Sample records for microbial inactivation properties

  1. Microbial Inactivation by Ultrasound Assisted Supercritical Fluids

    NASA Astrophysics Data System (ADS)

    Benedito, Jose; Ortuño, Carmen; Castillo-Zamudio, Rosa Isela; Mulet, Antonio

    A method combining supercritical carbon dioxide (SC-CO2) and high power ultrasound (HPU) has been developed and tested for microbial/enzyme inactivation purposes, at different process conditions for both liquid and solid matrices. In culture media, using only SC-CO2, the inactivation rate of E. coli and S. cerevisiae increased with pressure and temperature; and the total inactivation (7-8 log-cycles) was attained after 25 and 140 min of SC-CO2 (350 bar, 36 °C) treatment, respectively. Using SC-CO2+HPU, the time for the total inactivation of both microorganisms was reduced to only 1-2 min, at any condition selected. The SC-CO2+HPU inactivation of both microorganisms was slower in juices (avg. 4.9 min) than in culture media (avg. 1.5 min). In solid samples (chicken, turkey ham and dry-cured pork cured ham) treated with SC-CO2 and SC-CO2+HPU, the inactivation rate of E. coli increased with temperature. The application of HPU to the SC-CO2 treatments accelerated the inactivation rate of E. coli and that effect was more pronounced in treatments with isotonic solution surrounding the solid food samples. The application of HPU enhanced the SC-CO2 inactivation mechanisms of microorganisms, generating a vigorous agitation that facilitated the CO2 solubilization and the mass transfer process. The cavitation generated by HPU could damage the cell walls accelerating the extraction of vital constituents and the microbial death. Thus, using the combined technique, reasonable industrial processing times and mild process conditions could be used which could result into a cost reduction and lead to the minimization in the food nutritional and organoleptic changes.

  2. Stochastic and deterministic model of microbial heat inactivation.

    PubMed

    Corradini, Maria G; Normand, Mark D; Peleg, Micha

    2010-03-01

    Microbial inactivation is described by a model based on the changing survival probabilities of individual cells or spores. It is presented in a stochastic and discrete form for small groups, and as a continuous deterministic model for larger populations. If the underlying mortality probability function remains constant throughout the treatment, the model generates first-order ("log-linear") inactivation kinetics. Otherwise, it produces survival patterns that include Weibullian ("power-law") with upward or downward concavity, tailing with a residual survival level, complete elimination, flat "shoulder" with linear or curvilinear continuation, and sigmoid curves. In both forms, the same algorithm or model equation applies to isothermal and dynamic heat treatments alike. Constructing the model does not require assuming a kinetic order or knowledge of the inactivation mechanism. The general features of its underlying mortality probability function can be deduced from the experimental survival curve's shape. Once identified, the function's coefficients, the survival parameters, can be estimated directly from the experimental survival ratios by regression. The model is testable in principle but matching the estimated mortality or inactivation probabilities with those of the actual cells or spores can be a technical challenge. The model is not intended to replace current models to calculate sterility. Its main value, apart from connecting the various inactivation patterns to underlying probabilities at the cellular level, might be in simulating the irregular survival patterns of small groups of cells and spores. In principle, it can also be used for nonthermal methods of microbial inactivation and their combination with heat.

  3. Bioinactivation: Software for modelling dynamic microbial inactivation.

    PubMed

    Garre, Alberto; Fernández, Pablo S; Lindqvist, Roland; Egea, Jose A

    2017-03-01

    This contribution presents the bioinactivation software, which implements functions for the modelling of isothermal and non-isothermal microbial inactivation. This software offers features such as user-friendliness, modelling of dynamic conditions, possibility to choose the fitting algorithm and generation of prediction intervals. The software is offered in two different formats: Bioinactivation core and Bioinactivation SE. Bioinactivation core is a package for the R programming language, which includes features for the generation of predictions and for the fitting of models to inactivation experiments using non-linear regression or a Markov Chain Monte Carlo algorithm (MCMC). The calculations are based on inactivation models common in academia and industry (Bigelow, Peleg, Mafart and Geeraerd). Bioinactivation SE supplies a user-friendly interface to selected functions of Bioinactivation core, namely the model fitting of non-isothermal experiments and the generation of prediction intervals. The capabilities of bioinactivation are presented in this paper through a case study, modelling the non-isothermal inactivation of Bacillus sporothermodurans. This study has provided a full characterization of the response of the bacteria to dynamic temperature conditions, including confidence intervals for the model parameters and a prediction interval of the survivor curve. We conclude that the MCMC algorithm produces a better characterization of the biological uncertainty and variability than non-linear regression. The bioinactivation software can be relevant to the food and pharmaceutical industry, as well as to regulatory agencies, as part of a (quantitative) microbial risk assessment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Application of response surface methodology to optimise microbial inactivation of shrimp and conch by supercritical carbon dioxide.

    PubMed

    Chen, Manhua; Sui, Xiao; Ma, Xixiu; Feng, Xiaomei; Han, Yuqian

    2015-03-30

    Supercritical carbon dioxide (SC-CO2 ) has been shown to have a good pasteurising effect on food. However, very few research papers have investigated the possibility to exploit this treatment for solid foods, particularly for seafood. Considering the microbial safety of raw seafood consumption, the study aimed to explore the feasibility of microbial inactivation of shrimp (Metapenaeus ensis) and conch (Rapana venosa) by SC-CO2 treatment. Response surface methodology (RSM) models were established to predict and analyse the SC-CO2 process. A 3.69-log reduction in the total aerobic plate count (TPC) of shrimp was observed by SC-CO2 treatment at 53°C, 15 MPa for 40 min, and the logarithmic reduction in TPC of conch was 3.31 at 55°C, 14 MPa for 42 min. Sensory scores of the products achieved approximately 8 (desirable). The optimal parameters for microbial inactivation of shrimp and conch by SC-CO2 might be 55°C, 15 MPa and 40 min. SC-CO2 exerted a strong bactericidal effect on the TPC of shrimp and conch, and the products maintained good organoleptic properties. This study verified the feasibility of microbial inactivation of shrimp and conch by SC-CO2 treatment. © 2014 Society of Chemical Industry.

  5. In situ studies of microbial inactivation during high pressure processing

    NASA Astrophysics Data System (ADS)

    Maldonado, Jose Antonio; Schaffner, Donald W.; Cuitiño, Alberto M.; Karwe, Mukund V.

    2016-01-01

    High pressure processing (HPP) has been shown to reduce microbial concentration in foods. The mechanisms of microbial inactivation by HPP have been associated with damage to cell membranes. The real-time response of bacteria to HPP was measured to elucidate the mechanisms of inactivation, which can aid in designing more effective processes. Different pressure cycling conditions were used to expose Enterobacter aerogenes cells to HPP. Propidium iodide (PI) was used as a probe, which fluoresces after penetrating cells with damaged membranes and binding with nucleic acids. A HPP vessel with sapphire windows was used for measuring fluorescence in situ. Membrane damage was detected during pressurization and hold time, but not during depressurization. The drop in fluorescence was larger than expected after pressure cycles at higher pressure and longer times. This indicated possible reversible disassociation of ribosomes resulting in additional binding of PI to exposed RNA under pressure and its release after depressurization.

  6. Instrument for Study of Microbial Thermal Inactivation

    PubMed Central

    Dickerson, R. W.; Read, R. B.

    1968-01-01

    An instrument was designed for the study of thermal inactivation of microorganisms using heating times of less than 1 sec. The instrument operates on the principle of rapid automatic displacement of the microorganism to and from a saturated steam atmosphere, and the operating temperature range is 50 to 90 C. At a temperature of 70 C, thermometric lag (time required to respond to 63.2% of a step change) of the fluid sample containing microorganisms was 0.12 sec. Heating time required to heat the sample to within 0.1 C of the exposure temperature was less than 1 sec, permitting exposure periods as brief as 1 sec, provided the proper corrections are made for the lethal effect of heating. The instrument is most useful for heat exposure periods of less than 5 min, and, typically, more than 500 samples can be processed for microbial inactivation determinations within an 8-hr period. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 7 Fig. 8 PMID:4874466

  7. Electron beam inactivation of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge.

    PubMed

    Praveen, Chandni; Jesudhasan, Palmy R; Reimers, Robert S; Pillai, Suresh D

    2013-09-01

    Microbial pathogens in municipal sewage sludges need to be inactivated prior to environmental disposal. The efficacy of high energy (10 MeV) e-beam irradiation to inactivate a variety of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge was evaluated. Both bacterial and viral pathogens and indicator organisms are susceptible to e-beam irradiation. However, as expected there was a significant difference in their respective e-beam irradiation sensitivity. Somatic coliphages, bacterial endospores and enteric viruses were more resistant compared to bacterial pathogens. The current US EPA mandated 10 kGy minimum dose was capable of achieving significant reduction of both bacterial and viral pathogens. Somatic coliphages can be used as a microbial indicator for monitoring e-beam processes in terms of pathogen inactivation in sewage sludges. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Inactivation of pathogenic bacteria in food matrices: high pressure processing, photodynamic inactivation and pressure-assisted photodynamic inactivation

    NASA Astrophysics Data System (ADS)

    Cunha, A.; Couceiro, J.; Bonifácio, D.; Martins, C.; Almeida, A.; Neves, M. G. P. M. S.; Faustino, M. A. F.; Saraiva, J. A.

    2017-09-01

    Traditional food processing methods frequently depend on the application of high temperature. However, heat may cause undesirable changes in food properties and often has a negative impact on nutritional value and organoleptic characteristics. Therefore, reducing the microbial load without compromising the desirable properties of food products is still a technological challenge. High-pressure processing (HPP) can be classified as a cold pasteurization technique, since it is a non-thermal food preservation method that uses hydrostatic pressure to inactivate spoilage microorganisms. At the same time, it increases shelf life and retains the original features of food. Photodynamic inactivation (PDI) is also regarded as promising approach for the decontamination of food matrices. In this case, the inactivation of bacterial cells is achieved by the cytotoxic effects of reactive oxygens species (ROS) produced from the combined interaction of a photosensitizer molecule, light and oxygen. This short review examines some recent developments on the application of HPP and PDI with food-grade photosensitizers for the inactivation of listeriae, taken as a food pathogen model. The results of a proof-of-concept trial of the use of high-pressure as a coadjutant to increase the efficiency of photodynamic inactivation of bacterial endospores is also addressed.

  9. Effects of microbial loading and sporulation temperature on atmospheric plasma inactivation of Bacillus subtilis spores

    NASA Astrophysics Data System (ADS)

    Deng, X. T.; Shi, J. J.; Shama, G.; Kong, M. G.

    2005-10-01

    Current inactivation studies of Bacillus subtilis spores using atmospheric-pressure glow discharges (APGD) do not consider two important factors, namely microbial loading at the surface of a substrate and sporulation temperature. Yet these are known to affect significantly microbial resistance to heat and hydrogen peroxide. This letter investigates effects of microbial loading and sporulation temperature on spore resistance to APGD. It is shown that microbial loading can lead to a stacking structure as a protective shield against APGD treatment and that high sporulation temperature increases spore resistance by altering core water content and cross-linked muramic acid content of B. subtilis spores.

  10. Microbial Properties Database Editor Tutorial

    EPA Science Inventory

    A Microbial Properties Database Editor (MPDBE) has been developed to help consolidate microbial-relevant data to populate a microbial database and support a database editor by which an authorized user can modify physico-microbial properties related to microbial indicators and pat...

  11. Microbial properties database editor tutorial

    USDA-ARS?s Scientific Manuscript database

    A Microbial Properties Database Editor (MPDBE) has been developed to help consolidate microbialrelevant data to populate a microbial database and support a database editor by which an authorized user can modify physico-microbial properties related to microbial indicators and pathogens. Physical prop...

  12. Microbial Inactivation for Safe and Rapid Diagnostics of Infectious Samples ▿ †

    PubMed Central

    Sagripanti, Jose-Luis; Hülseweh, Birgit; Grote, Gudrun; Voß, Luzie; Böhling, Katrin; Marschall, Hans-Jürgen

    2011-01-01

    The high risk associated with biological threat agents dictates that any suspicious sample be handled under strict surety and safety controls and processed under high-level containment in specialized laboratories. This study attempted to find a rapid, reliable, and simple method for the complete inactivation of a wide range of pathogens, including spores, vegetative bacteria, and viruses, while preserving microbial nucleic acid fragments suitable for PCRs and proteinaceous epitopes for detection by immunoassays. Formaldehyde, hydrogen peroxide, and guanidium thiocyanate did not completely inactivate high titers of bacterial spores or viruses after 30 min at 21°C. Glutaraldehyde and sodium hypochlorite showed high microbicidal activity but obliterated the PCR or enzyme-linked immunosorbent assay (ELISA) detection of bacterial spores or viruses. High-level inactivation (more than 6 log10) of bacterial spores (Bacillus atrophaeus), vegetative bacteria (Pseudomonas aeruginosa), an RNA virus (the alphavirus Pixuna virus), or a DNA virus (the orthopoxvirus vaccinia virus) was attained within 30 min at 21°C by treatment with either peracetic acid or cupric ascorbate with minimal hindrance of subsequent PCR tests and immunoassays. The data described here should provide the basis for quickly rendering field samples noninfectious for further analysis under lower-level containment and considerably lower cost. PMID:21856830

  13. Molecular Viability Testing of UV-Inactivated Bacteria.

    PubMed

    Weigel, Kris M; Nguyen, Felicia K; Kearney, Moira R; Meschke, John S; Cangelosi, Gerard A

    2017-05-15

    PCR is effective in detecting bacterial DNA in samples, but it is unable to differentiate viable bacteria from inactivated cells or free DNA fragments. New PCR-based analytical strategies have been developed to address this limitation. Molecular viability testing (MVT) correlates bacterial viability with the ability to rapidly synthesize species-specific rRNA precursors (pre-rRNA) in response to brief nutritional stimulation. Previous studies demonstrated that MVT can assess bacterial inactivation by chlorine, serum, and low-temperature pasteurization. Here, we demonstrate that MVT can detect inactivation of Escherichia coli , Aeromonas hydrophila , and Enterococcus faecalis cells by UV irradiation. Some UV-inactivated E. coli cells transiently retained the ability to synthesize pre-rRNA postirradiation (generating false-positive MVT results), but this activity ceased within 1 h following UV exposure. Viable but transiently undetectable (by culture) E. coli cells were consistently detected by MVT. An alternative viability testing method, viability PCR (vPCR), correlates viability with cell envelope integrity. This method did not distinguish viable bacteria from UV-inactivated bacteria under some conditions, indicating that the inactivated cells retained intact cell envelopes. MVT holds promise as a means to rapidly assess microbial inactivation by UV treatment. IMPORTANCE UV irradiation is increasingly being used to disinfect water, food, and other materials for human use. Confirming the effectiveness of UV disinfection remains a challenging task. In particular, microbiological methods that rely on rapid detection of microbial DNA can yield misleading results, due to the detection of remnant DNA associated with dead microbial cells. This report describes a novel method that rapidly distinguishes living microbial cells from dead microbial cells after UV disinfection. Copyright © 2017 American Society for Microbiology.

