic metamaterial with nested resonator

    NASA Astrophysics Data System (ADS)

    Gao, Nansha; Hou, Hong; Zhang, Yanni; Wu, Jiu Hui

    2018-02-01

    This study designs and investigates high-efficiency sound absorption of new oblique-section nested resonators. Impedance tube experiment results show that different combinations of oblique-section nest resonators have tunable low-frequency bandwidth characteristics. The sound absorption mechanism is due to air friction losses in the slotted region and the sample structure resonance. The acousto-electric analogy model demonstrates that the sound absorption peak and bandwidth can be modulated over an even wider frequency range by changing the geometric size and combinations of structures. The proposed structure can be easily fabricated and used in low-frequency sound absorption applications.

  5. Can a Red Wood-Ant Nest Be Associated with Fault-Related CH4 Micro-Seepage? A Case Study from Continuous Short-Term In-Situ Sampling

    PubMed Central

    Grumpe, Arne; Becker, Adrian; Wöhler, Christian

    2018-01-01

    Simple Summary Methane (CH4) is common on Earth but its natural sources are not well-characterized. We investigated concentrations of CH4 and its stable carbon isotope (δ13C-CH4) within a red wood-ant (RWA; Formica polyctena) nest in the Neuwied Basin, a part of the East Eifel Volcanic Field (EEVF), and tested for associations between methane concentration and RWA activity patterns, earthquakes, and earth tides. Methane degassing was not synchronized with earth tides, nor was it influenced by a micro-earthquake or RWA activity. Elevated CH4 concentrations in nest gas appear to result from a combination of microbial activity and fault-related emissions. The latter could result from micro-seepage of methane derived from low-temperature gas-water-rock reactions that subsequently moves via fault networks through the RWA nest or from overlapping micro-seepage of magmatic CH4 from the Eifel plume. Given the abundance of RWA nests on the landscape, their role as sources of microbial CH4 and biological indicators for abiotically-derived CH4 should be included in estimations of methane emissions that are contributing to climatic change. Abstract We measured methane (CH4) and stable carbon isotope of methane (δ13C-CH4) concentrations in ambient air and within a red wood-ant (RWA; Formica polyctena) nest in the Neuwied Basin (Germany) using high-resolution in-situ sampling to detect microbial, thermogenic, and abiotic fault-related micro-seepage of CH4. Methane degassing from RWA nests was not synchronized with earth tides, nor was it influenced by micro-earthquake degassing or concomitantly measured RWA activity. Two δ13C-CH4 signatures were identified in nest gas: −69‰ and −37‰. The lower peak was attributed to microbial decomposition of organic matter within the RWA nest, in line with previous observations that RWA nests are hot-spots of microbial CH4. The higher peak has not been reported in previous studies. We attribute this peak to fault-related CH4 emissions moving via fault networks into the RWA nest, which could originate either from thermogenic or abiotic CH4 formation. Sources of these micro-seepages could be Devonian schists, iron-bearing “Klerf Schichten”, or overlapping micro-seepage of magmatic CH4 from the Eifel plume. Given the abundance of RWA nests on the landscape, their role as sources of microbial CH4 and biological indicators for abiotically-derived CH4 should be included in estimation of methane emissions that are contributing to climatic change. PMID:29597318

  6. Influence of nesting shell size on brightness longevity and resistance to ultrasound-induced dissolution during enhanced B-mode contrast imaging.

    PubMed

    Wallace, N; Dicker, S; Lewin, P; Wrenn, S P

    2014-12-01

    This study aims to bridge the gap between transport mechanisms of an improved ultrasound contrast agent (UCA) and its resulting behavior in a clinical imaging study. Phospholipid-shelled microbubbles nested within the aqueous core of a polymer microcapsule are examined for their use and feasibility as an improved UCA. The nested formulation provides contrast comparable to traditional formulations, specifically an SF6 microbubble coated by a DSPC PEG-3000 monolayer, with the advantage that contrast persists at least nine times longer in a mock clinical, in vitro setting. The effectiveness of the sample was measured using a contrast ratio in units of decibels (dB) which compares the brightness of the nested microbubbles to a reference value of a phantom tissue mimic. During a 40min imaging study, six nesting formulations with average outer capsule diameters of 1.95, 2.53, 5.55, 9.95, 14.95, and 20.51μm reached final contrast ratio values of 0.25, 2.35, 3.68, 4.51, 5.93, and 8.00dB, respectively. The starting contrast ratio in each case was approximately 8dB and accounts for the brightness attributed to the nesting shell. As compared with empty microcapsules (no microbubbles nested within), enhancement of the initial contrast ratio increased systematically with decreasing microcapsule size. The time required to reach a steady state in the temporal contrast ratio profile also varied with microcapsule diameter and was found to be 420s for each of the four smallest shell diameters and 210s and 150s, respectively, for the largest two shell diameters. All nested formulations were longer-lived and gave higher final contrast ratios than a control sample comprising un-nested, but otherwise equivalent, microbubbles. Specifically, the contrast ratio of the un-nested microbubbles decreased to a negative value after 4min of continuous ultrasound exposure with complete disappearance of the microbubbles after 15min whereas all nested formulations maintained positive contrast ratio values for the duration of the 40min trial. The results are consistent with two distinct stages of gas transport: in the first stage, passive diffusion occurs under ambient conditions across the microbubble monolayer within the first few minutes after formulation until the aqueous interior of the microcapsule is saturated with gas; in the second stage ultrasound drives additional gas dissolution even further due to pressure modulation. It is important to understand the chemistry and transport mechanisms of this contrast agent under the influence of ultrasound to attain better perspicacity for enhanced applications in imaging. Results from this study will facilitate future preclinical studies and clinical applications of nested microbubbles for therapeutic and diagnostic imaging. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Assessment of Provisional MODIS-derived Surfaces Related to the Global Carbon Cycle

    NASA Astrophysics Data System (ADS)

    Cohen, W. B.; Maiersperger, T. K.; Turner, D. P.; Gower, S. T.; Kennedy, R. E.; Running, S. W.

    2002-12-01

    The global carbon cycle is one of the most important foci of an emerging global biosphere monitoring system. A key component of such a system is the MODIS sensor, onboard the Terra satellite platform. Biosphere monitoring requires an integrated program of satellite observations, Earth-system models, and in situ data. Related to the carbon cycle, MODIS science teams routinely develop a variety of global surfaces such as land cover, leaf area index, and net primary production using MODIS data and functional algorithms. The quality of these surfaces must be evaluated to determine their effectiveness for global biosphere monitoring. A project called BigFoot (http://www.fsl.orst.edu/larse/bigfoot/) is an organized effort across nine biomes to assess the quality of the abovementioned surfaces: (1) Arctic tundra; (2) boreal evergreen needle-leaved forest; temperate (3) cropland, (4) grassland, (5) evergreen needle-leaved forest, and (6) deciduous broad-leaved forest; desert (7) grassland and (8) shrubland; and (9) tropical evergreen broad-leaved forest. Each biome is represented by a site that has an eddy-covariance flux tower that measures water vapor and CO2 fluxes. Flux tower footprints are relatively small-approximately 1 km2. BigFoot characterizes 25 km2 around each tower, using field data, Landsat ETM+ image data, and ecosystem process models. Our innovative field sampling design incorporates a nested spatial series to facilitate geostatistical analyses, samples the ecological variability at a site, and is logistically efficient. Field data are used both to develop site-specific algorithms for mapping/modeling the variables of interest and to characterize the errors in derived BigFoot surfaces. Direct comparisons of BigFoot- and MODIS-derived surfaces are made to help understand the sources of error in MODIS-derived surfaces and to facilitate improvements to MODIS algorithms. Results from four BigFoot sites will be presented.

  8. Nested PCR for specific diagnosis of Taenia solium taeniasis.

    PubMed

    Mayta, Holger; Gilman, Robert H; Prendergast, Emily; Castillo, Janeth P; Tinoco, Yeny O; Garcia, Hector H; Gonzalez, Armando E; Sterling, Charles R

    2008-01-01

    Taeniasis due to Taenia solium is a disease with important public health consequences, since the larval stage is not exclusive to the animal intermediate, the pig, but also infects humans, causing neurocysticercosis. Early diagnosis and treatment of T. solium tapeworm carriers is important to prevent human cysticercosis. Current diagnosis based on microscopic observation of eggs lacks both sensitivity and specificity. In the present study, a nested-PCR assay targeting the Tso31 gene was developed for the specific diagnosis of taeniasis due to T. solium. Initial specificity and sensitivity testing was performed using stored known T. solium-positive and -negative samples. The assay was further analyzed under field conditions by conducting a case-control study of pretreatment stool samples collected from a population in an area of endemicity. Using the archived samples, the assay showed 97% (31/32) sensitivity and 100% (123/123) specificity. Under field conditions, the assay had 100% sensitivity and specificity using microscopy/enzyme-linked immunosorbent assay coproantigen testing as the gold standards. The Tso31 nested PCR described here might be a useful tool for the early diagnosis and prevention of taeniasis/cysticercosis.

  9. Avifauna associated with early growth vegetation on clearcuts in the Oregon coast ranges.

    Treesearch

    Michael L. Morrison; E. Charles Meslow

    1983-01-01

    This paper provides estimates of bird density, diversity, and evenness on 13 clearcut units of the Siuslaw National Forest in the Coast Ranges of Oregon, sampled during 1979,1980, and 1981. Total density of nesting birds ranged from 322 to 588 per 40.5 hectares (100 acres); there were 15 to 19 species nesting on each site.

  10. The Empty Nest Syndrome: Ways to Enhance Quality of Life

    ERIC Educational Resources Information Center

    Chen, Dianbing; Yang, Xinxiao; Aagard, Steve Dale

    2012-01-01

    Empty nest syndrome occurs as a result of urbanization and loosened relationships among family members. It may threaten the life quality of older adults and stability of society as a whole. This survey was designed to investigate the situation and factors that influence the life quality of a sample of older adults in a western state. Thirty-five…

  11. Optimal Design in Three-Level Block Randomized Designs with Two Levels of Nesting: An ANOVA Framework with Random Effects

    ERIC Educational Resources Information Center

    Konstantopoulos, Spyros

    2013-01-01

    Large-scale experiments that involve nested structures may assign treatment conditions either to subgroups such as classrooms or to individuals such as students within subgroups. Key aspects of the design of such experiments include knowledge of the variance structure in higher levels and the sample sizes necessary to reach sufficient power to…

  12. [Iditification of five imported cases of Plasmodium ovale wallikeri infection in Zhejiang Province].

    PubMed

    Zhang, Ling-ling; Ruan, Wei; Chen, Hua-liang; Lu, Qiao-yi; Yao, Li-nong

    2014-10-01

    To identify and analyze Plasmodium ovale wallikeri in 5 imported malaria cases, who were detected positive by microscopy and negative by conventional PCR. Epidemiological information and blood samples were collected from the five patients. The detection was conducted by microscopy, Rapid Diagnostic Test (RDT) and nested PCR with Plasmodium genus-specific, species-specific and Plasmodium ovale wallikeri-specific primers. The amplified products were sequenced and Blast analysis was performed on line in NCBI. The five patients returned from Africa, and all had a history of malaria. They were microscopically positive for Plasmodium sp., and two cases showed Pan positive RDT result. All blood samples were negative for four Plasmodium spp. by conventional nested PCR, but positive by nested PCR with Plasmodium ovale wallikeri-specific primers. Blast analysis showed that the amplified sequences of the five cases had complete homology with P. ovale wallikeri clone RSH10 18S ribosomal RNA gene (Accession No. KF219561.1). The five cases which classified as positive by microscopy while negative by conventional PCR have been confirmed as Plasmodium ovale wallikeri infection by nested PCR with P. ovale wallikeri-specific primers.

  13. Early diagnosis of typhoid fever by nested PCR for flagellin gene of Salmonella enterica serotype Typhi.

    PubMed

    Khan, S; Harish, B N; Menezes, G A; Acharya, N S; Parija, S C

    2012-11-01

    Typhoid fever caused by Salmonella Typhi continues to be a major health problem in spite of the use of antibiotics and the development of newer antibacterial drugs. Inability to make an early laboratory diagnosis and resort to empirical therapy, often lead to increased morbidity and mortality in cases of typhoid fever. This study was aimed to optimize a nested PCR for early diagnosis of typhoid fever and using it as a diagnostic tool in culture negative cases of suspected typhoid fever. Eighty patients with clinical diagnosis of typhoid fever and 40 controls were included in the study. The blood samples collected were subjected to culture, Widal and nested PCR targeting the flagellin gene of S. Typhi. The sensitivity of PCR on blood was found to be 100 per cent whereas the specificity was 76.9 per cent. The positive predictive value (PPV) of PCR was calculated to be 76.9 per cent with an accuracy of 86 per cent. None of the 40 control samples gave a positive PCR. Due to its high sensitivity and specificity nested PCR can be used as a useful tool to diagnose clinically suspected, culture negative cases of typhoid fever.

  14. Possibility expectation and its decision making algorithm

    NASA Technical Reports Server (NTRS)

    Keller, James M.; Yan, Bolin

    1992-01-01

    The fuzzy integral has been shown to be an effective tool for the aggregation of evidence in decision making. Of primary importance in the development of a fuzzy integral pattern recognition algorithm is the choice (construction) of the measure which embodies the importance of subsets of sources of evidence. Sugeno fuzzy measures have received the most attention due to the recursive nature of the fabrication of the measure on nested sequences of subsets. Possibility measures exhibit an even simpler generation capability, but usually require that one of the sources of information possess complete credibility. In real applications, such normalization may not be possible, or even desirable. In this report, both the theory and a decision making algorithm for a variation of the fuzzy integral are presented. This integral is based on a possibility measure where it is not required that the measure of the universe be unity. A training algorithm for the possibility densities in a pattern recognition application is also presented with the results demonstrated on the shuttle-earth-space training and testing images.

  15. Inferring Foraging Areas of Nesting Loggerhead Turtles Using Satellite Telemetry and Stable Isotopes

    PubMed Central

    Ceriani, Simona A.; Roth, James D.; Evans, Daniel R.; Weishampel, John F.; Ehrhart, Llewellyn M.

    2012-01-01

    In recent years, the use of intrinsic markers such as stable isotopes to link breeding and foraging grounds of migratory species has increased. Nevertheless, several assumptions still must be tested to interpret isotopic patterns found in the marine realm. We used a combination of satellite telemetry and stable isotope analysis to (i) identify key foraging grounds used by female loggerheads nesting in Florida and (ii) examine the relationship between stable isotope ratios and post-nesting migration destinations. We collected tissue samples for stable isotope analysis from 14 females equipped with satellite tags and an additional 57 untracked nesting females. Telemetry identified three post-nesting migratory pathways and associated non-breeding foraging grounds: (1) a seasonal continental shelf–constrained migratory pattern along the northeast U.S. coastline, (2) a non-breeding residency in southern foraging areas and (3) a residency in the waters adjacent to the breeding area. Isotopic variability in both δ13C and δ15N among individuals allowed identification of three distinct foraging aggregations. We used discriminant function analysis to examine how well δ13C and δ15N predict female post-nesting migration destination. The discriminant analysis classified correctly the foraging ground used for all but one individual and was used to predict putative feeding areas of untracked turtles. We provide the first documentation that the continental shelf of the Mid- and South Atlantic Bights are prime foraging areas for a large number (61%) of adult female loggerheads from the largest loggerhead nesting population in the western hemisphere and the second largest in the world. Our findings offer insights for future management efforts and suggest that this technique can be used to infer foraging strategies and residence areas in lieu of more expensive satellite telemetry, enabling sample sizes that are more representative at the population level. PMID:23028943

  16. Assessment of Contaminant Exposure and Effects on Ospreys Nesting along the Lower Duwamish River, Washington, 2006-07

    USGS Publications Warehouse

    Johnson, Branden L.; Henny, Charles J.; Kaiser, James L.; Davis, Jay W.; Schulz, Edmund P.

    2009-01-01

    We evaluated the effects of contaminants on osprey (Pandion haliaetus) nesting along the lower Duwamish River (LDR), Washington, and used the upper reach of the Willamette River (WR), Oregon, as a reference site. Osprey eggs and nestling blood (plasma) were collected at nests along the LDR (11 eggs, 7 plasmas) and WR (10 eggs, 6 plasmas) in 2006-07 and analyzed for contaminants. Additionally, hematology and serum chemistries were determined in the blood/plasma samples of nestlings (about 35-45 days old) and were used as potential indicators of stress induced by contaminant exposure. Detailed foraging information for ospreys nesting along the LDR was collected and evaluated to better understand contaminant profiles observed in the eggs and plasma. Additional residue data from 26 osprey eggs collected and analyzed in 2002-03 from nests along the LDR, Snohomish River Estuary (SRE) and Lake Washington (LW) in the Puget Sound (PS) region also were evaluated.

  17. Relatedness predicts multiple measures of investment in cooperative nest construction in sociable weavers

    PubMed Central

    Leighton, Gavin M.; Echeverri, Sebastian; Heinrich, Dirk; Kolberg, Holger

    2015-01-01

    Although communal goods are often critical to society, they are simultaneously susceptible to exploitation and are evolutionarily stable only if mechanisms exist to curtail exploitation. Mechanisms such as punishment and kin selection have been offered as general explanations for how communal resources can be maintained. Evidence for these mechanisms comes largely from humans and social insects, leaving their generality in question. To assess how communal resources are maintained, we observed cooperative nest construction in sociable weavers (Philetairus socius). The communal nest of sociable weavers provides thermal benefits for all individuals but requires continual maintenance. We observed cooperative nest construction and also recorded basic morphological characteristics. We also collected blood samples, performed next-generation sequencing, and isolated 2358 variable single nucleotide polymorphisms (SNPs) to estimate relatedness. We find that relatedness predicts investment in cooperative nest construction, while no other morphological characters significantly explain cooperative output. We argue that indirect benefits are a critical fitness component for maintaining the cooperative behavior that maintains the communal good. PMID:26726282

  18. A distributed algorithm to maintain and repair the trail networks of arboreal ants.

    PubMed

    Chandrasekhar, Arjun; Gordon, Deborah M; Navlakha, Saket

    2018-06-18

    We study how the arboreal turtle ant (Cephalotes goniodontus) solves a fundamental computing problem: maintaining a trail network and finding alternative paths to route around broken links in the network. Turtle ants form a routing backbone of foraging trails linking several nests and temporary food sources. This species travels only in the trees, so their foraging trails are constrained to lie on a natural graph formed by overlapping branches and vines in the tangled canopy. Links between branches, however, can be ephemeral, easily destroyed by wind, rain, or animal movements. Here we report a biologically feasible distributed algorithm, parameterized using field data, that can plausibly describe how turtle ants maintain the routing backbone and find alternative paths to circumvent broken links in the backbone. We validate the ability of this probabilistic algorithm to circumvent simulated breaks in synthetic and real-world networks, and we derive an analytic explanation for why certain features are crucial to improve the algorithm's success. Our proposed algorithm uses fewer computational resources than common distributed graph search algorithms, and thus may be useful in other domains, such as for swarm computing or for coordinating molecular robots.

  19. Effect of Methodological and Ecological Approaches on Heterogeneity of Nest-Site Selection of a Long-Lived Vulture

    PubMed Central

    Moreno-Opo, Rubén; Fernández-Olalla, Mariana; Margalida, Antoni; Arredondo, Ángel; Guil, Francisco

    2012-01-01

    The application of scientific-based conservation measures requires that sampling methodologies in studies modelling similar ecological aspects produce comparable results making easier their interpretation. We aimed to show how the choice of different methodological and ecological approaches can affect conclusions in nest-site selection studies along different Palearctic meta-populations of an indicator species. First, a multivariate analysis of the variables affecting nest-site selection in a breeding colony of cinereous vulture (Aegypius monachus) in central Spain was performed. Then, a meta-analysis was applied to establish how methodological and habitat-type factors determine differences and similarities in the results obtained by previous studies that have modelled the forest breeding habitat of the species. Our results revealed patterns in nesting-habitat modelling by the cinereous vulture throughout its whole range: steep and south-facing slopes, great cover of large trees and distance to human activities were generally selected. The ratio and situation of the studied plots (nests/random), the use of plots vs. polygons as sampling units and the number of years of data set determined the variability explained by the model. Moreover, a greater size of the breeding colony implied that ecological and geomorphological variables at landscape level were more influential. Additionally, human activities affected in greater proportion to colonies situated in Mediterranean forests. For the first time, a meta-analysis regarding the factors determining nest-site selection heterogeneity for a single species at broad scale was achieved. It is essential to homogenize and coordinate experimental design in modelling the selection of species' ecological requirements in order to avoid that differences in results among studies would be due to methodological heterogeneity. This would optimize best conservation and management practices for habitats and species in a global context. PMID:22413023

  20. Contaminant exposure and reproductive success of Ospreys (Pandion haliaetus) nesting in Chesapeake Bay regions of concern

    USGS Publications Warehouse

    Rattner, B.A.; McGowan, P.C.; Golden, N.H.; Hatfield, J.S.; Toschik, P.C.; Lukei, R.F.; Hale, R.C.; Schmitz-Afonso, I.; Rice, C.P.

    2004-01-01

    The Chesapeake Bay osprey population has more than doubled in size since restrictions were placed on the production and use of DDT and other toxic organochlorine contaminants in the 1970s. Ospreys are now nesting in the most highly polluted portions of the Bay. In 2000 and 2001, contaminant exposure and reproduction were monitored in ospreys nesting in regions of concern, including Baltimore Harbor and the Patapsco River, the Anacostia and middle Potomac rivers, and the Elizabeth River, and a presumed reference site consisting of the South, West, and Rhode rivers. A 'sample egg' from each study nest was collected for contaminant analysis, and the fate of eggs remaining in each nest (n = 14-16/site) was monitored at 7- to 10-day intervals from egg incubation through fledging of young. Ospreys fledged young in regions of concern (observed success: 0.88 -1.53 fledglings/active nest), although productivity was marginal for sustaining local populations in Baltimore Harbor and the Patapsco River and in the Anacostia and middle Potomac rivers. Concentrations of p,p'DDE and many other organochlorine pesticides or metabolites, total PCBs, some arylhydrocarbon receptor-active PCB congeners and polybrominated diphenyl ether congeners, and perfluorooctanesulfonate were often greater in sample eggs from regions of concern compared to the reference site. Nonetheless, logistic regression analyses did not provide evidence linking marginal productivity to p,p' -DDE, total PCBs, or arylhydrocarbon receptor-active PCB congener exposure in regions of concern. In view of the moderate concentrations of total PCBs in eggs from the reference site, concerns related to new and emerging toxicants, and the absence of ecotoxicological data for terrestrial vertebrates in many Bay tributaries, a more thorough spatial evaluation of contaminant exposure in ospreys throughout the Chesapeake may be warranted.

  1. Analyzing gene perturbation screens with nested effects models in R and bioconductor.

    PubMed

    Fröhlich, Holger; Beissbarth, Tim; Tresch, Achim; Kostka, Dennis; Jacob, Juby; Spang, Rainer; Markowetz, F

    2008-11-01

    Nested effects models (NEMs) are a class of probabilistic models introduced to analyze the effects of gene perturbation screens visible in high-dimensional phenotypes like microarrays or cell morphology. NEMs reverse engineer upstream/downstream relations of cellular signaling cascades. NEMs take as input a set of candidate pathway genes and phenotypic profiles of perturbing these genes. NEMs return a pathway structure explaining the observed perturbation effects. Here, we describe the package nem, an open-source software to efficiently infer NEMs from data. Our software implements several search algorithms for model fitting and is applicable to a wide range of different data types and representations. The methods we present summarize the current state-of-the-art in NEMs. Our software is written in the R language and freely avail-able via the Bioconductor project at http://www.bioconductor.org.

  2. Global optimization of additive potential energy functions: Predicting binary Lennard-Jones clusters

    NASA Astrophysics Data System (ADS)

    Kolossváry, István; Bowers, Kevin J.

    2010-11-01

    We present a method for minimizing additive potential-energy functions. Our hidden-force algorithm can be described as an intricate multiplayer tug-of-war game in which teams try to break an impasse by randomly assigning some players to drop their ropes while the others are still tugging until a partial impasse is reached, then, instructing the dropouts to resume tugging, for all teams to come to a new overall impasse. Utilizing our algorithm in a non-Markovian parallel Monte Carlo search, we found 17 new putative global minima for binary Lennard-Jones clusters in the size range of 90-100 particles. The method is efficient enough that an unbiased search was possible; no potential-energy surface symmetries were exploited. All new minima are comprised of three nested polyicosahedral or polytetrahedral shells when viewed as a nested set of Connolly surfaces (though the shell structure has previously gone unscrutinized, known minima are often qualitatively similar). Unlike known minima, in which the outer and inner shells are comprised of the larger and smaller atoms, respectively, in 13 of the new minima, the atoms are not as clearly separated by size. Furthermore, while some known minima have inner shells stabilized by larger atoms, four of the new minima have outer shells stabilized by smaller atoms.

  3. A test of the nest sanitation hypothesis for the evolution of foreign egg rejection in an avian brood parasite rejecter host species

    NASA Astrophysics Data System (ADS)

    Luro, Alec B.; Hauber, Mark E.

    2017-04-01

    Hosts of avian brood parasites have evolved diverse defenses to avoid the costs associated with raising brood parasite nestlings. In egg ejection, the host recognizes and removes foreign eggs laid in its nest. Nest sanitation, a behavior similar in motor pattern to egg ejection, has been proposed repeatedly as a potential pre-adaptation to egg ejection. Here, we separately placed blue 3D-printed, brown-headed cowbird ( Molothrus ater) eggs known to elicit interindividual variation in ejection responses and semi-natural leaves into American robins' ( Turdus migratorius) nests to test proximate predictions that (1) rejecter hosts should sanitize debris from nests more frequently and consistently than accepter hosts and (2) hosts that sanitize their nests of debris prior to the presentation of a foreign egg will be more likely to eject the foreign egg. Egg ejection responses were highly repeatable within individuals yet variable between them, but were not influenced by prior exposure to debris, nor related to sanitation tendencies as a whole, because nearly all individuals sanitized their nests. Additionally, we collected published data for eight different host species to test for a potential positive correlation between sanitation and egg ejection. We found no significant correlation between nest sanitation and egg ejection rates; however, our comparative analysis was limited to a sample size of 8, and we advise that more data from additional species are necessary to properly address interspecific tests of the pre-adaptation hypothesis. In lack of support for the nest sanitation hypothesis, our study suggests that, within individuals, foreign egg ejection is distinct from nest sanitation tendencies, and sanitation and foreign egg ejection may not correlate across species.

  4. A test of the nest sanitation hypothesis for the evolution of foreign egg rejection in an avian brood parasite rejecter host species.

    PubMed

    Luro, Alec B; Hauber, Mark E

    2017-04-01

    Hosts of avian brood parasites have evolved diverse defenses to avoid the costs associated with raising brood parasite nestlings. In egg ejection, the host recognizes and removes foreign eggs laid in its nest. Nest sanitation, a behavior similar in motor pattern to egg ejection, has been proposed repeatedly as a potential pre-adaptation to egg ejection. Here, we separately placed blue 3D-printed, brown-headed cowbird (Molothrus ater) eggs known to elicit interindividual variation in ejection responses and semi-natural leaves into American robins' (Turdus migratorius) nests to test proximate predictions that (1) rejecter hosts should sanitize debris from nests more frequently and consistently than accepter hosts and (2) hosts that sanitize their nests of debris prior to the presentation of a foreign egg will be more likely to eject the foreign egg. Egg ejection responses were highly repeatable within individuals yet variable between them, but were not influenced by prior exposure to debris, nor related to sanitation tendencies as a whole, because nearly all individuals sanitized their nests. Additionally, we collected published data for eight different host species to test for a potential positive correlation between sanitation and egg ejection. We found no significant correlation between nest sanitation and egg ejection rates; however, our comparative analysis was limited to a sample size of 8, and we advise that more data from additional species are necessary to properly address interspecific tests of the pre-adaptation hypothesis. In lack of support for the nest sanitation hypothesis, our study suggests that, within individuals, foreign egg ejection is distinct from nest sanitation tendencies, and sanitation and foreign egg ejection may not correlate across species.

  5. Pre- and post-experimental manipulation assessments confirm the increase in number of birds due to the addition of nest boxes

    PubMed Central

    Cuatianquiz Lima, Cecilia

    2016-01-01

    Secondary cavity nesting (SCN) birds breed in holes that they do not excavate themselves. This is possible where there are large trees whose size and age permit the digging of holes by primary excavators and only rarely happens in forest plantations, where we expected a deficit of both breeding holes and SCN species. We assessed whether the availability of tree cavities influenced the number of SCNs in two temperate forest types, and evaluated the change in number of SCNs after adding nest boxes. First, we counted all cavities within each of our 25-m radius sampling points in mature and young forest plots during 2009. We then added nest boxes at standardised locations during 2010 and 2011 and conducted fortnightly bird counts (January–October 2009–2011). In 2011 we added two extra plots of each forest type, where we also conducted bird counts. Prior to adding nest boxes, counts revealed more SCNs in mature than in young forest. Following the addition of nest boxes, the number of SCNs increased significantly in the points with nest boxes in both types of forest. Counts in 2011 confirmed the increase in number of birds due to the addition of nest boxes. Given the likely benefits associated with a richer bird community we propose that, as is routinely done in some countries, forest management programs preserve old tree stumps and add nest boxes to forest plantations in order to increase bird numbers and bird community diversity. PMID:26998410

  6. Diagnosis of neonatal group B Streptococcus sepsis by nested-PCR of residual urine samples

    PubMed Central

    Cezarino, Bruno Nicolino; Yamamoto, Lidia; Del Negro, Gilda Maria Barbaro; Rocha, Daisy; Okay, Thelma Suely

    2008-01-01

    Group B streptococcus (GBS) remains the most common cause of early-onset sepsis in newborns. Laboratory gold-standard, broth culture methods are highly specific, but lack sensitivity. The aim of this study was to validate a nested-PCR and to determine whether residue volumes of urine samples obtained by non invasive, non sterile methods could be used to confirm neonatal GBS sepsis. The nested-PCR was performed with primers of the major GBS surface antigen. Unavailability of biological samples to perform life supporting exams, as well as others to elucidate the etiology of infections is a frequent problem concerning newborn patients. Nevertheless, we decided to include cases according to strict criteria: newborns had to present with signs and symptoms compatible with GBS infection; at least one of the following biological samples had to be sent for culture: blood, urine, or cerebrospinal fluid; availability of residue volumes of the samples sent for cultures, or of others collected on the day of hospitalization, prior to antibiotic therapy prescription, to be analyzed by PCR; favorable outcome after GBS empiric treatment. In only one newborn GBS infection was confirmed by cultures, while infection was only presumptive in the other three patients (they fulfilled inclusion criteria but were GBS-culture negative). From a total of 12 biological samples (5 blood, 3 CSF and 4 urine specimen), eight were tested by culture methods (2/8 were positive), and 8 were tested by PCR (7/8 were positive), and only 4 samples were simultaneously tested by both methods (1 positive by culture and 3 by PCR). In conclusion, although based on a restricted number of neonates and samples, our results suggest that the proposed nested-PCR might be used to diagnose GBS sepsis as it has successfully amplified the three types of biological samples analyzed (blood, urine and cerebrospinal fluid), and was more sensitive than culture methods as PCR in urine confirmed diagnosis in all four patients. Moreover, PCR has enabled us to use residue volumes of urine samples collected by non invasive, non sterile methods, what is technically adequate as GBS is not part of the normal urine flora, thus avoiding invasive procedures such as suprapubic bladder punction or transurethral catheterization. At the same time, the use of urine instead of blood samples could help preventing newborns blood spoliation. PMID:24031170

  7. Food provisioning and parental status in songbirds: can occupancy models be used to estimate nesting performance?

    PubMed

    Corbani, Aude Catherine; Hachey, Marie-Hélène; Desrochers, André

    2014-01-01

    Indirect methods to estimate parental status, such as the observation of parental provisioning, have been problematic due to potential biases associated with imperfect detection. We developed a method to evaluate parental status based on a novel combination of parental provisioning observations and hierarchical modeling. In the summers of 2009 to 2011, we surveyed 393 sites, each on three to four consecutive days at Forêt Montmorency, Québec, Canada. We assessed parental status of 2331 adult songbirds based on parental food provisioning. To account for imperfect detection of parental status, we applied MacKenzie et al.'s (2002) two-state hierarchical model to obtain unbiased estimates of the proportion of sites with successfully nesting birds, and the proportion of adults with offspring. To obtain an independent evaluation of detection probability, we monitored 16 active nests in 2010 and conducted parental provisioning observations away from them. The probability of detecting food provisioning was 0.31 when using nest monitoring, a value within the 0.11 to 0.38 range that was estimated by two-state models. The proportion of adults or sites with broods approached 0.90 and varied depending on date during the sampling season and year, exemplifying the role of eastern boreal forests as highly productive nesting grounds for songbirds. This study offers a simple and effective sampling design for studying avian reproductive performance that could be implemented in national surveys such as breeding bird atlases.

  8. Food Provisioning and Parental Status in Songbirds: Can Occupancy Models Be Used to Estimate Nesting Performance?

    PubMed Central

    Corbani, Aude Catherine; Hachey, Marie-Hélène; Desrochers, André

    2014-01-01

    Indirect methods to estimate parental status, such as the observation of parental provisioning, have been problematic due to potential biases associated with imperfect detection. We developed a method to evaluate parental status based on a novel combination of parental provisioning observations and hierarchical modeling. In the summers of 2009 to 2011, we surveyed 393 sites, each on three to four consecutive days at Forêt Montmorency, Québec, Canada. We assessed parental status of 2331 adult songbirds based on parental food provisioning. To account for imperfect detection of parental status, we applied MacKenzie et al.'s (2002) two-state hierarchical model to obtain unbiased estimates of the proportion of sites with successfully nesting birds, and the proportion of adults with offspring. To obtain an independent evaluation of detection probability, we monitored 16 active nests in 2010 and conducted parental provisioning observations away from them. The probability of detecting food provisioning was 0.31 when using nest monitoring, a value within the 0.11 to 0.38 range that was estimated by two-state models. The proportion of adults or sites with broods approached 0.90 and varied depending on date during the sampling season and year, exemplifying the role of eastern boreal forests as highly productive nesting grounds for songbirds. This study offers a simple and effective sampling design for studying avian reproductive performance that could be implemented in national surveys such as breeding bird atlases. PMID:24999969

  9. Luminescence dating of anthropogenic features of the San Luis Valley, Colorado: from stone huts to stone walls

    USGS Publications Warehouse

    Mahan, Shannon; Donlan, Rebecca A.; Kardos, Barbara Maat

    2015-01-01

    The Snake Nest Wall site and the Crestone Stone Huts are in the northern San Luis Valley, Colorado, and provide a unique opportunity to date high-altitude archeological sites of unknown age and origin using optically stimulated luminescence (OSL). We sampled sediment underlying foundation stones of these structures to establish a chronological framework for each site's construction. OSL dating of the quartz grains directly under the Snake Nest Wall suggest that the stones and, therefore, the structure was most recently emplaced between 1855 and 1890 A.D. Dating of the sediment beneath the Crestone Stone Huts suggests the construction time of these huts is between 1860 and 1890 A.D. Analysis of the equivalent dose (DE) dispersion of the OSL samples at Snake Nest Wall and the Crestone Huts shows that the majority of sediments were fully bleached prior to deposition and the low scatter suggests that short-term or shallow alluvial processes were the dominant transport for sediments. In both cases, the OSL ages show that the construction was during very recent historical times, although it is likely that the Snake Nest Wall was rebuilt in the late 19th century. Further study is warranted at the Snake Nest Wall since it shows signs of greater antiquity and a continued presence of human use. The Crestone Huts are shown to be a product of railroad building during the boomtown days of Lucky and Crestone.