  14. Bacterial spore inactivation induced by cold plasma.

    PubMed

    Liao, Xinyu; Muhammad, Aliyu Idris; Chen, Shiguo; Hu, Yaqin; Ye, Xingqian; Liu, Donghong; Ding, Tian

    2018-04-05

    Cold plasma has emerged as a non-thermal technology for microbial inactivation in the food industry over the last decade. Spore-forming microorganisms pose challenges for microbiological safety and for the prevention of food spoilage. Inactivation of spores induced by cold plasma has been reported by several studies. However, the exact mechanism of spore deactivation by cold plasma is poorly understood; therefore, it is difficult to control this process and to optimize cold plasma processing for efficient spore inactivation. In this review, we summarize the factors that affect the resistance of spores to cold plasma, including processing parameters, environmental elements, and spore properties. We then describe possible inactivation targets in spore cells (e.g., outer structure, DNA, and metabolic proteins) that associated with inactivation by cold plasma according to previous studies. Kinetic models of the sporicidal activity of cold plasma have also been described here. A better understanding of the interaction between spores and cold plasma is essential for the development and optimization of cold plasma technology in food the industry.

  15. Feasibility of utilizing bioindicators for testing microbial inactivation in sweetpotato purees processed with a continuous-flow microwave system.

    PubMed

    Brinley, T A; Dock, C N; Truong, V-D; Coronel, P; Kumar, P; Simunovic, J; Sandeep, K P; Cartwright, G D; Swartzel, K R; Jaykus, L-A

    2007-06-01

    Continuous-flow microwave heating has potential in aseptic processing of various food products, including purees from sweetpotatoes and other vegetables. Establishing the feasibility of a new processing technology for achieving commercial sterility requires evaluating microbial inactivation. This study aimed to assess the feasibility of using commercially available plastic pouches of bioindicators containing spores of Geobacillius stearothermophilus ATCC 7953 and Bacillus subtilis ATCC 35021 for evaluating the degree of microbial inactivation achieved in vegetable purees processed in a continuous-flow microwave heating unit. Sweetpotato puree seeded with the bioindicators was subjected to 3 levels of processing based on the fastest particles: undertarget process (F(0) approximately 0.65), target process (F(0) approximately 2.8), and overtarget process (F(0) approximately 10.10). After initial experiments, we found it was necessary to engineer a setup with 2 removable tubes connected to the continuous-flow microwave system to facilitate the injection of indicators into the unit without interrupting the puree flow. Using this approach, 60% of the indicators injected into the system could be recovered postprocess. Spore survival after processing, as evaluated by use of growth indicator dyes and standard plating methods, verified inactivation of the spores in sweetpotato puree. The log reduction results for B. subtilis were equivalent to the predesigned degrees of sterilization (F(0)). This study presents the first report suggesting that bioindicators such as the flexible, food-grade plastic pouches can be used for microbial validation of commercial sterilization in aseptic processing of foods using a continuous-flow microwave system.

  16. Impact of cold plasma on Citrobacter freundii in apple juice: inactivation kinetics and mechanisms.

    PubMed

    Surowsky, Björn; Fröhling, Antje; Gottschalk, Nathalie; Schlüter, Oliver; Knorr, Dietrich

    2014-03-17

    Various studies have shown that cold plasma is capable of inactivating microorganisms located on a variety of food surfaces, food packaging materials and process equipment under atmospheric pressure conditions; however, less attention has been paid to the impact of cold plasma on microorganisms in liquid foodstuffs. The present study investigates cold plasma's ability to inactivate Citrobacter freundii in apple juice. Optical emission spectroscopy (OES) and temperature measurements were performed to characterise the plasma source. The plasma-related impact on microbial loads was evaluated by traditional plate count methods, while morphological changes were determined using scanning electron microscopy (SEM). Physiological property changes were obtained through flow cytometric measurements (membrane integrity, esterase activity and membrane potential). In addition, mathematical modelling was performed in order to achieve a reliable prediction of microbial inactivation and to establish the basis for possible industrial implementation. C. freundii loads in apple juice were reduced by about 5 log cycles after a plasma exposure of 480s using argon and 0.1% oxygen plus a subsequent storage time of 24h. The results indicate that a direct contact between bacterial cells and plasma is not necessary for achieving successful inactivation. The plasma-generated compounds in the liquid, such as H2O2 and most likely hydroperoxy radicals, are particularly responsible for microbial inactivation. Copyright © 2014. Published by Elsevier B.V.

  17. Optimization of supercritical carbon dioxide treatment for the inactivation of the natural microbial flora in cubed cooked ham.

    PubMed

    Ferrentino, Giovanna; Balzan, Sara; Spilimbergo, Sara

    2013-02-15

    This study aims to investigate the effects of supercritical carbon dioxide (SC-CO₂) treatment on the inactivation of the natural microbial flora in cubed cooked ham. Response surface methodology with a central composite design was applied to determine the optimal process conditions and investigate the effect of three independent variables (pressure, temperature and treatment time). Additionally, analyses of texture, pH and color together with a storage study of the product were performed to determine its microbial and qualitative stability. Response surface analysis revealed that 12 MPa, 50 °C, 5 min were the optimal conditions to obtain about 3.0, 1.6, and 2.5 Log(CFU/g) reductions of mesophilic aerobic bacteria, psychrophilic bacteria and lactic acid bacteria respectively. Inactivation to undetectable levels of yeasts and molds and coliforms was also obtained. A storage study of 30 days at 4 °C was carried out on the treated product (12 MPa, 50 °C, 5 min) monitoring microbial growth, pH, texture, and color parameters (L*, a*, b* and ΔE). Microbial loads slightly increased and after 30 days of storage reached the same levels detected in the fresh product. Color parameters (L*, a*, b*) showed slight variations while pH and texture did not change significantly. On the basis of the results obtained, SC-CO₂ can be considered a promising technique to microbiologically stabilize cubed cooked ham and, in general, cut/sliced meat products without affecting its quality attributes. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. High pressure, temperature and time-dependent effects on enzymatic and microbial properties of fresh sugarcane juice.

    PubMed

    Chauhan, O P; Ravi, N; Roopa, N; Kumar, Sumeet; Raju, P S

    2017-11-01

    Efficacy of variable high pressure, temperature and time on the browning causing enzymes and microbial activities, which are major spoilage factors during preservation of sugarcane juice, was studied. The juice was processed at 200-600 MPa pressure for 2-8 min at 40 and 60 °C and their effect on polyphenol oxidase and peroxidase as well as microbiological quality in terms of total plate count, yeast and molds and total coliforms was studied. Application of high pressures were found to cause significant decrease in enzymatic and microbial activities. The effects were found to be significantly more pronounced at 60 °C as compared to 40 °C. Process time also caused significant ( p  < 0.05) negative effect on microbial and enzyme activities. The sugarcane juice treated at 600 MPa for 6 min at 60 °C was found sufficient to inactivate the microbial counts completely. Whereas, enzymes were found to be completely inactivated in the samples processed at 600 MPa for 8 min at 60 °C. A pressure of 600 MPa at 60 °C for 8 min could be applied during commercial preservation of sugarcane juice for getting complete inactivation of browning causing enzymes and spoilage causing microorganisms.

  19. Two-Phase Slug Flow Heat Exchanger for Microbial Thermal Inactivation Research

    PubMed Central

    Stroup, W. H.; Dickerson, R. W.; Read, R. B.

    1969-01-01

    A continuous two-phase (air-liquid), slug flow, tubular heat exchanger was developed for microbial thermal inactivation research to give exposure times and temperatures in the range of high-temperature, short-time milk pasteurization as well as heat-treated sample volumes of at least 2 ml. The use of air to compartmentalize the liquid in the capillary tubing prevented the development of laminar flow, which enabled precise identification of the residence time of the fastest flowing particles in the heating, holding, and cooling sections of the instrument. Residence time distributions were quantitated by measuring the degree of spreading of radioactive tracers for water, whole milk, chocolate milk, cream, and ice-cream mix with holding temperatures from 50 to 72 C, holding times from 2 to 60 sec, and heating and cooling times of 1.7 sec each. For a holding time of 60 sec and a fastest particle velocity of 10.2 cm/sec, the velocity ratios of the fastest moving particle to the median particle were 1.05, 1.05, 1.10, and 1.13 for whole milk, chocolate milk, cream, and ice-cream mix, respectively. With shorter holding times, these velocity ratios were even closer to unity. These velocity ratios indicated that the instrument would be an effective tool for thermal inactivation research on microorganisms suspended in homogeneous fluids with a viscosity of 15 centipoises or less at the exposure temperature. PMID:5395711

  20. Protocol for Determining Ultraviolet Light Emitting Diode (UV-LED) Fluence for Microbial Inactivation Studies.

    PubMed

    Kheyrandish, Ataollah; Mohseni, Madjid; Taghipour, Fariborz

    2018-06-15

    Determining fluence is essential to derive the inactivation kinetics of microorganisms and to design ultraviolet (UV) reactors for water disinfection. UV light emitting diodes (UV-LEDs) are emerging UV sources with various advantages compared to conventional UV lamps. Unlike conventional mercury lamps, no standard method is available to determine the average fluence of the UV-LEDs, and conventional methods used to determine the fluence for UV mercury lamps are not applicable to UV-LEDs due to the relatively low power output, polychromatic wavelength, and specific radiation profile of UV-LEDs. In this study, a method was developed to determine the average fluence inside a water suspension in a UV-LED experimental setup. In this method, the average fluence was estimated by measuring the irradiance at a few points for a collimated and uniform radiation on a Petri dish surface. New correction parameters were defined and proposed, and several of the existing parameters for determining the fluence of the UV mercury lamp apparatus were revised to measure and quantify the collimation and uniformity of the radiation. To study the effect of polychromatic output and radiation profile of the UV-LEDs, two UV-LEDs with peak wavelengths of 262 and 275 nm and different radiation profiles were selected as the representatives of typical UV-LEDs applied to microbial inactivation. The proper setup configuration for microorganism inactivation studies was also determined based on the defined correction factors.

  1. Inactivation of possible microorganism food contaminants on packaging foils using nonthermal plasma and hydrogen peroxide

    NASA Astrophysics Data System (ADS)

    Scholtz, V.; Khun, J.; Soušková, H.; Čeřovský, M.

    2015-07-01

    The inactivation effect of nonthermal plasma generated in electric discharge burning in air atmosphere with water or hydrogen peroxide aerosol for the application to the microbial decontamination of packaging foils is studied. The microbial inactivation is studied on two bacterial, two yeasts, and two filamentous micromycete species. The inactivation of all contaminating microorganisms becomes on the area of full 8.5 cm in diameter circular sample after short times of several tens of seconds. Described apparatus may present a possible alternative method of microbial decontamination of food packaging material or other thermolabile materials.

  2. Inactivation of possible microorganism food contaminants on packaging foils using nonthermal plasma and hydrogen peroxide

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

    Scholtz, V., E-mail: Vladimir.Scholtz@vscht.cz; Khun, J.; Soušková, H.

    The inactivation effect of nonthermal plasma generated in electric discharge burning in air atmosphere with water or hydrogen peroxide aerosol for the application to the microbial decontamination of packaging foils is studied. The microbial inactivation is studied on two bacterial, two yeasts, and two filamentous micromycete species. The inactivation of all contaminating microorganisms becomes on the area of full 8.5 cm in diameter circular sample after short times of several tens of seconds. Described apparatus may present a possible alternative method of microbial decontamination of food packaging material or other thermolabile materials.