  10. Genetic structure of Florida green turtle rookeries as indicated by mitochondrial DNA control region sequences

    USGS Publications Warehouse

    Shamblin, Brian M.; Bagley, Dean A.; Ehrhart, Llewellyn M.; Desjardin, Nicole A.; Martin, R. Erik; Hart, Kristen M.; Naro-Maciel, Eugenia; Rusenko, Kirt; Stiner, John C.; Sobel, Debra; Johnson, Chris; Wilmers, Thomas; Wright, Laura J.; Nairn, Campbell J.

    2014-01-01

    Green turtle (Chelonia mydas) nesting has increased dramatically in Florida over the past two decades, ranking the Florida nesting aggregation among the largest in the Greater Caribbean region. Individual beaches that comprise several hundred kilometers of Florida’s east coast and Keys support tens to thousands of nests annually. These beaches encompass natural to highly developed habitats, and the degree of demographic partitioning among rookeries was previously unresolved. We characterized the genetic structure of ten Florida rookeries from Cape Canaveral to the Dry Tortugas through analysis of 817 base pair mitochondrial DNA (mtDNA) control region sequences from 485 nesting turtles. Two common haplotypes, CM-A1.1 and CM-A3.1, accounted for 87 % of samples, and the haplotype frequencies were strongly partitioned by latitude along Florida’s Atlantic coast. Most genetic structure occurred between rookeries on either side of an apparent genetic break in the vicinity of the St. Lucie Inlet that separates Hutchinson Island and Jupiter Island, representing the finest scale at which mtDNA structure has been documented in marine turtle rookeries. Florida and Caribbean scale analyses of population structure support recognition of at least two management units: central eastern Florida and southern Florida. More thorough sampling and deeper sequencing are necessary to better characterize connectivity among Florida green turtle rookeries as well as between the Florida nesting aggregation and others in the Greater Caribbean region.

  11. Clinical Usefulness of a One-Tube Nested Reverse Transcription Quantitative Polymerase Chain Reaction Assay for Evaluating Human Epidermal Growth Factor Receptor 2 mRNA Overexpression in Formalin-Fixed and Paraffin-Embedded Breast Cancer Tissue Samples.

    PubMed

    Wang, Hye-Young; Ahn, Sungwoo; Park, Sunyoung; Kim, SeungIl; Lee, Hyeyoung

    2017-01-01

    Currently, the two main methods used to analyze human epidermal growth factor receptor 2 (HER2) amplification or overexpression have a limited accuracy and high costs. These limitations can be overcome by the development of complementary quantitative methods. In this study, we analyzed HER2 mRNA expression in clinical formalin-fixed and paraffin-embedded (FFPE) samples using a one-tube nested reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay. We measured expression relative to 3 reference genes and compared the results to those obtained by conventional immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) assays with 226 FFPE breast cancer tissue samples. The one-tube nested RT-qPCR assay proved to be highly sensitive and specific based on comparisons with IHC (96.9 and 97.7%, respectively) and FISH (92.4 and 92.9%, respectively) obtained with the validation set. Comparisons with clinicopathological data revealed significant associations between HER2 overexpression and TNM stage (p < 0.01), histological type (p < 0.01), ER status (p < 0.001), PR status (p < 0.05), HER2 status (p < 0.001), and molecular subtypes (p < 0.001). Based on these findings, our one-tube nested RT-qPCR assay is a potentially useful and complementary screening tool for the detection of HER2 mRNA overexpression. © 2016 S. Karger AG, Basel.

  12. Functional Additive Mixed Models

    PubMed Central

    Scheipl, Fabian; Staicu, Ana-Maria; Greven, Sonja

    2014-01-01

    We propose an extensive framework for additive regression models for correlated functional responses, allowing for multiple partially nested or crossed functional random effects with flexible correlation structures for, e.g., spatial, temporal, or longitudinal functional data. Additionally, our framework includes linear and nonlinear effects of functional and scalar covariates that may vary smoothly over the index of the functional response. It accommodates densely or sparsely observed functional responses and predictors which may be observed with additional error and includes both spline-based and functional principal component-based terms. Estimation and inference in this framework is based on standard additive mixed models, allowing us to take advantage of established methods and robust, flexible algorithms. We provide easy-to-use open source software in the pffr() function for the R-package refund. Simulations show that the proposed method recovers relevant effects reliably, handles small sample sizes well and also scales to larger data sets. Applications with spatially and longitudinally observed functional data demonstrate the flexibility in modeling and interpretability of results of our approach. PMID:26347592

  13. Evaluation of ERTS multispectral signatures in relation to ground control signatures using a nested-sampling approach

    NASA Technical Reports Server (NTRS)

    Lyon, R. J. P. (Principal Investigator)

    1975-01-01

    The author has identified the following significant results. Ground measured spectral signatures of wavelength bands matching ERTS MSS were collected using a radiometer at several Californian and Nevadan sites, and directly compared with similar data from ERTS CCTs. The comparison was tested at the highest possible spatial resolution for ERTS, using deconvoluted MSS data, and contrasted with that of ground measured spectra, originally from 1 meter squares. In the mobile traverses of the grassland sites, these one meter fields of view were integrated into eighty meter transects along the five km track across four major rock/soil types. Suitable software was developed to read the MSS CCT tapes, to shadeprint individual bands with user-determined greyscale stretching. Four new algorithms for unsupervised and supervised, normalized and unnormalized clustering were developed, into a program termed STANSORT. Parallel software allowed the field data to be calibrated, and by using concurrently continuously collected, upward- and downward-viewing, 4 band radiometers, bidirectional reflectances could be calculated.

  14. Functional Additive Mixed Models.

    PubMed

    Scheipl, Fabian; Staicu, Ana-Maria; Greven, Sonja

    2015-04-01

    We propose an extensive framework for additive regression models for correlated functional responses, allowing for multiple partially nested or crossed functional random effects with flexible correlation structures for, e.g., spatial, temporal, or longitudinal functional data. Additionally, our framework includes linear and nonlinear effects of functional and scalar covariates that may vary smoothly over the index of the functional response. It accommodates densely or sparsely observed functional responses and predictors which may be observed with additional error and includes both spline-based and functional principal component-based terms. Estimation and inference in this framework is based on standard additive mixed models, allowing us to take advantage of established methods and robust, flexible algorithms. We provide easy-to-use open source software in the pffr() function for the R-package refund. Simulations show that the proposed method recovers relevant effects reliably, handles small sample sizes well and also scales to larger data sets. Applications with spatially and longitudinally observed functional data demonstrate the flexibility in modeling and interpretability of results of our approach.

  15. Contribution of ants in modifying of soil acidity and particle size distribution

    NASA Astrophysics Data System (ADS)

    Morgun, Alexandra; Golichenkov, Maxim

    2015-04-01

    Being a natural body, formed by the influence of biota on the upper layers of the Earth's crust, the soil is the most striking example of biogenic-abiogenic interactions in the biosphere. Invertebrates (especially ants that build soil nests) are important agents that change soil properties in well developed terrestrial ecosystems. Impact of soil microorganisms on soil properties is particularly described in numerous literature and concerns mainly chemical properties and general indicators of soil biological activity. Influence of ants (as representatives of the soil mesofauna) mostly appears as mechanical movement of soil particles and aggregates, and chemical effects caused by concentration of organic matter within the ant's nest. The aim of this research was to evaluate the effect of ants on physical and chemical soil attributes such as particle size distribution and soil acidity. The samples were taken from aerial parts of Lasius niger nests, selected on different elements of the relief (summit position, slope, terrace and floodplain) in the Arkhangelsk region (north of the European part of Russia) and compared with the specimens of the upper horizons of the reference soils. Particle size distribution was determined by laser diffraction method using laser diffraction particle size analyzer «Analysette 22 comfort» (FRITSCH, Germany). The acidity (pH) was determined by potentiometry in water suspension. Particle size distribution of the samples from the nests is more variable as compared to the control samples. For example, the content of 5-10 μm fraction ranges from 9% to 12% in reference soils, while in the anthill samples the variation is from 8% to 15%. Similarly, for 50-250 μm fraction - it ranges from 15% to 18% in reference soils, whereas in anthills - from 6% to 29%. The results of particle size analysis showed that the reference sample on the terrace has silty loam texture and nests soil L. niger are medium loam. The reference soil on the slope is characterized as medium loam, and ant's nest material has silty loam texture. The control samples of soil and ants nests on the summit position are similar and have medium loam texture. Generally we outline that the particle size distribution of anthill samples shows more variability. We assume that ants operate with small soil aggregates, in which fine fractions may link together coarser particles. pH measurements show that the reference soils have a strongly acidic reaction on the summit position (pH 4.6), slightly acidic on the slope (pH 5.5) and neutral on the terrace and on the floodplain (pH 7.2). While the material of the anthills tends to be slightly alkalinized on the summit (pH 4.8) and alkalinized on the slope (pH 7.2), but acidified to neutral on the floodplain and terrace (pH 6.4 and 5.7). Therefore, the ants form specific physico-chemical conditions that are different from the surrounding (native) soil, significantly increasing the complexity of soil cover structure. This is a clear example of ecosystem engineering functions of ants in nature. Increased complexity of soil pattern is the result of variations in pH and particle size distribution. Both cause the preconditions for the formation of new environmental niches and enhance biodiversity in natural ecosystems.

  16. Data preprocessing for determining outer/inner parallelization in the nested loop problem using OpenMP

    NASA Astrophysics Data System (ADS)

    Handhika, T.; Bustamam, A.; Ernastuti, Kerami, D.

    2017-07-01

    Multi-thread programming using OpenMP on the shared-memory architecture with hyperthreading technology allows the resource to be accessed by multiple processors simultaneously. Each processor can execute more than one thread for a certain period of time. However, its speedup depends on the ability of the processor to execute threads in limited quantities, especially the sequential algorithm which contains a nested loop. The number of the outer loop iterations is greater than the maximum number of threads that can be executed by a processor. The thread distribution technique that had been found previously only be applied by the high-level programmer. This paper generates a parallelization procedure for low-level programmer in dealing with 2-level nested loop problems with the maximum number of threads that can be executed by a processor is smaller than the number of the outer loop iterations. Data preprocessing which is related to the number of the outer loop and the inner loop iterations, the computational time required to execute each iteration and the maximum number of threads that can be executed by a processor are used as a strategy to determine which parallel region that will produce optimal speedup.

  17. Bayesian data analysis tools for atomic physics

    NASA Astrophysics Data System (ADS)

    Trassinelli, Martino

    2017-10-01

    We present an introduction to some concepts of Bayesian data analysis in the context of atomic physics. Starting from basic rules of probability, we present the Bayes' theorem and its applications. In particular we discuss about how to calculate simple and joint probability distributions and the Bayesian evidence, a model dependent quantity that allows to assign probabilities to different hypotheses from the analysis of a same data set. To give some practical examples, these methods are applied to two concrete cases. In the first example, the presence or not of a satellite line in an atomic spectrum is investigated. In the second example, we determine the most probable model among a set of possible profiles from the analysis of a statistically poor spectrum. We show also how to calculate the probability distribution of the main spectral component without having to determine uniquely the spectrum modeling. For these two studies, we implement the program Nested_fit to calculate the different probability distributions and other related quantities. Nested_fit is a Fortran90/Python code developed during the last years for analysis of atomic spectra. As indicated by the name, it is based on the nested algorithm, which is presented in details together with the program itself.

  18. Solving very large, sparse linear systems on mesh-connected parallel computers

    NASA Technical Reports Server (NTRS)

    Opsahl, Torstein; Reif, John

    1987-01-01

    The implementation of Pan and Reif's Parallel Nested Dissection (PND) algorithm on mesh connected parallel computers is described. This is the first known algorithm that allows very large, sparse linear systems of equations to be solved efficiently in polylog time using a small number of processors. How the processor bound of PND can be matched to the number of processors available on a given parallel computer by slowing down the algorithm by constant factors is described. Also, for the important class of problems where G(A) is a grid graph, a unique memory mapping that reduces the inter-processor communication requirements of PND to those that can be executed on mesh connected parallel machines is detailed. A description of an implementation on the Goodyear Massively Parallel Processor (MPP), located at Goddard is given. Also, a detailed discussion of data mappings and performance issues is given.

  19. Flag-based detection of weak gas signatures in long-wave infrared hyperspectral image sequences

    NASA Astrophysics Data System (ADS)

    Marrinan, Timothy; Beveridge, J. Ross; Draper, Bruce; Kirby, Michael; Peterson, Chris

    2016-05-01

    We present a flag manifold based method for detecting chemical plumes in long-wave infrared hyperspectral movies. The method encodes temporal and spatial information related to a hyperspectral pixel into a flag, or nested sequence of linear subspaces. The technique used to create the flags pushes information about the background clutter, ambient conditions, and potential chemical agents into the leading elements of the flags. Exploiting this temporal information allows for a detection algorithm that is sensitive to the presence of weak signals. This method is compared to existing techniques qualitatively on real data and quantitatively on synthetic data to show that the flag-based algorithm consistently performs better on data when the SINRdB is low, and beats the ACE and MF algorithms in probability of detection for low probabilities of false alarm even when the SINRdB is high.

  20. Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems.

    PubMed

    Ximenes, Camila; Brandão, Eduardo; Oliveira, Paula; Rocha, Abraham; Rego, Tamisa; Medeiros, Rafael; Aguiar-Santos, Ana; Ferraz, João; Reis, Christian; Araujo, Paulo; Carvalho, Luiz; Melo, Fabio L

    2014-12-01

    The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

  1. Serological and molecular diagnosis of Toxoplasma gondii in patients with schizophrenia.

    PubMed

    Ebrahimzadeh, Adel; Shahraki, Mehdi Khoshsima; Mohammadi, Azad

    2018-06-01

    Schizophrenia is a persistent neuropsychiatric syndrome of uncertain source. Toxoplasmosis is the most prevalent parasitic protozoan infecting one-third of the worldwide human population. Infectious agents such as toxoplasma are the probable cause of schizophrenia. This study was aimed to evaluate the association between schizophrenia and toxoplasmosis using SAG1 and B1 Target gene. During February to December 2016, 92 patients with schizophrenia are imported in our study. All cases were assessed by serological (IgG and IgM antibodies) and molecular examinations. ELISA was performed by Commercial kits according to manufactures procedure. DNA was extracted and nested PCR was done using two pairs of primers. From 92 patients, 59 (64.13%) cases were positive for toxoplasmosis by serological examinations (14 samples positive for IgM and IgG, 40 samples positive for only IgG and 5 samples Positive for only IgM) and 58 (63.04%) were positive by Nested PCR technique. Based on the nested PCR method, 68.47 and 47.82% of samples were positive by B1 and SAG1 genes, respectively. Our results showed the importance of use both serological and molecular diagnostic methods for accurate recognition of T . gondii in patients with schizophrenia. Moreover our results indicated that B1 gene is more sensitive than SAG1 gene.

  2. Drought-caused delay in nesting of Sonoran Desert birds and its facilitation of parasite- and predator-mediated variation in reproductive success

    USGS Publications Warehouse

    McCreedy, Chris; van Riper, Charles

    2015-01-01

    As our understanding of climate change has increased, so has our awareness of the impacts of these changes on biotic systems. Climate models are nearly unanimous in their predictions for increased drought frequency in southwestern North America, and delays in nest initiation due to drought may influence nesting success and productivity for many Sonoran Desert bird species. In southeastern California and western Arizona in 2004–2009, we found negative correlations for 13 of 13 species between nest initiation date and rainfall accumulation during the preceding 4-month winter rainy season. Nesting was delayed more than 3 weeks for some species during extreme droughts in 2006 and 2007. During 2004–2009, we found a significant negative effect of nest initiation date on nest survival probability (β̂ = −0.031 ± 0.005 SE, P < 0.001) for the four species of greatest sample size. To investigate the role of nesting delay in nesting success and productivity, in 2010 we conducted a manipulative experiment with Black-tailed Gnatcatchers (Polioptila melanura; BTGN) and Verdins (Auriparus flaviceps; VERD). Following a wet winter, we delayed clutch initiation dates for treatment pairs to match first-egg dates that we observed during droughts in 2006 and 2007. Nest initiation date had a significant negative effect on nest survival of both species (BTGN: β̂ = −1.18 ± 0.27 SE, P < 0.001; VERD: β̂ = −2.33 ± 0.51 SE, P = 0.003). Treatment pairs were unable to overcome the lost period of high productivity in March and early April, and had lower productivity than control pairs over the entire breeding season. As nest predation and Brown-headed Cowbird (Molothrus ater) parasitism were the most common causes of nest failure, we conclude that the impacts of climate change–caused drought on annual reproductive output in the Sonoran Desert will be further compounded by parasitism and predation for Black-tailed Gnatcatchers and by predation for Verdins.

  3. Inter-assemblage facilitation: the functional diversity of cavity-producing beetles drives the size diversity of cavity-nesting bees.

    PubMed

    Sydenham, Markus A K; Häusler, Lise D; Moe, Stein R; Eldegard, Katrine

    2016-01-01

    Inter-specific interactions are important drivers and maintainers of biodiversity. Compared to trophic and competitive interactions, the role of non-trophic facilitation among species has received less attention. Cavity-nesting bees nest in old beetle borings in dead wood, with restricted diameters corresponding to the body size of the bee species. The aim of this study was to test the hypothesis that the functional diversity of cavity-producing wood boring beetles - in terms of cavity diameters - drives the size diversity of cavity-nesting bees. The invertebrate communities were sampled in 30 sites, located in forested landscapes along an elevational gradient. We regressed the species richness and abundance of cavity nesting bees against the species richness and abundance of wood boring beetles, non-wood boring beetles and elevation. The proportion of cavity nesting bees in bee species assemblage was regressed against the species richness and abundance of wood boring beetles. We also tested the relationships between the size diversity of cavity nesting bees and wood boring beetles. The species richness and abundance of cavity nesting bees increased with the species richness and abundance of wood boring beetles. No such relationship was found for non-wood boring beetles. The abundance of wood boring beetles was also related to an increased proportion of cavity nesting bee individuals. Moreover, the size diversity of cavity-nesting bees increased with the functional diversity of wood boring beetles. Specifically, the mean and dispersion of bee body sizes increased with the functional dispersion of large wood boring beetles. The positive relationships between cavity producing bees and cavity nesting bees suggest that non-trophic facilitative interactions between species assemblages play important roles in organizing bee species assemblages. Considering a community-wide approach may therefore be required if we are to successfully understand and conserve wild bee species assemblages in forested landscapes.

  4. Towards Artificial Speech Therapy: A Neural System for Impaired Speech Segmentation.

    PubMed

    Iliya, Sunday; Neri, Ferrante

    2016-09-01

    This paper presents a neural system-based technique for segmenting short impaired speech utterances into silent, unvoiced, and voiced sections. Moreover, the proposed technique identifies those points of the (voiced) speech where the spectrum becomes steady. The resulting technique thus aims at detecting that limited section of the speech which contains the information about the potential impairment of the speech. This section is of interest to the speech therapist as it corresponds to the possibly incorrect movements of speech organs (lower lip and tongue with respect to the vocal tract). Two segmentation models to detect and identify the various sections of the disordered (impaired) speech signals have been developed and compared. The first makes use of a combination of four artificial neural networks. The second is based on a support vector machine (SVM). The SVM has been trained by means of an ad hoc nested algorithm whose outer layer is a metaheuristic while the inner layer is a convex optimization algorithm. Several metaheuristics have been tested and compared leading to the conclusion that some variants of the compact differential evolution (CDE) algorithm appears to be well-suited to address this problem. Numerical results show that the SVM model with a radial basis function is capable of effective detection of the portion of speech that is of interest to a therapist. The best performance has been achieved when the system is trained by the nested algorithm whose outer layer is hybrid-population-based/CDE. A population-based approach displays the best performance for the isolation of silence/noise sections, and the detection of unvoiced sections. On the other hand, a compact approach appears to be clearly well-suited to detect the beginning of the steady state of the voiced signal. Both the proposed segmentation models display outperformed two modern segmentation techniques based on Gaussian mixture model and deep learning.

  5. Comparison between single PCR and nested PCR in detection of human papilloma viruses in paraffin-embedded OSCC and fresh oral mucosa.

    PubMed

    Jalouli, Miranda; Jalouli, Jamshid; Ibrahim, Salah O; Hirsch, Jan-Michaél; Sand, Lars

    2015-01-01

    Infection with human papilloma virus (HPV) has been implicated as one of the risk factors for the development of oropharyngeal cancer. Many different HPV tests exist, and information regarding their specific technical, analytical, and clinical properties is increasing. This study aimed to compare the level of detection of HPV using two reliable polymerase chain reaction (PCR) methods, nested PCR (NPCR) and single PCR (SPCR), in archival paraffin-embedded oral squamous cell carcinoma (OSCC) samples and fresh oral mucosa specimens. The presence of HPV genome in two groups of tissue samples was analyzed: (i) 57 paraffin-embedded OSCC samples from Sudan and (ii) eight healthy fresh oral mucosal samples from Swedish volunteers. The specimens were tested by SPCR with primer pair MY9/MY11 and NPCR using GP5+/GP6+ primer sets. Eighteen (32%) out of the 57 paraffin-embedded OSCC samples, and five (62%) out of the eight fresh clinically healthy samples were found to be HPV-positive with NPCR. With SPCR, four (7%) out of the paraffin-embedded OSCC samples were HPV-positive. A statistically significant difference between HPV-positive and -negative samples was found when comparing NPCR and SPCR in OSCC and fresh oral mucosa (p<0.0001). The comparative test between SPCR and NPCR showed 100% sensitivity and 69% specificity for OSCC. The use of the GP5+/GP6+ nested PCR increased the positivity rate, efficiency rate and sensitivity of HPV detection in oral samples significantly and should be considered as the method of choice. Copyright © 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  6. Area and Family Effects on the Psychopathology of the Millennium Cohort Study Children and Their Older Siblings

    ERIC Educational Resources Information Center

    Flouri, Eirini; Tzavidis, Nikos; Kallis, Constantinos

    2010-01-01

    Background: To model and compare contextual (area and family) effects on the psychopathology of children nested in families nested in areas. Method: Data from the first two sweeps of the UK's Millennium Cohort Study were used. The final study sample was 9,630 children clustered in 6,052 families clustered in 1,681 Lower-layer Super Output Areas.…

  7. Detection of Pneumocystis jirovecii dihydropteroate synthase polymorphisms in patients with Pneumocystis pneumonia.

    PubMed

    Costa, M C; Gaspar, J; Mansinho, K; Esteves, F; Antunes, F; Matos, O

    2005-01-01

    In the present study, in order to improve the detection of Pneumocystis jirovecii dihydropteroate synthase (DHPS) mutations in pulmonary specimens of HIV-infected patients with P. jirovecii pneumonia, we evaluated a microfiltration procedure for the removal of human cell contamination and a nested-PCR method, for amplification in specimens with low parasite load. In the studied population, PCR amplification of the DHPS gene was more successful in unfiltered than in filtered specimens, with both touchdown-PCR and nested-PCR procedures (p<0.05 and p<0.001, respectively), but the amount of host DNA in the samples analysed seems to be inversely related with the successful PCR parasite detection. Amplification of P. jirovecii DHPS gene with nested-PCR was achieved in 77.5% of the specimens studied, demonstrating that this is a useful method for the identification of mutations in pulmonary specimens, including samples with low parasite loads, and will facilitate the evaluation of the relationship between the P. jirovecii DHPS polymorphisms and clinical resistance to sulfa drugs.

  8. Evidence of simian retrovirus type D by polymerase chain reaction.

    PubMed

    Hwa, Christian Z R; Tsai, Sheung Pun; Yee, JoAnn L; Van Rompay, Koen K; Roberts, Jeffrey A

    2017-06-01

    Over the past few years, there have been reports of finding Simian retrovirus type D (SRV) in macaque colonies where some animals were characterized as antibody positive but virus negative raising questions about how SRV was transmitted or whether there is a variant strain detected by antibody but not polymerase chain reaction (PCR) in current use. We developed a three-round nested PCR assay using degenerate primers targeting the pol gene to detect for SRV serotypes 1-5 and applied this newly validated PCR assay to test macaque DNA samples collected in China from 2010 to 2015. Using the nested PCR assay validated in this study, we found 0.15% of the samples archived on FTA ® cards were positive. The source of SRV infection identified within domestic colonies might have originated from imported macaques. The multiplex nested PCR assay developed here may supplement the current assays for SRV. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Food habits of the southwestern willow flycatcher during the nesting season

    USGS Publications Warehouse

    Drost, Charles A.; Paxton, Eben H.; Sogge, Mark K.; Whitfield, Mary J.

    2003-01-01

    The food habits and prey base of the endangered Southwestern Willow Flycatcher (Empidonax traillii extimus) are not well known. We analyzed prey remains in 59 fecal samples from an intensively-studied population of this flycatcher at the Kern River Preserve in southern California. These samples were collected during the nesting season in 1996 and 1997 from adults caught in mist nets, and from nestlings temporarily removed from the nest for banding. A total of 379 prey individuals were identified in the samples. Dominant prey taxa, both in total numbers and in percent occurrence, were true bugs (Hemiptera), flies (Diptera), and beetles (Coleoptera). Leafhoppers (Homoptera: Cicadellidae), spiders (Araneae), bees and wasps (Hymenoptera), and dragonflies and damselflies (Odonata) were also common items. Diet composition was significantly different between years, due to a large difference in the numbers of spiders between 1996 and 1997. There was also a significant difference between the diet of young and adults, with the diet of young birds having significantly higher numbers of odonates and beetles. There was a trend toward diet differences between males and females, but this was not significant at the P = 0.05 level.

  10. Characterisation of microbial communities in Chinese liquor fermentation starters Daqu using nested PCR-DGGE.

    PubMed

    Zhang, Liqiang; Wu, Chongde; Ding, Xiaofei; Zheng, Jia; Zhou, Rongqing

    2014-12-01

    In this study, characterises of the microbial community structures of three typical Chinese liquor Daqu, as well as different kinds of light flavour Daqu were investigated using nested PCR-denaturing gradient gel electrophoresis (DGGE). The results showed that microbial diversity was considerably different, and the microfloral compositions were highly variable among various Daqu. Lactic acid bacteria, which accounted for 30.95 % of all identified bacteria, were dominant in all Daqu samples, whereas Bacillus species were also predominant in the Luzhou (14.8 %) and Langjiu Daqu (18.2 %). Citrobacter and Burkholderia were first identified in light flavour Daqu. Aspergillus was the dominant moulds, and the non-Saccharomyces yeast species, Saccharomycopsis fibuligera, Wallemia sebi, Wallemia muriae, and Pichia subpelliculosa, were the dominant yeasts. Rasamsonia, Galactomyces, Geotrichum and Wallemia were first identified using nested PCR-DGGE. Cluster analysis indicated that the microbial community structures of different Daqu samples exhibited some differences. These may be ascribed to the different peak production temperatures, raw material constituents and microhabitats around the liquor enterprises. The current study provides insights into the microbial community structures of three typical Daqu samples, and may facilitate the development of starter cultures for manufacturing Chinese liquor.

  11. Contextualising the Last Survivors: Population Structure of Marine Turtles in the Dominican Republic.

    PubMed

    Carreras, Carlos; Godley, Brendan J; León, Yolanda M; Hawkes, Lucy A; Revuelta, Ohiana; Raga, Juan A; Tomás, Jesús

    2013-01-01

    Nesting by three species of marine turtles persists in the Dominican Republic, despite historic threats and long-term population decline. We conducted a genetic survey of marine turtles in the Dominican Republic in order to link them with other rookeries around the Caribbean. We sequenced a 740bp fragment of the control region of the mitochondrial DNA of 92 samples from three marine turtle species [hawksbill (n = 48), green (n = 2) and leatherback (n = 42)], and incorporated published data from other nesting populations and foraging grounds. The leatherback turtle (Dermochelys coriacea) in the Dominican Republic appeared to be isolated from Awala-Yalimapo, Cayenne, Trinidad and St. Croix but connected with other Caribbean populations. Two distinct nesting populations of hawksbill turtles (Eremochelys imbricata) were detected in the Dominican Republic and exhibited interesting patterns of connectivity with other nesting sites and juvenile and adult male foraging aggregations. The green sea turtle (Chelonia mydas) has almost been extirpated from the Dominican Republic and limited inference could be made from our samples. Finally, results were compared with Lagrangian drifting buoys and published Lagrangian virtual particles that travelled through the Dominican Republic and Caribbean waters. Conservation implications of sink-source effects or genetic isolation derived from these complex inter-connections are discussed for each species and population.

  12. Contextualising the Last Survivors: Population Structure of Marine Turtles in the Dominican Republic

    PubMed Central

    Carreras, Carlos; Godley, Brendan J.; León, Yolanda M.; Hawkes, Lucy A.; Revuelta, Ohiana; Raga, Juan A.; Tomás, Jesús

    2013-01-01

    Nesting by three species of marine turtles persists in the Dominican Republic, despite historic threats and long-term population decline. We conducted a genetic survey of marine turtles in the Dominican Republic in order to link them with other rookeries around the Caribbean. We sequenced a 740bp fragment of the control region of the mitochondrial DNA of 92 samples from three marine turtle species [hawksbill (n = 48), green (n = 2) and leatherback (n = 42)], and incorporated published data from other nesting populations and foraging grounds. The leatherback turtle (Dermochelys coriacea) in the Dominican Republic appeared to be isolated from Awala-Yalimapo, Cayenne, Trinidad and St. Croix but connected with other Caribbean populations. Two distinct nesting populations of hawksbill turtles (Eremochelys imbricata) were detected in the Dominican Republic and exhibited interesting patterns of connectivity with other nesting sites and juvenile and adult male foraging aggregations. The green sea turtle (Chelonia mydas) has almost been extirpated from the Dominican Republic and limited inference could be made from our samples. Finally, results were compared with Lagrangian drifting buoys and published Lagrangian virtual particles that travelled through the Dominican Republic and Caribbean waters. Conservation implications of sink-source effects or genetic isolation derived from these complex inter-connections are discussed for each species and population. PMID:23840394

  13. Temporal flexibility of reproduction in temperate-breeding dabbling ducks

    USGS Publications Warehouse

    Krapu, Gary L.

    2000-01-01

    I compared nesting intervals during three consecutive years in five species of temperate-nesting dabbling ducks (Mallard [Anas platyrhynchos], Northern Pintail [Anas acuta], Northern Shoveler [Anas clypeata], Blue-winged Teal [Anas discors], Gadwall [Anas strepera]) and assessed whether differences existed in timing of refractoriness. Most nesting by females of all five species ended by the summer solstice. Nesting ended earliest for Northern Shovelers and Northern Pintails and latest for Gadwalls. Some Mallards, Blue-winged Teal, and Gadwalls continued to nest into mid- and late summer, whereas Northern Shovelers and Northern pintails did not. Mallards, Blue-winged Teal, and Gadwalls accounted for 99% (81 of 82) of flightless broods resulting from nests initiated during mid- or late summer in North Dakota and 98% (58 of 59) of flightless juveniles shot on or after 1 October by a random sample of duck hunters from across the United States. Early cessation of breeding by Northern Shovelers may have evolved in response to the species' limited flexibility in diet. Photorefractory mechanisms that limit most breeding to spring presumably evolved in response to severe constraints on reproductive success when nesting continued through summer(e.g. mortality of late-hatched young and molting females due to low temperatures). Interspecific differences in photosensitivity may account for variation in timing of cessation of nesting in late spring, but controlled experiments are needed to assess the possible role of non-photic influences. My results suggest that the refractory mechanisms controlling length of the breeding in temperate-nesting dabbling ducks are more varied and complex than previously thought, with non-photic influences (e.g. water conditions, food availability, food quality) having a larger role than indicated by earlier research.

  14. Mixed-stock analysis reveals the migrations of juvenile hawksbill turtles (Eretmochelys imbricata) in the Caribbean Sea.

    PubMed

    Bowen, B W; Grant, W S; Hillis-Starr, Z; Shaver, D J; Bjorndal, K A; Bolten, A B; Bass, A L

    2007-01-01

    Hawksbill turtles (Eretmochelys imbricata) migrate between nesting beaches and feeding habitats that are often associated with tropical reefs, but it is uncertain which nesting colonies supply which feeding habitats. To address this gap in hawksbill biology, we compile previously published and new mitochondrial DNA (mtDNA) haplotype data for 10 nesting colonies (N = 347) in the western Atlantic and compare these profiles to four feeding populations and four previously published feeding samples (N = 626). Nesting colonies differ significantly in mtDNA haplotype frequencies (Phi(ST) = 0.588, P < 0.001), corroborating earlier conclusions of nesting site fidelity and setting the stage for mixed-stock analysis. Feeding aggregations show lower but significant structure (Phi(ST) = 0.089, P < 0.001), indicating that foraging populations are not homogenous across the Caribbean Sea. Bayesian mixed-stock estimates of the origins of juveniles in foraging areas show a highly significant, but shallow, correlation with nesting population size (r = 0.378, P = 0.004), supporting the premise that larger rookeries contribute more juveniles to feeding areas. A significant correlation between the estimated contribution and geographical distance from nesting areas (r = -0.394, P = 0.003) demonstrates the influence of proximity on recruitment to feeding areas. The influence of oceanic currents is illustrated by pelagic stage juveniles stranded in Texas, which are assigned primarily (93%) to the upstream rookery in Yucatan. One juvenile had a haplotype previously identified only in the eastern Atlantic, invoking rare trans-oceanic migrations. The mixed-stock analysis demonstrates that harvests in feeding habitats will impact nesting colonies throughout the region, with the greatest detriment to nearby nesting populations.

  15. Relationships of environmental contaminants to reproductive success in red-breasted mergansers (Mergus serrator) from Lake Michigan

    USGS Publications Warehouse

    Heinz, G.H.; Haseltine, S.D.; Reichel, W.L.; Hensler, G.L.

    1983-01-01

    In 1977 and 1978, we studied red-breasted mergansers Mergus serrator nesting on islands in northwestern Lake Michigan to determine whether environmental contaminants were having effects on reproduction. Seventeen contaminants were measured in randomly chosen eggs from 206 nests under study. Using a variety of statistical approaches, we looked for effects of individual contaminants and combinations of contaminants on reproductive measurements such as nest desertion, failure of eggs to hatch, death of newly hatched ducklings, percentage hatching success, number of ducklings leaving the nest and eggshell thickness. We also looked for relationships between the levels of some contaminants in blood samples of 39 incubating females and reproductive success. A small degree of eggshell thinning was attributed to DDE and a few other statistical tests were significant, but no contaminant or combination of contaminants we measured seemed to have a pronounced effect on the aspects of reproduction we followed.