  3. Regression model for estimating inactivation of microbial aerosols by solar radiation.

    PubMed

    Ben-David, Avishai; Sagripanti, Jose-Luis

    2013-01-01

    The inactivation of pathogenic aerosols by solar radiation is relevant to public health and biodefense. We investigated whether a relatively simple method to calculate solar diffuse and total irradiances could be developed and used in environmental photobiology estimations instead of complex atmospheric radiative transfer computer programs. The second-order regression model that we developed reproduced 13 radiation quantities calculated for equinoxes and solstices at 35(°) latitude with a computer-intensive and rather complex atmospheric radiative transfer program (MODTRAN) with a mean error <6% (2% for most radiation quantities). Extending the application of the regression model from a reference latitude and date (chosen as 35° latitude for 21 March) to different latitudes and days of the year was accomplished with variable success: usually with a mean error <15% (but as high as 150% for some combination of latitudes and days of year). This accuracy of the methodology proposed here compares favorably to photobiological experiments where the microbial survival is usually measured with an accuracy no better than ±0.5 log10 units. The approach and equations presented in this study should assist in estimating the maximum time during which microbial pathogens remain infectious after accidental or intentional aerosolization in open environments. © Published 2013. This article is a U.S. Government work and is in the public domain in the USA. Photochemistry and Photobiology © 2013 The American Society of Photobiology.

  4. Effects of Bacterial Inactivation Methods on Downstream Proteomic Analysis

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

    Lin, Andy; Merkley, Eric D.; Clowers, Brian H.

    2015-05-01

    Inactivation of pathogenic microbial samples is often necessary for the protection of researchers and to comply with local and federal regulations. By its nature, biological inactivation causes changes to microbial samples, potentially affecting observed experimental results. While inactivation induced damage to materials such as DNA has been evaluated, the effect of various inactivation strategies on proteomic data, to our knowledge, has not been discussed. To this end, we inactivated samples of Yersinia pestis and Escherichia coli by autoclave, ethanol, or irradiation treatment to determine how inactivation changes liquid chromatography tandem mass spectrometry data quality as well as apparent protein contentmore » of cells. Proteomic datasets obtained from aliquots of samples inactivated by different methods were highly similar, with Pearson correlation coefficients ranging from 0.822 to 0.985 and 0.816 to 0.985 for E. coli and Y. pestis, respectively, suggesting that inactivation had only slight impacts on the set of proteins identified. In addition, spectral quality metrics such as distributions of various database search algorithm scores remained constant across inactivation methods, indicating that inactivation does not appreciably degrade spectral quality. Though overall changes resulting from inactivation were small, there were detectable trends. For example, one-sided Fischer exact tests determined that periplasmic proteins decrease in observed abundance after sample inactivation by autoclaving (α = 1.71x10-2 for E. coli, α = 4.97x10-4 for Y. pestis) and irradiation (α = 9.43x10-7 for E. coli, α = 1.21x10-5 for Y. pestis) when compared to controls that were not inactivated. Based on our data, if sample inactivation is necessary, we recommend inactivation with ethanol treatment with secondary preference given to irradiation.« less

  5. Kinetics of Hydrothermal Inactivation of Endotoxins ▿

    PubMed Central

    Li, Lixiong; Wilbur, Chris L.; Mintz, Kathryn L.

    2011-01-01

    A kinetic model was established for the inactivation of endotoxins in water at temperatures ranging from 210°C to 270°C and a pressure of 6.2 × 106 Pa. Data were generated using a bench scale continuous-flow reactor system to process feed water spiked with endotoxin standard (Escherichia coli O113:H10). Product water samples were collected and quantified by the Limulus amebocyte lysate assay. At 250°C, 5-log endotoxin inactivation was achieved in about 1 s of exposure, followed by a lower inactivation rate. This non-log-linear pattern is similar to reported trends in microbial survival curves. Predictions and parameters of several non-log-linear models are presented. In the fast-reaction zone (3- to 5-log reduction), the Arrhenius rate constant fits well at temperatures ranging from 120°C to 250°C on the basis of data from this work and the literature. Both biphasic and modified Weibull models are comparable to account for both the high and low rates of inactivation in terms of prediction accuracy and the number of parameters used. A unified representation of thermal resistance curves for a 3-log reduction and a 3 D value associated with endotoxin inactivation and microbial survival, respectively, is presented. PMID:21193667

  6. C. botulinum inactivation kinetics implemented in a computational model of a high-pressure sterilization process.

    PubMed

    Juliano, Pablo; Knoerzer, Kai; Fryer, Peter J; Versteeg, Cornelis

    2009-01-01

    High-pressure, high-temperature (HPHT) processing is effective for microbial spore inactivation using mild preheating, followed by rapid volumetric compression heating and cooling on pressure release, enabling much shorter processing times than conventional thermal processing for many food products. A computational thermal fluid dynamic (CTFD) model has been developed to model all processing steps, including the vertical pressure vessel, an internal polymeric carrier, and food packages in an axis-symmetric geometry. Heat transfer and fluid dynamic equations were coupled to four selected kinetic models for the inactivation of C. botulinum; the traditional first-order kinetic model, the Weibull model, an nth-order model, and a combined discrete log-linear nth-order model. The models were solved to compare the resulting microbial inactivation distributions. The initial temperature of the system was set to 90 degrees C and pressure was selected at 600 MPa, holding for 220 s, with a target temperature of 121 degrees C. A representation of the extent of microbial inactivation throughout all processing steps was obtained for each microbial model. Comparison of the models showed that the conventional thermal processing kinetics (not accounting for pressure) required shorter holding times to achieve a 12D reduction of C. botulinum spores than the other models. The temperature distribution inside the vessel resulted in a more uniform inactivation distribution when using a Weibull or an nth-order kinetics model than when using log-linear kinetics. The CTFD platform could illustrate the inactivation extent and uniformity provided by the microbial models. The platform is expected to be useful to evaluate models fitted into new C. botulinum inactivation data at varying conditions of pressure and temperature, as an aid for regulatory filing of the technology as well as in process and equipment design.

  7. Wastewater disinfection by peracetic acid: assessment of models for tracking residual measurements and inactivation.

    PubMed

    Santoro, Domenico; Gehr, Ronald; Bartrand, Timothy A; Liberti, Lorenzo; Notarnicola, Michele; Dell'Erba, Adele; Falsanisi, Dario; Haas, Charles N

    2007-07-01

    With its potential for low (if any) disinfection byproduct formation and easy retrofit for chlorine contactors, peracetic acid (PAA) or use of PAA in combination with other disinfectant technologies may be an attractive alternative to chlorine-based disinfection. Examples of systems that might benefit from use of PAA are water reuse schemes or plants discharging to sensitive receiving water bodies. Though PAA is in use in numerous wastewater treatment plants in Europe, its chemical kinetics, microbial inactivation rates, and mode of action against microorganisms are not thoroughly understood. This paper presents results from experimental studies of PAA demand, PAA decay, and microbial inactivation, with a complementary modeling analysis. Model results are used to evaluate techniques for measurement of PAA concentration and to develop hypotheses regarding the mode of action of PAA in bacterial inactivation. Kinetic and microbial inactivation rate data were collected for typical wastewaters and may be useful for engineers in evaluating whether to convert from chlorine to PAA disinfection.

  8. Plasma inactivation of microorganisms on sprout seeds in a dielectric barrier discharge.

    PubMed

    Butscher, Denis; Van Loon, Hanne; Waskow, Alexandra; Rudolf von Rohr, Philipp; Schuppler, Markus

    2016-12-05

    Fresh produce is frequently contaminated by microorganisms, which may lead to spoilage or even pose a threat to human health. In particular sprouts are considered to be among the most risky foods sold at retail since they are grown in an environment practically ideal for growth of bacteria and usually consumed raw. Because heat treatment has a detrimental effect on the germination abilities of sprout seeds, alternative treatment technologies need to be developed for microbial inactivation purposes. In this study, non-thermal plasma decontamination of sprout seeds is evaluated as a promising option to enhance food safety while maintaining the seed germination capabilities. In detail, investigations focus on understanding the efficiency of non-thermal plasma inactivation of microorganisms as influenced by the type of microbial contamination, substrate surface properties and moisture content, as well as variations in the power input to the plasma device. To evaluate the impact of these parameters, we studied the reduction of native microbiota or artificially applied E. coli on alfalfa, onion, radish and cress seeds exposed to non-thermal plasma in an atmospheric pressure pulsed dielectric barrier discharge streamed with argon. Plasma treatment resulted in a maximum reduction of 3.4 logarithmic units for E. coli on cress seeds. A major challenge in plasma decontamination of granular food products turned out to be the complex surface topology, where the rough surface with cracks and crevices can shield microorganisms from plasma-generated reactive species, thus reducing the treatment efficiency. However, improvement of the inactivation efficiency was possible by optimizing substrate characteristics such as the moisture level and by tuning the power supply settings (voltage, frequency) to increase the production of reactive species. While the germination ability of alfalfa seeds was considerably decreased by harsh plasma treatment, enhanced germination was observed under

  9. Inactivation of Lactobacillus rhamnosus GG by fixation modifies its probiotic properties.

    PubMed

    Markowicz, C; Kubiak, P; Grajek, W; Schmidt, M T

    2016-01-01

    Probiotics are microorganisms that have beneficial effects on the host and are safe for oral intake in a suitable dose. However, there are situations in which the administration of living microorganisms poses a risk for immunocompromised host. The objective of this study was to evaluate the influence of several fixation methods on selected biological properties of Lactobacillus rhamnosus GG that are relevant to its probiotic action. Fixation of the bacterial cells with ethanol, 2-propanol, glutaraldehyde, paraformaldehyde, and heat treatment resulted in a significant decrease of alkaline phosphatase, peroxidase, and β-galactosidase activities. Most of the fixation procedures reduced bacterial cell hydrophobicity and increased adhesion capacity. The fixation procedures resulted in a different perception of the bacterial cells by enterocytes, which was shown as changes in gene expression in enterocytes. The results show that some procedures of inactivation allow a fraction of the enzymatic activity to be maintained. The adhesion properties of the bacterial cells were enhanced, but the response of enterocytes to fixed cells was different than to live bacteria. Inactivation allows maintenance and modification of some of the properties of the bacterial cells.

  10. Microbial Decontamination of Dried Alaska Pollock Shreds Using Corona Discharge Plasma Jet: Effects on Physicochemical and Sensory Characteristics.

    PubMed

    Choi, Soee; Puligundla, Pradeep; Mok, Chulkyoon

    2016-04-01

    Nonthermal techniques for microbial decontamination are becoming more common for ensuring food safety. In this study, a corona discharge plasma jet (CDPJ) was used for inactivation of microbial contaminants of dried Alaska pollock shreds. Corona plasma jet was generated at a current strength of 1.5 A, and a span length of 25 mm was maintained between the electrode tip and the sample. Upon the CDPJ treatment (0 to 3 min) of dried shreds, microbial contaminants namely aerobic and marine bacteria, and Staphylococcus aureus were inactivated by 2.5, 1.5, and >1.0 log units, respectively. Also, a one-log reduction of molds and yeasts contaminants was observed. The inactivation patterns are fitted well to the pseudo-first-order kinetics or Singh-Heldman model. The CDPJ treatment did not exert statistically significant (P > 0.05) changes in physicochemical properties, namely color characteristics, volatile basic nitrogen, and peroxide value of dried fish shreds, with some exceptions, as compared to untreated controls. Furthermore, CDPJ treatment had no significant impact on the sensory characteristics of dried fish shreds. © 2016 Institute of Food Technologists®

  11. Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components. [bibliography

    NASA Technical Reports Server (NTRS)

    Reyes, A. L.; Campbell, J. E.

    1976-01-01

    Almost 600 articles and books published since 1960 about microbial and viral inactivation are listed. This bibliography is presented to facilitate literature reviews on chemical, heat, and radiation inactivation of microorganisms and viral particles.

  12. Effects of Background Fluid on the Efficiency of Inactivating Yeast with Non-Thermal Atmospheric Pressure Plasma

    PubMed Central

    Ryu, Young-Hyo; Kim, Yong-Hee; Lee, Jin-Young; Shim, Gun-Bo; Uhm, Han-Sup; Park, Gyungsoon; Choi, Eun Ha

    2013-01-01

    Non-thermal plasma at atmospheric pressure has been actively applied to sterilization. However, its efficiency for inactivating microorganisms often varies depending on microbial species and environments surrounding the microorganisms. We investigated the influence of environmental factors (surrounding media) on the efficiency of microbial inactivation by plasma using an eukaryotic model microbe, Saccharomyces cerevisiae, to elucidate the mechanisms for differential efficiency of sterilization by plasma. Yeast cells treated with plasma in water showed the most severe damage in viability and cell morphology as well as damage to membrane lipids, and genomic DNA. Cells in saline were less damaged compared to those in water, and those in YPD (Yeast extract, Peptone, Dextrose) were least impaired. HOG1 mitogen activated protein kinase was activated in cells exposed to plasma in water and saline. Inactivation of yeast cells in water and saline was due to the acidification of the solutions by plasma, but higher survival of yeast cells treated in saline may have resulted from the additional effect related to salt strength. Levels of hydroxyl radical (OH.) produced by plasma were the highest in water and the lowest in YPD. This may have resulted in differential inactivation of yeast cells in water, saline, and YPD by plasma. Taken together, our data suggest that the surrounding media (environment) can crucially affect the outcomes of yeast cell plasma treatment because plasma modulates vital properties of media, and the toxic nature of plasma can also be altered by the surrounding media. PMID:23799081

  13. Inactivation of Escherichia coli by citral.

    PubMed

    Somolinos, M; García, D; Condón, S; Mackey, B; Pagán, R

    2010-06-01

    The aim was to evaluate (i) the resistance of Escherichia coli BJ4 to citral in a buffer system as a function of citral concentration, treatment medium pH, storage time and initial inoculum size, (ii) the role of the sigma factor RpoS on citral resistance of E. coli, (iii) the role of the cell envelope damage in the mechanism of microbial inactivation by citral and (iiii) possible synergistic effects of mild heat treatment and pulsed electric fields (PEF) treatment combined with citral. The initial inoculum size greatly affected the efficacy of citral against E. coli cells. Exposure to 200 microl l(-1) of citral at pH 4.0 for 24 h at 20 degrees C caused the inactivation of more than 5 log(10) cycles of cells starting at an inoculum size of 10(6) or 10(7) CFU ml(-1), whereas increasing the cell concentration to 10(9) CFU ml(-1) caused <1 log(10) cycle of inactivation. Escherichia coli showed higher resistance to citral at pH 4.0 than pH 7.0. The rpoS null mutant strain E. coli BJ4L1 was less resistant to citral than the wild-type strain. Occurrence of sublethal injury to both the cytoplasmic and outer membranes was demonstrated by adding sodium chloride or bile salts to the recovery media. The majority of sublethally injured cells by citral required energy and lipid synthesis for repair. A strongly synergistic lethal effect was shown by mild heat treatment combined with citral but the presence of citral during the application of a PEF treatment did not show any advantage. This work confirms that cell envelope damage is an important event in citral inactivation of bacteria, and it describes the key factors on the inactivation of E. coli cells by citral. Knowledge about the mechanism of microbial inactivation by citral helps establish successful combined preservation treatments.

  14. Effect of extrusion conditions and lipoxygenase inactivation treatment on the physical and nutritional properties of corn/cowpea (Vigna unguiculata) blends.

    PubMed

    Sosa-Moguel, Odri; Ruiz-Ruiz, Jorge; Martínez-Ayala, Alma; González, Rolando; Drago, Silvina; Betancur-Ancona, David; Chel-Guerrero, Luis

    2009-01-01

    The influence of lipoxygenase inactivation and extrusion cooking on the physical and nutritional properties of corn/cowpea (Vigna unguiculata) blends was studied. Corn was blended in an 80:15 proportion with cowpea flour treated to inactivate lipoxygenase (CI) or non-inactivated cowpea flour (CNI). Extrusion variables were temperature (150 degrees C, 165 degrees C and 180 degrees C) and moisture (15%, 17% and 19%). Based on their physical properties, the 165 degrees C/15% corn:CNI, and 165 degrees C/15% corn:CI, and 150 degrees C/15% corn:CI blends were chosen for nutritional quality analysis. Extrudate chemical composition indicated high crude protein levels compared with standard corn-based products. With the exception of lysine, essential amino acids content in the three treatments met FAO requirements. Extrusion and lipoxygenase inactivation are promising options for developing corn/cowpea extruded snack products with good physical properties and nutritional quality.

  15. Chlorophyll mediated photodynamic inactivation of blue laser on Streptococcus mutans

    NASA Astrophysics Data System (ADS)

    Astuti, Suryani Dyah; Zaidan, A.; Setiawati, Ernie Maduratna; Suhariningsih

    2016-03-01

    Photodynamic inactivation is an inactivation method in microbial pathogens that utilize light and photosensitizer. This study was conducted to investigate photodynamic inactivation effects of low intensity laser exposure with various dose energy on Streptococcus mutans bacteria. The photodynamic inactivation was achieved with the addition of chlorophyll as photosensitizers. To determine the survival percentage of Streptococcus mutans bacteria after laser exposure, the total plate count method was used. For this study, the wavelength of the laser is 405 nm and variables of energy doses are 1.44, 2.87, 4.31, 5.74, 7.18, and 8.61 in J/cm2. The results show that exposure to laser with energy dose of 7.18 J/cm2 has the best photodynamic inactivation with a decrease of 78% in Streptococcus

  16. Physicochemical changes and microbial inactivation after high-intensity ultrasound processing of prebiotic whey beverage applying different ultrasonic power levels.

    PubMed

    Guimarães, Jonas T; Silva, Eric Keven; Alvarenga, Verônica O; Costa, Ana Letícia R; Cunha, Rosiane L; Sant'Ana, Anderson S; Freitas, Monica Q; Meireles, M Angela A; Cruz, Adriano G

    2018-06-01

    In this work, we investigated the effects of the ultrasonic power (0, 200, 400 and 600 W) on non-thermal processing of an inulin-enriched whey beverage. We studied the effects of high-intensity ultrasound (HIUS) on microbial inactivation (aerobic mesophilic heterotrophic bacteria (AMHB), total and thermotolerant coliforms and yeasts and molds), zeta potential, microstructure (optical microscopy, particle size distribution), rheology, kinetic stability and color. The non-thermal processing applying 600 W of ultrasonic power was comparable to high-temperature short-time (HTST) treatment (75 °C for 15 s) concerning the inactivation of AMHB and yeasts and molds (2 vs 2 log and 0.2 vs 0.4 log, respectively), although HIUS has reached a lower output temperature (53 ± 3 °C). The HIUS was better than HTST to improve beverage kinetic stability, avoiding phase separation, which was mainly attributed to the decrease of particles size, denaturation of whey proteins and gelation of polysaccharides (inulin and gellan gum). Thus, non-thermal processing by HIUS seems to be an interesting technology for prebiotic dairy beverages production. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. The efficacy of preservation methods to inactivate foodborne viruses.

    PubMed

    Baert, L; Debevere, J; Uyttendaele, M

    2009-05-31

    During the last decade an increased incidence of infections and outbreaks attributed to foodborne viruses, in particular noroviruses (NoV), was observed world wide. The awareness of the presence of viruses on food emphasized the need to acquire knowledge regarding the effect of preservation methods upon viruses. Most foodborne viruses cannot be cultured in the laboratory, which hinders studies of their stability in food. Cultivable surrogate viruses, genetically related to the human infecting strains, are taken as a substitute to define inactivation rates. The last years, the number of survival and inactivation studies using various surrogate viruses increased. In this review, state-of-the-art information regarding the efficacy of preservation methods to reduce the level of viruses on food is compiled. In the first place, the effect of preservation methods establishing microbial growth inhibition (chilling, freezing, acidification, reduced water activity and modified atmosphere packaging) upon foodborne viruses is described. Secondly, the use of preservation methods establishing microbial inactivation such as heat treatment, high hydrostatic pressure processing and irradiation to eliminate viruses is discussed. In the third place, the efficacy of decontamination methods on fresh produce and purification procedures applied on live bivalve shellfish to reduce the viral load is included. These studies indicate that viruses persist well on chilled, acidified, frozen foods and foods packed under modified atmosphere or in dried conditions. Intervention strategies inducing microbial inactivation are required to achieve a 3 log reduction of the level of viruses. Decontamination of fresh produce reduces viruses with a maximum of 1 to 2 log while purification of live bivalves is not adequate to prevent viral outbreaks. It was noted that the effect of a particular food preservation method is dependent upon the virus tested and type of food.

  18. Comfort and microbial barrier properties of garments worn next to the skin

    NASA Astrophysics Data System (ADS)

    Kopitar, D.; Rogina-Car, B.; Skenderi, Z.

    2017-10-01

    Compared with viscose fibre, modal fibre is characterized by some advantageous properties such as higher dry and wet tenacities, higher wet modulus, lower water retention capacity and lower level of swelling. Impact of different knitted fabric structure made of cotton and 97 % CMD/3 % EL fibres on thermo-physiological comfort and microbial barrier properties were investigated. All knitted fabrics have very good physiological properties. The microbial barrier permeability of knitted fabric after extreme contamination with bacterial spores in dry state showed that double jersey offered more effective microbial barrier than the single jersey knitted fabrics respectively the greater thickness of double jersey knitted fabric provide more difficult barrier to bacterial spores to pass. In wet state all knitted fabrics have more effective microbial barrier which could be explained by cellulose fibres swelling. In wet state 97 % CMD/3 % EL single jersey knitted fabric have more effective microbial barrier then cotton double and single jersey knitted fabrics.

  19. Bacteria, mould and yeast spore inactivation studies by scanning electron microscope observations.

    PubMed

    Rozali, Siti N M; Milani, Elham A; Deed, Rebecca C; Silva, Filipa V M

    2017-12-18

    Spores are the most resistant form of microbial cells, thus difficult to inactivate. The pathogenic or food spoilage effects of certain spore-forming microorganisms have been the primary basis of sterilization and pasteurization processes. Thermal sterilization is the most common method to inactivate spores present on medical equipment and foods. High pressure processing (HPP) is an emerging and commercial non-thermal food pasteurization technique. Although previous studies demonstrated the effectiveness of thermal and non-thermal spore inactivation, the in-depth mechanisms of spore inactivation are as yet unclear. Live and dead forms of two food spoilage bacteria, a mould and a yeast were examined using scanning electron microscopy before and after the inactivation treatment. Alicyclobacillus acidoterrestris and Geobacillus stearothermophilus bacteria are indicators of acidic foods pasteurization and sterilization processes, respectively. Neosartorya fischeri is a phyto-pathogenic mould attacking fruits. Saccharomyces cerevisiae is a yeast with various applications for winemaking, brewing, baking and the production of biofuel from crops (e.g. sugar cane). Spores of the four microbial species were thermally inactivated. Spores of S. cerevisiae were observed in the ascus and free form after thermal and HPP treatments. Different forms of damage and cell destruction were observed for each microbial spore. Thermal treatment inactivated bacterial spores of A. acidoterrestris and G. stearothermophilus by attacking the inner core of the spore. The heat first altered the membrane permeability allowing the release of intracellular components. Subsequently, hydration of spores, physicochemical modifications of proteins, flattening and formation of indentations occurred, with subsequent spore death. Regarding N. fischeri, thermal inactivation caused cell destruction and leakage of intracellular components. Both thermal and HPP treatments of S. cerevisiae free spores attacked

  20. Inactivation of Escherichia coli, Saccharomyces cerevisiae, and Lactobacillus brevis in Low-fat Milk by Pulsed Electric Field Treatment: A Pilot-scale Study

    PubMed Central

    Han, Bok Kung; Choi, Hyuk Joon; Kang, Shin Ho; Baick, Seung Chun

    2015-01-01

    We investigated the effects of a pulsed electric field (PEF) treatment on microbial inactivation and the physical properties of low-fat milk. Milk inoculated with Escherichia coli, Saccharomyces cerevisiae, or Lactobacillus brevis was supplied to a pilot-scale PEF treatment system at a flow rate of 30 L/h. Pulses with an electric field strength of 10 kV/cm and a pulse width of 30 μs were applied to the milk with total pulse energies of 50-250 kJ/L achieved by varying the pulse frequency. The inactivation curves of the test microorganisms were biphasic with an initial lag phase (or shoulder) followed by a phase of rapid inactivation. PEF treatments with a total pulse energy of 200 kJ/L resulted in a 4.5-log reduction in E. coli, a 4.4-log reduction in L. brevis, and a 6.0-log reduction in S. cerevisiae. Total pulse energies of 200 and 250 kJ/L resulted in greater than 5-log reductions in microbial counts in stored PEF-treated milk, and the growth of surviving microorganisms was slow during storage for 15 d at 4℃. PEF treatment did not change milk physical properties such as pH, color, or particle-size distribution (p<0.05). These results indicate that a relatively low electric-field strength of 10 kV/cm can be used to pasteurize low-fat milk. PMID:26877640

  1. Inactivation of Escherichia coli, Saccharomyces cerevisiae, and Lactobacillus brevis in Low-fat Milk by Pulsed Electric Field Treatment: A Pilot-scale Study.

    PubMed

    Lee, Gun Joon; Han, Bok Kung; Choi, Hyuk Joon; Kang, Shin Ho; Baick, Seung Chun; Lee, Dong-Un

    2015-01-01

    We investigated the effects of a pulsed electric field (PEF) treatment on microbial inactivation and the physical properties of low-fat milk. Milk inoculated with Escherichia coli, Saccharomyces cerevisiae, or Lactobacillus brevis was supplied to a pilot-scale PEF treatment system at a flow rate of 30 L/h. Pulses with an electric field strength of 10 kV/cm and a pulse width of 30 μs were applied to the milk with total pulse energies of 50-250 kJ/L achieved by varying the pulse frequency. The inactivation curves of the test microorganisms were biphasic with an initial lag phase (or shoulder) followed by a phase of rapid inactivation. PEF treatments with a total pulse energy of 200 kJ/L resulted in a 4.5-log reduction in E. coli, a 4.4-log reduction in L. brevis, and a 6.0-log reduction in S. cerevisiae. Total pulse energies of 200 and 250 kJ/L resulted in greater than 5-log reductions in microbial counts in stored PEF-treated milk, and the growth of surviving microorganisms was slow during storage for 15 d at 4℃. PEF treatment did not change milk physical properties such as pH, color, or particle-size distribution (p<0.05). These results indicate that a relatively low electric-field strength of 10 kV/cm can be used to pasteurize low-fat milk.