  16. Localized Fault Recovery for Nested Fork-Join Programs

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kestor, Gokcen; Krishnamoorthy, Sriram; Ma, Wenjing

    Nested fork-join programs scheduled using work stealing can automatically balance load and adapt to changes in the execution environment. In this paper, we design an approach to efficiently recover from faults encountered by these programs. Specifically, we focus on localized recovery of the task space in the presence of fail-stop failures. We present an approach to efficiently track, under work stealing, the relationships between the work executed by various threads. This information is used to identify and schedule the tasks to be re-executed without interfering with normal task execution. The algorithm precisely computes the work lost, incurs minimal re-execution overhead,more » and can recover from an arbitrary number of failures. Experimental evaluation demonstrates low overheads in the absence of failures, recovery overheads on the same order as the lost work, and much lower recovery costs than alternative strategies.« less

  17. Multi-Generational Kinship, Multiple Mating, and Flexible Modes of Parental Care in a Breeding Population of the Veery (Catharus fuscescens), a Trans-Hemispheric Migratory Songbird

    PubMed Central

    Kalavacharla, Venugopal

    2016-01-01

    We discovered variable modes of parental care in a breeding population of color-banded Veeries (Catharus fuscescens), a Nearctic-Neotropical migratory songbird, long thought to be socially monogamous, and performed a multi-locus DNA microsatellite analysis to estimate parentage and kinship in a sample of 37 adults and 21 offspring. We detected multiple mating in both sexes, and four modes of parental care that varied in frequency within and between years including multiple male feeders at some nests, and males attending multiple nests in the same season, each with a different female. Unlike other polygynandrous systems, genetic evidence indicates that multi-generational patterns of kinship occur among adult Veeries at our study site, and this was corroborated by the capture of an adult male in 2013 that had been banded as a nestling in 2011 at a nest attended by multiple male feeders. All genotyped adults (n = 37) were related to at least one other bird in the sample at the cousin level or greater (r ≥ 0.125), and 81% were related to at least one other bird at the half-sibling level or greater (r ≥ 0.25, range 0.25–0.60). Although our sample size is small, it appears that the kin structure is maintained by natal philopatry in both sexes, and that Veeries avoid mating with close genetic kin. At nests where all adult feeders were genotyped (n = 9), the male(s) were unrelated to the female (mean r = -0.11 ± 0.15), whereas genetic data suggest close kinship (r = 0.254) between two male co-feeders at the nests of two females in 2011, and among three of four females that were mated to the same polygynous male in 2012. To our knowledge, this is the first evidence of polygynandry occurring among multiple generations of close genetic kin on the breeding ground of a Nearctic-Neotropical migratory songbird. PMID:27331399

  18. Natural cavities used by wood ducks in north-central Minnesota

    USGS Publications Warehouse

    Gilmer, D.S.; Ball, I.J.; Cowardin, L.M.; Mathisen, J.

    1978-01-01

    Radio telemetry was used to locate 31 wood duck (Aix sponsa) nest cavity sites in 16 forest stands. Stands were of 2 types: (1) mature (mean = 107 years) northern hardwoods (10 nest sites), and (2) mature (mean = 68 years) quaking aspen (Populus tremuloides) (21 nest sites). Aspen was the most important cavity-producing tree used by wood ducks and accounted for 57 percent of 28 cavities inspected. In stands used by wood ducks, the average density of suitable cavities was about 4 per hectare. Trees containing nests were closer to water areas (P < 0.05) and the nearest forest canopy openings (P < 0.01) than was a random sample of trees from the same stands. A significant (P < 0.005) relationship existed between the orientation of the cavity entrance and the nearest canopy opening. Potential wood duck cavities usually were clustered within a stand rather than randomly distributed. Selection of trees by woodpeckers for nest hole construction probably influenced the availability of cavities used by wood ducks. A plan for managing forests to benefit wood ducks and other wildlife dependent on old-growth timber is discussed.

  19. Compilation of 1990 annual reports of the Navy ELF communications system ecological monitoring program. Volume 2: Tabs C thru F

    NASA Astrophysics Data System (ADS)

    Zapotosky, J. E.

    1991-08-01

    This portion of the report includes monitoring of and data for arthropoda and earthworms; pollinating insects; and small mammals and nesting birds. During the 1990 growing season the ELF antenna was operated more frequently than in prior years. This provides 2 years of intermittent ELF exposure for the biological systems to react to the radiation, one year of very limited exposure and greater exposure in 1990. Arthropod and earthworm sampling was conducted at intervals of two weeks from early May to late October. High voltage transmission lines and magnetic fields have been shown to affect honeybee reproduction, survival, orientation, and nest structure. ELF EM fields could have similar effects on native megachild bees. Changes in cell length, number of cells per nest, number of leaver per cell, orientation of nest entrances, and time to collect a round leaf pierce to cap a cell were monitored. We have not detected significant changes that could be attributed to ELF EM fields. Small mammal and nesting bird biological studies in the western Upper Peninsula of Michigan for the year 1990 are reported.

  20. Lyme disease with facial nerve palsy: rapid diagnosis using a nested polymerase chain reaction-restriction fragment length polymorphism analysis.

    PubMed

    Hashimoto, Y; Takahashi, H; Kishiyama, K; Sato, Y; Nakao, M; Miyamoto, K; Iizuka, H

    1998-02-01

    A 64-year-old woman with Lyme disease and manifesting facial nerve palsy had been bitten by a tick on the left frontal scalp 4 weeks previously. Erythema migrans appeared on the left forehead, accompanied by left facial paralysis. Nested polymerase chain reaction-restriction fragment length polymorphism analysis (nested PCR-RFLP) was performed on DNA extracted from a skin biopsy of the erythema on the left forehead. Borrelia flagellin gene DNA was detected and its RFLP pattern indicated that the organism was B. garinii, Five weeks later, B. garinii was isolated by conventional culture from the erythematous skin lesion, but not from the cerebrospinal fluid. After treatment with ceftriaxone intravenously for 10 days and oral administration of minocycline for 7 days, both the erythema and facial nerve palsy improved significantly. Nested PCR and culture taken after the lesion subsided, using skin samples obtained from a site adjacent to the original biopsy, were both negative. We suggest that nested PCR-RFLP analysis might be useful for the rapid diagnosis of Lyme disease and for evaluating therapy.

  1. A Molecular Approach to Nested RT-PCR Using a New Set of Primers for the Detection of the Human Immunodeficiency Virus Protease Gene.

    PubMed

    Zarei, Mohammad; Ravanshad, Mehrdad; Bagban, Ashraf; Fallahi, Shahab

    2016-07-01

    The human immunodeficiency virus (HIV-1) is the etiologic agent of AIDS. The disease can be transmitted via blood in the window period prior to the development of antibodies to the disease. Thus, an appropriate method for the detection of HIV-1 during this window period is very important. This descriptive study proposes a sensitive, efficient, inexpensive, and easy method to detect HIV-1. In this study 25 serum samples of patients under treatment and also 10 positive and 10 negative control samples were studied. Twenty-five blood samples were obtained from HIV-1-infected individuals who were receiving treatment at the acquired immune deficiency syndrome (AIDS) research center of Imam Khomeini hospital in Tehran. The identification of HIV-1-positive samples was done by using reverse transcription to produce copy deoxyribonucleic acid (cDNA) and then optimizing the nested polymerase chain reaction (PCR) method. Two pairs of primers were then designed specifically for the protease gene fragment of the nested real time-PCR (RT-PCR) samples. Electrophoresis was used to examine the PCR products. The results were analyzed using statistical tests, including Fisher's exact test, and SPSS17 software. The 325 bp band of the protease gene was observed in all the positive control samples and in none of the negative control samples. The proposed method correctly identified HIV-1 in 23 of the 25 samples. These results suggest that, in comparison with viral cultures, antibody detection by enzyme linked immunosorbent assay (ELISAs), and conventional PCR methods, the proposed method has high sensitivity and specificity for the detection of HIV-1.

  2. Using variance components to estimate power in a hierarchically nested sampling design improving monitoring of larval Devils Hole pupfish

    USGS Publications Warehouse

    Dzul, Maria C.; Dixon, Philip M.; Quist, Michael C.; Dinsomore, Stephen J.; Bower, Michael R.; Wilson, Kevin P.; Gaines, D. Bailey

    2013-01-01

    We used variance components to assess allocation of sampling effort in a hierarchically nested sampling design for ongoing monitoring of early life history stages of the federally endangered Devils Hole pupfish (DHP) (Cyprinodon diabolis). Sampling design for larval DHP included surveys (5 days each spring 2007–2009), events, and plots. Each survey was comprised of three counting events, where DHP larvae on nine plots were counted plot by plot. Statistical analysis of larval abundance included three components: (1) evaluation of power from various sample size combinations, (2) comparison of power in fixed and random plot designs, and (3) assessment of yearly differences in the power of the survey. Results indicated that increasing the sample size at the lowest level of sampling represented the most realistic option to increase the survey's power, fixed plot designs had greater power than random plot designs, and the power of the larval survey varied by year. This study provides an example of how monitoring efforts may benefit from coupling variance components estimation with power analysis to assess sampling design.

  3. Foraging ecology of least terns and piping plovers nesting on Central Platte River sandpits and sandbars

    USGS Publications Warehouse

    Sherfy, Mark H.; Anteau, Michael J.; Shaffer, Terry L.; Sovada, Marsha A.; Stucker, Jennifer H.

    2012-01-01

    Federally listed least terns (Sternula antillarum) and piping plovers (Charadrius melodus) nest on riverine sandbars on many major midcontinent river systems. On the Central Platte River, availability of sandbar habitat is limited, and both species nest on excavated sandpits in the river's floodplain. However, the extent to which sandpit-nesting birds use riverine habitats for foraging is unknown. We evaluated use of foraging habitats by least terns and piping plovers by collecting data on movements, behavior, foraging habitat, and productivity. We radiomarked 16 piping plovers and 23 least terns in 2009-2010 and monitored their movements using a network of fixed telemetry dataloggers. Piping plovers were detected primarily by the datalogger located in their nesting sandpit, whereas least terns were more frequently detected on dataloggers outside of the nesting sandpit. Telemetry data and behavioral observations showed that least terns tended to concentrate at the Kearney Canal Diversion Gates, where forage fish were apparently readily available. Fish sampling data suggested that forage fish were more abundant in riverine than in sandpit habitats, and behavioral observations showed that least terns foraged more frequently in riverine than in sandpit habitats. Piping plovers tended to forage in wet substrates along sandpit shorelines, but also used dry substrates and sandpit interior habitats. The greater mobility of least terns makes a wider range of potential foraging habitats available during brood rearing, making them able to exploit concentrations of fish outside the nesting colony. Thus, our data suggest that different spatial scales should be considered in managing nesting and foraging habitat complexes for piping plovers and least terns.

  4. Use of mammal manure by nesting burrowing owls: a test of four functional hypotheses

    USGS Publications Warehouse

    Smith, M.D.; Conway, C.J.

    2007-01-01

    Animals have evolved an impressive array of behavioural traits to avoid depredation. Olfactory camouflage of conspicuous odours is a strategy to avoid depredation that has been implicated only in a few species of birds. Burrowing owls, Athene cunicularia, routinely collect dried manure from mammals and scatter it in their nest chamber, in the tunnel leading to their nest and at the entrance to their nesting burrow. This unusual behaviour was thought to reduce nest depredation by concealing the scent of adults and juveniles, but a recent study suggests that manure functions to attract arthropod prey. However, burrowing owls routinely scatter other materials in the same way that they scatter manure, and this fact seems to be at odds with both of these hypotheses. Thus, we examined the function of this behaviour by testing four alternative hypotheses. We found no support for the widely cited olfactory-camouflage hypothesis (manure did not lower the probability of depredation), or for the mate-attraction hypothesis (males collected manure after, not before, pair formation). Predictions of the burrow-occupied hypothesis (manure indicates occupancy to conspecifics and thereby reduces agonistic interactions) were supported, but results were not statistically significant. Our results also supported several predictions of the prey-attraction hypothesis. Pitfall traps at sampling sites with manure collected more arthropod biomass (of taxa common in the diet of burrowing owls) than pitfall traps at sampling sites without manure. Scattering behaviour of burrowing owls appears to function to attract arthropod prey, but may also signal occupancy of a burrow to conspecifics. ?? 2006 The Association for the Study of Animal Behaviour.

  5. Prevalence and molecular characterization of Giardia duodenalis in dogs in Aydin, Turkey.

    PubMed

    Gultekin, Mehmet; Ural, Kerem; Aysul, Nuran; Ayan, Adnan; Balikci, Canberk; Akyildiz, Gurkan

    2017-06-01

    The purpose of the present study was to investigate the prevalence and molecular characterization of Giardia duodenalis among dogs in Aydin, Turkey. A total of 473 faecal samples from dogs were collected. The overall prevalence of G. duodenalis was 18.8%. Higher infection rates were observed in dogs younger than three months old, from shelters, and with diarrhoea. Faecal samples of 89 dogs, diagnosed Giardia-positive by microscopy, were found positive by nested PCR. The β-giardin nested PCR assay revealed assemblage B in all samples (100%), whereas 38 of the samples were mixed with assemblage A (42%). Sequence analysis of isolates indicated sub-genotypes A3 and B4 which have been previously detected in human isolates from Turkey. The results of the present study indicated the relatively high prevalence of giardiasis and the presence of the zoonotic sub-genotypes suggesting the important role of dogs as potential reservoirs of human infections.

  6. A novel loop-mediated isothermal amplification-based test for detecting Neospora caninum DNA.

    PubMed

    Ramos, Andrea Estefanía; Muñoz, Marina; Cortés-Vecino, Jesús Alfredo; Barato, Paola; Patarroyo, Manuel Alfonso

    2017-11-29

    Neospora caninum is a cyst-forming, coccidian parasite which is known to cause neurological disorders in dogs and abortion and neonatal mortality in cows and other livestock. This study reports the development of a loop-mediated isothermal amplification (LAMP) assay based on the Neospora caninum Nc-5 gene and compares its efficacy for detecting DNA to that of a semi-nested PCR test. Six primers were designed based on the Nc-5 repeat region of N. caninum. Specific LAMP primers led to successful amplification of N. caninum DNA at 63 °C in 30 min. The LAMP assay was highly specific (i.e. it did not reveal cross-reactivity with other parasite species) and had a low N. caninum plasmid DNA limit of detection (1 fg), which is ten times higher than that for the semi-nested PCR. LAMP applicability was evaluated using a set of naturally-infected samples (59 from canine faeces and five from bovine abortions). Thirty-nine percent (25/64) of the naturally-infected samples were positive for N. caninum DNA by LAMP and 36% (23/64) by semi-nested PCR. However, the LAMP assay is much faster to perform than semi-nested PCR and provides results in 30 min. The optimized reaction conditions described in this study resulted in a sensitive, specific and rapid technique for detecting N. caninum DNA. Considering the advantages of LAMP for detecting N. caninum DNA, further assays aimed at testing its usefulness on a wider range of field samples are recommended.

  7. Physiological ecology of overwintering in the hatchling painted turtle: multiple-scale variation in response to environmental stress.

    PubMed

    Costanzo, Jon P; Dinkelacker, Stephen A; Iverson, John B; Lee, Richard E

    2004-01-01

    We integrated field and laboratory studies in an investigation of water balance, energy use, and mechanisms of cold-hardiness in hatchling painted turtles (Chrysemys picta) indigenous to west-central Nebraska (Chrysemys picta bellii) and northern Indiana (Chrysemys picta marginata) during the winters of 1999-2000 and 2000-2001. We examined 184 nests, 80 of which provided the hatchlings (n=580) and/or samples of soil used in laboratory analyses. Whereas winter 1999-2000 was relatively dry and mild, the following winter was wet and cold; serendipitously, the contrast illuminated a marked plasticity in physiological response to environmental stress. Physiological and cold-hardiness responses of turtles also varied between study locales, largely owing to differences in precipitation and edaphics and the lower prevailing and minimum nest temperatures (to -13.2 degrees C) encountered by Nebraska turtles. In Nebraska, winter mortality occurred within 12.5% (1999-2000) and 42.3% (2000-2001) of the sampled nests; no turtles died in the Indiana nests. Laboratory studies of the mechanisms of cold-hardiness used by hatchling C. picta showed that resistance to inoculative freezing and capacity for freeze tolerance increased as winter approached. However, the level of inoculation resistance strongly depended on the physical characteristics of nest soil, as well as its moisture content, which varied seasonally. Risk of inoculative freezing (and mortality) was greatest in midwinter when nest temperatures were lowest and soil moisture and activity of constituent organic ice nuclei were highest. Water balance in overwintering hatchlings was closely linked to dynamics of precipitation and soil moisture, whereas energy use and the size of the energy reserve available to hatchlings in spring depended on the winter thermal regime. Acute chilling resulted in hyperglycemia and hyperlactemia, which persisted throughout winter; this response may be cryoprotective. Some physiological characteristics and cold-hardiness attributes varied between years, between study sites, among nests at the same site, and among siblings sharing nests. Such variation may reflect adaptive phenotypic plasticity, maternal or paternal influence on an individual's response to environmental challenge, or a combination of these factors. Some evidence suggests that life-history traits, such as clutch size and body size, have been shaped by constraints imposed by the harsh winter environment.

  8. Uniform Atmospheric Retrievals of Ultracool Late-T and Early-Y dwarfs

    NASA Astrophysics Data System (ADS)

    Garland, Ryan; Irwin, Patrick

    2018-01-01

    A significant number of ultracool (<600K) extrasolar objects have been unearthed in the past decade thanks to wide-field surveys such as WISE. These objects present a perfect testbed for examining the evolution of atmospheric structure as we transition from typically hot extrasolar temperatures to the temperatures found within our Solar System.By examining these types of objects with a uniform retrieval method, we hope to elucidate any trends and (dis)similarities found in atmospheric parameters, such as chemical abundances, temperature-pressure profile, and cloud structure, for a sample of 7 ultracool brown dwarfs as we transition from hotter (~700K) to colder objects (~450K).We perform atmospheric retrievals on two late-T and five early-Y dwarfs. We use the NEMESIS atmospheric retrieval code coupled to a Nested Sampling algorithm, along with a standard uniform model for all of our retrievals. The uniform model assumes the atmosphere is described by a gray radiative-convective temperature profile, (optionally) a self-consistent Mie scattering cloud, and a number of relevant gases. We first verify our methods by comparing it to a benchmark retrieval for Gliese 570D, which is found to be consistent. Furthermore, we present the retrieved gaseous composition, temperature structure, spectroscopic mass and radius, cloud structure and the trends associated with decreasing temperature found in this small sample of objects.

  9. Challenging loop-mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii.

    PubMed

    Fallahi, Shirzad; Mazar, Zahra Arab; Ghasemian, Mehrdad; Haghighi, Ali

    2015-05-01

    To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA. The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions of T. gondii DNA ranging from 1 ng to 0.01 fg. DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays. After testing LAMP and nested-PCR in duplicate, the detection limit of RE-LAMP, B1-LAMP, RE-nested PCR and B1-nested PCR assays was one fg, 100 fg, 1 pg and 10 pg of T. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection of T. gondii DNA. The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T. gondii. Furthermore, these findings indicate that primers based on the RE are more suitable than those based on the B1 gene. However, the B1-LAMP assay has potential as a diagnostic tool for detection of T. gondii. Copyright © 2015 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.

  10. Sporadic nesting reveals long distance colonisation in the philopatric loggerhead sea turtle (Caretta caretta).

    PubMed

    Carreras, Carlos; Pascual, Marta; Tomás, Jesús; Marco, Adolfo; Hochscheid, Sandra; Castillo, Juan José; Gozalbes, Patricia; Parga, Mariluz; Piovano, Susanna; Cardona, Luis

    2018-01-23

    The colonisation of new suitable habitats is crucial for species survival at evolutionary scale under changing environmental conditions. However, colonisation potential may be limited by philopatry that facilitates exploiting successful habitats across generations. We examine the mechanisms of long distance dispersal of the philopatric loggerhead sea turtle (Caretta caretta) by analysing 40 sporadic nesting events in the western Mediterranean. The analysis of a fragment of the mitochondrial DNA and 7 microsatellites of 121 samples from 18 of these nesting events revealed that these nests were colonising events associated with juveniles from distant populations feeding in nearby foraging grounds. Considering the temperature-dependent sex determination of the species, we simulated the effect of the incubation temperature and propagule pressure on a potential colonisation scenario. Our results indicated that colonisation will succeed if warm temperature conditions, already existing in some of the beaches in the area, extend to the whole western Mediterranean. We hypothesize that the sporadic nesting events in developmental foraging grounds may be a mechanism to overcome philopatry limitations thus increasing the dispersal capabilities of the species and the adaptability to changing environments. Sporadic nesting in the western Mediterranean can be viewed as potential new populations in a scenario of rising temperatures.

  11. Photometric classification and redshift estimation of LSST Supernovae

    NASA Astrophysics Data System (ADS)

    Dai, Mi; Kuhlmann, Steve; Wang, Yun; Kovacs, Eve

    2018-07-01

    Supernova (SN) classification and redshift estimation using photometric data only have become very important for the Large Synoptic Survey Telescope (LSST), given the large number of SNe that LSST will observe and the impossibility of spectroscopically following up all the SNe. We investigate the performance of an SN classifier that uses SN colours to classify LSST SNe with the Random Forest classification algorithm. Our classifier results in an area-under-the-curve of 0.98 which represents excellent classification. We are able to obtain a photometric SN sample containing 99 per cent SNe Ia by choosing a probability threshold. We estimate the photometric redshifts (photo-z) of SNe in our sample by fitting the SN light curves using the SALT2 model with nested sampling. We obtain a mean bias (⟨zphot - zspec⟩) of 0.012 with σ (z_phot-z_spec/1+z_spec) = 0.0294 without using a host-galaxy photo-z prior, and a mean bias (⟨zphot - zspec⟩) of 0.0017 with σ (z_phot-z_spec/1+z_spec) = 0.0116 using a host-galaxy photo-z prior. Assuming a flat ΛCDM model with Ωm = 0.3, we obtain Ωm of 0.305 ± 0.008 (statistical errors only), using the simulated LSST sample of photometric SNe Ia (with intrinsic scatter σint = 0.11) derived using our methodology without using host-galaxy photo-z prior. Our method will help boost the power of SNe from the LSST as cosmological probes.

  12. Prevalence of mip virulence gene and PCR-base sequence typing of Legionella pneumophila from cooling water systems of two cities in Iran.

    PubMed

    Ahmadrajabi, Roya; Shakibaie, Mohammad Reza; Iranmanesh, Zahra; Mollaei, Hamid Reza; Sobhanipoor, Mohammad Hossein

    2016-07-03

    Legionella pneumophila is the primary respiratory pathogen and mostly transmitted to human through water cooling systems and cause mild to severe pneumonia with high mortality rate especially in elderly both in hospitals and community. However, current Legionella risk assessments may be compromised by uncertainties in Legionella detection methods. Here, we investigated the presence of L. pneumophila mip gene in water samples collected from different hospitals cooling towers, nursing homes and building/hotels water coolants from two geographical locations of Iran (Kerman and Bam cities) during summer season of 2015 by both nested and real-time PCR methods. Analysis of the 128 water samples for presence of the mip gene by nested-PCR revealed, 18 (23%) positive cases in Kerman and 7(14%) in Bam. However, when samples were tested by real-time PCR, we identified 4 more new cases of L. pneumophila in the hospitals as well as nursing homes water systems that were missed by nested-PCR. The highest rate of contamination was detected in water obtained from hospitals cooling towers in both the cities (p≤0.05). Dendrogram analysis and clonal relationship by PCR-base sequence typing (SBT) of the L. pneumophila genomic DNAs in Kerman water samples showed close clonal similarities among the isolates, in contrast, isolates identified from Bam city demonstrated two fingerprint patterns. The clones from hospital water samples were more related to the L. pneumophila serogroup- 1.

  13. Prevalence of mip virulence gene and PCR-base sequence typing of Legionella pneumophila from cooling water systems of two cities in Iran

    PubMed Central

    Ahmadrajabi, Roya; Shakibaie, Mohammad Reza; Iranmanesh, Zahra; Mollaei, Hamid Reza; Sobhanipoor, Mohammad Hossein

    2016-01-01

    ABSTRACT Legionella pneumophila is the primary respiratory pathogen and mostly transmitted to human through water cooling systems and cause mild to severe pneumonia with high mortality rate especially in elderly both in hospitals and community. However, current Legionella risk assessments may be compromised by uncertainties in Legionella detection methods. Here, we investigated the presence of L. pneumophila mip gene in water samples collected from different hospitals cooling towers, nursing homes and building/hotels water coolants from two geographical locations of Iran (Kerman and Bam cities) during summer season of 2015 by both nested and real-time PCR methods. Analysis of the 128 water samples for presence of the mip gene by nested-PCR revealed, 18 (23%) positive cases in Kerman and 7(14%) in Bam. However, when samples were tested by real-time PCR, we identified 4 more new cases of L. pneumophila in the hospitals as well as nursing homes water systems that were missed by nested-PCR. The highest rate of contamination was detected in water obtained from hospitals cooling towers in both the cities (p≤0.05). Dendrogram analysis and clonal relationship by PCR-base sequence typing (SBT) of the L. pneumophila genomic DNAs in Kerman water samples showed close clonal similarities among the isolates, in contrast, isolates identified from Bam city demonstrated two fingerprint patterns. The clones from hospital water samples were more related to the L. pneumophila serogroup- 1. PMID:27028760

  14. Development of a reverse transcription polymerase chain reaction method for yellow fever virus detection.

    PubMed

    Méndez, María C; Domingo, Cristina; Tenorio, Antonio; Pardo, Lissethe C; Rey, Gloria J; Méndez, Jairo A

    2013-09-01

    Yellow fever is considered a re-emerging disease and is endemic in tropical regions of Africa and South America. At present, there are no standardized or commercialized kits available for yellow fever virus detection. Therefore, diagnosis must be made by time-consuming routine techniques, and sometimes, the virus or its proteins are not detected. Furthermore, co-circulation with other flaviviruses, including dengue virus, increases the difficulty of diagnosis. To develop a specific reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR-based assay to improve the detection and diagnosis of yellow fever virus using both serum and fresh tissue samples. RT-PCR primers were designed to amplify a short fragment of all yellow fever virus genotypes reported. A second set of primers was used in a nested PCR to increase sensitivity. Thirty-three clinical samples were tested with the standardized reaction. The expected amplicon was obtained in 25 out of 33 samples analyzed using this approach, and 2 more samples tested positive after a subsequent nested PCR approach. This improved technique not only ensures the specific detection of a wide range of yellow fever virus genotypes but also may increase the sensitivity of detection by introducing a second round of amplification, allowing a rapid differential diagnosis between dengue and yellow fever infection, which is required for effective surveillance and opportune epidemiologic measures.

  15. Comparison of Microscopy, Nested-PCR, and Real-Time-PCR Assays Using High-Throughput Screening of Pooled Samples for Diagnosis of Malaria in Asymptomatic Carriers from Areas of Endemicity in Myanmar

    PubMed Central

    Wang, Bo; Han, Soe-Soe; Cho, Cho; Han, Jin-Hee; Cheng, Yang; Lee, Seong-Kyun; Galappaththy, Gawrie N. L; Thimasarn, Krongthong; Soe, Myat Thu; Oo, Htet Wai; Kyaw, Myat Phone

    2014-01-01

    Asymptomatic infection is an important obstacle for controlling disease in countries where malaria is endemic. Because asymptomatic carriers do not seek treatment for their infections, they can have high levels of gametocytes and constitute a reservoir available for new infection. We employed a sample pooling/PCR-based molecular detection strategy for screening malaria infection in residents from areas of Myanmar where malaria is endemic. Blood samples (n = 1,552) were collected from residents in three areas of malaria endemicity (Kayin State, Bago, and Tanintharyi regions) of Myanmar. Two nested PCR and real-time PCR assays showed that asymptomatic infection was detected in about 1.0% to 9.4% of residents from the surveyed areas. The sensitivities of the two nested PCR and real-time PCR techniques were higher than that of microscopy examination (sensitivity, 100% versus 26.4%; kappa values, 0.2 to 0.5). Among the three regions, parasite-positive samples were highly detected in subjects from the Bago and Tanintharyi regions. Active surveillance of residents from regions of intense malaria transmission would reduce the risk of morbidity and mitigate transmission to the population in these areas of endemicity. Our data demonstrate that PCR-based molecular techniques are more efficient than microscopy for nationwide surveillance of malaria in countries where malaria is endemic. PMID:24648557

  16. A novel and highly sensitive real-time nested RT-PCR assay in a single closed tube for detection of enterovirus.

    PubMed

    Shen, Xin-Xin; Qiu, Fang-Zhou; Zhao, Huai-Long; Yang, Meng-Jie; Hong, Liu; Xu, Song-Tao; Zhou, Shuai-Feng; Li, Gui-Xia; Feng, Zhi-Shan; Ma, Xue-Jun

    2018-03-01

    The sensitivity of qRT-PCR assay is not adequate for the detection of the samples with lower viral load, particularly in the cerebrospinal fluid (CSF) of patients. Here, we present the development of a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of human enterovirus (HEV). The clinical performance of both RTN RT-PCR and qRT-PCR was also tested and compared using 140 CSF and fecal specimens. The sensitivities of RTN RT-PCR assay for EV71, Coxsackievirus A (CVA)16, CVA6 and CVA10 achieved 10 -8 dilution with a corresponding Ct value of 38.20, 36.45, 36.75, and 36.45, respectively, which is equal to traditional two-step nested RT-PCR assay and approximately 2-10-fold lower than that of qRT-PCR assay. The specificity of RTN RT-PCR assay was extensively analyzed insilico and subsequently verified using the reference isolates and clinical samples. Sixteen qRT-PCR-negative samples were detected by RTN RT-PCR and a variety of enterovirus serotypes was identified by sequencing of inner PCR products. We conclude RTN RT-PCR is more sensitive than qRT-PCR for the detection of HEV in clinical samples. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Differentiating Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii using nested polymerase chain reaction (PCR) in rural communities in Malaysia

    PubMed Central

    2012-01-01

    Background In this study, a total of 426 human faecal samples were examined for the presence of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii infection via a combination of microscopic examination and nested polymerase chain reaction (PCR) targeting 16S ribosomal RNA of Entamoeba species. Methods Faecal sample were collected from 426 participants in five rural villages in Peninsular Malaysia. The faecal samples were processed by direct wet smear and formalin ethyl acetate concentration technique followed by iodine staining and examined via microscopy for the presence of Entamoeba species and other intestinal parasites. Microscopically positive samples for Entamoeba species cysts were further characterized using a Nested Polymerase Chain Reaction (Nested-PCR) targeting 16S-like ribosomal RNA gene. The data entry and analysis was carried out using the SPSS software (Statistical Package for the Social Sciences) program for Windows version 17 (SPSS, Chicago, IL, USA). Results Based on single faecal examination, overall prevalence of Entamoeba infection was 17.6% (75/426). Females (19.1%) were more commonly infected compared to males (15.9%). Comparison by age groups showed that adults (23.9%) had higher infection rates than children (15.3%). The PCR results showed that 52 out of 75 microscopy positive samples successfully generated species-specific amplicons. The infection with E. histolytica (75.0%; 39/52) was the most common, followed by E. dispar (30.8%; 18/52) and E. moshkovskii (5.8%; 3/52). Of these, 33 (63.5%) were shown to contain only E. histolytica, 10 (19.2%) contained E. dispar and 3 (5.8%) contained only E. moshkovskii. Mixed infection with E. histolytica and E. dispar was found in 6 (11.5%) samples. Conclusions The present study essentially emphasized the benefit of molecular techniques in discriminating the pathogenic Entamoeba species from the non-pathogenic for accurate diagnosis and better management of amoebiasis. The presence of E. moshkovskii is of great public health concern as it was the first time it has been reported in Malaysia. PMID:22947430

  18. Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR. I. Standardization of methods.

    PubMed

    Wójcik-Fatla, Angelina; Stojek, Nimfa Maria; Dutkiewicz, Jacek

    2012-01-01

    The aim of the present study was: - to compare methods for concentration and isolation of Legionella DNA from water; - to examine the efficacy of various modifications of PCR test (PCR, semi-nested PCR, and real-time PCR) for the detection of known numbers of Legionella pneumophila in water samples artificially contaminated with the strain of this bacterium and in randomly selected samples of environmental water, in parallel with examination by culture. It was found that filtration is much more effective than centrifugation for the concentration of DNA in water samples, and that the Qiamp DNA Mini-Kit is the most efficient for isolation of Legionella DNA from water. The semi-nested PCR and real-time PCR proved to be the most sensitive methods for detection of Legionella DNA in water samples. Both PCR modifications showed a high correlation with recovery of Legionella by culture (p<0.01), while no correlation occurred between the results of one-stage PCR and culture (p>0.1).

  19. Occurrence of Giardia intestinalis and Cryptosporidium sp. in wastewater samples from São Paulo State, Brazil, and Lima, Peru.

    PubMed

    Ulloa-Stanojlović, Francisco Miroslav; Aguiar, Bruna; Jara, Luis M; Sato, Maria Inês Zanoli; Guerrero, Juana Arzola; Hachich, Elayse; Matté, Glavur Rogério; Dropa, Milena; Matté, Maria Helena; de Araújo, Ronalda Silva

    2016-11-01

    The objectives of the study were to detect and genotype Cryptosporidium spp. and Giardia intestinalis in wastewater samples obtained from five cities with high transit of people in the State of São Paulo, Brazil, and at the entrance of a Wastewater Treatment Plant (WWTP) in Lima, Peru. Samples were collected and concentrated by centrifugation. The genomic DNA was extracted for molecular characterization by nested PCR for Cryptosporidium and double nested PCR for Giardia, followed by sequencing and phylogenetic analysis. G. intestinalis was found in 63.6 % of the samples, and the human assemblages A and B were identified. Cryptosporidium sp. was found in 36.4 % of the samples, and the species were corresponding to Cryptosporidium hominis, Cryptosporidium cuniculus, and Cryptosporidium muris. Results revealed the presence of human pathogenic Cryptosporidium species and G. intestinalis human pathogenic assemblages. Molecular tools highlight the importance to map the genetic diversity of these parasites, as well as to detect their epidemiological circulation pathway in the environment.

  20. Inertial cavitation threshold of nested microbubbles.

    PubMed

    Wallace, N; Dicker, S; Lewin, Peter; Wrenn, S P

    2015-04-01

    Cavitation of ultrasound contrast agents (UCAs) promotes both beneficial and detrimental bioeffects in vivo (Radhakrishnan et al., 2013) [1]. The ability to determine the inertial cavitation threshold of UCA microbubbles has potential application in contrast imaging, development of therapeutic agents, and evaluation of localized effects on the body (Ammi et al., 2006) [2]. This study evaluates a novel UCA and its inertial cavitation behavior as determined by a home built cavitation detection system. Two 2.25 MHz transducers are placed at a 90° angle to one another where one transducer is driven by a high voltage pulser and the other transducer receives the signal from the oscillating microbubble. The sample chamber is placed in the overlap of the focal region of the two transducers where the microbubbles are exposed to a pulser signal consisting of 600 pulse trains per experiment at a pulse repetition frequency of 5 Hz where each train has four pulses of four cycles. The formulation being analyzed is comprised of an SF6 microbubble coated by a DSPC PEG-3000 monolayer nested within a poly-lactic acid (PLA) spherical shell. The effect of varying shell diameters and microbubble concentration on cavitation threshold profile for peak negative pressures ranging from 50 kPa to 2 MPa are presented and discussed in this paper. The nesting shell decreases inertial cavitation events from 97.96% for an un-nested microbubble to 19.09% for the same microbubbles nested within a 2.53 μm shell. As shell diameter decreases, the percentage of inertially cavitating microbubbles also decreases. For nesting formulations with average outer capsule diameters of 20.52, 14.95, 9.95, 5.55, 2.53, and 1.95 μm, the percentage of sample destroyed at 1 MPa was 51.02, 38.94, 33.25, 25.27, 19.09, and 5.37% respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Molecular detection of Toxoplasma gondii in water samples from Scotland and a comparison between the 529bp real-time PCR and ITS1 nested PCR.

    PubMed

    Wells, Beth; Shaw, Hannah; Innocent, Giles; Guido, Stefano; Hotchkiss, Emily; Parigi, Maria; Opsteegh, Marieke; Green, James; Gillespie, Simon; Innes, Elisabeth A; Katzer, Frank

    2015-12-15

    Waterborne transmission of Toxoplasma gondii is a potential public health risk and there are currently no agreed optimised methods for the recovery, processing and detection of T. gondii oocysts in water samples. In this study modified methods of T. gondii oocyst recovery and DNA extraction were applied to 1427 samples collected from 147 public water supplies throughout Scotland. T. gondii DNA was detected, using real time PCR (qPCR) targeting the 529bp repeat element, in 8.79% of interpretable samples (124 out of 1411 samples). The samples which were positive for T. gondii DNA originated from a third of the sampled water sources. The samples which were positive by qPCR and some of the negative samples were reanalysed using ITS1 nested PCR (nPCR) and results compared. The 529bp qPCR was the more sensitive technique and a full analysis of assay performance, by Bayesian analysis using a Markov Chain Monte Carlo method, was completed which demonstrated the efficacy of this method for the detection of T. gondii in water samples. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. PIRLS 2011 User Guide for the International Database. Supplement 4: PIRLS 2011 Sampling Stratification Information

    ERIC Educational Resources Information Center

    Foy, Pierre, Ed.; Drucker, Kathleen T., Ed.