  2. Inactivation of Template-Directed Misfolding of Infectious Prion Protein by Ozone

    PubMed Central

    Ding, Ning; Price, Luke M.; Braithwaite, Shannon L.; Balachandran, Aru; Belosevic, Miodrag

    2012-01-01

    Misfolded prions (PrPSc) are well known for their resistance to conventional decontamination processes. The potential risk of contamination of the water environment, as a result of disposal of specified risk materials (SRM), has raised public concerns. Ozone is commonly utilized in the water industry for inactivation of microbial contaminants and was tested in this study for its ability to inactivate prions (263K hamster scrapie = PrPSc). Treatment variables included initial ozone dose (7.6 to 25.7 mg/liter), contact time (5 s and 5 min), temperature (4°C and 20°C), and pH (pH 4.4, 6.0, and 8.0). Exposure of dilute suspensions of the infected 263K hamster brain homogenates (IBH) (0.01%) to ozone resulted in the in vitro destruction of the templating properties of PrPSc, as measured by the protein misfolding cyclic amplification (PMCA) assay. The highest levels of prion inactivation (≥4 log10) were observed with ozone doses of 13.0 mg/liter, at pH 4.4 and 20°C, resulting in a CT (the product of residual ozone concentration and contact time) value as low as 0.59 mg · liter−1 min. A comparison of ozone CT requirements among various pathogens suggests that prions are more susceptible to ozone degradation than some model bacteria and protozoa and that ozone treatment may be an effective solution for inactivating prions in water and wastewater. PMID:22138993

  3. Mycobacteria inactivation using Engineered Water Nanostructures (EWNS).

    PubMed

    Pyrgiotakis, Georgios; McDevitt, James; Gao, Ya; Branco, Alan; Eleftheriadou, Mary; Lemos, Bernardo; Nardell, Edward; Demokritou, Philip

    2014-08-01

    Airborne transmitted pathogens such as Mycobacterium tuberculosis (Mtb) cause serious, often fatal infectious disease with enormous global health implications. Due to their unique cell wall and slow growth, mycobacteria are among the most resilient microbial forms. Herein we evaluate the ability of an emerging, chemical-free, nanotechnology-based method to inactivate M. parafortuitum (Mtb surrogate). This method is based on the transformation of atmospheric water vapor into engineered water nano-structures (EWNS) via electrospray. We demonstrate that the EWNS can interact with and inactivate airborne mycobacteria, reducing their concentration levels significantly. Additionally, EWNS can inactivate M. parafortuitum on surfaces eight times faster than the control. The mechanism of mycobacteria inactivation was also investigated in this study. It was demonstrated that the EWNS effectively deliver the reactive oxygen species, encapsulated during the electrospray process, to the bacteria oxidizing their cell membrane resulting into inactivation. Overall, this is a method with the potential to become an effective intervention technology in the battle against airborne infections. This study demonstrates the feasibility of mycobacterium inactivation in airborne form or on contact surfaces using electrospray activated water nano-structures. Given that the method is free of toxic chemicals, this might become an important tool in the prevention of mycobacterial infections, which are notoriously hard to treat. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Antimicrobial blue light: a drug-free approach for inactivating pathogenic microbes

    NASA Astrophysics Data System (ADS)

    Wang, Ying; Dai, Tianhong

    2018-02-01

    Due to the growing global threat of antibiotic resistance, there is a critical need for the development of alternative therapeutics for infectious diseases. Antimicrobial blue light (aBL), as an innovative non-antibiotic approach, has attracted increasing attention. This paper discussed the basic concepts of aBL and recent findings in the studies of aBL. It is commonly hypothesized that the antimicrobial property of aBL is attributed to the presence of endogenous photosensitizing chromophores in microbial cells, which produce cytotoxic reactive oxygen species upon light irradiation. A wide range of important microbes are found to be susceptible to aBL inactivation. Studies have also shown there exist therapeutic windows where microbes are selectively inactivated by aBL while host cells are preserved. The combination of aBL with some other agents result in synergistically improved antimicrobial efficacy. Future efforts should be exerted on the standardization of study design for evaluating aBL efficacy, further elucidation of the mechanism of action, optimization of the technical parameters, and translation of this technique to clinic.

  5. Modeling of inactivation of surface borne microorganisms occurring on seeds by cold atmospheric plasma (CAP)

    NASA Astrophysics Data System (ADS)

    Mitra, Anindita; Li, Y.-F.; Shimizu, T.; Klämpfl, Tobias; Zimmermann, J. L.; Morfill, G. E.

    2012-10-01

    Cold Atmospheric Plasma (CAP) is a fast, low cost, simple, easy to handle technology for biological application. Our group has developed a number of different CAP devices using the microwave technology and the surface micro discharge (SMD) technology. In this study, FlatPlaSter2.0 at different time intervals (0.5 to 5 min) is used for microbial inactivation. There is a continuous demand for deactivation of microorganisms associated with raw foods/seeds without loosing their properties. This research focuses on the kinetics of CAP induced microbial inactivation of naturally growing surface microorganisms on seeds. The data were assessed for log- linear and non-log-linear models for survivor curves as a function of time. The Weibull model showed the best fitting performance of the data. No shoulder and tail was observed. The models are focused in terms of the number of log cycles reduction rather than on classical D-values with statistical measurements. The viability of seeds was not affected for CAP treatment times up to 3 min with our device. The optimum result was observed at 1 min with increased percentage of germination from 60.83% to 89.16% compared to the control. This result suggests the advantage and promising role of CAP in food industry.

  6. Time-dependent effect of composted tannery sludge on the chemical and microbial properties of soil.

    PubMed

    de Sousa, Ricardo Silva; Santos, Vilma Maria; de Melo, Wanderley Jose; Nunes, Luis Alfredo Pinheiro Leal; van den Brink, Paul J; Araújo, Ademir Sérgio Ferreira

    2017-12-01

    Composting has been suggested as an efficient method for tannery sludge recycling before its application to the soil. However, the application of composted tannery sludge (CTS) should be monitored to evaluate its effect on the chemical and microbial properties of soil. This study evaluated the time-dependent effect of CTS on the chemical and microbial properties of soil. CTS was applied at 0, 2.5, 5, 10, and 20 Mg ha -1 and the soil chemical and microbial properties were evaluated at 0, 45, 75, 150, and 180 days. Increased CTS rates increased the levels of Ca, Cr, and Mg. While Soil pH, organic C, and P increased with the CTS rates initially, this effect decreased over time. Soil microbial biomass, respiration, metabolic quotient, and dehydrogenase increased with the application of CTS, but decreased over time. Analysis of the Principal Response Curve showed a significant effect of CTS rate on the chemical and microbial properties of the soil over time. The weight of each variable indicated that all soil properties, except β-glucosidase, dehydrogenase and microbial quotient, increased due to the CTS application. However, the highest weights were found for Cr, pH, Ca, P, phosphatase and total organic C. The application of CTS in the soil changed the chemical and microbial properties over time, indicating Cr, pH, Ca, phosphatase, and soil respiration as the more responsive chemical and microbial variables by CTS application.

  7. Characterization of Mechanical Properties of Microbial Biofilms

    NASA Astrophysics Data System (ADS)

    Callison, Elizabeth; Gose, James; Perlin, Marc; Ceccio, Steven

    2017-11-01

    The physical properties of microbial biofilms grown subject to shear flows determine the form and mechanical characteristics of the biofilm structure, and consequently, the turbulent interactions over and through the biofilm. These biofilms - sometimes referred to as slime - are comprised of microbial cells and extracellular polymeric substance (EPS) matrices that surround the multicellular communities. Some of the EPSs take the form of streamers that tend to oscillate in flows, causing increased turbulent mixing and drag. As the presence of EPS governs the compliance and overall stability of the filamentous streamers, investigation of the mechanical properties of biofilms may also inform efforts to understand hydrodynamic performance of fouled systems. In this study, a mixture of four diatom genera was grown under turbulent shear flow on test panels. The mechanical properties and hydrodynamic performance of the biofilm were investigated using rheology and turbulent flow studies in the Skin-Friction Flow Facility at the University of Michigan. The diatoms in the mixture of algae were identified, and the elastic and viscous moduli were determined from small-amplitude oscillations, while a creep test was used to evaluate the biofilm compliance.

  8. Combined Effect of Ultrasound and Mild Temperatures on the Inactivation of E. coli in Fresh Carrot Juice and Changes on its Physicochemical Characteristics.

    PubMed

    Pokhrel, Prashant Raj; Bermúdez-Aguirre, Daniela; Martínez-Flores, Héctor E; Garnica-Romo, M Guadalupe; Sablani, Shyam; Tang, Juming; Barbosa-Cánovas, Gustavo V

    2017-10-01

    The combination of ultrasound and mild temperatures to process fruits and vegetables juices is a novel approach that is showing promising results for microbial inactivation and preservation of bioactive compounds and sensory attributes. This study centers on investigating the inactivation of Escherichia coli (ATCC 11755) in carrot juice as a result of the combined effect of ultrasound (24 kHz frequency, 120 μm, and 400 W) with temperature (50, 54, and 58 °C) and processing time (0 to 10 min). In addition, the possible changes in physicochemical properties and the retention of bioactive compounds after processing were analyzed. Microbial inactivation with ultrasound treatment at 50 °C resulted in 3.5 log reduction after 10 min, whereas at 54 °C almost 5 log reduction was attained in the same period of time; meanwhile, for treatment at 58 °C, no viable cells were detected (>5 log reduction) after 2 min. There was no significant difference (P > 0.05) on pH (6.80 to 6.82), °Brix (8.0 to 8.5), titratable acidity (0.29% to 0.30%), total carotenoid (1774 to 1835 μg/100 mL), phenolic compounds (20.19 to 20.63 μg/mL), ascorbic acid (4.8 mg/100 mL), and color parameters between fresh and ultrasound treated samples at the studied temperatures. To predict the inactivation patterns, observed values were tested using 3 different general models: first-order, Weibull distribution, and biphasic. The Weibull and biphasic models show good correlation for inactivation under all processing conditions. Results show ultrasound in combination with mild temperature could be effectively used to process fresh carrot juice providing a safe product without affecting physicochemical characteristics. The combination of ultrasound and mild temperatures is effective in reducing microbial load in carrot juice to safe levels. This combination would be beneficial in the industrial processing of carrot juice without altering the quality attributes or bioactive compounds. © 2017 Institute of

  9. Cationic antimicrobial peptides inactivate Shiga toxin-encoding bacteriophages

    NASA Astrophysics Data System (ADS)

    Del Cogliano, Manuel E.; Hollmann, Axel; Martinez, Melina; Semorile, Liliana; Ghiringhelli, Pablo D.; Maffía, Paulo C.; Bentancor, Leticia V.

    2017-12-01

    Shiga toxin (Stx) is the principal virulence factor during Shiga toxin-producing Escherichia coli (STEC) infections. We have previously reported the inactivation of bacteriophage encoding Stx after treatment with chitosan, a linear polysaccharide polymer with cationic properties. Cationic antimicrobial peptides (cAMPs) are short linear aminoacidic sequences, with a positive net charge, which display bactericidal or bacteriostatic activity against a wide range of bacterial species. They are promising novel antibiotics since they have shown bactericidal effects against multiresistant bacteria. To evaluate whether cationic properties are responsible for bacteriophage inactivation, we tested seven cationic peptides with proven antimicrobial activity as anti-bacteriophage agents, and one random sequence cationic peptide with no antimicrobial activity as a control. We observed bacteriophage inactivation after incubation with five cAMPs, but no inactivating activity was observed with the random sequence cationic peptide or with the non alpha helical cAMP Omiganan. Finally, to confirm peptide-bacteriophage interaction, zeta potential was analyzed by following changes on bacteriophage surface charges after peptide incubation. According to our results we could propose that: 1) direct interaction of peptides with phage is a necessary step for bacteriophage inactivation, 2) cationic properties are necessary but not sufficient for bacteriophage inactivation, and 3) inactivation by cationic peptides could be sequence (or structure) specific. Overall our data suggest that these peptides could be considered a new family of molecules potentially useful to decrease bacteriophage replication and Stx expression.

  10. Microbial and chemical properties of log ponds along the Oregon Coast.

    Treesearch

    Iwan Ho; Ching Yan Li

    1987-01-01

    The microbial and chemical properties of log ponds along the Oregon coast were investigated. The log ponds were highly eutrophic, containing high concentrations of ammonium and nitrate nitrogen, phosphate, and organic compounds. Because of large microbial populations, the biochemical oxygen demand was high and dissolved oxygen was low. Bacterial species in log ponds...

  11. Low Stress Mechanical Properties of Plasma-Treated Cotton Fabric Subjected to Zinc Oxide-Anti-Microbial Treatment.

    PubMed

    Kan, Chi-Wai; Lam, Yin-Ling

    2013-01-22

    Cotton fabrics are highly popular because of their excellent properties such as regeneration, bio-degradation, softness, affinity to skin and hygroscopic properties. When in contact with the human body, cotton fabrics offer an ideal environment for microbial growth due to their ability to retain oxygen, moisture and warmth, as well as nutrients from spillages and body sweat. Therefore, an anti-microbial coating formulation (Microfresh and Microban together with zinc oxide as catalyst) was developed for cotton fabrics to improve treatment effectiveness. In addition, plasma technology was employed in the study which roughened the surface of the materials, improving the loading of zinc oxides on the surface. In this study, the low stress mechanical properties of plasma pre-treated and/or anti-microbial-treated cotton fabric were studied. The overall results show that the specimens had improved bending properties when zinc oxides were added in the anti-microbial coating recipe. Also, without plasma pre-treatment, anti-microbial-treatment of cotton fabric had a positive effect only on tensile resilience, shear stress at 0.5° and compressional energy, while plasma-treated specimens had better overall tensile properties even after anti-microbial treatment.

  12. Low Stress Mechanical Properties of Plasma-Treated Cotton Fabric Subjected to Zinc Oxide-Anti-Microbial Treatment

    PubMed Central

    Kan, Chi-Wai; Lam, Yin-Ling

    2013-01-01

    Cotton fabrics are highly popular because of their excellent properties such as regeneration, bio-degradation, softness, affinity to skin and hygroscopic properties. When in contact with the human body, cotton fabrics offer an ideal environment for microbial growth due to their ability to retain oxygen, moisture and warmth, as well as nutrients from spillages and body sweat. Therefore, an anti-microbial coating formulation (Microfresh and Microban together with zinc oxide as catalyst) was developed for cotton fabrics to improve treatment effectiveness. In addition, plasma technology was employed in the study which roughened the surface of the materials, improving the loading of zinc oxides on the surface. In this study, the low stress mechanical properties of plasma pre-treated and/or anti-microbial-treated cotton fabric were studied. The overall results show that the specimens had improved bending properties when zinc oxides were added in the anti-microbial coating recipe. Also, without plasma pre-treatment, anti-microbial-treatment of cotton fabric had a positive effect only on tensile resilience, shear stress at 0.5° and compressional energy, while plasma-treated specimens had better overall tensile properties even after anti-microbial treatment. PMID:28809311

  13. Inactivation of Salmonella spp. in ground chicken using High Pressure Processing

    USDA-ARS?s Scientific Manuscript database

    High pressure processing (HPP) is a safe and effective process for improving the microbial safety and shelf-life of foods. Salmonella is a common contaminant in poultry meat and is frequently responsible for foodborne illness associated with contaminated poultry meat. In this study the inactivation...

  14. Evaluation of Different Dose-Response Models for High Hydrostatic Pressure Inactivation of Microorganisms

    PubMed Central

    2017-01-01

    Modeling of microbial inactivation by high hydrostatic pressure (HHP) requires a plot of the log microbial count or survival ratio versus time data under a constant pressure and temperature. However, at low pressure and temperature values, very long holding times are needed to obtain measurable inactivation. Since the time has a significant effect on the cost of HHP processing it may be reasonable to fix the time at an appropriate value and quantify the inactivation with respect to pressure. Such a plot is called dose-response curve and it may be more beneficial than the traditional inactivation modeling since short holding times with different pressure values can be selected and used for the modeling of HHP inactivation. For this purpose, 49 dose-response curves (with at least 4 log10 reduction and ≥5 data points including the atmospheric pressure value (P = 0.1 MPa), and with holding time ≤10 min) for HHP inactivation of microorganisms obtained from published studies were fitted with four different models, namely the Discrete model, Shoulder model, Fermi equation, and Weibull model, and the pressure value needed for 5 log10 (P5) inactivation was calculated for all the models above. The Shoulder model and Fermi equation produced exactly the same parameter and P5 values, while the Discrete model produced similar or sometimes the exact same parameter values as the Fermi equation. The Weibull model produced the worst fit (had the lowest adjusted determination coefficient (R2adj) and highest mean square error (MSE) values), while the Fermi equation had the best fit (the highest R2adj and lowest MSE values). Parameters of the models and also P5 values of each model can be useful for the further experimental design of HHP processing and also for the comparison of the pressure resistance of different microorganisms. Further experiments can be done to verify the P5 values at given conditions. The procedure given in this study can also be extended for enzyme

  15. Toward Linking Aboveground Vegetation Properties and Soil Microbial Communities Using Remote Sensing

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

    Hamada, Yuki; Gilbert, Jack A.; Larsen, Peter E.

    2014-04-01

    Despite their vital role in terrestrial ecosystem function, the distributions and dynamics of soil microbial communities (SMCs) are poorly understood. Vegetation and soil properties are the primary factors that influence SMCs. This paper discusses the potential effectiveness of remote sensing science and technologies for mapping SMC biogeography by characterizing surface biophysical properties (e.g., plant traits and community composition) strongly correlated with SMCs. Using remotely sensed biophysical properties to predict SMC distributions is extremely challenging because of the intricate interactions between biotic and abiotic factors and between above- and belowground ecosystems. However, the integration of biophysical and soil remote sensing withmore » geospatial information about the e nvironment holds great promise for mapping SMC biogeography. Additional research needs invol ve microbial taxonomic definition, soil environmental complexity, and scaling strategies. The collaborative effort of experts from diverse disciplines is essential to linking terrestrial surface biosphere observations with subsurface microbial community distributions using remote sensing.« less

  16. Toward Linking Aboveground Vegetation Properties and Soil Microbial Communities Using Remote Sensing

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

    Hamada, Yuki; Gilbert, Jack A.; Larsen, Peter E.

    2014-04-01

    Despite their vital role in terrestrial ecosystem function, the distributions and dynamics of soil microbial communities (SMCs) are poorly understood. Vegetation and soil properties are the primary factors that influence SMCs. This paper discusses the potential effectiveness of remote sensing science and technologies for mapping SMC biogeography by characterizing surface biophysical properties (e.g., plant traits and community composition) strongly correlated with SMCs. Using remotely sensed biophysical properties to predict SMC distributions is extremely challenging because of the intricate interactions between biotic and abiotic factors and between above- and below-ground ecosystems. However, the integration of biophysical and soil remote sensing withmore » geospatial information about the environment holds great promise for mapping SMC biogeography. Additional research needs involve microbial taxonomic definition, soil environmental complexity, and scaling strategies. The collaborative effort of experts from diverse disciplines is essential to linking terrestrial surface biosphere observations with subsurface microbial community distributions using remote sensing.« less

  17. Optimising the inactivation of grape juice spoilage organisms by pulse electric fields.

    PubMed

    Marsellés-Fontanet, A Robert; Puig, Anna; Olmos, Paola; Mínguez-Sanz, Santiago; Martín-Belloso, Olga

    2009-04-15

    The effect of some pulsed electric field (PEF) processing parameters (electric field strength, pulse frequency and treatment time), on a mixture of microorganisms (Kloeckera apiculata, Saccharomyces cerevisiae, Lactobacillus plantarum, Lactobacillus hilgardii and Gluconobacter oxydans) typically present in grape juice and wine were evaluated. An experimental design based on response surface methodology (RSM) was used and results were also compared with those of a factorially designed experiment. The relationship between the levels of inactivation of microorganisms and the energy applied to the grape juice was analysed. Yeast and bacteria were inactivated by the PEF treatments, with reductions that ranged from 2.24 to 3.94 log units. All PEF parameters affected microbial inactivation. Optimal inactivation of the mixture of spoilage microorganisms was predicted by the RSM models at 35.0 kV cm(-1) with 303 Hz pulse width for 1 ms. Inactivation was greater for yeasts than for bacteria, as was predicted by the RSM. The maximum efficacy of the PEF treatment for inactivation of microorganisms in grape juice was observed around 1500 MJ L(-1) for all the microorganisms investigated. The RSM could be used in the fruit juice industry to optimise the inactivation of spoilage microorganisms by PEF.

  18. Seasonal variation in functional properties of microbial communities in beech forest soil

    PubMed Central

    Koranda, Marianne; Kaiser, Christina; Fuchslueger, Lucia; Kitzler, Barbara; Sessitsch, Angela; Zechmeister-Boltenstern, Sophie; Richter, Andreas

    2013-01-01

    Substrate quality and the availability of nutrients are major factors controlling microbial decomposition processes in soils. Seasonal alteration in resource availability, which is driven by plants via belowground C allocation, nutrient uptake and litter fall, also exerts effects on soil microbial community composition. Here we investigate if seasonal and experimentally induced changes in microbial community composition lead to alterations in functional properties of microbial communities and thus microbial processes. Beech forest soils characterized by three distinct microbial communities (winter and summer community, and summer community from a tree girdling plot, in which belowground carbon allocation was interrupted) were incubated with different 13C-labeled substrates with or without inorganic N supply and analyzed for substrate use and various microbial processes. Our results clearly demonstrate that the three investigated microbial communities differed in their functional response to addition of various substrates. The winter communities revealed a higher capacity for degradation of complex C substrates (cellulose, plant cell walls) than the summer communities, indicated by enhanced cellulase activities and reduced mineralization of soil organic matter. In contrast, utilization of labile C sources (glucose) was lower in winter than in summer, demonstrating that summer and winter community were adapted to the availability of different substrates. The saprotrophic community established in girdled plots exhibited a significantly higher utilization of complex C substrates than the more plant root associated community in control plots if additional nitrogen was provided. In this study we were able to demonstrate experimentally that variation in resource availability as well as seasonality in temperate forest soils cause a seasonal variation in functional properties of soil microorganisms, which is due to shifts in community structure and physiological adaptations

  19. Inactivation disinfection property of Moringa Oleifera seed extract: optimization and kinetic studies

    NASA Astrophysics Data System (ADS)

    Idris, M. A.; Jami, M. S.; Hammed, A. M.

    2017-05-01

    This paper presents the statistical optimization study of disinfection inactivation parameters of defatted Moringa oleifera seed extract on Pseudomonas aeruginosa bacterial cells. Three level factorial design was used to estimate the optimum range and the kinetics of the inactivation process was also carried. The inactivation process involved comparing different disinfection models of Chicks-Watson, Collins-Selleck and Homs models. The results from analysis of variance (ANOVA) of the statistical optimization process revealed that only contact time was significant. The optimum disinfection range of the seed extract was 125 mg/L, 30 minutes and 120rpm agitation. At the optimum dose, the inactivation kinetics followed the Collin-Selleck model with coefficient of determination (R2) of 0.6320. This study is the first of its kind in determining the inactivation kinetics of pseudomonas aeruginosa using the defatted seed extract.

  20. Effects of Soil Organic Matter Properties and Microbial Community Composition on Enzyme Activities in Cryoturbated Arctic Soils

    PubMed Central

    Schnecker, Jörg; Wild, Birgit; Hofhansl, Florian; Eloy Alves, Ricardo J.; Bárta, Jiří; Čapek, Petr; Fuchslueger, Lucia; Gentsch, Norman; Gittel, Antje; Guggenberger, Georg; Hofer, Angelika; Kienzl, Sandra; Knoltsch, Anna; Lashchinskiy, Nikolay; Mikutta, Robert; Šantrůčková, Hana; Shibistova, Olga; Takriti, Mounir; Urich, Tim; Weltin, Georg; Richter, Andreas

    2014-01-01

    Enzyme-mediated decomposition of soil organic matter (SOM) is controlled, amongst other factors, by organic matter properties and by the microbial decomposer community present. Since microbial community composition and SOM properties are often interrelated and both change with soil depth, the drivers of enzymatic decomposition are hard to dissect. We investigated soils from three regions in the Siberian Arctic, where carbon rich topsoil material has been incorporated into the subsoil (cryoturbation). We took advantage of this subduction to test if SOM properties shape microbial community composition, and to identify controls of both on enzyme activities. We found that microbial community composition (estimated by phospholipid fatty acid analysis), was similar in cryoturbated material and in surrounding subsoil, although carbon and nitrogen contents were similar in cryoturbated material and topsoils. This suggests that the microbial community in cryoturbated material was not well adapted to SOM properties. We also measured three potential enzyme activities (cellobiohydrolase, leucine-amino-peptidase and phenoloxidase) and used structural equation models (SEMs) to identify direct and indirect drivers of the three enzyme activities. The models included microbial community composition, carbon and nitrogen contents, clay content, water content, and pH. Models for regular horizons, excluding cryoturbated material, showed that all enzyme activities were mainly controlled by carbon or nitrogen. Microbial community composition had no effect. In contrast, models for cryoturbated material showed that enzyme activities were also related to microbial community composition. The additional control of microbial community composition could have restrained enzyme activities and furthermore decomposition in general. The functional decoupling of SOM properties and microbial community composition might thus be one of the reasons for low decomposition rates and the persistence of 400 Gt

  1. Inactivation credit of UV radiation for viruses, bacteria and protozoan (oo)cysts in water: a review.

    PubMed

    Hijnen, W A M; Beerendonk, E F; Medema, G J

    2006-01-01

    UV disinfection technology is of growing interest in the water industry since it was demonstrated that UV radiation is very effective against (oo)cysts of Cryptosporidium and Giardia, two pathogenic micro-organisms of major importance for the safety of drinking water. Quantitative Microbial Risk Assessment, the new concept for microbial safety of drinking water and wastewater, requires quantitative data of the inactivation or removal of pathogenic micro-organisms by water treatment processes. The objective of this study was to review the literature on UV disinfection and extract quantitative information about the relation between the inactivation of micro-organisms and the applied UV fluence. The quality of the available studies was evaluated and only high-quality studies were incorporated in the analysis of the inactivation kinetics. The results show that UV is effective against all waterborne pathogens. The inactivation of micro-organisms by UV could be described with first-order kinetics using fluence-inactivation data from laboratory studies in collimated beam tests. No inactivation at low fluences (offset) and/or no further increase of inactivation at higher fluences (tailing) was observed for some micro-organisms. Where observed, these were included in the description of the inactivation kinetics, even though the cause of tailing is still a matter of debate. The parameters that were used to describe inactivation are the inactivation rate constant k (cm(2)/mJ), the maximum inactivation demonstrated and (only for bacterial spores and Acanthamoeba) the offset value. These parameters were the basis for the calculation of the microbial inactivation credit (MIC="log-credits") that can be assigned to a certain UV fluence. The most UV-resistant organisms are viruses, specifically Adenoviruses, and bacterial spores. The protozoon Acanthamoeba is also highly UV resistant. Bacteria and (oo)cysts of Cryptosporidium and Giardia are more susceptible with a fluence

  2. Properties of an intermediate-duration inactivation process of the voltage-gated sodium conductance in rat hippocampal CA1 neurons.