    2013-01-01

    This supplement contains documentation on the explicit and implicit stratification variables included in the PIRLS 2011 data files. The explicit strata are smaller sampling frames, created from the national sampling frames, from which national samples of schools were drawn. The implicit strata are nested within the explicit strata, and were used…

  3. TIMSS 2011 User Guide for the International Database. Supplement 4: TIMSS 2011 Sampling Stratification Information

    ERIC Educational Resources Information Center

    Foy, Pierre, Ed.; Arora, Alka, Ed.; Stanco, Gabrielle M., Ed.

    2013-01-01

    This supplement contains documentation on the explicit and implicit stratification variables included in the TIMSS 2011 data files. The explicit strata are smaller sampling frames, created from the national sampling frames, from which national samples of schools were drawn. The implicit strata are nested within the explicit strata, and were used…

  4. Molecular detection of a novel paramyxovirus in fruit bats from Indonesia

    PubMed Central

    2012-01-01

    Background Fruit bats are known to harbor zoonotic paramyxoviruses including Nipah, Hendra, and Menangle viruses. The aim of this study was to detect the presence of paramyxovirus RNA in fruit bats from Indonesia. Methods RNA samples were obtained from the spleens of 110 fruit bats collected from four locations in Indonesia. All samples were screened by semi-nested broad spectrum reverse transcription PCR targeting the paramyxovirus polymerase (L) genes. Results Semi-nested reverse transcription PCR detected five previously unidentified paramyxoviruses from six fruit bats. Phylogenetic analysis showed that these virus sequences were related to henipavirus or rubulavirus. Conclusions This study indicates the presence of novel paramyxoviruses among fruit bat populations in Indonesia. PMID:23082748

  5. On estimation of linear transformation models with nested case–control sampling

    PubMed Central

    Liu, Mengling

    2011-01-01

    Nested case–control (NCC) sampling is widely used in large epidemiological cohort studies for its cost effectiveness, but its data analysis primarily relies on the Cox proportional hazards model. In this paper, we consider a family of linear transformation models for analyzing NCC data and propose an inverse selection probability weighted estimating equation method for inference. Consistency and asymptotic normality of our estimators for regression coefficients are established. We show that the asymptotic variance has a closed analytic form and can be easily estimated. Numerical studies are conducted to support the theory and an application to the Wilms’ Tumor Study is also given to illustrate the methodology. PMID:21912975

  6. Soil hydric characteristics and environmental ice nuclei influence supercooling capacity of hatchling painted turtles Chrysemys picta.

    PubMed

    Costanzo, J P; Litzgus, J D; Iverson, J B; Lee, R E

    1998-11-01

    Hatchling painted turtles (Chrysemys picta) hibernate in their shallow natal nests where temperatures occasionally fall below -10 C during cold winters. Because the thermal limit of freeze tolerance in this species is approximately -4 C, hatchlings rely on supercooling to survive exposure to extreme cold. We investigated the influence of environmental ice nuclei on susceptibility to inoculative freezing in hatchling C. picta indigenous to the Sandhills of west-central Nebraska. In the absence of external ice nuclei, hatchlings cooled to -14.6 1.9 C (mean s.e.m.; N=5) before spontaneously freezing. Supercooling capacity varied markedly among turtles cooled in physical contact with sandy soil collected from nesting locales or samples of the native soil to which water-binding agents (clay or peat) had been added, despite the fact that all substrata contained the same amount of moisture (7.5 % moisture, w/w). The temperature of crystallization (Tc) of turtles exposed to frozen native soil was -1.6 0.4 C (N=5), whereas turtles exposed to frozen soil/clay and soil/peat mixtures supercooled extensively (mean Tc values approximately -13 C). Hatchlings cooled in contact with drier (less than or equal to 4 % moisture) native soil also supercooled extensively. Thus, inoculative freezing is promoted by exposure to sandy soils containing abundant moisture and little clay or organic matter. Soil collected at turtle nesting locales in mid and late winter contained variable amounts of moisture (4-15 % w/w) and organic matter (1-3 % w/w). In addition to ice, the soil at turtle nesting locales may harbor inorganic and organic ice nuclei that may also seed the freezing of hatchlings. Bulk samples of native soil, which were autoclaved to destroy any organic nuclei, nucleated aqueous solutions at approximately -7 C (Tc range -6.1 to -8.2 C). Non-autoclaved samples contained water-extractable, presumably organic, ice nuclei (Tc range -4.4 to -5.3 C). Ice nuclei of both classes varied in potency among turtle nesting locales. Interaction with ice nuclei in the winter microenvironment determines whether hatchling C. picta remain supercooled or freeze and may ultimately account for differential mortality in nests at a given locale and for variation in winter survival rates among populations.

  7. A binary search approach to whole-genome data analysis.

    PubMed

    Brodsky, Leonid; Kogan, Simon; Benjacob, Eshel; Nevo, Eviatar

    2010-09-28

    A sequence analysis-oriented binary search-like algorithm was transformed to a sensitive and accurate analysis tool for processing whole-genome data. The advantage of the algorithm over previous methods is its ability to detect the margins of both short and long genome fragments, enriched by up-regulated signals, at equal accuracy. The score of an enriched genome fragment reflects the difference between the actual concentration of up-regulated signals in the fragment and the chromosome signal baseline. The "divide-and-conquer"-type algorithm detects a series of nonintersecting fragments of various lengths with locally optimal scores. The procedure is applied to detected fragments in a nested manner by recalculating the lower-than-baseline signals in the chromosome. The algorithm was applied to simulated whole-genome data, and its sensitivity/specificity were compared with those of several alternative algorithms. The algorithm was also tested with four biological tiling array datasets comprising Arabidopsis (i) expression and (ii) histone 3 lysine 27 trimethylation CHIP-on-chip datasets; Saccharomyces cerevisiae (iii) spliced intron data and (iv) chromatin remodeling factor binding sites. The analyses' results demonstrate the power of the algorithm in identifying both the short up-regulated fragments (such as exons and transcription factor binding sites) and the long--even moderately up-regulated zones--at their precise genome margins. The algorithm generates an accurate whole-genome landscape that could be used for cross-comparison of signals across the same genome in evolutionary and general genomic studies.

  8. Identification of hierarchical chromatin domains

    PubMed Central

    Weinreb, Caleb; Raphael, Benjamin J.

    2016-01-01

    Motivation: The three-dimensional structure of the genome is an important regulator of many cellular processes including differentiation and gene regulation. Recently, technologies such as Hi-C that combine proximity ligation with high-throughput sequencing have revealed domains of self-interacting chromatin, called topologically associating domains (TADs), in many organisms. Current methods for identifying TADs using Hi-C data assume that TADs are non-overlapping, despite evidence for a nested structure in which TADs and sub-TADs form a complex hierarchy. Results: We introduce a model for decomposition of contact frequencies into a hierarchy of nested TADs. This model is based on empirical distributions of contact frequencies within TADs, where positions that are far apart have a greater enrichment of contacts than positions that are close together. We find that the increase in contact enrichment with distance is stronger for the inner TAD than for the outer TAD in a TAD/sub-TAD pair. Using this model, we develop the TADtree algorithm for detecting hierarchies of nested TADs. TADtree compares favorably with previous methods, finding TADs with a greater enrichment of chromatin marks such as CTCF at their boundaries. Availability and implementation: A python implementation of TADtree is available at http://compbio.cs.brown.edu/software/ Contact: braphael@cs.brown.edu Supplementary information: Supplementary data are available at Bioinformatics online. PMID:26315910

  9. Predation of artificial ground nests on white-tailed prairie dog colonies

    USGS Publications Warehouse

    Baker, B.W.; Stanley, T.R.; Sedgwick, J.A.

    1999-01-01

    Prairie dog (Cynomys spp.) colonies are unique to prairie and shrub-steppe landscapes. However, widespread eradication, habitat loss, and sylvatic plague (Yersinia pestis) have reduced their numbers by 98% since historical times. Birds associated with prairie dogs also are declining. Potential nest predators, such as coyotes (Canis latrans), swift foxes (Vulpes velox), and badgers (Taxidea taxus), may be attracted to colonies where a high concentration of prairie dogs serve as available prey. Increased abundance of small mammals, including prairie dogs, also may increase the risk of predation for birds nesting on colonies. Finally, because grazing by prairie dogs may decrease vegetation height and canopy cover, bird nests may be easier for predators to locate. In this study, we placed 1,444 artificial ground nests on and off 74 white-tailed prairie dog (C. leucurus) colonies to test the hypothesis that nest predation rates are higher on colonies than at nearby off sites (i.e., uncolonized habitat). We sampled colonies from 27 May to 16 July 1997 at the following 3 complexes: Coyote Basin, Utah and Colorado; Moxa Arch, Wyoming; and Shirley Basin, Wyoming. Differences in daily predation rates between colonies and paired off sites averaged 1.0% (P = 0.060). When converted to a typical 14-day incubation period, predation rates averaged 14% higher on colonies (57.7 ?? 2.7%; ?? ?? SE) than at off sites (50.4 ?? 3.1%). Comparisons of habitat variables on colonies to off sites showed percent canopy cover of vegetation was similar (P = 0.114), percent bare ground was higher on colonies (P 0.288). Although we found the risk of nest predation was higher on white-tailed prairie dog colonies than at off sites, fitness of birds nesting on colonies might depend on other factors that influence foraging success, reproductive success, or nestling survival.

  10. Association of empty nest with depressive symptom in a Chinese elderly population: A cross-sectional study.

    PubMed

    Zhai, Yujia; Yi, Huaiming; Shen, Wei; Xiao, Yuanyuan; Fan, Haixiao; He, Fan; Li, Fudong; Wang, Xinyi; Shang, Xiaopeng; Lin, Junfen

    2015-11-15

    Empty nest has been becoming the main family pattern among old people, and influencing the traditional family providing for the aged in China. This study aimed to investigate the association of empty nest with depressive symptom in a Chinese elderly population. Our study was based on the baseline investigation of Zhejiang Major Public Health Surveillance which was conducted in 2014. The final analyzed sample consisted of 9215 participants aged no less than 60 years. Subjects completed a questionnaire including demographic characteristics, living arrangements, behavioral risk factors, health status, subjective assessments, and Patient Health Questionnaire-9 scale. More than half of the participants were empty nest elders (57.4%). The overall prevalence of depressive symptom was 10.3%, while it was higher in empty nest group than in non-empty nest group (11.6% vs. 8.6%, P<0.001). A positive association was found between empty nest and depressive symptom, the OR (95%CI) was 1.223(1.045, 1.431). After stratified by gender/age/education level/employment status, this association was just prominent in participants of male, 70 years old and above, primary school education, and retirees. No matter living with spouse or living alone, empty nesters were more likely to have depressive symptoms than non-empty nesters. Cross-sectional study could not make a causation conclusion. The social supports of participants were not been investigated in detail. Empty nest elders, especially those who are male, 70 years old and above, primary school education, and retired, are more vulnerable to depressive symptom. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. [Reproductive activity of Chelonia mydas (Testudines: Cheloniidae) in Isla de Aves, Venezuela (2001-2008)].

    PubMed

    Vera, Vicente; Buitrago, Joaquín

    2012-06-01

    The second major nesting-site for green turtles in the Caribbean is Isla de Aves, an island protected as a wildlife refuge since 1972, located at 650km Northeast from La Guaira, Venezuela. In this island, the nesting population monitoring started in 1972 and in a more continuous way after 1978, when a Scientific-Naval Station was established and scientific observations started. Since historical data show that female captures had severely affected population levels in this island before 1978, this study aim to describe recent reproductive activities. For this, during the nesting seasons of 2001-2002 and 2005-2008, nesting females were measured and tagged using metal flipper tags. A total of 458 nights were sampled observing 5 154 female emergences, with a maximum of 53 in a single night. Non-observed emergences were calculated fitting the temporal distribution of observed emergences to a normal curve. Total emergences estimated varied from X=637.1+/-106.6 in 2001 to X =2 853+/-42.5 in 2008 (ANOVA F(6.5df)=60.37, p<0.0001). Internesting interval in the same season was estimated in X=10.71+/-1.32 days. Clutch frequency in a nesting season was calculated as X=1.71+/-1.6 times per female and season. Estimated number of nesting females per year varied from X=373+/-12.5 females in 2001 to X=l 669+/-56.1 females in 2008 (ANOVA F 55.6df)=89.42, p<0.0001); with a positive and significant trend (r=0.842, p=0.036). Results show that nesting females numbers are increasing. We suggest that the protection of the nesting area for more than 30 years, has contributed with this population increase.

  12. Crucial nesting habitat for gunnison sage-grouse: A spatially explicit hierarchical approach

    USGS Publications Warehouse

    Aldridge, Cameron L.; Saher, D.J.; Childers, T.M.; Stahlnecker, K.E.; Bowen, Z.H.

    2012-01-01

    Gunnison sage-grouse (Centrocercus minimus) is a species of special concern and is currently considered a candidate species under Endangered Species Act. Careful management is therefore required to ensure that suitable habitat is maintained, particularly because much of the species' current distribution is faced with exurban development pressures. We assessed hierarchical nest site selection patterns of Gunnison sage-grouse inhabiting the western portion of the Gunnison Basin, Colorado, USA, at multiple spatial scales, using logistic regression-based resource selection functions. Models were selected using Akaike Information Criterion corrected for small sample sizes (AIC c) and predictive surfaces were generated using model averaged relative probabilities. Landscape-scale factors that had the most influence on nest site selection included the proportion of sagebrush cover >5%, mean productivity, and density of 2 wheel-drive roads. The landscape-scale predictive surface captured 97% of known Gunnison sage-grouse nests within the top 5 of 10 prediction bins, implicating 57% of the basin as crucial nesting habitat. Crucial habitat identified by the landscape model was used to define the extent for patch-scale modeling efforts. Patch-scale variables that had the greatest influence on nest site selection were the proportion of big sagebrush cover >10%, distance to residential development, distance to high volume paved roads, and mean productivity. This model accurately predicted independent nest locations. The unique hierarchical structure of our models more accurately captures the nested nature of habitat selection, and allowed for increased discrimination within larger landscapes of suitable habitat. We extrapolated the landscape-scale model to the entire Gunnison Basin because of conservation concerns for this species. We believe this predictive surface is a valuable tool which can be incorporated into land use and conservation planning as well the assessment of future land-use scenarios. ?? 2011 The Wildlife Society.

  13. Nest poaching in Neotropical parrots

    USGS Publications Warehouse

    Wright, T.F.; Toft, C.A.; Enkerlin-Hoeflich, E.; Gonzalez-Elizondo, J.; Albornoz, M.; Rodriguez-Ferraro, A.; Rojas-Suarez, F.; Sanz, V.; Trujillo, A.; Beissinger, S.R.; Berovides A., V.; Galvez A., X.; Brice, A.T.; Joyner, K.; Eberhard, J.; Gilardi, J.; Koenig, S.E.; Stoleson, S.; Martuscelli, P.; Meyers, J.M.; Renton, K.; Rodriguez, A.M.; Sosa-Asanza, A.C.; Vilella, F.J.; Wiley, J.W.

    2001-01-01

    Although the poaching of nestlings for the pet trade is thought to contribute to the decline of many species of parrots, its effects have been poorly demonstrated. We calculated rates of mortality due to nest poaching in 23 studies of Neotropical parrots, representing 4024 nesting attempts in 21 species and 14 countries. We also examined how poaching rates vary with geographic region, presence of active protection programs, conservation status and economic value of a species, and passage of the U.S. Wild Bird Conservation Act. The average poaching rate across all studies was 30% of all nests observed. Thirteen studies reported poaching rates of >20%, and four reported rates of >70%. Only six studies documented no nest poaching. Of these, four were conducted on islands in the Caribbean region, which had significantly lower poaching rates than the mainland Neotropics. The other two studies that showed no poaching were conducted on the two species with the lowest economic value in our sample (U.S. retail price). In four studies that allowed direct comparison between poaching at sites with active nest protection versus that at unprotected sites, poaching rates were significantly lower at protected sites, suggesting that active protection efforts can be effective in reducing nest poaching. In those studies conducted both before and after the passage of the U.S. Wild Bird Conservation Act, poaching rates were found to be significantly lower following its enactment than in the period before. This result supports the hypothesis that the legal and illegal parrot trades are positively related, rather than inversely related as has been suggested by avicultural interests. Overall, our study indicates that poaching of parrot nestlings for economic gain is a widespread and biologically significant source of nest mortality in Neotropical parrots.

  14. Rapid identification of HPV 16 and 18 by multiplex nested PCR-immunochromatographic test.

    PubMed

    Kuo, Yung-Bin; Li, Yi-Shuan; Chan, Err-Cheng

    2015-02-01

    Human papillomavirus (HPV) types 16 and 18 are known to be high-risk viruses that cause cervical cancer. An HPV rapid testing kit that could help physicians to make early and more informed decisions regarding patient care is needed urgently but not yet available. This study aimed to develop a multiplex nested polymerase chain reaction-immunochromatographic test (PCR-ICT) for the rapid identification of HPV 16 and 18. A multiplex nested PCR was constructed to amplify the HPV 16 and 18 genotype-specific L1 gene fragments and followed by ICT which coated with antibodies to identify rapidly the different PCR products. The type-specific gene regions of high-risk HPV 16 and 18 could be amplified successfully by multiplex nested PCR at molecular sizes of approximately 99 and 101bp, respectively. The capture antibodies raised specifically against the moleculars labeled on the PCR products could be detected simultaneously both HPV 16 and 18 in one strip. Under optimal conditions, this PCR-ICT assay had the capability to detect HPV in a sample with as low as 100 copies of HPV viral DNA. The PCR-ICT system has the advantage of direct and simultaneous detection of two high-risk HPV 16 and 18 DNA targets in one sample, which suggested a significant potential of this assay for clinical application. Copyright © 2014. Published by Elsevier B.V.

  15. Egg laying sequence influences egg mercury concentrations and egg size in three bird species: Implications for contaminant monitoring programs

    USGS Publications Warehouse

    Ackerman, Joshua T.; Eagles-Smith, Collin A.; Herzog, Mark P.; Yee, Julie L.; Hartman, C. Alex

    2016-01-01

    Bird eggs are commonly used in contaminant monitoring programs and toxicological risk assessments, but intra-clutch variation and sampling methodology could influence interpretability. We examined the influence of egg laying sequence on egg mercury concentrations and burdens in American avocets, black-necked stilts, and Forster's terns. The average decline in mercury concentrations between the first and last egg laid was 33% for stilts, 22% for terns, and 11% for avocets, and most of this decline occurred between the first and second eggs laid (24% for stilts, 18% for terns, and 9% for avocets). Trends in egg size with egg laying order were inconsistent among species and overall differences in egg volume, mass, length, and width were <3%. We summarized the literature and, among 17 species studied, mercury concentrations generally declined by 16% between the first and second eggs laid. Despite the strong effect of egg laying sequence, most of the variance in egg mercury concentrations still occurred among clutches (75%-91%) rather than within clutches (9%-25%). Using simulations, we determined that to accurately estimate a population's mean egg mercury concentration using only a single random egg from a subset of nests, it would require sampling >60 nests to represent a large population (10% accuracy) or ≥14 nests to represent a small colony that contained <100 nests (20% accuracy).

  16. Detection of Herpesvirus anguillae (AngHV-1) in European eel Anguilla anguilla (L.) originating from northern Poland-assessment of suitability of selected diagnostic methods.

    PubMed

    Nguyen, Tuan Thuc; Jin, Yeonhwa; Kiełpińska, Jolanta; Bergmann, Sven M; Lenk, Matthias; Panicz, Remigiusz

    2017-11-01

    The Community Action Plan requests EU member states to implement measures that ensure the recovery of the severely depleted European eel stocks. One of the main threats is posed by Anguillid herpesvirus 1 (AngHV-1) leading to increased mortality in both wild and farmed eels. Following recommendations of the OIE to minimize the risk of obtaining false-negative results, the main aim of the study was to optimize diagnostic methods for AngHV-1 detection using conventional PCR, nested PCR and in situ hybridization assay. While 53.3% of the individual organ samples were tested positive for AngHV-1 by PCR, the additional virus analysis via nested PCR revealed that the actual prevalence was 93.3%. In the cell cultivation passages, a cytopathic effect was hardly found in the first two rounds. In the third passage onto cell cultures, a lytic CPE was detected. The identification and confirmation of the viruses obtained from cell cultures as well as directly from the organ tissues were proceeded by PCR, nested PCR and sequencing of the PCR products. While no positive signal was detectable in the first round by PCR using samples from the third cell culture passages, the nested PCR provided weak but visible positive signals. © 2017 John Wiley & Sons Ltd.

  17. Mercury in blood and eggs of the sea turtle Lepidochelys olivacea from a nesting colony in Oaxaca, Mexico.

    PubMed

    Páez-Osuna, F; Calderón-Campuzano, M F; Soto-Jiménez, M F; Ruelas-Inzunza, J

    2011-06-01

    Mercury concentrations were assessed in the sea turtle Lepidochelys olivacea from a nesting colony of Oaxaca, Mexico; 25 female turtles were sampled, a total of 250 eggs were collected during the season 2005-2006. Higher concentrations were found in yolk fraction, while in blood and albumen mean levels were below of 0.0010μg g(-1) dry wt. On the basis of one nesting season, the maternal transfer of Hg via eggs-laying was estimated in 2.0±1.1%. According to international norms, the health of this population and its habitats is acceptable for Hg and corresponds to baseline levels of a nearly pristine environment. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Breeding sex ratios in adult leatherback turtles (Dermochelys coriacea) may compensate for female-biased hatchling sex ratios.

    PubMed

    Stewart, Kelly R; Dutton, Peter H

    2014-01-01

    For vertebrates with temperature-dependent sex determination, primary (or hatchling) sex ratios are often skewed, an issue of particular relevance to concerns over effects of climate change on populations. However, the ratio of breeding males to females, or the operational sex ratio (OSR), is important to understand because it has consequences for population demographics and determines the capacity of a species to persist. The OSR also affects mating behaviors and mate choice, depending on the more abundant sex. For sea turtles, hatchling and juvenile sex ratios are generally female-biased, and with warming nesting beach temperatures, there is concern that populations may become feminized. Our purpose was to evaluate the breeding sex ratio for leatherback turtles at a nesting beach in St. Croix, USVI. In 2010, we sampled nesting females and later sampled their hatchlings as they emerged from nests. Total genomic DNA was extracted and all individuals were genotyped using 6 polymorphic microsatellite markers. We genotyped 662 hatchlings from 58 females, matching 55 females conclusively to their nests. Of the 55, 42 females mated with one male each, 9 mated with 2 males each and 4 mated with at least 3 males each, for a multiple paternity rate of 23.6%. Using GERUD1.0, we reconstructed parental genotypes, identifying 47 different males and 46 females for an estimated breeding sex ratio of 1.02 males for every female. Thus we demonstrate that there are as many actively breeding males as females in this population. Concerns about female-biased adult sex ratios may be premature, and mate choice or competition may play more of a role in sea turtle reproduction than previously thought. We recommend monitoring breeding sex ratios in the future to allow the integration of this demographic parameter in population models.

  19. Wintering area DDE source to migratory white-faced ibis revealed by satellite telemetry and prey sampling.

    PubMed

    Yates, Michael A; Fuller, Mark R; Henny, Charles J; Seegar, William S; Garcia, Jaqueline

    2010-01-01

    Locations of contaminant exposure for nesting migratory species are difficult to fully understand because of possible additional sources encountered during migration or on the wintering grounds. A portion of the migratory white-faced ibis (Plegadis chihi) nesting at Carson Lake, Nevada continues to be exposed to dichloro-diphenyldichloro-ethylene (DDE) with no change, which is unusual, observed in egg concentrations between 1985 and 2000. About 45-63% of the earliest nesting segment shows reduced reproductive success correlated with elevated egg concentrations of >4 microg/g wet weight (ww). Local prey (primarily earthworms) near nests contained little DDE so we tracked the migration and wintering movements of 20 adult males during 2000-2004 to determine the possible source. At various wintering sites, we found a correlation (r (2) = 0.518, P = 0.0125, N = 11) between DDE in earthworm composites and DDE in blood plasma of white-faced ibis wintering there, although the plasma was collected on their breeding grounds soon after arrival. The main source of DDE was wintering areas in the Mexicali Valley of Baja California Norte, Mexico, and probably the adjacent Imperial Valley, California, USA. This unusual continuing DDE problem for white-faced ibis is associated with: the long-term persistence in soil of DDE; the earthworms' ability to bioconcentrate DDE from soil; the proclivity of white-faced ibis to feed on earthworms in agricultural fields; the species's extreme sensitivity to DDE in their eggs; and perhaps its life history strategy of being a "capital breeder". We suggest surveying and sampling white-faced ibis eggs at nesting colonies, especially at Carson Lake, to monitor the continuing influence of DDE.

  20. Development of a highly sensitive real-time nested RT-PCR assay in a single closed tube for detection of enterovirus 71 in hand, foot, and mouth disease.

    PubMed

    Niu, Peihua; Qi, Shunxiang; Yu, Benzhang; Zhang, Chen; Wang, Ji; Li, Qi; Ma, Xuejun

    2016-11-01

    Enterovirus 71 (EV71) is one of the major causative agents of outbreaks of hand, foot, and mouth disease (HFMD). A commercial TaqMan probe-based real-time PCR assay has been widely used for the differential detection of EV71 despite its relatively high cost and failure to detect samples with a low viral load (Ct value > 35). In this study, a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of EV71 in HFMD was developed. The sensitivity and specificity of this assay were evaluated using a reference EV71 stock and a panel of controls consisting of coxsackievirus A16 (CVA16) and common respiratory viruses, respectively. The clinical performance of this assay was evaluated and compared with those of a commercial TaqMan probe-based real-time PCR (qRT-PCR) assay and a traditional two-step nested RT-PCR assay. The limit of detection for the RTN RT-PCR assay was 0.01 TCID50/ml, with a Ct value of 38.3, which was the same as that of the traditional two-step nested RT-PCR assay and approximately tenfold lower than that of the qRT-PCR assay. When testing the reference strain EV71, this assay showed favorable detection reproducibility and no obvious cross-reactivity. The testing results of 100 clinical throat swabs from HFMD-suspected patients revealed that 41 samples were positive for EV71 by both RTN RT-PCR and traditional two-step nested RT-PCR assays, whereas only 29 were EV71 positive by qRT-PCR assay.

  1. Nested PCR detection of malaria directly using blood filter paper samples from epidemiological surveys.

    PubMed

    Li, Peipei; Zhao, Zhenjun; Wang, Ying; Xing, Hua; Parker, Daniel M; Yang, Zhaoqing; Baum, Elizabeth; Li, Wenli; Sattabongkot, Jetsumon; Sirichaisinthop, Jeeraphat; Li, Shuying; Yan, Guiyun; Cui, Liwang; Fan, Qi

    2014-05-08

    Nested PCR is considered a sensitive and specific method for detecting malaria parasites and is especially useful in epidemiological surveys. However, the preparation of DNA templates for PCR is often time-consuming and costly. A simplified PCR method was developed to directly use a small blood filter paper square (2 × 2 mm) as the DNA template after treatment with saponin. This filter paper-based nested PCR method (FP-PCR) was compared to microscopy and standard nested PCR with DNA extracted by using a Qiagen DNA mini kit from filter paper blood spots of 204 febrile cases. The FP-PCR technique was further applied to evaluate malaria infections in 1,708 participants from cross-sectional epidemiological surveys conducted in Myanmar and Thailand. The FP-PCR method had a detection limit of ~0.2 parasites/μL blood, estimated using cultured Plasmodium falciparum parasites. With 204 field samples, the sensitivity of the FP-PCR method was comparable to that of the standard nested PCR method, which was significantly higher than that of microscopy. Application of the FP-PCR method in large cross-sectional studies conducted in Myanmar and Thailand detected 1.9% (12/638) and 6.2% (66/1,070) asymptomatic Plasmodium infections, respectively, as compared to the detection rates of 1.3% (8/638) and 0.04% (4/1,070) by microscopy. This FP-PCR method was much more sensitive than microscopy in detecting Plasmodium infections. It drastically increased the detection sensitivity of asymptomatic infections in cross-sectional surveys conducted in Thailand and Myanmar, suggesting that this FP-PCR method has a potential for future applications in malaria epidemiology studies.

  2. Wintering area DDE source to migratory white-faced ibis revealed by satellite telemetry and prey sampling

    USGS Publications Warehouse

    Yates, M.A.; Fuller, M.R.; Henny, C.J.; Seegar, W.S.; Garcia, Jorge H.

    2010-01-01

    Locations of contaminant exposure for nesting migratory species are difficult to fully understand because of possible additional sources encountered during migration or on the wintering grounds. A portion of the migratory white-faced ibis (Plegadis chihi) nesting at Carson Lake, Nevada continues to be exposed to dichloro-diphenyldichloro-ethylene (DDE) with no change, which is unusual, observed in egg concentrations between 1985 and 2000. About 45-63% of the earliest nesting segment shows reduced reproductive success correlated with elevated egg concentrations of >4 ??g/g wet weight (ww). Local prey (primarily earthworms) near nests contained little DDE so we tracked the migration and wintering movements of 20 adult males during 2000-2004 to determine the possible source. At various wintering sites, we found a correlation (r 2 = 0.518, P = 0.0125, N = 11) between DDE in earthworm composites and DDE in blood plasma of white-faced ibis wintering there, although the plasma was collected on their breeding grounds soon after arrival. The main source of DDE was wintering areas in the Mexicali Valley of Baja California Norte, Mexico, and probably the adjacent Imperial Valley, California, USA. This unusual continuing DDE problem for white-faced ibis is associated with: the long-term persistence in soil of DDE; the earthworms' ability to bioconcentrate DDE from soil; the proclivity of white-faced ibis to feed on earthworms in agricultural fields; the species's extreme sensitivity to DDE in their eggs; and perhaps its life history strategy of being a "capital breeder". We suggest surveying and sampling white-faced ibis eggs at nesting colonies, especially at Carson Lake, to monitor the continuing influence of DDE. ?? 2009 Springer Science+Business Media, LLC.

  3. The efficacy of a nested PCR in detecting cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei var. Hominis for diagnosing scabies.

    PubMed

    Hahm, J E; Kim, C W; Kim, S S

    2018-04-06

    A widespread scabies infestation, associated to long-term residence in nursing homes, is becoming a serious issue in developed countries. Mineral oil examination is regarded as the gold standard in diagnosing scabies, but the sensitivity of this method is generally low-approximately 50%. Molecular tests may contribute to enhance the sensitivity of current tests for laboratory diagnosis of human scabies. In this study, we developed new primers for a nested PCR for the cytochrome c oxidase subunit 1 (cox1) gene of Sarcoptes scabiei var. hominis to increase the sensitivity of a previously developed conventional PCR. Clinically suspected scabies patients underwent dermoscopy-guided skin scraping with microscopic examination. The diagnosis was positive for scabies when mites or eggs were found under the microscope, and patients were then designated as 'microscopy-positive'. Patients in the 'microscopy-negative' group presented with negative microscopic results. Skin scrapings were collected from both groups for PCR. Of the total 63 samples, 28 were microscopy-positive and 35 were negative with no differences in sex and age between the two groups. All microscopically proven scabies cases were positive with the cox1 nested PCR. Among microscopy-negative ones, S. scabiei DNA was detected in 9 samples. If sensitivity of the cox1 nested PCR is considered 100% (95% CI, 90.51-100), then sensitivity of microscopy is 75.68% (95% CI, 58.80-88.23; P = 0.004). Nested PCR can be successfully used as an alternative method for diagnosing suspected scabies patient. Therefore, infection control measures and treatments can be initiated before significant transmission occurs, minimizing the risk of outbreaks. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  4. Chemosterilization of the sea lamprey (Petromyzon marinus)

    USGS Publications Warehouse

    Hanson, Lee H.; Manion, Patrick J.

    1978-01-01

    The chemical, P,P-bis(1-aziridinyl)-N-methylphosphinothioic amide (bisazir), was found in laboratory studies to be an effective sterilant for both sexes of adult sea lampreys (Petromyzon marinus) when given intraperitoneally at a dosage of 100 mg per kilogram of body weight. A total of 300 normal spawning-run sea lampreys and 300 injected with bisazir were released into the Big Garlic River, Marquette County, Michigan, (a small stream divided into five sections by natural barriers), to determine the effect of bisazir on the nesting and spawning behavior of the adults and on the production of larvae. The lampreys constructed and spawned in 95 nests. Sterile adults showed no abnormal nest building or spawning behavior. Sterile males competed effectively with normal males for females. Egg samples taken from nests indicated that eggs in nests where sterile males spawned with sterile or normal females did not hatch, although some embryonic development occurred. Extensive surveys with electric shockers produced no larvae in stream sections where sterile males spawned, but yielded numerous larvae in sections where normal males spawned with normal females. These findings suggest that the release of sterile males may be an effective tool in an integrated approach to control of sea lampreys in the Great Lakes.

  5. Nest predation and circulating corticosterone levels within and among species

    USGS Publications Warehouse

    Fontaine, Joseph J.; Arriero, Elena; Schwabl, Hubert; Martin, Thomas E.

    2011-01-01

    Variation in the risk of predation to offspring can influence the expression of reproductive strategies both within and among species. Appropriate expression of reproductive strategies in environments that differ in predation risk can have clear advantages for fitness. Although adult-predation risk appears to influence glucocorticosteroid levels, leading to changes in behavioral and life-history strategies, the influence of offspring-predation risk on adult glucocorticosteroid levels remains unclear. We compared total baseline corticosterone concentrations in Gray-headed Juncos (Junco hyemalis dorsalis) nesting on plots with and without experimentally reduced risk of nest predation. Despite differences in risk between treatments, we failed to find differences in total baseline corticosterone concentrations. When we examined corticosterone concentrations across a suite of sympatric species, however, higher risk of nest predation correlated with higher total baseline corticosterone levels. As found previously, total baseline corticosterone was negatively correlated with body condition and positively correlated with date of sampling. However, we also found that corticosterone levels increased seasonally, independent of stage of breeding. Nest predation can alter the expression of birds' reproductive strategies, but our findings suggest that total baseline corticosterone is not the physiological mechanism regulating these responses.

  6. Nest predation and circulating corticosterone levels within and among species

    USGS Publications Warehouse

    Fontaine, J.J.; Arriero, E.; Schwabl, H.; Martin, T.E.

    2011-01-01

    Variation in the risk of predation to offspring can influence the expression of reproductive strategies both within and among species. Appropriate expression of reproductive strategies in environments that differ in predation risk can have clear advantages for fitness. Although adult-predation risk appears to influence glucocorticosteroid levels, leading to changes in behavioral and life-history strategies, the influence of offspring-predation risk on adult glucocorticosteroid levels remains unclear. We compared total baseline corticosterone concentrations in Gray-headed Juncos (Junco hyemalis dorsalis) nesting on plots with and without experimentally reduced risk of nest predation. Despite differences in risk between treatments, we failed to find differences in total baseline corticosterone concentrations. When we examined corticosterone concentrations across a suite of sympatric species, however, higher risk of nest predation correlated with higher total baseline corticosterone levels. As found previously, total baseline corticosterone was negatively correlated with body condition and positively correlated with date of sampling. However, we also found that corticosterone levels increased seasonally, independent of stage of breeding. Nest predation can alter the expression of birds' reproductive strategies, but our findings suggest that total baseline corticosterone is not the physiological mechanism regulating these responses. ?? The Cooper Ornithological Society 2011.