    PubMed

    French, Christopher R; Zeng, Zhen; Williams, David A; Hill-Yardin, Elisa L; O'Brien, Terence J

    2016-02-01

    Rapid transmembrane flow of sodium ions produces the depolarizing phase of action potentials (APs) in most excitable tissue through voltage-gated sodium channels (NaV). Macroscopic currents display rapid activation followed by fast inactivation (IF) within milliseconds. Slow inactivation (IS) has been subsequently observed in several preparations including neuronal tissues. IS serves important physiological functions, but the kinetic properties are incompletely characterized, especially the operative timescales. Here we present evidence for an "intermediate inactivation" (II) process in rat hippocampal CA1 neurons with time constants of the order of 100 ms. The half-inactivation potentials (V0.5) of steady-state inactivation curves were hyperpolarized by increasing conditioning pulse duration from 50 to 500 ms and could be described by a sum of Boltzmann relations. II state transitions were observed after opening as well as subthreshold potentials. Entry into II after opening was relatively insensitive to membrane potential, and recovery of II became more rapid at hyperpolarized potentials. Removal of fast inactivation with cytoplasmic papaine revealed time constants of INa decay corresponding to II and IS with long depolarizations. Dynamic clamp revealed attenuation of trains of APs over the 10(2)-ms timescale, suggesting a functional role of II in repetitive firing accommodation. These experimental findings could be reproduced with a five-state Markov model. It is likely that II affects important aspects of hippocampal neuron response and may provide a drug target for sodium channel modulation. Copyright © 2016 the American Physiological Society.

  3. Inactivation of bacteriophage MS2 with potassium ferrate(VI).

    PubMed

    Hu, Lanhua; Page, Martin A; Sigstam, Therese; Kohn, Tamar; Mariñas, Benito J; Strathmann, Timothy J

    2012-11-06

    Ferrate [Fe(VI); FeO(4)(2-)] is an emerging oxidizing agent capable of controlling chemical and microbial water contaminants. Here, inactivation of MS2 coliphage by Fe(VI) was examined. The inactivation kinetics observed in individual batch experiments was well described by a Chick-Watson model with first-order dependences on disinfectant and infective phage concentrations. The inactivation rate constant k(i) at a Fe(VI) dose of 1.23 mgFe/L (pH 7.0, 25 °C) was 2.27(±0.05) L/(mgFe × min), corresponding to 99.99% inactivation at a Ct of ~4 (mgFe × min)/L. Measured k(i) values were found to increase with increasing applied Fe(VI) dose (0.56-2.24 mgFe/L), increasing temperature (5-30 °C), and decreasing pH conditions (pH 6-11). The Fe(VI) dose effect suggested that an unidentified Fe byproduct also contributed to inactivation. Temperature dependence was characterized by an activation energy of 39(±6) kJ mol(-1), and k(i) increased >50-fold when pH decreased from 11 to 6. The pH effect was quantitatively described by parallel reactions with HFeO(4)(-) and FeO(4)(2-). Mass spectrometry and qRT-PCR analyses demonstrated that both capsid protein and genome damage increased with the extent of inactivation, suggesting that both may contribute to phage inactivation. Capsid protein damage, localized in the two regions containing oxidant-sensitive cysteine residues, and protein cleavage in one of the two regions may facilitate genome damage by increasing Fe(VI) access to the interior of the virion.

  4. Quantitative analysis of wet-heat inactivation in bovine spongiform encephalopathy

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

    Matsuura, Yuichi; Ishikawa, Yukiko; Bo, Xiao

    2013-03-01

    Highlights: ► We quantitatively analyzed wet-heat inactivation of the BSE agent. ► Infectivity of the BSE macerate did not survive 155 °C wet-heat treatment. ► Once the sample was dehydrated, infectivity was observed even at 170 °C. ► A quantitative PMCA assay was used to evaluate the degree of BSE inactivation. - Abstract: The bovine spongiform encephalopathy (BSE) agent is resistant to conventional microbial inactivation procedures and thus threatens the safety of cattle products and by-products. To obtain information necessary to assess BSE inactivation, we performed quantitative analysis of wet-heat inactivation of infectivity in BSE-infected cattle spinal cords. Using amore » highly sensitive bioassay, we found that infectivity in BSE cattle macerates fell with increase in temperatures from 133 °C to 150 °C and was not detected in the samples subjected to temperatures above 155 °C. In dry cattle tissues, infectivity was detected even at 170 °C. Thus, BSE infectivity reduces with increase in wet-heat temperatures but is less affected when tissues are dehydrated prior to the wet-heat treatment. The results of the quantitative protein misfolding cyclic amplification assay also demonstrated that the level of the protease-resistant prion protein fell below the bioassay detection limit by wet-heat at 155 °C and higher and could help assess BSE inactivation. Our results show that BSE infectivity is strongly resistant to wet-heat inactivation and that it is necessary to pay attention to BSE decontamination in recycled cattle by-products.« less

  5. Role of hydraulic retention time and granular medium in microbial removal in tertiary treatment reed beds.

    PubMed

    García, Joan; Vivar, Joan; Aromir, Maria; Mujeriego, Rafael

    2003-06-01

    The main objective of this paper is to evaluate the role of hydraulic retention time (HRT) and granular medium in faecal coliform (FC) and somatic coliphage (SC) removal in tertiary reed beds. Experiments were carried out in a pilot plant with four parallel reed beds (horizontal subsurface flow constructed wetlands), each one containing a different type of granular medium. This pilot plant is located in a wastewater treatment plant in Montcada i Reixac, near Barcelona, in northeastern Spain. The microbial inactivation ratios obtained in the different beds are compared as a function of three selected HRTs. Secondary effluent from the wastewater treatment plant was used as the influent of the pilot system. The microbial inactivation ratio ranged between 0.1 and 2.7 log-units for FC and from 0.5 to 1.7 log-units for SC in beds with coarser granular material (5-25mm), while it ranged between 0.7 and 3.4 log-units for FC and from 0.9 to 2.6 log-units for SC in the bed with finer material (2-13mm). HRT and granular medium are both key factors in microbial removal in the tertiary reed beds. The microbial inactivation ratio rises as the HRT increases until it reaches a saturation value (in general at an HRT of 3 days). The value of the microbial inactivation ratio at the saturation level depends on the granular medium contained in the bed. The specific surface area necessary to reach 2-3 log-units of FC and SC is approximately 3m(2)/person-equivalent.

  6. Inactivation of Prions and Amyloid Seeds with Hypochlorous Acid

    PubMed Central

    Kraus, Allison; Phillips, Katie; Contreras, Luis; Zanusso, Gianluigi; Caughey, Byron

    2016-01-01

    Hypochlorous acid (HOCl) is produced naturally by neutrophils and other cells to kill conventional microbes in vivo. Synthetic preparations containing HOCl can also be effective as microbial disinfectants. Here we have tested whether HOCl can also inactivate prions and other self-propagating protein amyloid seeds. Prions are deadly pathogens that are notoriously difficult to inactivate, and standard microbial disinfection protocols are often inadequate. Recommended treatments for prion decontamination include strongly basic (pH ≥~12) sodium hypochlorite bleach, ≥1 N sodium hydroxide, and/or prolonged autoclaving. These treatments are damaging and/or unsuitable for many clinical, agricultural and environmental applications. We have tested the anti-prion activity of a weakly acidic aqueous formulation of HOCl (BrioHOCl) that poses no apparent hazard to either users or many surfaces. For example, BrioHOCl can be applied directly to skin and mucous membranes and has been aerosolized to treat entire rooms without apparent deleterious effects. Here, we demonstrate that immersion in BrioHOCl can inactivate not only a range of target microbes, including spores of Bacillus subtilis, but also prions in tissue suspensions and on stainless steel. Real-time quaking-induced conversion (RT-QuIC) assays showed that BrioHOCl treatments eliminated all detectable prion seeding activity of human Creutzfeldt-Jakob disease, bovine spongiform encephalopathy, cervine chronic wasting disease, sheep scrapie and hamster scrapie; these findings indicated reductions of ≥103- to 106-fold. Transgenic mouse bioassays showed that all detectable hamster-adapted scrapie infectivity in brain homogenates or on steel wires was eliminated, representing reductions of ≥~105.75-fold and >104-fold, respectively. Inactivation of RT-QuIC seeding activity correlated with free chlorine concentration and higher order aggregation or destruction of proteins generally, including prion protein. Brio

  7. Catalytic power of enzymes decreases with temperature: New insights for understanding soil C cycling and microbial ecology under warming.

    PubMed

    Alvarez, Gaël; Shahzad, Tanvir; Andanson, Laurence; Bahn, Michael; Wallenstein, Matthew D; Fontaine, Sébastien

    2018-04-23

    Most current models of soil C dynamics predict that climate warming will accelerate soil C mineralization, resulting in a long-term CO 2 release and positive feedback to global warming. However, ecosystem warming experiments show that CO 2 loss from warmed soils declines to control levels within a few years. Here, we explore the temperature dependence of enzymatic conversion of polymerized soil organic C (SOC) into assimilable compounds, which is presumed the rate-limiting step of SOC mineralization. Combining literature review, modelling and enzyme assays, we studied the effect of temperature on activity of enzymes considering their thermal inactivation and catalytic activity. We defined the catalytic power of enzymes (E power ) as the cumulative amount of degraded substrate by one unit of enzyme until its complete inactivation. We show a universal pattern of enzyme's thermodynamic properties: activation energy of catalytic activity (EA cat ) < activation energy of thermal inactivation (EA inact ). By investing in stable enzymes (high EA inact ) having high catalytic activity (low EA cat ), microorganisms may maximize the E power of their enzymes. The counterpart of such EAs' hierarchical pattern is the higher relative temperature sensitivity of enzyme inactivation than catalysis, resulting in a reduction in E power under warming. Our findings could explain the decrease with temperature in soil enzyme pools, microbial biomass (MB) and carbon use efficiency (CUE) reported in some warming experiments and studies monitoring the seasonal variation in soil enzymes. They also suggest that a decrease in soil enzyme pools due to their faster inactivation under warming contributes to the observed attenuation of warming effect on soil C mineralization. This testable theory predicts that the ultimate response of SOC degradation to warming can be positive or negative depending on the relative temperature response of E power and microbial production of enzymes. © 2018 John

  8. High-pressure processing of apple juice: kinetics of pectin methyl esterase inactivation.

    PubMed

    Riahi, Esmaeil; Ramaswamy, Hosahalli S

    2003-01-01

    High-pressure (HP) inactivation kinetics of pectin methyl esterase (PME) in apple juice were evaluated. Commercial PME was dispensed in clarified apple juice, sealed in dual peel sterilizable plastic bags, and subjected to different high-pressure processing conditions (200-400 MPa, 0-180 min). Residual enzyme activity was determined by a titration method estimating the rate of free carboxyl group released by the enzyme acting on pectin substrate at pH 7.5 (30 degrees C). The effects of pressure level and pressure holding time on enzyme inactivation were significant (p < 0.05). PME from the microbial source was found to be more resistant (p < 0.05) to pressure inactivation than PME from the orange peel. Almost a full decimal reduction in the activity of commercial PME was achieved by HP treatment at 400 MPa for 25 min. Inactivation kinetics were evaluated on the basis of a dual effect model involving a pressure pulse effect and a first-order rate model, and the pressure sensitivity of rate constants was modeled by using the z-value concept.

  9. Effects of different soil management practices on soil properties and microbial diversity

    NASA Astrophysics Data System (ADS)

    Gajda, Anna M.; Czyż, Ewa A.; Dexter, Anthony R.; Furtak, Karolina M.; Grządziel, Jarosław; Stanek-Tarkowska, Jadwiga

    2018-01-01

    The effects of different tillage systems on the properties and microbial diversity of an agricultural soil was investigated. In doing so, soil physical, chemical and biological properties were analysed in 2013-2015, on a long-term field experiment on a loamy sand at the IUNG-PIB Experimental Station in Grabów, Poland. Winter wheat was grown under two tillage treatments: conventional tillage using a mouldboard plough and traditional soil tillage equipment, and reduced tillage based on soil crushing-loosening equipment and a rigid-tine cultivator. Chopped wheat straw was used as a mulch on both treatments. Reduced tillage resulted in increased water content throughout the whole soil profile, in comparison with conventional tillage. Under reduced tillage, the content of readily dispersible clay was also reduced, and, therefore, soil stability was increased in the toplayers, compared with conventional tillage. In addition, the beneficial effects of reduced tillage were reflected in higher soil microbial activity as measured with dehydrogenases and hydrolysis of fluorescein diacetate, compared with conventional tillage. Moreover, the polimerase chain reaction - denaturing gradient gel electrophoresis analysis showed that soil under reduced till-age had greater diversity of microbial communities, compared with conventionally-tilled soil. Finally, reduced tillage increased organic matter content, stability in water and microbial diversity in the top layer of the soil.