  7. Anti-parasite treatment, but not mercury burdens, influence nesting propensity dependent on arrival time or body condition in a marine bird.

    PubMed

    Provencher, J F; Forbes, M R; Mallory, M L; Wilson, S; Gilchrist, H G

    2017-01-01

    Arctic wildlife can be exposed to high mercury (Hg) levels, and are also naturally exposed to gastrointestinal parasites that can reduce condition and negatively affect reproductive output and/or survival in similar ways. Importantly, both Hg and parasites are increasing in wildlife in some Arctic regions. We studied the northern common eider duck (Somateria mollissima) to explore how Hg in association with both natural levels and experimentally reduced parasitic infections, affect reproduction and survival. Female eiders were measured, banded, and blood sampled to determine blood Hg burdens, prior to breeding. Propensity to nest, clutch size, nest survival, nest attendance, and return rates were assessed in relation to both Hg burden and parasite treatment. Neither reproduction nor return rates of females varied with Hg concentrations, but females arriving late to the colony, or in low body condition, showed increased nesting propensity when given the anti-parasite treatment as compared to placebo treatment. Our results suggest that parasites can play a critical role in decisions to invest in avian breeding annually, particularly among individuals with a late onset to breeding, and in poor condition. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  8. Compilation of 1990 annual reports of the Navy ELF Communications System Ecological-Monitoring Program. Volume 2. Tabs C thru F. Annual report, Jan-Dec 90

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zapotosky, J.E.

    1991-08-01

    This portion of the report includes monitoring of and data for arthropoda and earthworms; pollinating insects; and small mammals and nesting birds. During the 1990 growing season the ELF antenna was operated more frequently than in prior years. This provides 2 years of intermittent ELF exposure for the biological systems to react to the radiation, one year of very limited exposure and greater exposure in 1990. Arthropod and earthworm sampling was conducted at intervals of two weeks from early May to late October. High voltage transmission lines and magnetic fields have been shown to affect honeybee reproduction, survival, orientation, andmore » nest structure. ELF EM fields could have similar effects on native megachild bees. Changes in cell length, number of cells per nest, number of leaver per cell, orientation of nest entrances, and time to collect a round leaf pierce to cap a cell were monitored. We have not detected significant changes that could be attributed to ELF EM fields. Small mammal and nesting bird biological studies in the western Upper Peninsula of Michigan for the year 1990 are reported.« less

  9. Biomarker discovery study design for type 1 diabetes in The Environmental Determinants of Diabetes in the Young (TEDDY) study.

    PubMed

    Lee, Hye-Seung; Burkhardt, Brant R; McLeod, Wendy; Smith, Susan; Eberhard, Chris; Lynch, Kristian; Hadley, David; Rewers, Marian; Simell, Olli; She, Jin-Xiong; Hagopian, Bill; Lernmark, Ake; Akolkar, Beena; Ziegler, Anette G; Krischer, Jeffrey P

    2014-07-01

    The Environmental Determinants of Diabetes in the Young planned biomarker discovery studies on longitudinal samples for persistent confirmed islet cell autoantibodies and type 1 diabetes using dietary biomarkers, metabolomics, microbiome/viral metagenomics and gene expression. This article describes the details of planning The Environmental Determinants of Diabetes in the Young biomarker discovery studies using a nested case-control design that was chosen as an alternative to the full cohort analysis. In the frame of a nested case-control design, it guides the choice of matching factors, selection of controls, preparation of external quality control samples and reduction of batch effects along with proper sample allocation. Our design is to reduce potential bias and retain study power while reducing the costs by limiting the numbers of samples requiring laboratory analyses. It also covers two primary end points (the occurrence of diabetes-related autoantibodies and the diagnosis of type 1 diabetes). The resulting list of case-control matched samples for each laboratory was augmented with external quality control samples. Copyright © 2013 John Wiley & Sons, Ltd.

  10. Information flow, opinion polling and collective intelligence in house-hunting social insects.

    PubMed

    Franks, Nigel R; Pratt, Stephen C; Mallon, Eamonn B; Britton, Nicholas F; Sumpter, David J T

    2002-11-29

    The sharing and collective processing of information by certain insect societies is one of the reasons that they warrant the superlative epithet 'super-organisms' (Franks 1989, Am. Sci. 77, 138-145). We describe a detailed experimental and mathematical analysis of information exchange and decision-making in, arguably, the most difficult collective choices that social insects face: namely, house hunting by complete societies. The key issue is how can a complete colony select the single best nest-site among several alternatives? Individual scouts respond to the diverse information they have personally obtained about the quality of a potential nest-site by producing a recruitment signal. The colony then deliberates over (i.e. integrates) different incoming recruitment signals associated with different potential nest-sites to achieve a well-informed collective decision. We compare this process in honeybees and in the ant Leptothorax albipennis. Notwithstanding many differences - for example, honeybee colonies have 100 times more individuals than L. albipennis colonies - there are certain similarities in the fundamental algorithms these societies appear to employ when they are house hunting. Scout honeybees use the full power of the waggle dance to inform their nest-mates about the distance and direction of a potential nest-site (and they indicate the quality of a nest-site indirectly through the vigour of their dance), and yet individual bees perhaps only rarely make direct comparisons of such sites. By contrast, scouts from L. albipennis colonies often compare nest-sites, but they cannot directly inform one another of their estimation of the quality of a potential site. Instead, they discriminate between sites by initiating recruitment sooner to better ones. Nevertheless, both species do make use of forms of opinion polling. For example, scout bees that have formerly danced for a certain site cease such advertising and monitor the dances of others at random. That is, they act without prejudice. They neither favour nor disdain dancers that advocate the site they had formerly advertised or the alternatives. Thus, in general the bees are less well informed than they would be if they systematically monitored dances for alternative sites rather than spending their time reprocessing information they already have. However, as a result of their lack of prejudice, less time overall will be wasted in endless debate among stubborn and potentially biased bees. Among the ants, the opinions of nest-mates are also pooled effectively when scouts use a threshold population of their nest-mates present in a new nest-site as a cue to switch to more rapid recruitment. Furthermore, the ants' reluctance to begin recruiting to poor nest-sites means that more time is available for the discovery and direct comparison of alternatives. Likewise, the retirement of honeybee scouts from dancing for a given site allows more time for other scouts to find potentially better sites. Thus, both the ants and the bees have time-lags built into their decision-making systems that should facilitate a compromise between thorough surveys for good nest-sites and relatively rapid decisions. We have also been able to show that classical mathematical models can illuminate the processes by which colonies are able to achieve decisions that are relatively swift and very well informed.

  11. Information flow, opinion polling and collective intelligence in house-hunting social insects.

    PubMed Central

    Franks, Nigel R; Pratt, Stephen C; Mallon, Eamonn B; Britton, Nicholas F; Sumpter, David J T

    2002-01-01

    The sharing and collective processing of information by certain insect societies is one of the reasons that they warrant the superlative epithet 'super-organisms' (Franks 1989, Am. Sci. 77, 138-145). We describe a detailed experimental and mathematical analysis of information exchange and decision-making in, arguably, the most difficult collective choices that social insects face: namely, house hunting by complete societies. The key issue is how can a complete colony select the single best nest-site among several alternatives? Individual scouts respond to the diverse information they have personally obtained about the quality of a potential nest-site by producing a recruitment signal. The colony then deliberates over (i.e. integrates) different incoming recruitment signals associated with different potential nest-sites to achieve a well-informed collective decision. We compare this process in honeybees and in the ant Leptothorax albipennis. Notwithstanding many differences - for example, honeybee colonies have 100 times more individuals than L. albipennis colonies - there are certain similarities in the fundamental algorithms these societies appear to employ when they are house hunting. Scout honeybees use the full power of the waggle dance to inform their nest-mates about the distance and direction of a potential nest-site (and they indicate the quality of a nest-site indirectly through the vigour of their dance), and yet individual bees perhaps only rarely make direct comparisons of such sites. By contrast, scouts from L. albipennis colonies often compare nest-sites, but they cannot directly inform one another of their estimation of the quality of a potential site. Instead, they discriminate between sites by initiating recruitment sooner to better ones. Nevertheless, both species do make use of forms of opinion polling. For example, scout bees that have formerly danced for a certain site cease such advertising and monitor the dances of others at random. That is, they act without prejudice. They neither favour nor disdain dancers that advocate the site they had formerly advertised or the alternatives. Thus, in general the bees are less well informed than they would be if they systematically monitored dances for alternative sites rather than spending their time reprocessing information they already have. However, as a result of their lack of prejudice, less time overall will be wasted in endless debate among stubborn and potentially biased bees. Among the ants, the opinions of nest-mates are also pooled effectively when scouts use a threshold population of their nest-mates present in a new nest-site as a cue to switch to more rapid recruitment. Furthermore, the ants' reluctance to begin recruiting to poor nest-sites means that more time is available for the discovery and direct comparison of alternatives. Likewise, the retirement of honeybee scouts from dancing for a given site allows more time for other scouts to find potentially better sites. Thus, both the ants and the bees have time-lags built into their decision-making systems that should facilitate a compromise between thorough surveys for good nest-sites and relatively rapid decisions. We have also been able to show that classical mathematical models can illuminate the processes by which colonies are able to achieve decisions that are relatively swift and very well informed. PMID:12495514

  12. Distribution of Hepatitis C virus (HCV) genotypes in seropositive patients in the state of Alagoas, Brazil

    PubMed Central

    Gonzaga, Rosa Maria S.; Rodart, Itatiana F.; Reis, Mitermayer Galvão; Ramalho Neto, Cícero Eduardo; Silva, Denise Wanderlei

    2008-01-01

    We determined the frequency of hepatitis C virus (HCV) genotypes in anti-HCV seropositive patients in the state of Alagoas, Brazil, by means of nested-reverse transcription-polymerase chain reaction (RT-nested-PCR) followed by restriction fragment length polymorphism (RFLP) of amplified fragments of the 5´NCR. The nested-PCR with genotype-specific primers from the core region was carried out when detection was not possible by the first approach. Detectable HCV-RNA was present in 115 (74.7%) of 154 serum samples. Genotype 1 was the most frequent (77.4%), against 20.9% of genotype 3 and 0.8% of genotype 2. Subtype 1b was predominant (65.2%), followed by subtypes 1a (8.7%), and 3a (6.1%). Coinfection (1a/3a) was detected in 0.8% of the samples. Indeed, there was no significant differences in the prevalence of genotype 1 compared to what has been obtained from anti-HCV seropositive patients from other locations in Brazil. Here we report for the first time the genotype 2 in the state of Alagoas. PMID:24031281

  13. About improving efficiency of the P3 M algorithms when computing the inter-particle forces in beam dynamics

    NASA Astrophysics Data System (ADS)

    Kozynchenko, Alexander I.; Kozynchenko, Sergey A.

    2017-03-01

    In the paper, a problem of improving efficiency of the particle-particle- particle-mesh (P3M) algorithm in computing the inter-particle electrostatic forces is considered. The particle-mesh (PM) part of the algorithm is modified in such a way that the space field equation is solved by the direct method of summation of potentials over the ensemble of particles lying not too close to a reference particle. For this purpose, a specific matrix "pattern" is introduced to describe the spatial field distribution of a single point charge, so the "pattern" contains pre-calculated potential values. This approach allows to reduce a set of arithmetic operations performed at the innermost of nested loops down to an addition and assignment operators and, therefore, to decrease the running time substantially. The simulation model developed in C++ substantiates this view, showing the descent accuracy acceptable in particle beam calculations together with the improved speed performance.

  14. Inferences about nested subsets structure when not all species are detected

    USGS Publications Warehouse

    Cam, E.; Nichols, J.D.; Hines, J.E.; Sauer, J.R.

    2000-01-01

    Comparisons of species composition among ecological communities of different size have often provided evidence that the species in communities with lower species richness form nested subsets of the species in larger communities. In the vast majority of studies, the question of nested subsets has been addressed using information on presence-absence, where a '0' is interpreted as the absence of a given species from a given location. Most of the methodological discussion in earlier studies investigating nestedness concerns the approach to generation of model-based matrices. However, it is most likely that in many situations investigators cannot detect all the species present in the location sampled. The possibility that zeros in incidence matrices reflect nondetection rather than absence of species has not been considered in studies addressing nested subsets, even though the position of zeros in these matrices forms the basis of earlier inference methods. These sampling artifacts are likely to lead to erroneous conclusions about both variation over space in species richness and the degree of similarity of the various locations. Here we propose an approach to investigation of nestedness, based on statistical inference methods explicitly incorporating species detection probability, that take into account the probabilistic nature of the sampling process. We use presence-absence data collected under Pollock?s robust capture-recapture design, and resort to an estimator of species richness originally developed for closed populations to assess the proportion of species shared by different locations. We develop testable predictions corresponding to the null hypothesis of a nonnested pattern, and an alternative hypothesis of perfect nestedness. We also present an index for assessing the degree of nestedness of a system of ecological communities. We illustrate our approach using avian data from the North American Breeding Bird Survey collected in Florida Keys.

  15. Evaluation of four molecular methods to detect Leishmania infection in dogs.

    PubMed

    Albuquerque, Andreia; Campino, Lenea; Cardoso, Luís; Cortes, Sofia

    2017-03-13

    Canine leishmaniasis, a zoonotic disease caused by Leishmania infantum vectored by phlebotomine sand flies, is considered a relevant veterinary and public health problem in various countries, namely in the Mediterranean basin and Brazil, where dogs are considered the main reservoir hosts. Not only diseased dogs but also those subclinically infected play a relevant role in the transmission of L. infantum to vectors; therefore, early diagnosis is essential, under both a clinical and an epidemiological perspective. Molecular tools can be a more accurate and sensitive approach for diagnosis, with a wide range of protocols currently in use. The aim of the present report was to compare four PCR based protocols for the diagnosis of canine Leishmania infection in a cohort of dogs from the Douro region, Portugal. A total of 229 bone marrow samples were collected from dogs living in the Douro region, an endemic region for leishmaniasis. Four PCR protocols were evaluated for Leishmania DNA detection in canine samples, three single (ITS1-PCR, MC-PCR and Uni21/Lmj4-PCR) and one nested (nested SSU rRNA-PCR). Two of the protocols were based on nuclear targets and the other two on kinetoplastid targets. The higher overall percentage of infected dogs was detected with the nested SSU rRNA-PCR (37.6%), which also was able to detect Leishmania DNA in a higher number of samples from apparently healthy dogs (25.3%). The ITS1-PCR presented the lowest level of Leishmania detection. Nested SSU rRNA-PCR is an appropriate method to detect Leishmania infection in dogs. Accurate and early diagnosis in clinically suspect as well as apparently healthy dogs is essential, in order to treat and protect animals and public health and contribute to the control and awareness of the disease.

  16. Detection of Leishmania DNA in saliva among patients with HIV/AIDS in Trang Province, southern Thailand.

    PubMed

    Pandey, Netranapha; Siripattanapipong, Suradej; Leelayoova, Saovanee; Manomat, Jipada; Mungthin, Mathirut; Tan-Ariya, Peerapan; Bualert, Lertwut; Naaglor, Tawee; Siriyasatien, Padet; Phumee, Atchara; Piyaraj, Phunlerd

    2018-06-08

    Leishmaniasis is a neglected tropical disease causing opportunistic infection among patients with HIV/AIDS. The fatal form of this disease is visceral leishmaniasis (VL). DNA of Leishmania can be detected in saliva, for which the collection is noninvasive and requires little expertise. This study aimed to evaluate the sensitivity and specificity of a nested-PCR to amplify the Internal Transcribed Spacer 1 (ITS1) to detect Leishmania DNA in paired saliva and buffy coat samples of 305 Thai patients with HIV/AIDS in Trang Hospital, Trang Province, southern Thailand. For asymptomatic Leishmania infection among Thai patients with HIV/AIDS, the sensitivity and specificity of the nested-PCR-ITS1 in buffy coat were 73.9 and 100%, respectively. However, the sensitivity in saliva was 26.1% and specificity was 100%. Using the nested-PCR-ITS1, saliva and buffy coat samples showed positive agreement in only 52.0% of patients. Saliva tested results with the nested-PCR-ITS1 showed positive agreement with the Direct Agglutination Test (DAT) in 46.5% of patients. Only 12.1% of the samples showed positive agreement for Leishmania infection among all the three tests: saliva, buffy coat and DAT results. Using nucleotide sequencing, at least three species of Leishmania infection were identified in saliva, i.e., L. siamensis (n = 28), L. martiniquensis (n = 9), and L. donovani complex (n = 1). As a result, buffy coat still appears to be a better specimen to diagnose asymptomatic VL infection among individuals with HIV. However, the use of both buffy coat and saliva together as clinical specimens would increase the sensitivity of Leishmania detection. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. Asynchronous Distributed Flow Control Algorithms.

    DTIC Science & Technology

    1984-10-01

    all yeX, y <x. We may think of X as a "feasible" set. The fair allocation vector solves the following set of nested problems. The first problem is to...interval parameters that can be adjusted: the time between a succesful update and the next update attempt, and the time between an unsuccessful attempt...For years she took great delight in being able to tell people , quite truthfully, that she was from Mars. In 1970, she graduated from the University of

  18. OpenMP Parallelization and Optimization of Graph-based Machine Learning Algorithms

    DTIC Science & Technology

    2016-05-01

    composed of hyper - spectral video sequences recording the release of chemical plumes at the Dugway Proving Ground. We use the 329 frames of the...video. Each frame is a hyper - spectral image with dimension 128 × 320 × 129, where 129 is the dimension of the channel of each pixel. The total number of...j=1 . Then we use the nested for- loop to calculate the values of WXY by the formula (1). We then put the corresponding value in an array which

  19. Molecular surveillance of Theileria ovis, Theileria lestoquardi and Theileria annulata infection in sheep and ixodid ticks in Iran.

    PubMed

    Razmi, Gholamreza; Yaghfoori, Saeed

    2013-01-01

    A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulata in sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T.annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.

  20. Comparison of a conventional and nested PCR for diagnostic confirmation and genotyping of Orientia tsutsugamushi.

    PubMed

    Janardhanan, Jeshina; Prakash, John Antony Jude; Abraham, Ooriapadickal C; Varghese, George M

    2014-05-01

    A nested polymerase chain reaction (PCR) targeting the 56-kDa antigen gene is currently the most commonly used molecular technique for confirmation of scrub typhus and genotyping of Orientia tsutsugamushi. In this study, we have compared the commonly used nested PCR (N-PCR) with a single-step conventional PCR (C-PCR) for amplification and genotyping. Eschar samples collected from 24 patients with scrub typhus confirmed by IgM enzyme-linked immunosorbent assay were used for DNA extraction following which amplifications were carried out using nested and C-PCR methods. The amplicons were sequenced and compared to other sequences in the database using BLAST. Conventional PCR showed a high positivity rate of 95.8% compared to the 75% observed using N-PCR. On sequence analysis, the N-PCR amplified region showed more variation among strains than the C-PCR amplified region. The C-PCR, which is more economical, provided faster and better results compared to N-PCR. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Landscapes for Energy and Wildlife: Conservation Prioritization for Golden Eagles across Large Spatial Scales

    PubMed Central

    Tack, Jason D.; Fedy, Bradley C.

    2015-01-01

    Proactive conservation planning for species requires the identification of important spatial attributes across ecologically relevant scales in a model-based framework. However, it is often difficult to develop predictive models, as the explanatory data required for model development across regional management scales is rarely available. Golden eagles are a large-ranging predator of conservation concern in the United States that may be negatively affected by wind energy development. Thus, identifying landscapes least likely to pose conflict between eagles and wind development via shared space prior to development will be critical for conserving populations in the face of imposing development. We used publically available data on golden eagle nests to generate predictive models of golden eagle nesting sites in Wyoming, USA, using a suite of environmental and anthropogenic variables. By overlaying predictive models of golden eagle nesting habitat with wind energy resource maps, we highlight areas of potential conflict among eagle nesting habitat and wind development. However, our results suggest that wind potential and the relative probability of golden eagle nesting are not necessarily spatially correlated. Indeed, the majority of our sample frame includes areas with disparate predictions between suitable nesting habitat and potential for developing wind energy resources. Map predictions cannot replace on-the-ground monitoring for potential risk of wind turbines on wildlife populations, though they provide industry and managers a useful framework to first assess potential development. PMID:26262876

  2. Landscapes for energy and wildlife: conservation prioritization for golden eagles across large spatial scales

    USGS Publications Warehouse

    Tack, Jason D.; Fedy, Bradley C.

    2015-01-01

    Proactive conservation planning for species requires the identification of important spatial attributes across ecologically relevant scales in a model-based framework. However, it is often difficult to develop predictive models, as the explanatory data required for model development across regional management scales is rarely available. Golden eagles are a large-ranging predator of conservation concern in the United States that may be negatively affected by wind energy development. Thus, identifying landscapes least likely to pose conflict between eagles and wind development via shared space prior to development will be critical for conserving populations in the face of imposing development. We used publically available data on golden eagle nests to generate predictive models of golden eagle nesting sites in Wyoming, USA, using a suite of environmental and anthropogenic variables. By overlaying predictive models of golden eagle nesting habitat with wind energy resource maps, we highlight areas of potential conflict among eagle nesting habitat and wind development. However, our results suggest that wind potential and the relative probability of golden eagle nesting are not necessarily spatially correlated. Indeed, the majority of our sample frame includes areas with disparate predictions between suitable nesting habitat and potential for developing wind energy resources. Map predictions cannot replace on-the-ground monitoring for potential risk of wind turbines on wildlife populations, though they provide industry and managers a useful framework to first assess potential development.

  3. A survey of nested grid techniques and their potential for use within the MASS weather prediction model

    NASA Technical Reports Server (NTRS)

    Koch, Steven E.; Mcqueen, Jeffery T.

    1987-01-01

    A survey of various one- and two-way interactive nested grid techniques used in hydrostatic numerical weather prediction models is presented and the advantages and disadvantages of each method are discussed. The techniques for specifying the lateral boundary conditions for each nested grid scheme are described in detail. Averaging and interpolation techniques used when applying the coarse mesh grid (CMG) and fine mesh grid (FMG) interface conditions during two-way nesting are discussed separately. The survey shows that errors are commonly generated at the boundary between the CMG and FMG due to boundary formulation or specification discrepancies. Methods used to control this noise include application of smoothers, enhanced diffusion, or damping-type time integration schemes to model variables. The results from this survey provide the information needed to decide which one-way and two-way nested grid schemes merit future testing with the Mesoscale Atmospheric Simulation System (MASS) model. An analytically specified baroclinic wave will be used to conduct systematic tests of the chosen schemes since this will allow for objective determination of the interfacial noise in the kind of meteorological setting for which MASS is designed. Sample diagnostic plots from initial tests using the analytic wave are presented to illustrate how the model-generated noise is ascertained. These plots will be used to compare the accuracy of the various nesting schemes when incorporated into the MASS model.

  4. Landscapes for Energy and Wildlife: Conservation Prioritization for Golden Eagles across Large Spatial Scales.

    PubMed

    Tack, Jason D; Fedy, Bradley C

    2015-01-01

    Proactive conservation planning for species requires the identification of important spatial attributes across ecologically relevant scales in a model-based framework. However, it is often difficult to develop predictive models, as the explanatory data required for model development across regional management scales is rarely available. Golden eagles are a large-ranging predator of conservation concern in the United States that may be negatively affected by wind energy development. Thus, identifying landscapes least likely to pose conflict between eagles and wind development via shared space prior to development will be critical for conserving populations in the face of imposing development. We used publically available data on golden eagle nests to generate predictive models of golden eagle nesting sites in Wyoming, USA, using a suite of environmental and anthropogenic variables. By overlaying predictive models of golden eagle nesting habitat with wind energy resource maps, we highlight areas of potential conflict among eagle nesting habitat and wind development. However, our results suggest that wind potential and the relative probability of golden eagle nesting are not necessarily spatially correlated. Indeed, the majority of our sample frame includes areas with disparate predictions between suitable nesting habitat and potential for developing wind energy resources. Map predictions cannot replace on-the-ground monitoring for potential risk of wind turbines on wildlife populations, though they provide industry and managers a useful framework to first assess potential development.

  5. Nested generalized linear mixed model with ordinal response: Simulation and application on poverty data in Java Island

    NASA Astrophysics Data System (ADS)

    Widyaningsih, Yekti; Saefuddin, Asep; Notodiputro, Khairil A.; Wigena, Aji H.

    2012-05-01

    The objective of this research is to build a nested generalized linear mixed model using an ordinal response variable with some covariates. There are three main jobs in this paper, i.e. parameters estimation procedure, simulation, and implementation of the model for the real data. At the part of parameters estimation procedure, concepts of threshold, nested random effect, and computational algorithm are described. The simulations data are built for 3 conditions to know the effect of different parameter values of random effect distributions. The last job is the implementation of the model for the data about poverty in 9 districts of Java Island. The districts are Kuningan, Karawang, and Majalengka chose randomly in West Java; Temanggung, Boyolali, and Cilacap from Central Java; and Blitar, Ngawi, and Jember from East Java. The covariates in this model are province, number of bad nutrition cases, number of farmer families, and number of health personnel. In this modeling, all covariates are grouped as ordinal scale. Unit observation in this research is sub-district (kecamatan) nested in district, and districts (kabupaten) are nested in province. For the result of simulation, ARB (Absolute Relative Bias) and RRMSE (Relative Root of mean square errors) scale is used. They show that prov parameters have the highest bias, but more stable RRMSE in all conditions. The simulation design needs to be improved by adding other condition, such as higher correlation between covariates. Furthermore, as the result of the model implementation for the data, only number of farmer family and number of medical personnel have significant contributions to the level of poverty in Central Java and East Java province, and only district 2 (Karawang) of province 1 (West Java) has different random effect from the others. The source of the data is PODES (Potensi Desa) 2008 from BPS (Badan Pusat Statistik).

  6. Sedimentology, geochemistry and rock magnetic properties of beach sands in Galapagos Islands - implications for nesting marine turtles

    NASA Astrophysics Data System (ADS)

    Perez-Cruz, L.; Urrutia-Fucugauchi, J.; Vazquez-Gutierrez, F.; Carranza-Edwards, A.

    2007-12-01

    Marine turtles are well known for their navigation ability in the open ocean and fidelity to nesting beaches. Green turtle adult females migrate from foraging areas to island nesting beaches, traveling hundreds or thousands of kilometers each way. The marine turtle breeding in the Galapagos Islands is the Green Sea Turtle (Chelonia mydas agassisi); fairly common throughout the islands but with nesting sites located at Las Bachas (Santa Cruz), Barahona and Quinta Playa (Isabela), Salinas (Baltra), Gardner Bay (Española) and Bartolomé Islet. In order to characterize and to identify the geochemical signature of nesting marine turtle beaches in Galapagos Islands, sedimentological, geochemical and rock magnetic parameters are used. A total of one hundred and twenty sand samples were collected in four beaches to relate compositional characteristics between equivalent areas, these are: Las Bachas, Salinas, Barahona and Quinta Playa. Grain size is evaluated using laser particle analysis (Model Coulter LS 230). Bulk ICP-MS geochemical analysis is performed, following trace elements are analyzed: Al, V, Cr, Co, Ni, Cu, Zn, Cd, Ba, Pb, Fe, Mn, K, Na, Mg, Sr, Ca and Hg; and low-field magnetic susceptibility is measured in all samples at low and high frequencies. Granulometric analysis showed that Barahona and Quinta Playa are characterized for fine grained sands. In contrast, Salinas and Las Bachas exhibit medium to coarse sands. Trace metals concentrations and magnetic susceptibility show different distribution patterns in the beach sands. Calcium is the most abundant element in the samples. In particular, Co, K, and Na show similar concentrations in the four beaches. Las Bachas beach shows highest concentrations of Pb and Hg (maximum values 101.1 and 118.5 mg/kg, respectively), we suggest that the enrichment corresponds to an anthropogenic signal. Salinas beach samples show high concentrations of Fe, V, Cr, Zn, Mn and the highest values of magnetic susceptibility (maximum 932 10-6 SI), we propose, a human activity influence less evident than in Bachas beach that could overlap the contribution of continent source. Quinta Playa sands show the maximum concentration of calcium and also high concentration of Fe and Mg, and relatively high values of magnetic susceptibility. Ca results from marine biogenic carbonates (mainly coral reefs). Barahona also show high concentrations in calcium that could be correlated with the presence of biogenous source around the beach.

  7. Factors affecting hatch success of hawksbill sea turtles on Long Island, Antigua, West Indies.

    PubMed

    Ditmer, Mark Allan; Stapleton, Seth Patrick

    2012-01-01

    Current understanding of the factors influencing hawksbill sea turtle (Eretmochelys imbricata) hatch success is disparate and based on relatively short-term studies or limited sample sizes. Because global populations of hawksbills are heavily depleted, evaluating the parameters that impact hatch success is important to their conservation and recovery. Here, we use data collected by the Jumby Bay Hawksbill Project (JBHP) to investigate hatch success. The JBHP implements saturation tagging protocols to study a hawksbill rookery in Antigua, West Indies. Habitat data, which reflect the varied nesting beaches, are collected at egg deposition, and nest contents are exhumed and categorized post-emergence. We analyzed hatch success using mixed-model analyses with explanatory and predictive datasets. We incorporated a random effect for turtle identity and evaluated environmental, temporal and individual-based reproductive variables. Hatch success averaged 78.6% (SD: 21.2%) during the study period. Highly supported models included multiple covariates, including distance to vegetation, deposition date, individual intra-seasonal nest number, clutch size, organic content, and sand grain size. Nests located in open sand were predicted to produce 10.4 more viable hatchlings per clutch than nests located >1.5 m into vegetation. For an individual first nesting in early July, the fourth nest of the season yielded 13.2 more viable hatchlings than the initial clutch. Generalized beach section and inter-annual variation were also supported in our explanatory dataset, suggesting that gaps remain in our understanding of hatch success. Our findings illustrate that evaluating hatch success is a complex process, involving multiple environmental and individual variables. Although distance to vegetation and hatch success were inversely related, vegetation is an important component of hawksbill nesting habitat, and a more complete assessment of the impacts of specific vegetation types on hatch success and hatchling sex ratios is needed. Future research should explore the roles of sand structure, nest moisture, and local weather conditions.

  8. Reproductive biology of Pleurodema guayapae (Anura: Leptodactylidae: Leiuperinae).

    PubMed

    Valetti, Julián Alonso; Grenat, Pablo Raúl; Baraquet, Mariana; Martino, Adolfo Ludovico

    2014-03-01

    Pleurodema guayapae is a species that inhabits saline environments and semidesert zones from central Argentina. To date, the knowledge about the reproductive biology of this species is very poor, and our aim is to contribute to its knowledge with the description of some important reproductive aspects. For this, field work was undertaken in an area near to Patquia, La Rioja province. Sampling was undertaken during three summer periods (2006-2007; 2007-2008; 2008-2009) in Chamical-Patquia area, where we could find reproductively active populations. We observed and described breeding sites, type of clutch, process of foam nest construction, clutch and egg number and sizes, and hatching time and stage. Behaviour observations were performed from the time that males began to call until the pairs ended up the foam nests building, and layed the eggs. Additionally, one amplected pair was observed and filmed in the process of foam nest construction, and four amplectant pairs were collected and separatelly placed in plastic containers, for nests observations in the laboratory. Hatching time was based on three different foam nests of known age. We found that P. guayapae populations were acoustically active only after a rainfall. Its breeding sites were represented by ephemeral ponds of fresh water, product of rains. The males emitted their calls inside or outside these ponds. A detailed description of the foam nest construction process by both females and males was made. The clutches were in dome-shaped foam nest type of 6-9cm in diameter and 1-3cm in height, some of which were in communal nests. The nests had an average of 1 137 pigmented eggs. This species showed a short hatching time. Our results allow us to conclude that this species should be considered an extreme explosive breeder. Our results are discussed with others obtained for related species.

  9. Factors Affecting Hatch Success of Hawksbill Sea Turtles on Long Island, Antigua, West Indies

    PubMed Central

    Ditmer, Mark Allan; Stapleton, Seth Patrick

    2012-01-01

    Current understanding of the factors influencing hawksbill sea turtle (Eretmochelys imbricata) hatch success is disparate and based on relatively short-term studies or limited sample sizes. Because global populations of hawksbills are heavily depleted, evaluating the parameters that impact hatch success is important to their conservation and recovery. Here, we use data collected by the Jumby Bay Hawksbill Project (JBHP) to investigate hatch success. The JBHP implements saturation tagging protocols to study a hawksbill rookery in Antigua, West Indies. Habitat data, which reflect the varied nesting beaches, are collected at egg deposition, and nest contents are exhumed and categorized post-emergence. We analyzed hatch success using mixed-model analyses with explanatory and predictive datasets. We incorporated a random effect for turtle identity and evaluated environmental, temporal and individual-based reproductive variables. Hatch success averaged 78.6% (SD: 21.2%) during the study period. Highly supported models included multiple covariates, including distance to vegetation, deposition date, individual intra-seasonal nest number, clutch size, organic content, and sand grain size. Nests located in open sand were predicted to produce 10.4 more viable hatchlings per clutch than nests located >1.5 m into vegetation. For an individual first nesting in early July, the fourth nest of the season yielded 13.2 more viable hatchlings than the initial clutch. Generalized beach section and inter-annual variation were also supported in our explanatory dataset, suggesting that gaps remain in our understanding of hatch success. Our findings illustrate that evaluating hatch success is a complex process, involving multiple environmental and individual variables. Although distance to vegetation and hatch success were inversely related, vegetation is an important component of hawksbill nesting habitat, and a more complete assessment of the impacts of specific vegetation types on hatch success and hatchling sex ratios is needed. Future research should explore the roles of sand structure, nest moisture, and local weather conditions. PMID:22802928

  10. Hepatitis E Virus in Wild Boar in Northwest Poland: Sensitivity of Methods of Detection.

    PubMed

    Dorn-In, Samart; Schwaiger, Karin; Twarużek, Magdalena; Grajewski, Jan; Gottschalk, Christoph; Gareis, Manfred

    2017-02-01

    In northwest Poland, 163 blood and 53 fecal samples of wild boars were collected in winter 2012/13 and 2013/14. All blood samples were tested for the presence of hepatitis E virus (HEV) ribonucleic acid (RNA) by two reverse transcription-polymerase chain reaction (RT-PCR) based methods and by anti-HEV IgG enzyme-linked immunosorbent assay (ELISA). About 17.2% of blood samples were seropositive. One-step nested RT-PCR turned out to be too insensitive (11.6% were positive). Therefore a two-step nested RT-PCR was applied where 25.8% of the blood samples were tested positive for HEV RNA. About 50.0% of blood samples positive in ELISA were also positive in two-step nested RT-PCR. The prevalence of HEV RNA in feces was 9.4%. Based on the results of blood (ELISA, PCR) and fecal (PCR) tests, the overall prevalence of HEV in wild boars in northwest Poland was 36.8%. There was no correlation between the ELISA results and the presence of HEV RNA in plasma or in feces. According to the sequencing results of 348 bp PCR products of HEV, there were four different subtypes identified. Reports on the prevalence of HEV in wild boar populations are varying due to different sensitivities of the detection methods. However, this study reveals based on a highly sensitive method that HEV is widely spread in wild boar populations in the northwestern region of Poland and posing a potential risk to the consumer of game meat.

  11. Rapid Multiplex PCR Assay To Identify Respiratory Viral Pathogens: Moving Forward Diagnosing The Common Cold

    PubMed Central

    Gordon, Sarah M; Elegino-Steffens, Diane U; Agee, Willie; Barnhill, Jason; Hsue, Gunther

    2013-01-01

    Upper respiratory tract infections (URIs) can be a serious burden to the healthcare system. The majority of URIs are viral in etiology, but definitive diagnosis can prove difficult due to frequently overlapping clinical presentations of viral and bacterial infections, and the variable sensitivity, and lengthy turn-around time of viral culture. We tested new automated nested multiplex PCR technology, the FilmArray® system, in the TAMC department of clinical investigations, to determine the feasibility of replacing the standard viral culture with a rapid turn-around system. We conducted a feasibility study using a single-blinded comparison study, comparing PCR results with archived viral culture results from a convenience sample of cryopreserved archived nasopharyngeal swabs from acutely ill ED patients who presented with complaints of URI symptoms. A total of 61 archived samples were processed. Viral culture had previously identified 31 positive specimens from these samples. The automated nested multiplex PCR detected 38 positive samples. In total, PCR was 94.5% concordant with the previously positive viral culture results. However, PCR was only 63.4% concordant with the negative viral culture results, owing to PCR detection of 11 additional viral pathogens not recovered on viral culture. The average time to process a sample was 75 minutes. We determined that an automated nested multiplex PCR is a feasible alternative to viral culture in an acute clinical setting. We were able to detect at least 94.5% as many viral pathogens as viral culture is able to identify, with a faster turn-around time. PMID:24052914

  12. Detection of pathogenic Yersinia enterocolitica in foods and water by immunomagnetic separation, nested polymerase chain reactions, and colorimetric detection of amplified DNA.

    PubMed Central

    Kapperud, G; Vardund, T; Skjerve, E; Hornes, E; Michaelsen, T E

    1993-01-01

    A two-step polymerase chain reaction (PCR) procedure with two nested pairs of primers specific for the yadA gene of Yersinia enterocolitica was developed. The PCR assay identified all common pathogenic serogroups (O:3, O:5,27, O:8, O:9, O:13, and O:21) from three continents and differentiated pathogenic Y. enterocolitica from Y. pseudotuberculosis and from a variety of nonpathogenic yersiniae representing 25 serogroups and four species. The performance of the method was evaluated with seeded food and water samples. We compared two procedures for sample preparation prior to PCR: one was based on immunomagnetic separation of the target bacteria from the sample, using magnetic particles coated with immunoglobulin antibodies to Y. enterocolitica serogroup O:3, and the other method consisted of a series of centrifugation steps combined with proteinase treatment. Regardless of the method used, the PCR assay was capable of detecting 10 to 30 CFU/g of meat in 10(6)-fold excess of indigenous bacteria. When the samples were enriched overnight in a nonselective medium, the sensitivity was increased to approximately 2 CFU/g, except for samples with an extremely high background flora (> 10(7) CFU/g). We compared gel electrophoretic detection of PCR products with a colorimetric detection method designated DIANA (detection of immobilized amplified nucleic acids), which enabled easy visualization of amplified fragments in a microtiter plate format with an optical density reader. DIANA and gel electrophoresis showed complete concordance in their discrimination between positive and negative samples. The combination of immunomagnetic separation, nested PCR, and DIANA makes possible the development of a fully automated analytic process which requires a minimum of laboratory manipulations. Images PMID:8215366

  13. Anadromous sea lampreys (Petromyzon marinus) are ecosystem engineers in a spawning tributary

    USGS Publications Warehouse

    Hogg, Robert S.; Coghlan, Stephen M.; Zydlewski, Joseph D.; Simon, Kevin S.

    2014-01-01

    Sea lampreys (Petromyzon marinus) disturb the substratum during nest construction and alter the physical habitat, potentially affecting other stream organisms. We quantified differences in depth, velocity, fine-sediment coverage, embeddedness, intragravel permeability and benthic invertebrate assemblages (density and diversity) among nest mounds, nest pits and undisturbed reference locations over a 4-month period after June spawning. In 2010 and 2011, immediate and persistent effects of nest construction were assessed in summer (July) and in autumn (late September to early October), respectively. Randomly selected nests were sampled annually (25 each in summer and autumn). Nest construction increased stream-bed complexity by creating and juxtaposing shallow, swift, rocky habitat patches with deep, slow, sandy habitat patches. Mounds had a 50–143% less cover of fine sediment, and a 30–62% reduction in embeddedness, compared to pits and reference locations. These physical changes persisted into the autumn (almost 4 months). Five insect families contributed 74% of the benthic invertebrate abundance: Chironomidae (27%), Hydropsychidae (26%), Heptageniidae (8%), Philopotamidae (7%) and Ephemerellidae (6%). Densities of Hydropsychidae, Philopotamidae and Heptageniidae were up to 10 times greater in mounds than in pits and adjacent reference habitat. In summer, mounds had twice the density of Chironomidae than did pits, and 1.5 times more than reference habitats, but densities were similar among the habitats in autumn. These results suggest that spawning sea lampreys are ecosystem engineers. The physical disturbance caused by nest-building activity was significant and persistent, increasing habitat heterogeneity and favouring pollution-sensitive benthic invertebrates and, possibly, drift-feeding fish.

  14. A Fast Full Tensor Gravity computation algorithm for High Resolution 3D Geologic Interpretations

    NASA Astrophysics Data System (ADS)

    Jayaram, V.; Crain, K.; Keller, G. R.

    2011-12-01

    We present an algorithm to rapidly calculate the vertical gravity and full tensor gravity (FTG) values due to a 3-D geologic model. This algorithm can be implemented on single, multi-core CPU and graphical processing units (GPU) architectures. Our technique is based on the line element approximation with a constant density within each grid cell. This type of parameterization is well suited for high-resolution elevation datasets with grid size typically in the range of 1m to 30m. The large high-resolution data grids in our studies employ a pre-filtered mipmap pyramid type representation for the grid data known as the Geometry clipmap. The clipmap was first introduced by Microsoft Research in 2004 to do fly-through terrain visualization. This method caches nested rectangular extents of down-sampled data layers in the pyramid to create view-dependent calculation scheme. Together with the simple grid structure, this allows the gravity to be computed conveniently on-the-fly, or stored in a highly compressed format. Neither of these capabilities has previously been available. Our approach can perform rapid calculations on large topographies including crustal-scale models derived from complex geologic interpretations. For example, we used a 1KM Sphere model consisting of 105000 cells at 10m resolution with 100000 gravity stations. The line element approach took less than 90 seconds to compute the FTG and vertical gravity on an Intel Core i7 CPU at 3.07 GHz utilizing just its single core. Also, unlike traditional gravity computational algorithms, the line-element approach can calculate gravity effects at locations interior or exterior to the model. The only condition that must be met is the observation point cannot be located directly above the line element. Therefore, we perform a location test and then apply appropriate formulation to those data points. We will present and compare the computational performance of the traditional prism method versus the line element approach on different CPU-GPU system configurations. The algorithm calculates the expected gravity at station locations where the observed gravity and FTG data were acquired. This algorithm can be used for all fast forward model calculations of 3D geologic interpretations for data from airborne, space and submarine gravity, and FTG instrumentation.

  15. Enhanced detection of tuberculous mycobacteria in animal tissues using a semi-nested probe-based real-time PCR.

    PubMed

    Costa, Pedro; Ferreira, Ana S; Amaro, Ana; Albuquerque, Teresa; Botelho, Ana; Couto, Isabel; Cunha, Mónica V; Viveiros, Miguel; Inácio, João

    2013-01-01

    Bovine tuberculosis has been tackled for decades by costly eradication programs in most developed countries, involving the laboratory testing of tissue samples from allegedly infected animals for detection of Mycobacterium tuberculosis complex (MTC) members, namely Mycobacterium bovis. Definitive diagnosis is usually achieved by bacteriological culture, which may take up to 6-12 weeks, during which the suspect animal carcass and herd are under sanitary arrest. In this work, a user-friendly DNA extraction protocol adapted for tissues was coupled with an IS6110-targeted semi-nested duplex real-time PCR assay to enhance the direct detection of MTC bacteria in animal specimens, reducing the time to achieve a diagnosis and, thus, potentially limiting the herd restriction period. The duplex use of a novel β-actin gene targeted probe, with complementary targets in most mammals, allowed the assessment of amplification inhibitors in the tissue samples. The assay was evaluated with a group of 128 fresh tissue specimens collected from bovines, wild boars, deer and foxes. Mycobacterium bovis was cultured from 57 of these samples. Overall, the full test performance corresponds to a diagnostic sensitivity and specificity of 98.2% (CIP95% 89.4-99.9%) and 88.7% (CIP95% 78.5-94.7%), respectively. An observed kappa coefficient was estimated in 0.859 (CI P95% 0.771-0.948) for the overall agreement between the semi-nested PCR assay and the bacteriological culture. Considering only bovine samples (n = 69), the diagnostic sensitivity and specificity were estimated in 100% (CIP95% 84.0-100%) and 97.7% (CIP95% 86.2-99.9%), respectively. Eight negative culture samples exhibiting TB-like lesions were detected by the semi-nested real-time PCR, thus emphasizing the increased potential of this molecular approach to detect MTC-infected animal tissues. This novel IS6110-targeted assay allows the fast detection of tuberculous mycobacteria in animal specimens with very high sensitivity and specificity, being amenable and cost effective for use in the routine veterinary diagnostic laboratory with further automation possibilities.

  16. Loop mediated isothermal amplification (LAMP) accurately detects malaria DNA from filter paper blood samples of low density parasitaemias.

    PubMed

    Aydin-Schmidt, Berit; Xu, Weiping; González, Iveth J; Polley, Spencer D; Bell, David; Shakely, Delér; Msellem, Mwinyi I; Björkman, Anders; Mårtensson, Andreas

    2014-01-01

    Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6-782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94-99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0-4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1-98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2-99.9) and 76.9% (95%CI 46.2-95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1-100) in both study groups. Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings.

  17. Loop Mediated Isothermal Amplification (LAMP) Accurately Detects Malaria DNA from Filter Paper Blood Samples of Low Density Parasitaemias

    PubMed Central

    González, Iveth J.; Polley, Spencer D.; Bell, David; Shakely, Delér; Msellem, Mwinyi I.; Björkman, Anders; Mårtensson, Andreas

    2014-01-01

    Background Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Methods and Findings Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6–782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94–99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0–4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1–98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2–99.9) and 76.9% (95%CI 46.2–95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1–100) in both study groups. Conclusion Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings. PMID:25105591

  18. Mercury concentrations in tissues of Florida bald eagles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wood, P.B.; Wood, J.M.; White, J.H.

    1996-01-01

    We collected 48 blood and 61 feather samples from nestling bald eagles at 42 nests and adult feather samples from 20 nests in north and central Florida during 1991-93. We obtained 32 liver, 10 feather, and 5 blood samples from 33 eagle carcasses recovered in Florida during 1987-93. For nestlings, mercury concentrations in blood (GM = 0.16 ppm wet wt) and feather (GM = 3.23 ppm) samples were correlated (r = 0.69, P = 0.0001). Although nestlings had lower mercury concentrations in feathers than did adults (GM = 6.03 ppm), the feather mercury levels in nestlings and adults from themore » same nest were correlated (r = 0.63, P < 0.02). Mercury concentrations in blood of captive adult eagles (GM = 0.23 ppm) was similar to Florida nestlings but some Florida nestlings had blood mercury concentrations up to 0.61 ppm, more than twice as high as captive adults. Feather mercury concentrations in both nestlings and adults exceeded those in captive eagles, but concentrations in all tissues were similar to, or lower than, those in bald eagles from other wild populations. Although mercury concentrations in Florida eagles are below those that cause mortality, they are in the range of concentrations that can cause behavioral changes or reduce reproduction. We recommend periodic monitoring of mercury in Florida bald eagles for early detection of mercury increases before negative effects on reproduction occur. 26 refs., 5 figs., 2 tabs.« less

  19. [Detection of human parvovirus B19, human bocavirus and human parvovirus 4 infections in blood samples among 95 patients with liver disease in Nanjing by nested PCR].

    PubMed

    Tong, Rui; Zhou, Wei-Min; Liu, Xi-Jun; Wang, Yue; Lou, Yong-Liang; Tan, Wen-Jie

    2013-04-01

    To analyze the infection of human parvovirus B19, human bocavirus (HBoV) and human parvovirus 4 (PARV4) in blood samples among patients with liver disease in Nanjing by molecular detection. Nested PCR assays were designed and validated to detect B19, HBoV and PARV4, respectively. The assays were used to screen three parvoviruses in blood samples from 95 patients with different liver disease in Nanjing. The parvovirus infection was analyzed statistically. The detection limits were 10 copies of genomic DNA equivalents per reaction for each assays and the good specificity were observed. The frequency of B19 and HBoV were 2/95 (2.1%) and 9/95 (9.5%) in blood samples respectively. No PARV4 was detected. HBoV was detected in 3/5 patients with drug-induced hepatitis. Both B19 and HBoV infection were detected in blood from patients with liver disease.

  20. Detection of Bovine Leukemia Virus in Brains of Cattle with a Neurological Syndrome: Pathological and Molecular Studies

    PubMed Central

    D'Angelino, Rubens Henrique Ramos; Pituco, Edviges Maristela; Villalobos, Eliana Monteforte Cassaro; Harakava, Ricardo; Gregori, Fábio

    2013-01-01

    Bovine leukemia virus (BLV) was investigated in the central nervous system (CNS) of cattle with neurological syndrome. A total of 269 CNS samples were submitted to nested-PCR (BLV env gene gp51), and the viral genotypes were identified. The nested-PCR was positive in 4.8% (13/269) CNS samples, with 2.7% (2/74) presenting at histological examination lesions of nonpurulent meningoencephalitis (NPME), whereas 5.6% (11/195) not presenting NPME (P > 0.05). No samples presented lymphosarcoma. The PCR products (437 bp) were sequenced and submitted to phylogenetic analysis by neighbor-joining and maximum composite likelihood methods, and genotypes 1, 5, and 6 were detected, corroborating other South American studies. The genotype 6 barely described in Brazil and Argentina was more frequently detected in this study. The identity matrices showed maximum similarity (100%) among some samples of this study and one from Argentina (FJ808582), recovered from GenBank. There was no association among the genotypes and NPME lesions. PMID:23710448

  1. Molecular identification of Coccidioides spp. in soil samples from Brazil

    PubMed Central

    2011-01-01

    Background Since 1991 several outbreaks of acute coccidioidomycosis (CM) were diagnosed in the semi-arid Northeast of Brazil, mainly related to disturbance of armadillo burrows caused by hunters while digging them for the capture of these animals. This activity causes dust contaminated with arthroconidia of Coccidioides posadasii, which, once inhaled, cause the mycosis. We report on the identification of C. posadasii in soil samples related to outbreaks of CM. Results Twenty four soil samples had their DNA extracted and subsequently submitted to a semi-nested PCR technique using specific primers. While only 6 (25%) soil samples were positive for C. posadasii by mice inoculation, all (100%) were positive by the molecular tool. Conclusion This methodology represents a simple, sensitive and specific molecular technique to determine the environmental distribution of Coccidioides spp. in endemic areas, but cannot distinguish the species. Moreover, it may be useful to identify culture isolates. Key-words: 1. Coccidioidomycosis. 2. Coccidioides spp. 3. C. posadasii. 4. Semi-arid. 5. Semi-nested PCR PMID:21575248

  2. DEVELOPING ATMOSPHERIC RETRIEVAL METHODS FOR DIRECT IMAGING SPECTROSCOPY OF GAS GIANTS IN REFLECTED LIGHT. I. METHANE ABUNDANCES AND BASIC CLOUD PROPERTIES

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lupu, Roxana E.; Marley, Mark S.; Zahnle, Kevin

    Upcoming space-based coronagraphic instruments in the next decade will perform reflected light spectroscopy and photometry of cool directly imaged extrasolar giant planets. We are developing a new atmospheric retrieval methodology to help assess the science return and inform the instrument design for such future missions, and ultimately interpret the resulting observations. Our retrieval technique employs a geometric albedo model coupled with both a Markov chain Monte Carlo Ensemble Sampler ( emcee ) and a multimodal nested sampling algorithm ( MultiNest ) to map the posterior distribution. This combination makes the global evidence calculation more robust for any given model andmore » highlights possible discrepancies in the likelihood maps. As a proof of concept, our current atmospheric model contains one or two cloud layers, methane as a major absorber, and a H{sub 2}–He background gas. This 6-to-9 parameter model is appropriate for Jupiter-like planets and can be easily expanded in the future. In addition to deriving the marginal likelihood distribution and confidence intervals for the model parameters, we perform model selection to determine the significance of methane and cloud detection as a function of expected signal-to-noise ratio in the presence of spectral noise correlations. After internal validation, the method is applied to realistic spectra of Jupiter, Saturn, and HD 99492c, a model observing target. We find that the presence or absence of clouds and methane can be determined with high confidence, while parameter uncertainties are model dependent and correlated. Such general methods will also be applicable to the interpretation of direct imaging spectra of cloudy terrestrial planets.« less

  3. Developing Atmospheric Retrieval Methods for Direct Imaging Spectroscopy of Gas Giants in Reflected Light I: Methane Abundances and Basic Cloud Properties

    NASA Technical Reports Server (NTRS)

    Lupu, R. E.; Marley, M. S.; Lewis, N.; Line, M.; Traub, W.; Zahnle, K.

    2016-01-01

    Reflected light spectroscopy and photometry of cool, directly imaged extrasolar giant planets are expected to be performed in the next decade by space-based telescopes equipped with optical wavelength coronagraphs and integral field spectrographs, such as the Wide-Field Infrared Survey Telescope (WFIRST). We are developing a new atmospheric retrieval methodology to help assess the science return and inform the instrument design for such future missions, and ultimately interpret the resulting observations. Our retrieval technique employs an albedo model coupled with both a Markov chain Monte Carlo Ensemble Sampler (emcee) and a multimodal nested sampling algorithm (MultiNest) to map the posterior distribution. This combination makes the global evidence calculation more robust for any given model, and highlights possible discrepancies in the likelihood maps. Here we apply this methodology to simulated spectra of cool giant planets. As a proof-of-concept, our current atmospheric model contains 1 or 2 cloud layers, methane as a major absorber, and a H2-He background gas. This 6-to-9 parameter model is appropriate for Jupiter-like planets and can be easily expanded in the future. In addition to deriving the marginal likelihood distribution and confidence intervals for the model parameters, we perform model selection to determine the significance of methane and cloud detection as a function of expected signal-to-noise, in the presence of spectral noise correlations. After internal validation, the method is applied to realistic reflected-light spectra of Jupiter, Saturn, and HD 99492 c, a likely observing target. We find that the presence or absence of clouds and methane can be determined with high accuracy, while parameters uncertainties are model-dependent.

  4. An algorithm for automated layout of process description maps drawn in SBGN.

    PubMed

    Genc, Begum; Dogrusoz, Ugur

    2016-01-01

    Evolving technology has increased the focus on genomics. The combination of today's advanced techniques with decades of molecular biology research has yielded huge amounts of pathway data. A standard, named the Systems Biology Graphical Notation (SBGN), was recently introduced to allow scientists to represent biological pathways in an unambiguous, easy-to-understand and efficient manner. Although there are a number of automated layout algorithms for various types of biological networks, currently none specialize on process description (PD) maps as defined by SBGN. We propose a new automated layout algorithm for PD maps drawn in SBGN. Our algorithm is based on a force-directed automated layout algorithm called Compound Spring Embedder (CoSE). On top of the existing force scheme, additional heuristics employing new types of forces and movement rules are defined to address SBGN-specific rules. Our algorithm is the only automatic layout algorithm that properly addresses all SBGN rules for drawing PD maps, including placement of substrates and products of process nodes on opposite sides, compact tiling of members of molecular complexes and extensively making use of nested structures (compound nodes) to properly draw cellular locations and molecular complex structures. As demonstrated experimentally, the algorithm results in significant improvements over use of a generic layout algorithm such as CoSE in addressing SBGN rules on top of commonly accepted graph drawing criteria. An implementation of our algorithm in Java is available within ChiLay library (https://github.com/iVis-at-Bilkent/chilay). ugur@cs.bilkent.edu.tr or dogrusoz@cbio.mskcc.org Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press.

  5. An algorithm for automated layout of process description maps drawn in SBGN

    PubMed Central

    Genc, Begum; Dogrusoz, Ugur

    2016-01-01

    Motivation: Evolving technology has increased the focus on genomics. The combination of today’s advanced techniques with decades of molecular biology research has yielded huge amounts of pathway data. A standard, named the Systems Biology Graphical Notation (SBGN), was recently introduced to allow scientists to represent biological pathways in an unambiguous, easy-to-understand and efficient manner. Although there are a number of automated layout algorithms for various types of biological networks, currently none specialize on process description (PD) maps as defined by SBGN. Results: We propose a new automated layout algorithm for PD maps drawn in SBGN. Our algorithm is based on a force-directed automated layout algorithm called Compound Spring Embedder (CoSE). On top of the existing force scheme, additional heuristics employing new types of forces and movement rules are defined to address SBGN-specific rules. Our algorithm is the only automatic layout algorithm that properly addresses all SBGN rules for drawing PD maps, including placement of substrates and products of process nodes on opposite sides, compact tiling of members of molecular complexes and extensively making use of nested structures (compound nodes) to properly draw cellular locations and molecular complex structures. As demonstrated experimentally, the algorithm results in significant improvements over use of a generic layout algorithm such as CoSE in addressing SBGN rules on top of commonly accepted graph drawing criteria. Availability and implementation: An implementation of our algorithm in Java is available within ChiLay library (https://github.com/iVis-at-Bilkent/chilay). Contact: ugur@cs.bilkent.edu.tr or dogrusoz@cbio.mskcc.org Supplementary information: Supplementary data are available at Bioinformatics online. PMID:26363029

  6. K-Nearest Neighbor Algorithm Optimization in Text Categorization

    NASA Astrophysics Data System (ADS)

    Chen, Shufeng

    2018-01-01

    K-Nearest Neighbor (KNN) classification algorithm is one of the simplest methods of data mining. It has been widely used in classification, regression and pattern recognition. The traditional KNN method has some shortcomings such as large amount of sample computation and strong dependence on the sample library capacity. In this paper, a method of representative sample optimization based on CURE algorithm is proposed. On the basis of this, presenting a quick algorithm QKNN (Quick k-nearest neighbor) to find the nearest k neighbor samples, which greatly reduces the similarity calculation. The experimental results show that this algorithm can effectively reduce the number of samples and speed up the search for the k nearest neighbor samples to improve the performance of the algorithm.

  7. Efficient propagation-inside-layer expansion algorithm for solving the scattering from three-dimensional nested homogeneous dielectric bodies with arbitrary shape.

    PubMed

    Bellez, Sami; Bourlier, Christophe; Kubické, Gildas

    2015-03-01

    This paper deals with the evaluation of electromagnetic scattering from a three-dimensional structure consisting of two nested homogeneous dielectric bodies with arbitrary shape. The scattering problem is formulated in terms of a set of Poggio-Miller-Chang-Harrington-Wu integral equations that are afterwards converted into a system of linear equations (impedance matrix equation) by applying the Galerkin method of moments (MoM) with Rao-Wilton-Glisson basis functions. The MoM matrix equation is then solved by deploying the iterative propagation-inside-layer expansion (PILE) method in order to obtain the unknown surface current densities, which are thereafter used to handle the radar cross-section (RCS) patterns. Some numerical results for various structures including canonical geometries are presented and compared with those of the FEKO software in order to validate the PILE-based approach as well as to show its efficiency to analyze the full-polarized RCS patterns.

  8. Detection of Legionella spp. by a nested-PCR assay in air samples of a wastewater treatment plant and downwind distances in Isfahan

    PubMed Central

    Mirzaee, Seyyed Abbas; Nikaeen, Mahnaz; Hajizadeh, Yaghob; Nabavi, BiBi Fatemeh; Hassanzadeh, Akbar

    2015-01-01

    Background: Wastewater contains a variety of pathogens and bio -aerosols generated during the wastewater treatment process, which could be a potential health risk for exposed individuals. This study was carried out to detect Legionella spp. in the bio -aerosols generated from different processes of a wastewater treatment plant (WWTP) in Isfahan, Iran, and the downwind distances. Materials and Methods: A total of 54 air samples were collected and analyzed for the presence of Legionella spp. by a nested- polymerase chain reaction (PCR) assay. A liquid impingement biosampler was used to capture bio -aerosols. The weather conditions were also recorded. Results: Legionella were detected in 6% of the samples, including air samples above the aeration tank (1/9), belt filter press (1/9), and 250 m downwind (1/9). Conclusion: The result of this study revealed the presence of Legionella spp. in air samples of a WWTP and downwind distance, which consequently represent a potential health risk to the exposed individuals. PMID:25802817

  9. Comparison of milk culture, direct and nested polymerase chain reaction (PCR) with fecal culture based on samples from dairy herds infected with Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    Gao, Anli; Odumeru, Joseph; Raymond, Melinda; Hendrick, Steven; Duffield, Todd; Mutharia, Lucy

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in cattle and other farm ruminants, and is also a suspected pathogen of Crohn’s disease in humans. Development of diagnostic methods for MAP infection has been a challenge over the last few decades. The objective of this study was to investigate the relationship between different methods for detection of MAP in milk and fecal samples. A total of 134 milk samples and 110 feces samples were collected from 146 individual cows in 14 MAP-infected herds in southwestern Ontario. Culture, IS900 polymerase chain reaction (PCR) and nested PCR methods were used for detecting MAP in milk; results were compared with those of fecal culture. A significant relationship was found between milk culture, direct PCR, and nested PCR (P < 0.05). The fecal culture results were not related to any of the 3 assay methods used for the milk samples (P > 0.10). Although fecal culture showed a higher sensitivity than the milk culture method, the difference was not significant (P = 0.2473). The number of MAP colony-forming units (CFU) isolated by culture from fecal samples was, on average, higher than that isolated from milk samples (P = 0.0083). There was no significant correlation between the number of CFU cultured from milk and from feces (Pearson correlation coefficient = 0.1957, N = 63, P = 0.1243). The animals with high numbers of CFU in milk culture may not be detected by fecal culture at all, and vise versa. A significant proportion (29% to 41%) of the positive animals would be missed if only 1 culture method, instead of both milk and feces, were to be used for diagnosis. This suggests that the shedding of MAP in feces and milk is not synchronized. Most of the infected cows were low-level shedders. The proportion of low-level shedders may even be underestimated because MAP is killed during decontamination, thus reducing the chance of detection. Therefore, to identify suspected Johne’s-infected animals using the tests in this study, both milk and feces samples should be collected in duplicate to enhance the diagnostic rate. The high MAP kill rate identified in the culture methods during decontamination may be compensated for by using the nested PCR method, which had a higher sensitivity than the IS900 PCR method used. PMID:19337397

  10. Photometric classification and redshift estimation of LSST Supernovae

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dai, Mi; Kuhlmann, Steve; Wang, Yun

    Supernova (SN) classification and redshift estimation using photometric data only have become very important for the Large Synoptic Survey Telescope (LSST), given the large number of SNe that LSST will observe and the impossibility of spectroscopically following up all the SNe. We investigate the performance of an SN classifier that uses SN colours to classify LSST SNe with the Random Forest classification algorithm. Our classifier results in an area-under-the-curve of 0.98 which represents excellent classification. We are able to obtain a photometric SN sample containing 99 percent SNe Ia by choosing a probability threshold. We estimate the photometric redshifts (photo-z)more » of SNe in our sample by fitting the SN light curves using the SALT2 model with nested sampling. We obtain a mean bias (⟨zphot - zspec⟩) of 0.012 with σ(z phot -z spec 1+z spec )=0.0294 σ(zphot-zspec1+zspec)=0.0294 without using a host-galaxy photo-z prior, and a mean bias (⟨zphot - zspec⟩) of 0.0017 with σ(z phot -z spec 1+z spec )=0.0116 σ(zphot-zspec1+zspec)=0.0116 using a host-galaxy photo-z prior. Assuming a flat ΛCDM model with Ωm = 0.3, we obtain Ωm of 0.305 ± 0.008 (statistical errors only), using the simulated LSST sample of photometric SNe Ia (with intrinsic scatter σint = 0.11) derived using our methodology without using host-galaxy photo-z prior. Our method will help boost the power of SNe from the LSST as cosmological probes.« less

  11. [A novel TaqMan® MGB probe for specifically detecting Streptococcus mutans].

    PubMed

    Zheng, Hui; Lin, Jiu-Xiang; DU, Ning; Chen, Feng

    2013-10-18

    To design a new TaqMan® MGB probe for improving the specificity of Streptococcus mutans's detection. We extracted six DNA samples from different streptococcal strains for PCR reaction. Conventional nested PCR and TaqMan® MGB real-time PCR were applied independently. The first round of nested PCR was carried out with the bacterial universal primers, while a second PCR was conducted by using primers specific for the 16S rRNA gene of Streptococcus mutans. The TaqMan® MGB probe for Streptococcus mutans was designed from sequence analyses, and the primers were the same as nested PCR. Streptococcus mutans DNA with 2.5 mg/L was sequentially diluted at 5-fold intervals to 0.16 μg/L. Standard DNA samples were used to generate standard curves by TaqMan® MGB real-time PCR. In the nested PCR, the primers specific for Streptococcus mutans also detected Streptococcus gordonii with visible band of 282 bp, giving false-positive results. In the TaqMan® MGB real-time PCR reaction, only Streptococcus mutans was detected. The detection limitation of TaqMan® MGB real-time PCR for Streptococcus mutans 16S rRNA gene was 20 μg/L. We designed a new TaqMan® MGB probe, and successfully set up a PCR based method for detecting oral Streptococcus mutans. TaqMan® MGB real-time PCR is a both specific and sensitive bacterial detection method.

  12. Coevolutionary diversification creates nested-modular structure in phage–bacteria interaction networks

    PubMed Central

    Beckett, Stephen J.; Williams, Hywel T. P.

    2013-01-01

    Phage and their bacterial hosts are the most diverse and abundant biological entities in the oceans, where their interactions have a major impact on marine ecology and ecosystem function. The structure of interaction networks for natural phage–bacteria communities offers insight into their coevolutionary origin. At small phylogenetic scales, observed communities typically show a nested structure, in which both hosts and phages can be ranked by their range of resistance and infectivity, respectively. A qualitatively different multi-scale structure is seen at larger phylogenetic scales; a natural assemblage sampled from the Atlantic Ocean displays large-scale modularity and local nestedness within each module. Here, we show that such ‘nested-modular’ interaction networks can be produced by a simple model of host–phage coevolution in which infection depends on genetic matching. Negative frequency-dependent selection causes diversification of hosts (to escape phages) and phages (to track their evolving hosts). This creates a diverse community of bacteria and phage, maintained by kill-the-winner ecological dynamics. When the resulting communities are visualized as bipartite networks of who infects whom, they show the nested-modular structure characteristic of the Atlantic sample. The statistical significance and strength of this observation varies depending on whether the interaction networks take into account the density of the interacting strains, with implications for interpretation of interaction networks constructed by different methods. Our results suggest that the apparently complex community structures associated with marine bacteria and phage may arise from relatively simple coevolutionary origins. PMID:24516719

  13. What's in a Day? A Guide to Decomposing the Variance in Intensive Longitudinal Data

    PubMed Central

    de Haan-Rietdijk, Silvia; Kuppens, Peter; Hamaker, Ellen L.

    2016-01-01

    In recent years there has been a growing interest in the use of intensive longitudinal research designs to study within-person processes. Examples are studies that use experience sampling data and autoregressive modeling to investigate emotion dynamics and between-person differences therein. Such designs often involve multiple measurements per day and multiple days per person, and it is not clear how this nesting of the data should be accounted for: That is, should such data be considered as two-level data (which is common practice at this point), with occasions nested in persons, or as three-level data with beeps nested in days which are nested in persons. We show that a significance test of the day-level variance in an empty three-level model is not reliable when there is autocorrelation. Furthermore, we show that misspecifying the number of levels can lead to spurious or misleading findings, such as inflated variance or autoregression estimates. Throughout the paper we present instructions and R code for the implementation of the proposed models, which includes a novel three-level AR(1) model that estimates moment-to-moment inertia and day-to-day inertia. Based on our simulations we recommend model selection using autoregressive multilevel models in combination with the AIC. We illustrate this method using empirical emotion data from two independent samples, and discuss the implications and the relevance of the existence of a day level for the field. PMID:27378986

  14. Measuring the effect of mercury exposure on common loon productivity in Wisconsin

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Meyer, M.W.; Hartigan, J.; Woodford, J.

    1994-12-31

    In 1991, the Wisconsin DNR conducted a pilot study to determine the extent of mercury (Hg) exposure in common loons (Gavia immer) breeding in Wisconsin. Loons are at risk to elevated mercury exposure in Wisconsin because they often nest on acidified, low alkalinity lakes. Fish from these lakes bioaccumulate methyl-Hg to a greater extent than biota from neutral pH lakes. Using nightlighting techniques, 35 adult loons were captured on 22 northern Wisconsin lakes (pH = 5.0--8.7) in 1991. Blood and feather samples were collected for Hg analysis. There was a highly significant negative linear relationship between adult loon blood clotmore » Hg and lake pH (r{sup 2} = 0.38, F = 15.27, P < 0.001); indicating loons nesting on low pH lakes receive greater mercury exposure than loons nesting on neutral pH lakes. The relationship was greater amongst adult males (r{sup 2} = 0.56) than among adult females (r{sup 2} = 0.36). The research effort was expanded in 1992 and 1993; 253 loons have been captured to date on 62 lakes in northern Wisconsin. The mercury content of feather and blood samples is being measured. Individual loons were also fitted with unique colored leg bands. Reproductive performance, annual return rates, and nesting behavior of adult loons with the greatest Hg exposure is being compared to that of adult loons with the least Hg exposure. The preliminary results from 1991--93 field seasons will be presented.« less

  15. What's in a Day? A Guide to Decomposing the Variance in Intensive Longitudinal Data.

    PubMed

    de Haan-Rietdijk, Silvia; Kuppens, Peter; Hamaker, Ellen L

    2016-01-01

    In recent years there has been a growing interest in the use of intensive longitudinal research designs to study within-person processes. Examples are studies that use experience sampling data and autoregressive modeling to investigate emotion dynamics and between-person differences therein. Such designs often involve multiple measurements per day and multiple days per person, and it is not clear how this nesting of the data should be accounted for: That is, should such data be considered as two-level data (which is common practice at this point), with occasions nested in persons, or as three-level data with beeps nested in days which are nested in persons. We show that a significance test of the day-level variance in an empty three-level model is not reliable when there is autocorrelation. Furthermore, we show that misspecifying the number of levels can lead to spurious or misleading findings, such as inflated variance or autoregression estimates. Throughout the paper we present instructions and R code for the implementation of the proposed models, which includes a novel three-level AR(1) model that estimates moment-to-moment inertia and day-to-day inertia. Based on our simulations we recommend model selection using autoregressive multilevel models in combination with the AIC. We illustrate this method using empirical emotion data from two independent samples, and discuss the implications and the relevance of the existence of a day level for the field.

  16. [MicroRNA Target Prediction Based on Support Vector Machine Ensemble Classification Algorithm of Under-sampling Technique].

    PubMed

    Chen, Zhiru; Hong, Wenxue

    2016-02-01

    Considering the low accuracy of prediction in the positive samples and poor overall classification effects caused by unbalanced sample data of MicroRNA (miRNA) target, we proposes a support vector machine (SVM)-integration of under-sampling and weight (IUSM) algorithm in this paper, an under-sampling based on the ensemble learning algorithm. The algorithm adopts SVM as learning algorithm and AdaBoost as integration framework, and embeds clustering-based under-sampling into the iterative process, aiming at reducing the degree of unbalanced distribution of positive and negative samples. Meanwhile, in the process of adaptive weight adjustment of the samples, the SVM-IUSM algorithm eliminates the abnormal ones in negative samples with robust sample weights smoothing mechanism so as to avoid over-learning. Finally, the prediction of miRNA target integrated classifier is achieved with the combination of multiple weak classifiers through the voting mechanism. The experiment revealed that the SVM-IUSW, compared with other algorithms on unbalanced dataset collection, could not only improve the accuracy of positive targets and the overall effect of classification, but also enhance the generalization ability of miRNA target classifier.

  17. Molecular analysis of dolphin morbillivirus: A new sensitive detection method based on nested RT-PCR.

    PubMed

    Centelleghe, Cinzia; Beffagna, Giorgia; Zanetti, Rossella; Zappulli, Valentina; Di Guardo, Giovanni; Mazzariol, Sandro

    2016-09-01

    Cetacean Morbillivirus (CeMV) has been identified as the most pathogenic virus for cetaceans. Over the past three decades, this RNA virus has caused several outbreaks of lethal disease in odontocetes and mysticetes worldwide. Isolation and identification of CeMV RNA is very challenging in whales because of the poor preservation status frequently shown by tissues from stranded animals. Nested reverse transcription polymerase chain reaction (nested RT-PCR) is used instead of conventional RT-PCR when it is necessary to increase the sensitivity and the specificity of the reaction. This study describes a new nested RT-PCR technique useful to amplify small amounts of the cDNA copy of Cetacean morbillivirus (CeMV) when it is present in scant quantity in whales' biological specimens. This technique was used to analyze different tissues (lung, brain, spleen and other lymphoid tissues) from one under human care seal and seven cetaceans stranded along the Italian coastline between October 2011 and September 2015. A well-characterized, 200 base pair (bp) fragment of the dolphin Morbillivirus (DMV) haemagglutinin (H) gene, obtained by nested RT-PCR, was sequenced and used to confirm DMV positivity in all the eight marine mammals under study. In conclusion, this nested RT-PCR protocol can represent a sensitive detection method to identify CeMV-positive, poorly preserved tissue samples. Furthermore, this is also a rather inexpensive molecular technique, relatively easy to apply. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Development of an In-House Multiplex Nested RT-PCR Method for Detecting Acute HIV-1 Infection in High Risk Populations.

    PubMed

    Liu, Zhiying; Li, Wei; Xu, Meng; Sheng, Bo; Yang, Zixuan; Jiao, Yanmei; Zhang, Tong; Mou, Danlei; Chen, Dexi; Wu, Hao

    2015-01-01

    The detection of acute HIV infection (AHI) among high risk populations can help reduce secondary transmission of HIV. The nucleic acid testing (NAT) can shorten the test window period by up to 7-12 days. In this study, we describe an in-house NAT based on the multiplex nested RT-PCR method to detect the HIV RNA. We also evaluated it in a high risk cohort in Beijing. Four primer pairs were designed and evaluated for the detection of different HIV-1 subtypes in group M. Multiplex RT-PCR and nested PCR were performed. The sensitivity, specialty, primers compatibility among HIV subtypes were evaluated simultaneously. In an MSM cohort in Beijing during a 3-year period, a total of 11,808 blood samples that were negative by ELISA or indeterminate by Western blot were analyzed by this multiplex nested RT-PCR with pooling strategy. The multiplex nested RT-PCR was successfully applied for the detection of at least six HIV-1 subtypes. The sensitivity was 40 copies/ml and the specificity was 100%. A total of 29 people were tested HIV-1 positive with acute infection in a MSM cohort of Beijing during a 3 years period. This multiplex nested RT-PCR provides a useful tool for the rapid detection of acute HIV-1 infection. When used in combination with the 3(rd) generation ELISA, it can improve the detection rate of HIV infection, especially in the source limited regions.

  19. Compositional changes in bee and wasp communities along Neotropical mountain altitudinal gradient.

    PubMed

    Perillo, Lucas Neves; Neves, Frederico de Siqueira; Antonini, Yasmine; Martins, Rogério Parentoni

    2017-01-01

    Climate conditions tend to differ along an altitudinal gradient, resulting in some species groups' patterns of lower species richness with increasing altitude. While this pattern is well understood for tropical mountains, studies investigating possible determinants of variation in beta-diversity at its different altitudes are scarce. We sampled bee and wasp communities (Hymenoptera: Aculeata) along an altitudinal gradient (1,000-2,000 m.a.s.l.) in a tropical mountainous region of Brazil. Trap nests and Moericke traps were established at six sampling points, with 200 m difference in altitude between each point. We obtained average climate data (1970-2000) from Worldclim v2 for altitudes at each sampling site. Nest traps captured 17 bee and wasp species from six families, and Moericke traps captured 124 morphospecies from 13 families. We found a negative correlation between altitude and species richness and abundance. Temperature, precipitation, water vapor pressure, and wind speed influenced species richness and abundance, and were correlated with altitude. β-diversity was primarily determined by species turnover as opposed to nestedness, and Aculeate community similarity was higher for more similar altitudinal ranges. Moericke traps seem to be more efficient for altitudinal surveys compared to nest traps. We found high occurrence of singleton and doubleton species at all altitudes, highlighting the need for long-term studies to efficiently assess hymenopteran diversity in these environments.

  20. Detection of Helicobacter spp. DNA in the oral cavity of dogs.

    PubMed

    Recordati, Camilla; Gualdi, Valentina; Tosi, Sabrina; Facchini, Roberto Vailati; Pengo, Graziano; Luini, Mario; Simpson, Kenneth W; Scanziani, Eugenio

    2007-01-31

    The mode of acquisition of gastric Helicobacter spp. infection in dogs has not been determined. It is suspected that oral-oral and faecal-oral transmission may be involved. The present study sought to determine if Helicobacter spp. DNA is present in the oral cavity of healthy and vomiting dogs. Thirty-eight pet dogs (27 vomiting and 11 clinically healthy) were studied. The presence of Helicobacter spp. was determined by single and nested PCR evaluation of DNA extracted from saliva, dental plaque and gastric biopsy samples. Helicobacter spp. DNA was detected by nested PCR in 36 (94.7%) gastric biopsies, 17 (44.7%) dental plaque and 19 (50%) saliva samples out of the 38 dogs examined. Overall 27 (71.1%) dogs screened by nested PCR were found to harbour Helicobacter spp. DNA in the oral cavity (dental plaque and/or saliva). There was no significant difference in the prevalence of Helicobacter spp. DNA in the oral cavity of vomiting and healthy dogs, and the time from vomiting to oral sampling did not have significant impact. This study confirms the high prevalence of gastric Helicobacter spp. infection in dogs, and reveals that Helicobacter spp. DNA is detectable in the oral cavity of over 70% of dogs. These findings support the possibility of oral-oral transmission between dogs and that the canine oral cavity may act as source of non-pylori Helicobacter spp. infection for humans.

  1. Clinical Evaluation of a Loop-Mediated Amplification Kit for Diagnosis of Imported Malaria

    PubMed Central

    Polley, Spencer D.; González, Iveth J.; Mohamed, Deqa; Daly, Rosemarie; Bowers, Kathy; Watson, Julie; Mewse, Emma; Armstrong, Margaret; Gray, Christen; Perkins, Mark D.; Bell, David; Kanda, Hidetoshi; Tomita, Norihiro; Kubota, Yutaka; Mori, Yasuyoshi; Chiodini, Peter L.; Sutherland, Colin J.

    2013-01-01

    Background. Diagnosis of malaria relies on parasite detection by microscopy or antigen detection; both fail to detect low-density infections. New tests providing rapid, sensitive diagnosis with minimal need for training would enhance both malaria diagnosis and malaria control activities. We determined the diagnostic accuracy of a new loop-mediated amplification (LAMP) kit in febrile returned travelers. Methods. The kit was evaluated in sequential blood samples from returned travelers sent for pathogen testing to a specialist parasitology laboratory. Microscopy was performed, and then malaria LAMP was performed using Plasmodium genus and Plasmodium falciparum–specific tests in parallel. Nested polymerase chain reaction (PCR) was performed on all samples as the reference standard. Primary outcome measures for diagnostic accuracy were sensitivity and specificity of LAMP results, compared with those of nested PCR. Results. A total of 705 samples were tested in the primary analysis. Sensitivity and specificity were 98.4% and 98.1%, respectively, for the LAMP P. falciparum primers and 97.0% and 99.2%, respectively, for the Plasmodium genus primers. Post hoc repeat PCR analysis of all 15 tests with discrepant results resolved 4 results in favor of LAMP, suggesting that the primary analysis had underestimated diagnostic accuracy. Conclusions. Malaria LAMP had a diagnostic accuracy similar to that of nested PCR, with a greatly reduced time to result, and was superior to expert microscopy. PMID:23633403

  2. Methods of detecting and counting raptors: A review

    USGS Publications Warehouse

    Fuller, M.R.; Mosher, J.A.; Ralph, C. John; Scott, J. Michael

    1981-01-01

    Most raptors are wide-ranging, secretive, and occur at relatively low densities. These factors, in conjunction with the nocturnal activity of owls, cause the counting of raptors by most standard census and survey efforts to be very time consuming and expensive. This paper reviews the most common methods of detecting and counting raptors. It is hoped that it will be of use to the ever-increasing number of biologists, land-use planners, and managers that must determine the occurrence, density, or population dynamics of raptors. Road counts of fixed station or continuous transect design are often used to sample large areas. Detection of spontaneous or elicited vocalizations, especially those of owls, provides a means of detecting and estimating raptor numbers. Searches for nests are accomplished from foot surveys, observations from automobiles and boats, or from aircraft when nest structures are conspicuous (e.g., Osprey). Knowledge of nest habitat, historic records, and inquiries of local residents are useful for locating nests. Often several of these techniques are combined to help find nest sites. Aerial searches have also been used to locate or count large raptors (e.g., eagles), or those that may be conspicuous in open habitats (e.g., tundra). Counts of birds entering or leaving nest colonies or colonial roosts have been attempted on a limited basis. Results from Christmas Bird Counts have provided an index of the abundance of some species. Trapping and banding generally has proven to be an inefficient method of detecting raptors or estimating their populations. Concentrations of migrants at strategically located points around the world afford the best opportunity to count many rap tors in a relatively short period of time, but the influence of many unquantified variables has inhibited extensive interpretation of these counts. Few data exist to demonstrate the effectiveness of these methods. We believe more research on sampling techniques, rather than complete counts or intensive searches, will provide adequate yet affordable estimates of raptor numbers in addition to providing methods for detecting the presence of raptors on areas of interest to researchers and managers.

  3. Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis.

    PubMed

    Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui; Hu, Guohua

    2017-01-01

    Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis. The PCR product was detected by a lateral flow strip assay, which provided a basis for migration of the test to a point-of-care (POC) microfluidic format. The developed assay had an improved sensitivity compared with traditional PCR, and the limit of detection was up to 1 fg DNA isolated from M. tuberculosis. The assay was also specific for M. tuberculosis, and no cross-reactions were found in other non-target bacteria. The application of this technique to clinical samples was successfully evaluated, and OTNPCR-LFST showed 89% overall sensitivity and 100% specificity for TBM patients. This one-tube nested PCR-lateral flow strip assay is useful for detecting M. tuberculosis in TBM due to its rapidity, high sensitivity and simple manipulation.

  4. Development of a novel single tube nested PCR for enhanced detection of cytomegalovirus DNA from dried blood spots.

    PubMed

    Atkinson, C; Emery, V C; Griffiths, P D

    2014-02-01

    Newborn screening for congenital cytomegalovirus (CCMV) using dried blood spots (DBS) has been proposed because many developed countries have DBS screening programmes in place for other diseases. The aim of this study was to develop a rapid, single tube nested polymerase chain reaction (PCR) method for enhanced detection of CMV from DBS compared to existing (single target) real time PCRs. The new method was compared with existing real time PCRs for sensitivity and specificity. Overall sensitivity of the single target PCR assays in both asymptomatic and symptomatic infants with laboratory confirmed congenital CMV was 69% (CMV PCR or culture positive before day 21 of life). In contrast, the single tube nested assay had an increased sensitivity of 81% with100% specificity. Overall the assay detected CMV from a DBS equivalent to an original blood sample which contained 500IU/ml. In conclusion this single tube nested methodology allows simultaneous amplification and detection of CMV DNA in 1.5h removing the associated contamination risk of a two step nested PCR. Owing to its increased sensitivity, it has the potential to be used as a screening assay and ultimately allow early identification and intervention for children with congenital CMV. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Detection of fungi by conventional methods and semi-nested PCR in patients with presumed fungal keratitis.

    PubMed

    Haghani, I; Amirinia, F; Nowroozpoor-Dailami, K; Shokohi, T

    2015-06-01

    Fungal keratitis is a suppurative, ulcerative, and sight-threatening infection of the cornea that sometimes leads to blindness. The aims of this study were: recuperating facilities for laboratory diagnosis, determining the causative microorganisms, and comparing conventional laboratory diagnostic tools and semi-nested PCR. Sampling was conducted in patients with suspected fungal keratitis. Two corneal scrapings specimens, one for direct smear and culture and the other for semi- nested PCR were obtained. Of the 40 expected cases of mycotic keratitis, calcofluor white staining showed positivity in 25%, culture in 17.5%, KOH in 10%, and semi-nested PCR in 27.5%. The sensitivities of semi-nested PCR, KOH, and CFW were 57.1%, 28.5%, and 42% while the specificities were 78.7%, 94%, and 78.7%, respectively. The time taken for PCR assay was 4 to 8 hours, whereas positive fungal cultures took at least 5 to 7 days. Due to the increasing incidence of fungal infections in people with weakened immune systems, uninformed using of topical corticosteroids and improper use of contact lens, fast diagnosis and accurate treatment of keratomycosis seems to be essential . Therefore, according to the current study, molecular methods can detect mycotic keratitis early and correctly leading to appropriate treatment.

  6. Lack of spatial genetic structure among nesting and wintering King Eiders

    USGS Publications Warehouse

    Pearce, J.M.; Talbot, S.L.; Pierson, Barbara J.; Petersen, M.R.; Scribner, K.T.; Dickson, D.L.; Mosbech, A.

    2004-01-01

    The King Eider (Somateria spectabilis) has been delineated into two broadly distributed breeding populations in North America (the western and eastern Arctic) on the basis of banding data and their use of widely separated Pacific and Atlantic wintering areas. Little is known about the level of gene flow between these two populations. Also unknown is whether behavioral patterns common among migratory waterfowl, such as site fidelity to wintering areas and pair formation at these sites, have existed for sufficient time to create a population structure defined by philopatry to wintering rather than to nesting locations. We used six nuclear microsatellite DNA loci and cytochrome b mitochondrial DNA sequence data to estimate the extent of spatial genetic differentiation among nesting and wintering areas of King Eiders across North America and adjacent regions. Estimates of interpopulation variance in microsatellite allele and mtDNA haplotype frequency were both low and nonsignificant based on samples from three wintering and four nesting areas. Results from nested clade analysis, mismatch distributions, and coalescent-based analyses suggest historical population growth and gene flow that collectively may have homogenized gene frequencies. The presence of several unique mtDNA haplotypes among birds wintering near Greenland suggests that gene flow may now be more limited between the western and eastern Arctic, which is consistent with banding data.

  7. Use of the superpopulation approach to estimate breeding population size: An example in asynchronously breeding birds

    USGS Publications Warehouse

    Williams, K.A.; Frederick, P.C.; Nichols, J.D.

    2011-01-01

    Many populations of animals are fluid in both space and time, making estimation of numbers difficult. Much attention has been devoted to estimation of bias in detection of animals that are present at the time of survey. However, an equally important problem is estimation of population size when all animals are not present on all survey occasions. Here, we showcase use of the superpopulation approach to capture-recapture modeling for estimating populations where group membership is asynchronous, and where considerable overlap in group membership among sampling occasions may occur. We estimate total population size of long-legged wading bird (Great Egret and White Ibis) breeding colonies from aerial observations of individually identifiable nests at various times in the nesting season. Initiation and termination of nests were analogous to entry and departure from a population. Estimates using the superpopulation approach were 47-382% larger than peak aerial counts of the same colonies. Our results indicate that the use of the superpopulation approach to model nesting asynchrony provides a considerably less biased and more efficient estimate of nesting activity than traditional methods. We suggest that this approach may also be used to derive population estimates in a variety of situations where group membership is fluid. ?? 2011 by the Ecological Society of America.

  8. Ecosystem engineering of harvester ants: effects on vegetation in a sagebrush-steppe ecosystem

    USGS Publications Warehouse

    Gosselin, Elyce N; Holbrook, Joseph D.; Huggler, Katey; Brown, Emily; Vierling, Kerri T.; Arkle, Robert; Pilliod, David S.

    2016-01-01

    Harvester ants are influential in many ecosystems because they distribute and consume seeds, remove vegetation, and redistribute soil particles and nutrients. Understanding the interaction between harvester ants and plant communities is important for management and restoration efforts, particularly in systems altered by fire and invasive species such as the sagebrush-steppe. Our objective was to evaluate how vegetation cover changed as a function of distance from Owyhee harvester ant (Pogonomyrmex salinus) nests within a sagebrush-steppe ecosystem. We sampled 105 harvester ant nests within southern Idaho, USA, that occurred in different habitats: annual grassland, perennial grassland, and native shrubland. The influence of Owyhee harvester ants on vegetation was larger at the edge of ant nests, but the relationship was inconsistent among plant species. Percent cover was positively associated with distance from harvester ant nests for plant species that were considered undesirable food sources and were densely distributed. However, percent cover was negatively associated with distance-from-nests for patchily distributed and desirable plant species. For some plant species, there was no change in cover associated with distance-from-nests. Total vegetation cover was associated with distance-from-nests in the shrubland habitat but not in the 2 grasslands. The dominant plant species in the shrubland habitat was a densely distributed shrub (winterfat, Krascheninnikovia lanata) that was defoliated by harvester ants. Our results suggest that Owyhee harvester ants increase spatial heterogeneity in plant communities through plant clearing, but the direction and magnitude of effect will likely be contingent on the dominant vegetation groups. This information may inform future management and plant restoration efforts in sagebrush-steppe by directly considering the islands of influence associated with harvester ant engineering.

  9. Osprey: worldwide sentinel species for assessing and monitoring environmental contamination in rivers, lakes, reservoirs, and estuaries.

    PubMed

    Grove, Robert A; Henny, Charles J; Kaiser, James L

    2009-01-01

    In the United States, many fish and wildlife species have been used nationwide to monitor environmental contaminant exposure and effects, including carcasses of the bald eagle (Haliaeetus leucocephalus), the only top avian predator regularly used in the past. Unfortunately, bald eagles are sensitive to investigator intrusion at the nest. Thus, the osprey (Pandion haliaetus) is evaluated as a potential sentinel species for aquatic ecosystems. Several characteristics support the choice of the osprey as a sentinel species, including: (1) fish-eating diet atop the aquatic food web, (2) long-lived with strong nest fidelity, (3) adapts to human landscapes (potentially the most contaminated), (4) tolerates short-term nest disturbance, (5) nests spatially distributed at regular intervals, (6) highly visible nests easily located for study, (7) ability to accumulate most, if not all, lipophilic contaminants, (8) known sensitivity to many contaminants, and (9) nearly a worldwide distribution. These osprey traits have been instrumental in successfully using the species to understand population distribution, abundance, and changes over time; the effects of various contaminants on reproductive success; how contaminants in prey (fish on biomass basis) contribute to egg concentrations (i.e., biomagnification factors); and spatial residue patterns. Data summarized include nesting population surveys, detailed nesting studies, and chemical analyses of osprey egg, organ, blood, and feather samples for contaminants that bioaccumulate and/or biomagnify in aquatic food webs; and biochemical evaluations of blood and various organs. Studies in the United States, Canada, Mexico, Europe, and elsewhere have shown the osprey to be a useful sentinel species for monitoring selected environmental contaminants, including some emerging contaminants in lakes, reservoirs, rivers, and estuaries.

  10. Metal concentrations, foraging distances, and fledging success of great blue herons nesting along the Hanford Reach of the Columbia River.

    PubMed

    Tiller, Brett L; Marco, J D; Rickard, W H

    2005-05-01

    An ecological risk assessment of the spatial distribution of metal concentrations along the Hanford Reach of the free-flowing Columbia River in southcentral Washington, identified great blue herons, Ardea herodias, at potential risk through the ingestion of contaminated riverine biota, especially fish. We measured metal concentrations in livers of pre-flight herons from the Hanford Reach and excrement samples taken from the same nests. Nests were distributed among three colonies situated upstream and downstream from nine retired plutonium production reactors along the river where metals in reactor coolant waters had been released directly into the river or disposed to shoreline retention basins and ditches. Distances traveled by parent herons to foraging areas along the river shore were determined by visually tracking parent birds as they flew from nests to upriver and downriver foraging sites. Foraging flight distances varied between colonies with mean distances ranging between 0.7 and 3.1 km. Cadmium, Cr, and Pb concentrations were higher in excrement than in the livers of pre-flight herons but the opposite was noted for Cu, Hg, and Zn. Highest metal concentrations of Cr, Cu, Zn, Cd, and Pb, were measured in excrement taken from heron nests at the colony located upstream from all reactors. These results were consistent with metal concentrations reported in river sediment from the same regions, indicating excrement from the heron nests may be a useful indicator of dietary uptake of metals by herons. Fledging success and eggshell thickness measurements were used as an index of health of the local heron population. The results indicate that the reproductive health of great blue herons nesting along the Hanford Reach is among the highest reported in the continental United States.

  11. Use of apparent thickness for preprocessing of low-frequency electromagnetic data in inversion-based multibarrier evaluation workflow

    NASA Astrophysics Data System (ADS)

    Omar, Saad; Omeragic, Dzevat

    2018-04-01

    The concept of apparent thicknesses is introduced for the inversion-based, multicasing evaluation interpretation workflow using multifrequency and multispacing electromagnetic measurements. A thickness value is assigned to each measurement, enabling the development of two new preprocessing algorithms to remove casing collar artifacts. First, long-spacing apparent thicknesses are used to remove, from the pipe sections, artifacts ("ghosts") caused by the transmitter crossing a casing collar or corrosion. Second, a collar identification, localization, and assignment algorithm is developed to enable robust inversion in collar sections. Last, casing eccentering can also be identified on the basis of opposite deviation of short-spacing phase and magnitude apparent thicknesses from the nominal value. The proposed workflow can handle an arbitrary number of nested casings and has been validated on synthetic and field data.

  12. Belted kingfishers: Under surveillance and sampled in the privacy of their own burrow

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Baron, L.A.; Ashwood, T.L.

    The belted kingfisher, a common piscivore of the Oak Ridge Reservation (ORR), bioaccumulates contaminants from consumption of aquatic prey. Three kingfisher carcasses found near contaminated streams on the ORR were analyzed. Mercury, cadmium, and selenium bioaccumulated within the liver, kidney, and feathers. Additionally, PCB-1254 accumulated in muscle and lipid tissue, while Cesium-137 accumulated within the muscle and whole body. These contaminant levels have been shown to produce a variety of toxicological effects (i.e., reproductive impairment, central nervous system dysfunction) within other species of birds. In addition to use of this data for ecological risk assessment, kingfishers can also be monitoredmore » as a viable bioindicator species reflecting environmental contaminant levels over time. However, current sampling methods of burrow excavation or the use of mist nets can be detrimental to the reproductive success of the birds. The authors present a method for obtaining adequate samples of feathers and other remnants (i.e., egg shells, dried regurgitant) found in the burrow during or following the nesting season. The collection of samples following surveillance of the burrow and its contents was performed with the use of a 15 ft-long flexible, portable probe containing a video camera. Once sighted with the probe, contents of the burrow were collected by insertion of an additional tube attached to a hand held vacuum cleaner (Dirt Devil{reg_sign}). Feathers collected from a nest at an uncontaminated site contained selenium, lead and mercury. Cesium-137 was found in an egg shell collected from a nest at a radiologically contaminated site. Close-up photos of a kingfisher mother incubating her eggs and nestlings within two burrows will also be shown. This surveillance and sampling technique can also be used for monitoring other burrowing terrestrial species.« less

  13. A survey of microbial community diversity in marine sediments impacted by petroleum hydrocarbons from the Gulf of Mexico and Atlantic shorelines, Texas to Florida

    USGS Publications Warehouse

    Lisle, John T.; Stellick, Sarah H.

    2011-01-01

    Microbial community genomic DNA was extracted from sediment samples collected along the Gulf of Mexico and Atlantic coasts from Texas to Florida. Sample sites were identified as being ecologically sensitive and (or) as having high potential of being impacted by Macondo-1 (M-1) well oil from the Deepwater Horizon blowout. The diversity within the microbial communities associated with the collected sediments provides a baseline dataset to which microbial community-diversity data from impacted sites could be compared. To determine the microbial community diversity in the samples, genetic fingerprints were generated and compared. Specific sequences within the community genomic DNA were first amplified using the polymerase chain reaction (PCR) with a primer set that provides possible resolution to the species level. A second nested PCR was performed on the primary PCR products using a primer set on which a GC-clamp was attached to one of the primers. The nested PCR products were separated using denaturing-gradient gel electrophoresis (DGGE) that resolves the nested PCR products based on sequence dissimilarities (or similarities), forming a genomic fingerprint of the microbial diversity within the respective samples. Samples with similar fingerprints were grouped and compared to oil-fingerprint data from the same sites (Rosenbauer and others, 2011). The microbial community fingerprints were generally grouped into sites that had been shown to contain background concentrations of non-Deepwater Horizon oil. However, these groupings also included sites where no oil signature was detected. This report represents some of the first information on naturally occurring microbial communities in sediment from shorelines along the Gulf of Mexico and Atlantic coasts from Texas to Florida.

  14. Isotope analysis reveals foraging area dichotomy for atlantic leatherback turtles.

    PubMed

    Caut, Stéphane; Fossette, Sabrina; Guirlet, Elodie; Angulo, Elena; Das, Krishna; Girondot, Marc; Georges, Jean-Yves

    2008-03-26

    The leatherback turtle (Dermochelys coriacea) has undergone a dramatic decline over the last 25 years, and this is believed to be primarily the result of mortality associated with fisheries bycatch followed by egg and nesting female harvest. Atlantic leatherback turtles undertake long migrations across ocean basins from subtropical and tropical nesting beaches to productive frontal areas. Migration between two nesting seasons can last 2 or 3 years, a time period termed the remigration interval (RI). Recent satellite transmitter data revealed that Atlantic leatherbacks follow two major dispersion patterns after nesting season, through the North Gulf Stream area or more eastward across the North Equatorial Current. However, information on the whole RI is lacking, precluding the accurate identification of feeding areas where conservation measures may need to be applied. Using stable isotopes as dietary tracers we determined the characteristics of feeding grounds of leatherback females nesting in French Guiana. During migration, 3-year RI females differed from 2-year RI females in their isotope values, implying differences in their choice of feeding habitats (offshore vs. more coastal) and foraging latitude (North Atlantic vs. West African coasts, respectively). Egg-yolk and blood isotope values are correlated in nesting females, indicating that egg analysis is a useful tool for assessing isotope values in these turtles, including adults when not available. Our results complement previous data on turtle movements during the first year following the nesting season, integrating the diet consumed during the year before nesting. We suggest that the French Guiana leatherback population segregates into two distinct isotopic groupings, and highlight the urgent need to determine the feeding habitats of the turtle in the Atlantic in order to protect this species from incidental take by commercial fisheries. Our results also emphasize the use of eggs, a less-invasive sampling material than blood, to assess isotopic data and feeding habits for adult female leatherbacks.

  15. Maternal health status correlates with nest success of leatherback sea turtles (Dermochelys coriacea) from Florida.

    PubMed

    Perrault, Justin R; Miller, Debra L; Eads, Erica; Johnson, Chris; Merrill, Anita; Thompson, Larry J; Wyneken, Jeanette

    2012-01-01

    Of the seven sea turtle species, the critically endangered leatherback sea turtle (Dermochelys coriacea) exhibits the lowest and most variable nest success (i.e., hatching success and emergence success) for reasons that remain largely unknown. In an attempt to identify or rule out causes of low reproductive success in this species, we established the largest sample size (n = 60-70 for most values) of baseline blood parameters (protein electrophoresis, hematology, plasma biochemistry) for this species to date. Hematologic, protein electrophoretic and biochemical values are important tools that can provide information regarding the physiological condition of an individual and population health as a whole. It has been proposed that the health of nesting individuals affects their reproductive output. In order to establish correlations with low reproductive success in leatherback sea turtles from Florida, we compared maternal health indices to hatching success and emergence success of their nests. As expected, hatching success (median = 57.4%) and emergence success (median = 49.1%) in Floridian leatherbacks were low during the study period (2007-2008 nesting seasons), a trend common in most nesting leatherback populations (average global hatching success = ∼50%). One protein electrophoretic value (gamma globulin protein) and one hematologic value (red blood cell count) significantly correlated with hatching success and emergence success. Several maternal biochemical parameters correlated with hatching success and/or emergence success including alkaline phosphatase activity, blood urea nitrogen, calcium, calcium:phosphorus ratio, carbon dioxide, cholesterol, creatinine, and phosphorus. Our results suggest that in leatherbacks, physiological parameters correlate with hatching success and emergence success of their nests. We conclude that long-term and comparative studies are needed to determine if certain individuals produce nests with lower hatching success and emergence success than others, and if those individuals with evidence of chronic suboptimal health have lower reproductive success.

  16. Isotope Analysis Reveals Foraging Area Dichotomy for Atlantic Leatherback Turtles

    PubMed Central

    Angulo, Elena; Das, Krishna; Girondot, Marc

    2008-01-01

    Background The leatherback turtle (Dermochelys coriacea) has undergone a dramatic decline over the last 25 years, and this is believed to be primarily the result of mortality associated with fisheries bycatch followed by egg and nesting female harvest. Atlantic leatherback turtles undertake long migrations across ocean basins from subtropical and tropical nesting beaches to productive frontal areas. Migration between two nesting seasons can last 2 or 3 years, a time period termed the remigration interval (RI). Recent satellite transmitter data revealed that Atlantic leatherbacks follow two major dispersion patterns after nesting season, through the North Gulf Stream area or more eastward across the North Equatorial Current. However, information on the whole RI is lacking, precluding the accurate identification of feeding areas where conservation measures may need to be applied. Methodology/Principal Findings Using stable isotopes as dietary tracers we determined the characteristics of feeding grounds of leatherback females nesting in French Guiana. During migration, 3-year RI females differed from 2-year RI females in their isotope values, implying differences in their choice of feeding habitats (offshore vs. more coastal) and foraging latitude (North Atlantic vs. West African coasts, respectively). Egg-yolk and blood isotope values are correlated in nesting females, indicating that egg analysis is a useful tool for assessing isotope values in these turtles, including adults when not available. Conclusions/Significance Our results complement previous data on turtle movements during the first year following the nesting season, integrating the diet consumed during the year before nesting. We suggest that the French Guiana leatherback population segregates into two distinct isotopic groupings, and highlight the urgent need to determine the feeding habitats of the turtle in the Atlantic in order to protect this species from incidental take by commercial fisheries. Our results also emphasize the use of eggs, a less-invasive sampling material than blood, to assess isotopic data and feeding habits for adult female leatherbacks. PMID:18365003

  17. Maternal Health Status Correlates with Nest Success of Leatherback Sea Turtles (Dermochelys coriacea) from Florida

    PubMed Central

    Perrault, Justin R.; Miller, Debra L.; Eads, Erica; Johnson, Chris; Merrill, Anita; Thompson, Larry J.; Wyneken, Jeanette

    2012-01-01

    Of the seven sea turtle species, the critically endangered leatherback sea turtle (Dermochelys coriacea) exhibits the lowest and most variable nest success (i.e., hatching success and emergence success) for reasons that remain largely unknown. In an attempt to identify or rule out causes of low reproductive success in this species, we established the largest sample size (n = 60–70 for most values) of baseline blood parameters (protein electrophoresis, hematology, plasma biochemistry) for this species to date. Hematologic, protein electrophoretic and biochemical values are important tools that can provide information regarding the physiological condition of an individual and population health as a whole. It has been proposed that the health of nesting individuals affects their reproductive output. In order to establish correlations with low reproductive success in leatherback sea turtles from Florida, we compared maternal health indices to hatching success and emergence success of their nests. As expected, hatching success (median = 57.4%) and emergence success (median = 49.1%) in Floridian leatherbacks were low during the study period (2007–2008 nesting seasons), a trend common in most nesting leatherback populations (average global hatching success = ∼50%). One protein electrophoretic value (gamma globulin protein) and one hematologic value (red blood cell count) significantly correlated with hatching success and emergence success. Several maternal biochemical parameters correlated with hatching success and/or emergence success including alkaline phosphatase activity, blood urea nitrogen, calcium, calcium∶phosphorus ratio, carbon dioxide, cholesterol, creatinine, and phosphorus. Our results suggest that in leatherbacks, physiological parameters correlate with hatching success and emergence success of their nests. We conclude that long-term and comparative studies are needed to determine if certain individuals produce nests with lower hatching success and emergence success than others, and if those individuals with evidence of chronic suboptimal health have lower reproductive success. PMID:22359635

  18. Molecular characterization of Orientia tsutsugamushi serotypes causing scrub typhus outbreak in southern region of Andhra Pradesh, India.

    PubMed

    Usha, K; Kumar, E; Kalawat, Usha; Kumar, B Siddhartha; Chaudhury, A; Gopal, D V R Sai

    2016-10-01

    Scrub typhus is a vector-borne zoonotic infection caused by Orientiatsutsugamushi. Local epidemiology of the circulating serotypes of scrub typhus is not available from most parts of India. We conducted this study for the diagnosis of scrub typhus using IgM ELISA and to detect O. tsutsugamushi serotypes circulating in southern Andhra Pradesh, India. Samples were collected from patients clinically suspected to have scrub typhus and were subjected to IgM ELISA to measure IgM antibodies against O. tsutsugamushi. Nested polymerase chain reaction (PCR) was performed targeting strain-specific regions in ELISA-positive samples. Of a total of 663 samples, 258 (38.91%) were found to be positive by IgM ELISA. Serotypes could be detected in 230 (34.69%) samples only. Only two serotypes, Karp and Kawasaki, were found in the serum samples, with the former being predominant. The dual infection of Karp and Kawasaki serotypes was found in seven patients. Other serotypes such as Gilliam, Kuroki and Kato were not detected in the samples. The nested PCR products proved useful in presumptively identifying the endemic O. tsutsugamushi serotypes. The present study could be significant in understanding scrub typhus epidemiology in this region.

  19. Classification of Parkinson's disease utilizing multi-edit nearest-neighbor and ensemble learning algorithms with speech samples.

    PubMed

    Zhang, He-Hua; Yang, Liuyang; Liu, Yuchuan; Wang, Pin; Yin, Jun; Li, Yongming; Qiu, Mingguo; Zhu, Xueru; Yan, Fang

    2016-11-16

    The use of speech based data in the classification of Parkinson disease (PD) has been shown to provide an effect, non-invasive mode of classification in recent years. Thus, there has been an increased interest in speech pattern analysis methods applicable to Parkinsonism for building predictive tele-diagnosis and tele-monitoring models. One of the obstacles in optimizing classifications is to reduce noise within the collected speech samples, thus ensuring better classification accuracy and stability. While the currently used methods are effect, the ability to invoke instance selection has been seldomly examined. In this study, a PD classification algorithm was proposed and examined that combines a multi-edit-nearest-neighbor (MENN) algorithm and an ensemble learning algorithm. First, the MENN algorithm is applied for selecting optimal training speech samples iteratively, thereby obtaining samples with high separability. Next, an ensemble learning algorithm, random forest (RF) or decorrelated neural network ensembles (DNNE), is used to generate trained samples from the collected training samples. Lastly, the trained ensemble learning algorithms are applied to the test samples for PD classification. This proposed method was examined using a more recently deposited public datasets and compared against other currently used algorithms for validation. Experimental results showed that the proposed algorithm obtained the highest degree of improved classification accuracy (29.44%) compared with the other algorithm that was examined. Furthermore, the MENN algorithm alone was found to improve classification accuracy by as much as 45.72%. Moreover, the proposed algorithm was found to exhibit a higher stability, particularly when combining the MENN and RF algorithms. This study showed that the proposed method could improve PD classification when using speech data and can be applied to future studies seeking to improve PD classification methods.

  20. A sampling algorithm for segregation analysis

    PubMed Central

    Tier, Bruce; Henshall, John

    2001-01-01

    Methods for detecting Quantitative Trait Loci (QTL) without markers have generally used iterative peeling algorithms for determining genotype probabilities. These algorithms have considerable shortcomings in complex pedigrees. A Monte Carlo Markov chain (MCMC) method which samples the pedigree of the whole population jointly is described. Simultaneous sampling of the pedigree was achieved by sampling descent graphs using the Metropolis-Hastings algorithm. A descent graph describes the inheritance state of each allele and provides pedigrees guaranteed to be consistent with Mendelian sampling. Sampling descent graphs overcomes most, if not all, of the limitations incurred by iterative peeling algorithms. The algorithm was able to find the QTL in most of the simulated populations. However, when the QTL was not modeled or found then its effect was ascribed to the polygenic component. No QTL were detected when they were not simulated. PMID:11742631

  1. Mercury, cadmium, lead, and selenium in three waterbird species nesting in Galveston Bay, Texas, USA

    USGS Publications Warehouse

    King, K.A.; Cromartie, E.

    1986-01-01

    Heavy metal and selenium concentrations were determined in Olivaceous Cormorants (Phalacrocorax olivaceus ), Laughing gulls (Larus atricilla ), and Black Skimmers (Rynchops niger ) nesting in Galveston Bay, Texas, during 1980-81. Lead was detected at low levels in a small proportion of the liver samples. Mercury was present in all livers sampled, the highest levels being found in cormorants (7.8 ppm) and skimmers (16 ppm). Concentrations were considerably lower than those reported in birds from mercury-contaminated lakes in northern United States and Canada. Cadmium and selenium were detected in 93 and 95% of the kidneys. Cadmium was highest in gulls and skimmers with a maximum value of 16 ppm. Selenium levels were similar among species except for higher concentrations in gulls collected in 1981.

  2. Antagonistic interactions between garden yeasts and microfungal garden pathogens of leaf-cutting ants.

    PubMed

    Rodrigues, Andre; Cable, Rachel N; Mueller, Ulrich G; Bacci, Maurício; Pagnocca, Fernando C

    2009-10-01

    We investigate the diversity of yeasts isolated in gardens of the leafcutter ant Atta texana. Repeated sampling of gardens from four nests over a 1-year time period showed that gardens contain a diverse assemblage of yeasts. The yeast community in gardens consisted mostly of yeasts associated with plants or soil, but community composition changed between sampling periods. In order to understand the potential disease-suppressing roles of the garden yeasts, we screened isolates for antagonistic effects against known microfungal garden contaminants. In vitro assays revealed that yeasts inhibited the mycelial growth of two strains of Escovopsis (a specialized attine garden parasite), Syncephalastrum racemosum (a fungus often growing in gardens of leafcutter lab nests), and the insect pathogen Beauveria bassiana. These garden yeasts add to the growing list of disease-suppressing microbes in attine nests that may contribute synergistically, together with actinomycetes and Burkholderia bacteria, to protect the gardens and the ants against diseases. Additionally, we suggest that garden immunity against problem fungi may therefore derive not only from the presence of disease-suppressing Pseudonocardia actinomycetes, but from an enrichment of multiple disease-suppressing microorganisms in the garden matrix.

  3. Genetic variability in captive populations of the stingless bee Tetragonisca angustula.

    PubMed

    Santiago, Leandro R; Francisco, Flávio O; Jaffé, Rodolfo; Arias, Maria C

    2016-08-01

    Low genetic variability has normally been considered a consequence of animal husbandry and a major contributing factor to declining bee populations. Here, we performed a molecular analysis of captive and wild populations of the stingless bee Tetragonisca angustula, one of the most commonly kept species across South America. Microsatellite analyses showed similar genetic variability between wild and captive populations However, captive populations showed lower mitochondrial genetic variability. Male-mediated gene flow, transport and division of nests are suggested as the most probable explanations for the observed patterns of genetic structure. We conclude that increasing the number of colonies kept through nest divisions does not negatively affect nuclear genetic variability, which seems to be maintained by small-scale male dispersal and human-mediated nest transport. However, the transport of nests from distant localities should be practiced with caution given the high genetic differentiation observed between samples from western and eastern areas. The high genetic structure verified is the result of a long-term evolutionary process, and bees from distant localities may represent unique evolutionary lineages.

  4. PCDDs, PCDFs, PCBs, OC pesticides and mercury in fish and osprey eggs from Willamette River, Oregon (1993, 2001 and 2006) with calculated biomagnification factors

    USGS Publications Warehouse

    Henny, C.J.; Kaiser, J.L.; Grove, R.A.

    2009-01-01

    The osprey (Pandion haliaetus) population nesting along the main stem Willamette River and lower Santiam River was first studied to evaluate contaminants and reproductive rates in 1993 when 78 occupied nests were present. By 2001, the population increased to 234 occupied nests, a 13.7% annual rate of population increase. A sample egg was collected from each of a series of nests along the Upper River (river mile 55-187) in 1993, 2001 and 2006 to evaluate trends of persistent contaminants (organochlorine [OC] pesticides, polychlorinated biphenyls [PCBs], polychlorinated dibenzo-p-dioxins [PCDDs], and polychlorinated dibenzofurans [PCDFs]). Nearly all OC pesticide residues decreased significantly, e.g., p, p?-DDE (DDE) from 2,350 to 1,353 to 210 ??g/kg wet weight (ww). PCBs followed a similar pattern over time, e.g., ???PCBs 688 to 245 to 182 ??g/kg ww, while PCDDs and PCDFs showed a more precipitous decline (often 85-95%) between 1993 and 2001, with no egg analyses warranted in 2006. During 2001-2002, sample osprey eggs were also collected from nests at three Headwater Reservoirs and two lower reaches (Newberg Pool and Tidal Portland) of the Willamette River, as well as the lower portion of the Santiam River to evaluate spatial residue patterns. Significant differences were seldom detected among the different sampling areas for OC pesticides (probably due to small sample sizes), although higher concentrations were often seen in the lower reaches, e.g., DDE 901 ??g/kg ww (Headwater Reservoirs), 1,353 (Upper River), 1,384 (Newberg Pool) and 2,676 (Tidal Portland). PCB congener concentrations in eggs were usually higher in the Tidal Portland reach than at other locations and often significantly higher than at the Headwater Reservoirs or Upper River. Mercury (first analyzed in eggs in 2001), PCDDs and PCDFs were extremely low in 2001/2002 with no significant spatial patterns. Whole fish composite samples of largescale sucker (Catastomus macrocheilus) and northern pikeminnow (Ptychocheilus oregonensis), which account for about 90% of the biomass in the diet of this osprey population, were also collected from the Willamette River in 1993 and 2001 and analyzed for the same contaminants as osprey eggs. Contaminant residues in fish from the Upper River decreased between 1993 and 2001, paralleling findings for osprey eggs. Likewise, spatial patterns for fish residues paralleled findings for osprey eggs from the different reaches in 2001. A second empirical estimate of biomagnification factors (BMFs) from fish to osprey eggs for OC pesticides, PCBs, PCDDs and PCDFs (ww and lipid weight [lw] basis) was calculated based on residue data collected in 2001. The two independent BMF estimates (1993 and 2001) for each contaminant from the Upper River provide a measure of consistency, e.g., DDE (ww) 87 and 79, (lw) 103 and 112; ???PCBs (ww) 11 and 8.4, (lw) 13 and 12. Mercury did not biomagnify from fish to osprey eggs (BMF = 0.60). Legacy contaminants investigated had limited (perhaps only DDE), if any, effects on reproductive success of the increasing osprey population nesting along the Willamette River by 2001. ?? 2008 U.S. Government.

  5. Bloodmeal Host Congregation and Landscape Structure Impact the Estimation of Female Mosquito (Diptera: Culicidae) Abundance Using Dry Ice-Baited Traps

    PubMed Central

    THIEMANN, TARA; NELMS, BRITTANY; REISEN, WILLIAM K.

    2011-01-01

    Vegetation patterns and the presence of large numbers of nesting herons and egrets significantly altered the number of host-seeking Culex tarsalis Coquillett (Diptera: Culicidae) collected at dry ice-baited traps. The numbers of females collected per trap night at traps along the ecotone of Eucalyptus stands with and without a heron colony were always greater or equal to numbers collected at traps within or under canopy. No Cx. tarsalis were collected within or under Eucaplytus canopy during the peak heron nesting season, even though these birds frequently were infected with West Nile virus and large number of engorged females could be collected at resting boxes. These data indicate a diversion of host-seeking females from traps to nesting birds reducing sampling efficiency. PMID:21661310

  6. Reproductive success of Whiskered Tern Chlidonias hybrida in eastern Spain in relation to water level variation.

    PubMed

    Ortiz Lledó, Álvaro; Vidal Mateo, Javier; Urios Moliner, Vicente

    2018-01-01

    A study on the Whiskered Tern Chlidonias hybrida was carried out between 2002 and 2009 in wetlands of eastern Spain to evaluate how water level fluctuation affects its reproductive success (hatching, fledgling and breeding success). This species is catalogued as Vulnerable in Spain and has an unfavorable conservation status in Europe. Our study includes 18 sampling areas from five wetlands, covering a total of 663 nests, 1,618 eggs, 777 nestlings and 225 fledglings. The colonies were visited at least twice per week in breeding period. The number of eggs and/or nestlings present in each nest were annotated each time the colonies were visited with the aim to compare the evolution of these parameters with time. Hatching success was calculated as the proportion of egg that hatched successfully. Fledgling success and breeding success were calculated as the proportion of chicks that fledged successfully and the proportion of eggs that produced fledglings. We used the Kruskal-Wallis test to analyze the differences in the dependent variables hatching, fledgling and breeding success among the wetlands and the sampling areas. We explored the relationship between the different reproductive success with the average fluctuation rate and the anchoring depth of nests, using statistics of the linear regression. It was observed that the reproductive success varied significantly in the interaction among the different categories of water level fluctuation and the different areas (using the Kruskal-Wallis test). Our records showed that pronounced variations in water level destroyed several nests, which affected the Whiskered Tern reproductive success. Considering all events that occurred in 18 areas, the mean (±SD) of nests, eggs and nestlings that were lost after water level fluctuations were of 25.60 ± 21.79%, 32.06 ± 27.58% and 31.91 ± 21.28% respectively, also including the effects of rain and predation. Unfavorable climatic events, such as strong wind, rain or hail, also caused the loss of nests, eggs and nestlings, even when wetland water levels remained constant. The influence of the anchorage depth of the nest and the water level fluctuation rate were analyzed and did not provide statistically significant results. It was not possible to establish a clear pattern on these latter variables, so further studies are needed to obtain more significant results. We propose to undertake similar studies in wetlands where the water level can be regulated, with the range of nest anchorage depth on the emergent vegetation being between 30 and 60 cm, which could improve the reproductive success in this kind of habitats. As recommendation, in water level controlled wetlands (that use sluices), it should not vary more than ±6 cm in a short time (1-2 days) once the nests are established since it negatively affects their reproductive success.

  7. Clinical application of RT-nested PCR integrated with RFLP in Hantavirus detection and genotyping: a prospective study in Shandong Province, PR China.

    PubMed

    Liu, Yun-Xi; Zhao, Zhong-Tang; Cao, Wu-Chun; Xu, Xiao-Qun; Suo, Ji-Jiang; Xing, Yu-Bin; Jia, Ning; Du, Ming-Mei; Liu, Bo-Wei; Yao, Yuan

    2013-01-01

    The aim of the present study was to evaluate the clinical usefulness of applying RT-nested PCR along with RFLP as a method for diagnosis and genotypic differentiation of Hantavirus in the acute-stage sera of HFRS patients as compared to the ELISA technique. A prospective study of patients with suspected HFRS patients was carried out. Sera were collected for serological evaluation by ELISA and RT-nested PCR testing. Primers were selected from the published sequence of the S segment of HTNV strain 76-118 and SEOV strain SR-11, which made it possible to obtain an amplicon of 403 bp by RT-nested PCR. The genotypic differentiations of the RT-nested PCR amplicons were carried out by RFLP. Sequence analyses of the amplicons were used to confirm the accuracy of the results obtained by RFLP. Of the 48 acute-stage sera from suspected HFRS patients, 35 were ELISA-positive while 41 were positive by RT-nested PCR. With Hind III and Hinf I, RFLP profiles of the RT-nested PCR amplicons of the 41 positive sera exhibited two patterns. 33 had RFLP profiles similar to the reference strain R22, and thus belonged to the SEOV type. The other 8 samples which were collected during October-December had RFLP profiles similar to the reference strain 76-118, and thus belonged to the HTNV type. Sequence phylogenetic analysis of RT-nested PCR amplicons revealed sdp1, sdp2 YXL-2008, and sdp3 as close relatives of HTNV strain 76-118, while sdp22 and sdp37 as close relatives of SEOV strain Z37 and strain R22 located in two separate clusters in the phylogenetic tree. These results were identical to those acquired by RFLP. RT-nested PCR integrated with RFLP was a rapid, simple, accurate method for detecting and differentiating the genotypes of Hantavirus in the acute-stage sera of suspected HFRS patients. In Shandong province, the main genotypes of Hantavirus belonged to the SEOV types, while the HTNV types were observed during the autumn-winter season.

  8. Influence of food supply and chlorinated hydrocarbon contaminants on breeding success of bald eagles.

    PubMed

    Gill, Christopher E; Elliott, John E

    2003-01-01

    Food supply and contaminants were investigated as possible causes of low bald eagle productivity near a bleached kraft pulp and paper mill at Crofton on Vancouver Island, British Columbia. Over a seven year period, 1992-1998, average productivity of five eagle territories situated south of the pulp mill at Crofton was significantly lower (0.43 young/occupied territory) than six territories north of the mill (1.04 young/occupied territory). A reference population of 32 territories located in Barkley Sound on the west coast of Vancouver Island demonstrated intermediary mean productivity (0.75 young/occupied territory). Measures of prey biomass delivered to nests were lowest south of the mill, and correlated significantly with nesting success. On average, measures of energy delivered to nests and a parameter determined to be related to prey availability, adult nest attendance time, accounted for about 70% of variability in nest success. Contaminant concentrations, including pulp mill derived polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), as well as dichlorodiphenyldichloroethane (DDE), polychlorinated biphenyls (PCBs), and calculated tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) were significantly greater in plasma samples of nestlings from south of the mill compared to the other two sites, but did not correlate significantly with individual nest success data. Nests south of the mill concentrate around Maple Bay, which appears to be a deposition area for contaminants transported by tides and currents from sources such as the pulp mill. Concentrations of DDE and PCBs in plasma of nestling eagles from south of the mill were less than the critical values estimated to affect production of young. For TEQs, there are no published critical values for plasma by which to compare our results. We conclude that less than adequate energy provisioning to nests, presumably related to low prey availability, was likely the main cause of poor nest success south of the mill site at Crofton. However, higher concentrations of both DDE and PCDD/F derived TEQs may have acted in concert with food stress to further reduce bald eagle productivity.

  9. Implementation on a nonlinear concrete cracking algorithm in NASTRAN

    NASA Technical Reports Server (NTRS)

    Herting, D. N.; Herendeen, D. L.; Hoesly, R. L.; Chang, H.

    1976-01-01

    A computer code for the analysis of reinforced concrete structures was developed using NASTRAN as a basis. Nonlinear iteration procedures were developed for obtaining solutions with a wide variety of loading sequences. A direct access file system was used to save results at each load step to restart within the solution module for further analysis. A multi-nested looping capability was implemented to control the iterations and change the loads. The basis for the analysis is a set of mutli-layer plate elements which allow local definition of materials and cracking properties.

  10. Chimera states in networks of logistic maps with hierarchical connectivities

    NASA Astrophysics Data System (ADS)

    zur Bonsen, Alexander; Omelchenko, Iryna; Zakharova, Anna; Schöll, Eckehard

    2018-04-01

    Chimera states are complex spatiotemporal patterns consisting of coexisting domains of coherence and incoherence. We study networks of nonlocally coupled logistic maps and analyze systematically how the dilution of the network links influences the appearance of chimera patterns. The network connectivities are constructed using an iterative Cantor algorithm to generate fractal (hierarchical) connectivities. Increasing the hierarchical level of iteration, we compare the resulting spatiotemporal patterns. We demonstrate that a high clustering coefficient and symmetry of the base pattern promotes chimera states, and asymmetric connectivities result in complex nested chimera patterns.

  11. Improved pulse laser ranging algorithm based on high speed sampling

    NASA Astrophysics Data System (ADS)

    Gao, Xuan-yi; Qian, Rui-hai; Zhang, Yan-mei; Li, Huan; Guo, Hai-chao; He, Shi-jie; Guo, Xiao-kang

    2016-10-01

    Narrow pulse laser ranging achieves long-range target detection using laser pulse with low divergent beams. Pulse laser ranging is widely used in military, industrial, civil, engineering and transportation field. In this paper, an improved narrow pulse laser ranging algorithm is studied based on the high speed sampling. Firstly, theoretical simulation models have been built and analyzed including the laser emission and pulse laser ranging algorithm. An improved pulse ranging algorithm is developed. This new algorithm combines the matched filter algorithm and the constant fraction discrimination (CFD) algorithm. After the algorithm simulation, a laser ranging hardware system is set up to implement the improved algorithm. The laser ranging hardware system includes a laser diode, a laser detector and a high sample rate data logging circuit. Subsequently, using Verilog HDL language, the improved algorithm is implemented in the FPGA chip based on fusion of the matched filter algorithm and the CFD algorithm. Finally, the laser ranging experiment is carried out to test the improved algorithm ranging performance comparing to the matched filter algorithm and the CFD algorithm using the laser ranging hardware system. The test analysis result demonstrates that the laser ranging hardware system realized the high speed processing and high speed sampling data transmission. The algorithm analysis result presents that the improved algorithm achieves 0.3m distance ranging precision. The improved algorithm analysis result meets the expected effect, which is consistent with the theoretical simulation.

  12. Army ants algorithm for rare event sampling of delocalized nonadiabatic transitions by trajectory surface hopping and the estimation of sampling errors by the bootstrap method.

    PubMed

    Nangia, Shikha; Jasper, Ahren W; Miller, Thomas F; Truhlar, Donald G

    2004-02-22

    The most widely used algorithm for Monte Carlo sampling of electronic transitions in trajectory surface hopping (TSH) calculations is the so-called anteater algorithm, which is inefficient for sampling low-probability nonadiabatic events. We present a new sampling scheme (called the army ants algorithm) for carrying out TSH calculations that is applicable to systems with any strength of coupling. The army ants algorithm is a form of rare event sampling whose efficiency is controlled by an input parameter. By choosing a suitable value of the input parameter the army ants algorithm can be reduced to the anteater algorithm (which is efficient for strongly coupled cases), and by optimizing the parameter the army ants algorithm may be efficiently applied to systems with low-probability events. To demonstrate the efficiency of the army ants algorithm, we performed atom-diatom scattering calculations on a model system involving weakly coupled electronic states. Fully converged quantum mechanical calculations were performed, and the probabilities for nonadiabatic reaction and nonreactive deexcitation (quenching) were found to be on the order of 10(-8). For such low-probability events the anteater sampling scheme requires a large number of trajectories ( approximately 10(10)) to obtain good statistics and converged semiclassical results. In contrast by using the new army ants algorithm converged results were obtained by running 10(5) trajectories. Furthermore, the results were found to be in excellent agreement with the quantum mechanical results. Sampling errors were estimated using the bootstrap method, which is validated for use with the army ants algorithm. (c) 2004 American Institute of Physics.

  13. Towards high-throughput molecular detection of Plasmodium: new approaches and molecular markers

    PubMed Central

    Steenkeste, Nicolas; Incardona, Sandra; Chy, Sophy; Duval, Linda; Ekala, Marie-Thérèse; Lim, Pharath; Hewitt, Sean; Sochantha, Tho; Socheat, Doung; Rogier, Christophe; Mercereau-Puijalon, Odile; Fandeur, Thierry; Ariey, Frédéric

    2009-01-01

    Background Several strategies are currently deployed in many countries in the tropics to strengthen malaria control toward malaria elimination. To measure the impact of any intervention, there is a need to detect malaria properly. Mostly, decisions still rely on microscopy diagnosis. But sensitive diagnosis tools enabling to deal with a large number of samples are needed. The molecular detection approach offers a much higher sensitivity, and the flexibility to be automated and upgraded. Methods Two new molecular methods were developed: dot18S, a Plasmodium-specific nested PCR based on the 18S rRNA gene followed by dot-blot detection of species by using species-specific probes and CYTB, a Plasmodium-specific nested PCR based on cytochrome b gene followed by species detection using SNP analysis. The results were compared to those obtained with microscopic examination and the "standard" 18S rRNA gene based nested PCR using species specific primers. 337 samples were diagnosed. Results Compared to the microscopy the three molecular methods were more sensitive, greatly increasing the estimated prevalence of Plasmodium infection, including P. malariae and P. ovale. A high rate of mixed infections was uncovered with about one third of the villagers infected with more than one malaria parasite species. Dot18S and CYTB sensitivity outranged the "standard" nested PCR method, CYTB being the most sensitive. As a consequence, compared to the "standard" nested PCR method for the detection of Plasmodium spp., the sensitivity of dot18S and CYTB was respectively 95.3% and 97.3%. Consistent detection of Plasmodium spp. by the three molecular methods was obtained for 83% of tested isolates. Contradictory results were mostly related to detection of Plasmodium malariae and Plasmodium ovale in mixed infections, due to an "all-or-none" detection effect at low-level parasitaemia. Conclusion A large reservoir of asymptomatic infections was uncovered using the molecular methods. Dot18S and CYTB, the new methods reported herein are highly sensitive, allow parasite DNA extraction as well as genus- and species-specific diagnosis of several hundreds of samples, and are amenable to high-throughput scaling up for larger sample sizes. Such methods provide novel information on malaria prevalence and epidemiology and are suited for active malaria detection. The usefulness of such sensitive malaria diagnosis tools, especially in low endemic areas where eradication plans are now on-going, is discussed in this paper. PMID:19402894

  14. Identifying marker genes in transcription profiling data using a mixture of feature relevance experts.

    PubMed

    Chow, M L; Moler, E J; Mian, I S

    2001-03-08

    Transcription profiling experiments permit the expression levels of many genes to be measured simultaneously. Given profiling data from two types of samples, genes that most distinguish the samples (marker genes) are good candidates for subsequent in-depth experimental studies and developing decision support systems for diagnosis, prognosis, and monitoring. This work proposes a mixture of feature relevance experts as a method for identifying marker genes and illustrates the idea using published data from samples labeled as acute lymphoblastic and myeloid leukemia (ALL, AML). A feature relevance expert implements an algorithm that calculates how well a gene distinguishes samples, reorders genes according to this relevance measure, and uses a supervised learning method [here, support vector machines (SVMs)] to determine the generalization performances of different nested gene subsets. The mixture of three feature relevance experts examined implement two existing and one novel feature relevance measures. For each expert, a gene subset consisting of the top 50 genes distinguished ALL from AML samples as completely as all 7,070 genes. The 125 genes at the union of the top 50s are plausible markers for a prototype decision support system. Chromosomal aberration and other data support the prediction that the three genes at the intersection of the top 50s, cystatin C, azurocidin, and adipsin, are good targets for investigating the basic biology of ALL/AML. The same data were employed to identify markers that distinguish samples based on their labels of T cell/B cell, peripheral blood/bone marrow, and male/female. Selenoprotein W may discriminate T cells from B cells. Results from analysis of transcription profiling data from tumor/nontumor colon adenocarcinoma samples support the general utility of the aforementioned approach. Theoretical issues such as choosing SVM kernels and their parameters, training and evaluating feature relevance experts, and the impact of potentially mislabeled samples on marker identification (feature selection) are discussed.

  15. Cost-effective analysis of different algorithms for the diagnosis of hepatitis C virus infection.

    PubMed

    Barreto, A M E C; Takei, K; E C, Sabino; Bellesa, M A O; Salles, N A; Barreto, C C; Nishiya, A S; Chamone, D F

    2008-02-01

    We compared the cost-benefit of two algorithms, recently proposed by the Centers for Disease Control and Prevention, USA, with the conventional one, the most appropriate for the diagnosis of hepatitis C virus (HCV) infection in the Brazilian population. Serum samples were obtained from 517 ELISA-positive or -inconclusive blood donors who had returned to Fundação Pró-Sangue/Hemocentro de São Paulo to confirm previous results. Algorithm A was based on signal-to-cut-off (s/co) ratio of ELISA anti-HCV samples that show s/co ratio > or =95% concordance with immunoblot (IB) positivity. For algorithm B, reflex nucleic acid amplification testing by PCR was required for ELISA-positive or -inconclusive samples and IB for PCR-negative samples. For algorithm C, all positive or inconclusive ELISA samples were submitted to IB. We observed a similar rate of positive results with the three algorithms: 287, 287, and 285 for A, B, and C, respectively, and 283 were concordant with one another. Indeterminate results from algorithms A and C were elucidated by PCR (expanded algorithm) which detected two more positive samples. The estimated cost of algorithms A and B was US$21,299.39 and US$32,397.40, respectively, which were 43.5 and 14.0% more economic than C (US$37,673.79). The cost can vary according to the technique used. We conclude that both algorithms A and B are suitable for diagnosing HCV infection in the Brazilian population. Furthermore, algorithm A is the more practical and economical one since it requires supplemental tests for only 54% of the samples. Algorithm B provides early information about the presence of viremia.

  16. Breeding Sex Ratios in Adult Leatherback Turtles (Dermochelys coriacea) May Compensate for Female-Biased Hatchling Sex Ratios

    PubMed Central

    Stewart, Kelly R.; Dutton, Peter H.

    2014-01-01

    For vertebrates with temperature-dependent sex determination, primary (or hatchling) sex ratios are often skewed, an issue of particular relevance to concerns over effects of climate change on populations. However, the ratio of breeding males to females, or the operational sex ratio (OSR), is important to understand because it has consequences for population demographics and determines the capacity of a species to persist. The OSR also affects mating behaviors and mate choice, depending on the more abundant sex. For sea turtles, hatchling and juvenile sex ratios are generally female-biased, and with warming nesting beach temperatures, there is concern that populations may become feminized. Our purpose was to evaluate the breeding sex ratio for leatherback turtles at a nesting beach in St. Croix, USVI. In 2010, we sampled nesting females and later sampled their hatchlings as they emerged from nests. Total genomic DNA was extracted and all individuals were genotyped using 6 polymorphic microsatellite markers. We genotyped 662 hatchlings from 58 females, matching 55 females conclusively to their nests. Of the 55, 42 females mated with one male each, 9 mated with 2 males each and 4 mated with at least 3 males each, for a multiple paternity rate of 23.6%. Using GERUD1.0, we reconstructed parental genotypes, identifying 47 different males and 46 females for an estimated breeding sex ratio of 1.02 males for every female. Thus we demonstrate that there are as many actively breeding males as females in this population. Concerns about female-biased adult sex ratios may be premature, and mate choice or competition may play more of a role in sea turtle reproduction than previously thought. We recommend monitoring breeding sex ratios in the future to allow the integration of this demographic parameter in population models. PMID:24505403

  17. Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients.

    PubMed

    Jamshidi Makiani, Mahin; Davoodian, Parivash; Baghershiroodi, Mahnaz; Nejatizadeh, Abdol Azim; Fakkhar, Farideh; Zangeneh, Mehrangiz; Jahangiri, Nadia

    2016-08-01

    While tuberculosis (TB) can be diagnosed by microscopy and culture, the sensitivity of Ziehl-Neelsen staining is variable and culture results require 4 - 8 weeks to be determined. Polymerase chain reaction (PCR) and its modifications, including nested PCR, might be promising methods for the rapid diagnosis of TB. This study aimed to evaluate the performance of nested PCR on urine samples of human immunodeficiency virus (HIV)-positive and -negative patients with different manifestations of clinical TB. In a prospective study, three early-morning urine samples from 100 patients with pulmonary TB (PTB) or extrapulmonary TB (EPTB) were evaluated using a molecular target with insertion element IS6110, specific to the Mycobacterium tuberculosis genome, and nested PCR was performed. The results were analyzed with SPSS version 22. A total of 100 patients, including 74 (74%) with PTB and 26 (26%) with EPTB, were enrolled. Positive smears were seen in 38 patients (38%). Lymph nodes were the most commonly involved organ in 14 of the 26 (53.8%) EPTB patients (13.5%). Seven (23.1%) of the EPTB patients were HIV-positive. Urine PCR was positive in only 28 patients (28%). Seven HIV-positive patients with PTB showed positive urine PCR results. Moreover, PCR results were positive in only one of the seven HIV-positive subjects with EPTB. Positive PCR results were found in 20 of the 73 HIV-negative patients (27.4%) and in 8 of the 27 HIV-positive patients (29.6%). Therefore, there was no significant difference between the HIV-negative and HIV-positive patients for urine PCR (sensitivity 29.6%, specificity 72.6%; positive and negative predictive values 28% and 72%, respectively; P = 0.138). Nested PCR showed the same sensitivity in HIV-positive and HIV-negative patients. It can be applied as a rapid technique for the diagnosis of TB.

  18. Hawksbill turtle terra incognita: conservation genetics of eastern Pacific rookeries.

    PubMed

    Gaos, Alexander R; Lewison, Rebecca L; Liles, Michael J; Gadea, Velkiss; Altamirano, Eduardo; Henríquez, Ana V; Torres, Perla; Urteaga, José; Vallejo, Felipe; Baquero, Andres; LeMarie, Carolina; Muñoz, Juan Pablo; Chaves, Jaime A; Hart, Catherine E; Peña de Niz, Alejandro; Chácon, Didiher; Fonseca, Luis; Otterstrom, Sarah; Yañez, Ingrid L; LaCasella, Erin L; Frey, Amy; Jensen, Michael P; Dutton, Peter H

    2016-02-01

    Prior to 2008 and the discovery of several important hawksbill turtle (Eretmochelys imbricata) nesting colonies in the EP (Eastern Pacific), the species was considered virtually absent from the region. Research since that time has yielded new insights into EP hawksbills, salient among them being the use of mangrove estuaries for nesting. These recent revelations have raised interest in the genetic characterization of hawksbills in the EP, studies of which have remained lacking to date. Between 2008 and 2014, we collected tissue samples from 269 nesting hawksbills at nine rookeries across the EP and used mitochondrial DNA sequences (766 bp) to generate the first genetic characterization of rookeries in the region. Our results inform genetic diversity, population differentiation, and phylogeography of the species. Hawksbills in the EP demonstrate low genetic diversity: We identified a total of only seven haplotypes across the region, including five new and two previously identified nesting haplotypes (pooled frequencies of 58.4% and 41.6%, respectively), the former only evident in Central American rookeries. Despite low genetic diversity, we found strong stock structure between the four principal rookeries, suggesting the existence of multiple populations and warranting their recognition as distinct management units. Furthermore, haplotypes EiIP106 and EiIP108 are unique to hawksbills that nest in mangrove estuaries, a behavior found only in hawksbills along Pacific Central America. The detected genetic differentiation supports the existence of a novel mangrove estuary "reproductive ecotype" that may warrant additional conservation attention. From a phylogeographic perspective, our research indicates hawksbills colonized the EP via the Indo-Pacific, and do not represent relict populations isolated from the Atlantic by the rising of the Panama Isthmus. Low overall genetic diversity in the EP is likely the combined result of few rookeries, extremely small reproductive populations and evolutionarily recent colonization events. Additional research with larger sample sizes and variable markers will help further genetic understanding of hawksbill turtles in the EP.

  19. Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp.

    PubMed

    Nomura, Ryota; Ogaya, Yuko; Matayoshi, Saaya; Morita, Yumiko; Nakano, Kazuhiko

    2018-04-16

    Recently, dental pulp has been considered a possible source of infection of Helicobacter pylori (H. pylori) in children. We previously developed a novel PCR system for H. pylori detection with high specificity and sensitivity using primer sets constructed based on the complete genome information for 48 H. pylori strains. This PCR system showed high sensitivity with a detection limit of 1-10 cells when serial dilutions of H. pylori genomic DNA were used as templates. However, the detection limit was lower (10 2 -10 3 cells) when H. pylori bacterial DNA was detected from inflamed pulp specimens. Thus, we further refined the system using a nested PCR method, which was much more sensitive than the previous single PCR method. In addition, we examined the distribution and virulence of H. pylori in inflamed pulp tissue. Nested PCR system was constructed using primer sets designed from the complete genome information of 48 H. pylori strains. The detection limit of the nested PCR system was 1-10 cells using both H. pylori genomic DNA and bacterial DNA isolated from inflamed pulp specimens. Next, distribution of H. pylori was examined using 131 inflamed pulp specimens with the nested PCR system. In addition, association between the detection of H. pylori and clinical information regarding endodontic-infected teeth were investigated. Furthermore, adhesion property of H. pylori strains to human dental fibroblast cells was examined. H. pylori was present in 38.9% of inflamed pulp specimens using the nested PCR system. H. pylori was shown to be predominantly detected in primary teeth rather than permanent teeth. In addition, samplings of the inflamed pulp were performed twice from the same teeth at 1- or 2-week intervals, which revealed that H. pylori was detected in most specimens in both samplings. Furthermore, H. pylori strains showed adhesion property to human dental fibroblast cells. Our results suggest that H. pylori colonizes inflamed pulp in approximately 40% of all cases through adhesion to human dental fibroblast cells.

  20. Detection of Salmonella enterica Serovar Montevideo and Newport in Free-ranging Sea Turtles and Beach Sand in the Caribbean and Persistence in Sand and Seawater Microcosms.

    PubMed

    Ives, A-K; Antaki, E; Stewart, K; Francis, S; Jay-Russell, M T; Sithole, F; Kearney, M T; Griffin, M J; Soto, E

    2017-09-01

    Salmonellae are Gram-negative zoonotic bacteria that are frequently part of the normal reptilian gastrointestinal flora. The main objective of this project was to estimate the prevalence of non-typhoidal Salmonella enterica in the nesting and foraging populations of sea turtles on St. Kitts and in sand from known nesting beaches. Results suggest a higher prevalence of Salmonella in nesting leatherback sea turtles compared with foraging green and hawksbill sea turtles. Salmonella was cultured from 2/9 and identified by molecular diagnostic methods in 3/9 leatherback sea turtle samples. Salmonella DNA was detected in one hawksbill turtle, but viable isolates were not recovered from any hawksbill sea turtles. No Salmonella was detected in green sea turtles. In samples collected from nesting beaches, Salmonella was only recovered from a single dry sand sample. All recovered isolates were positive for the wzx gene, consistent with the O:7 serogroup. Further serotyping characterized serovars Montevideo and Newport present in cloacal and sand samples. Repetitive-element palindromic PCR (rep-PCR) fingerprint analysis and pulsed-field gel electrophoresis of the 2014 isolates from turtles and sand as well as archived Salmonella isolates recovered from leatherback sea turtles in 2012 and 2013, identified two distinct genotypes and four different pulsotypes, respectively. The genotyping and serotyping were directly correlated. To determine the persistence of representative strains of each serotype/genotype in these environments, laboratory-controlled microcosm studies were performed in water and sand (dry and wet) incubated at 25 or 35°C. Isolates persisted for at least 32 days in most microcosms, although there were significant decreases in culturable bacteria in several microcosms, with the greatest reduction in dry sand incubated at 35°C. This information provides a better understanding of the epizootiology of Salmonella in free-ranging marine reptiles and the potential public health risks associated with human interactions with these animals in the Caribbean. © 2016 Blackwell Verlag GmbH.

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