  10. CoBOP: Microbial Biofilms: A Parameter Altering the Apparent Optical Properties of Sediments, Seagrasses and Surfaces

    DTIC Science & Technology

    2002-09-30

    CoBOP: Microbial Biofilms: A Parameter Altering the Apparent Optical Properties of Sediments, Seagrasses and Surfaces Alan W. Decho Department...TITLE AND SUBTITLE CoBOP: Microbial Biofilms: A Parameter Altering the Apparent Optical Properties of Sediments, Seagrasses and Surfaces 5a. CONTRACT...structures produced by bacteria. Their growth appears to depend on biofilm processes and light distributions ( photosynthesis ). Therefore, the data acquired

  11. CoBOP: Microbial Biofilms: A Parameter Altering the Apparent Optical Properties of Sediments, Seagrasses and Surfaces

    DTIC Science & Technology

    2003-09-30

    CoBOP: MICROBIAL BIOFILMS: A PARAMETER ALTERING THE APPARENT OPTICAL PROPERTIES OF SEDIMENTS, SEAGRASSES AND SURFACES. Alan W. Decho Department...AND SUBTITLE CoBOP: Microbial Biofilms: A Parameter Altering The Apparent Optical Properties Of Sediments, Seagrasses And Surfaces 5a. CONTRACT...biofilm processes and light distributions ( photosynthesis ). Therefore, the data acquired from this project will be closely paired with results of

  12. Far-UVC light: A new tool to control the spread of airborne-mediated microbial diseases.

    PubMed

    Welch, David; Buonanno, Manuela; Grilj, Veljko; Shuryak, Igor; Crickmore, Connor; Bigelow, Alan W; Randers-Pehrson, Gerhard; Johnson, Gary W; Brenner, David J

    2018-02-09

    Airborne-mediated microbial diseases such as influenza and tuberculosis represent major public health challenges. A direct approach to prevent airborne transmission is inactivation of airborne pathogens, and the airborne antimicrobial potential of UVC ultraviolet light has long been established; however, its widespread use in public settings is limited because conventional UVC light sources are both carcinogenic and cataractogenic. By contrast, we have previously shown that far-UVC light (207-222 nm) efficiently inactivates bacteria without harm to exposed mammalian skin. This is because, due to its strong absorbance in biological materials, far-UVC light cannot penetrate even the outer (non living) layers of human skin or eye; however, because bacteria and viruses are of micrometer or smaller dimensions, far-UVC can penetrate and inactivate them. We show for the first time that far-UVC efficiently inactivates airborne aerosolized viruses, with a very low dose of 2 mJ/cm 2 of 222-nm light inactivating >95% of aerosolized H1N1 influenza virus. Continuous very low dose-rate far-UVC light in indoor public locations is a promising, safe and inexpensive tool to reduce the spread of airborne-mediated microbial diseases.

  13. Infectious Causes of Cholesteatoma and Treatment of Infected Ossicles prior to Reimplantation by Hydrostatic High-Pressure Inactivation

    PubMed Central

    Hinz, Rebecca

    2015-01-01

    Chronic inflammation, which is caused by recurrent infections, is one of the factors contributing to the pathogenesis of cholesteatoma. If reimplantation of autologous ossicles after a surgical intervention is intended, inactivation of planktonic bacteria and biofilms is desirable. High hydrostatic pressure treatment is a procedure, which has been used to inactivate cholesteatoma cells on ossicles. Here we discuss the potential inactivating effect of high hydrostatic pressure on microbial pathogens including biofilms. Recent experimental data suggest an incomplete inactivation at a pressure level, which is tolerable for the bone substance of ossicles and results at least in a considerable reduction of pathogen load. Further studies are necessary to access how far this quantitative reduction of pathogens is sufficient to prevent ongoing chronic infections, for example, due to forming of biofilms. PMID:25705686

  14. Inactivation kinetics and efficiencies of UV-LEDs against Pseudomonas aeruginosa, Legionella pneumophila, and surrogate microorganisms.

    PubMed

    Rattanakul, Surapong; Oguma, Kumiko

    2018-03-01

    To demonstrate the effectiveness of UV light-emitting diodes (UV-LEDs) to disinfect water, UV-LEDs at peak emission wavelengths of 265, 280, and 300 nm were adopted to inactivate pathogenic species, including Pseudomonas aeruginosa and Legionella pneumophila, and surrogate species, including Escherichia coli, Bacillus subtilis spores, and bacteriophage Qβ in water, compared to conventional low-pressure UV lamp emitting at 254 nm. The inactivation profiles of each species showed either a linear or sigmoidal survival curve, which both fit well with the Geeraerd's model. Based on the inactivation rate constant, the 265-nm UV-LED showed most effective fluence, except for with E. coli which showed similar inactivation rates at 265 and 254 nm. Electrical energy consumption required for 3-log 10 inactivation (E E,3 ) was lowest for the 280-nm UV-LED for all microbial species tested. Taken together, the findings of this study determined the inactivation profiles and kinetics of both pathogenic bacteria and surrogate species under UV-LED exposure at different wavelengths. We also demonstrated that not only inactivation rate constants, but also energy efficiency should be considered when selecting an emission wavelength for UV-LEDs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Physicochemical properties influencing denitrification rate and microbial activity in denitrification bioreactors

    NASA Astrophysics Data System (ADS)

    Schmidt, C. A.

    2012-12-01

    The use of N-based fertilizer will need to increase to meet future demands, yet existing applications have been implicated as the main source of coastal eutrophication and hypoxic zones. Producing sufficient crops to feed a growing planet will require efficient production in combination with sustainable treatment solutions. The long-term success of denitrification bioreactors to effectively remove nitrate (NO¬3), indicates this technology is a feasible treatment option. Assessing and quantifying the media properties that affect NO¬3 removal rate and microbial activity can improve predictions on bioreactor performance. It was hypothesized that denitrification rates and microbial biomass would be correlated with total C, NO¬3 concentration, metrics of organic matter quality, media surface area and laboratory measures of potential denitrification rate. NO¬3 removal rates and microbial biomass were evaluated in mesocosms filled with different wood treatments and the unique influence of these predictor variables was determined using a multiple linear regression analysis. NO3 reduction rates were independent of NO¬3 concentration indicating zero order reaction kinetics. Temperature was strongly correlated with denitrification rate (r2=0.87; Q10=4.7), indicating the variability of bioreactor performance in differing climates. Fiber quality, and media surface area were strong (R>0.50), unique predictors of rates and microbial biomass, although C:N ratio and potential denitrification rate did not predict actual denitrification rate or microbial biomass. Utilizing a stepwise multiple linear regression, indicates that the denitrification rate can be effectively (r2=0.56;p<0.0001) predicted if the groundwater temperature, neutral detergent fiber and surface area alone are quantified. These results will assist with the widespread implementation of denitrification bioreactors to achieve significant N load reductions in large watersheds. The nitrate reduction rate as a

  16. Microbial inactivation and cytotoxicity evaluation of UV irradiated coconut water in a novel continuous flow spiral reactor.

    PubMed

    Bhullar, Manreet Singh; Patras, Ankit; Kilanzo-Nthenge, Agnes; Pokharel, Bharat; Yannam, Sudheer Kumar; Rakariyatham, Kanyasiri; Pan, Che; Xiao, Hang; Sasges, Michael

    2018-01-01

    A continuous-flow UV reactor operating at 254nm wave-length was used to investigate inactivation of microorganisms including bacteriophage in coconut water, a highly opaque liquid food. UV-C inactivation kinetics of two surrogate viruses (MS2, T1UV) and three bacteria (E. coli ATCC 25922, Salmonella Typhimurium ATCC 13311, Listeria monocytogenes ATCC 19115) in buffer and coconut water were investigated (D 10 values ranging from 2.82 to 4.54mJ·cm -2 ). A series of known UV-C doses were delivered to the samples. Inactivation levels of all organisms were linearly proportional to UV-C dose (r 2 >0.97). At the highest dose of 30mJ·cm -2 , the three pathogenic organisms were inactivated by >5 log 10 (p<0.05). Results clearly demonstrated that UV-C irradiation effectively inactivated bacteriophage and pathogenic microbes in coconut water. The inactivation kinetics of microorganisms were best described by log linear model with a low root mean square error (RMSE) and high coefficient of determination (r 2 >0.97). Models for predicting log reduction as a function of UV-C irradiation dose were found to be significant (p<0.05) with low RMSE and high r 2 . The irradiated coconut water showed no cytotoxic effects on normal human intestinal cells and normal mouse liver cells. Overall, these results indicated that UV-C treatment did not generate cytotoxic compounds in the coconut water. This study clearly demonstrated that high levels of inactivation of pathogens can be achieved in coconut water, and suggested potential method for UV-C treatment of other liquid foods. This research paper provides scientific evidence of the potential benefits of UV-C irradiation in inactivating bacterial and viral surrogates at commercially relevant doses of 0-120mJ·cm -2 . The irradiated coconut water showed no cytotoxic effects on normal intestinal and healthy mice liver cells. UV-C irradiation is an attractive food preservation technology and offers opportunities for horticultural and food

  17. Changes in Microbial Community Structure and Soil Biological Properties in Mined Dune Areas During Re-vegetation.

    PubMed

    Escobar, Indra Elena C; Santos, Vilma M; da Silva, Danielle Karla A; Fernandes, Marcelo F; Cavalcante, Uided Maaze T; Maia, Leonor C

    2015-06-01

    The aim of this study was to describe the impact of re-vegetation on the restoration of microbial community structure and soil microbiological properties in sand dunes that had been affected by mining activity. Soil samples were collected during the dry and rainy seasons from a chronosequence (1, 9, 21 years) of re-vegetated dunes using a single preserved dune as a reference. The composition of the fatty acid methyl esters and soil microbial properties were evaluated. The results showed that the changes in microbial community structure were related to seasonal variations: biomarkers of Gram-positive bacteria were higher than Gram-negative bacteria during the dry season, showing that this group of organisms is more tolerant to these stressful conditions. The microbial community structure in the natural dune was less affected by seasonal variation compared to the re-vegetated areas, whereas the opposite was observed for microbiological properties. Thus, in general, the proportion of saprobic fungi was higher in the natural dune, whereas Gram-negative bacteria were proportionally more common in the younger areas. Although over time the re-vegetation allows the recovery of the microbial community and the soil functions, these communities and functions are different from those found in the undisturbed areas.

  18. Bacteria and fungi inactivation by photocatalysis under UVA irradiation: liquid and gas phase.

    PubMed

    Rodrigues-Silva, Caio; Miranda, Sandra M; Lopes, Filipe V S; Silva, Mário; Dezotti, Márcia; Silva, Adrián M T; Faria, Joaquim L; Boaventura, Rui A R; Vilar, Vítor J P; Pinto, Eugénia

    2017-03-01

    In the last decade, environmental risks associated with wastewater treatment plants (WWTPs) have become a concern in the scientific community due to the absence of specific legislation governing the occupational exposure limits (OEL) for microorganisms present in indoor air. Thus, it is necessary to develop techniques to effectively inactivate microorganisms present in the air of WWTPs facilities. In the present work, ultraviolet light A radiation was used as inactivation tool. The microbial population was not visibly reduced in the bioaerosol by ultraviolet light A (UVA) photolysis. The UVA photocatalytic process for the inactivation of microorganisms (bacteria and fungi, ATCC strains and isolates from indoor air samples of a WWTP) using titanium dioxide (TiO 2 P25) and zinc oxide (ZnO) was tested in both liquid-phase and airborne conditions. In the slurry conditions at liquid phase, P25 showed a better performance in inactivation. For this reason, gas-phase assays were performed in a tubular photoreactor packed with cellulose acetate monolithic structures coated with P25. The survival rate of microorganisms under study decreased with the catalyst load and the UVA exposure time. Inactivation of fungi was slower than resistant bacteria, followed by Gram-positive bacteria and Gram-negative bacteria. Graphical abstract Inactivation of fungi and bacteria in gas phase by photocatalitic process performed in a tubular photoreactor packed with cellulose acetate monolith structures coated with TiO 2 .

  19. Atmospheric-pressure cold plasma treatment of contaminated fresh fruit and vegetable slices: inactivation and physiochemical properties evaluation

    NASA Astrophysics Data System (ADS)

    Wang, R. X.; Nian, W. F.; Wu, H. Y.; Feng, H. Q.; Zhang, K.; Zhang, J.; Zhu, W. D.; Becker, K. H.; Fang, J.

    2012-10-01

    A direct-current, atmospheric-pressure air cold plasma microjet (PMJ) was applied to disinfect Salmonella directly deposited on fresh fruit and vegetable slices. Effective inactivation was achieved on sliced fruit and vegetables after 1 s plasma treatment. The physiochemical properties of the slices, such as water content, color parameters, and nutritional content were monitored before and after plasma treatment. It was found that the physiochemical properties changes caused by the plasma were within an acceptable range. Reactive oxygen species, which are believed to be the major bactericidal agents in the plasma, were detected by electron spin resonance spectroscopy and optical emission spectroscopy.

  20. Kinetics of Ozone Inactivation of Infectious Prion Protein

    PubMed Central

    Ding, Ning; Price, Luke M.; Braithwaite, Shannon L.; Balachandran, Aru; Mitchell, Gordon; Belosevic, Miodrag

    2013-01-01

    The kinetics of ozone inactivation of infectious prion protein (PrPSc, scrapie 263K) was investigated in ozone-demand-free phosphate-buffered saline (PBS). Diluted infectious brain homogenates (IBH) (0.01%) were exposed to a predetermined ozone dose (10.8 ± 2.0 mg/liter) at three pHs (pH 4.4, 6.0, and 8.0) and two temperatures (4°C and 20°C). The inactivation of PrPSc was quantified by determining the in vitro destruction of PrPSc templating properties using the protein misfolding cyclic amplification (PMCA) assay and bioassay, which were shown to correlate well. The inactivation kinetics were characterized by both Chick-Watson (CW) and efficiency factor Hom (EFH) models. It was found that the EFH model fit the experimental data more appropriately. The efficacy of ozone inactivation of PrPSc was both pH and temperature dependent. Based on the EFH model, CT (disinfectant concentration multiplied by contact time) values were determined for 2-log10, 3-log10, and 4-log10 inactivation at the conditions under which they were achieved. Our results indicated that ozone is effective for prion inactivation in ozone-demand-free water and may be applied for the inactivation of infectious prion in prion-contaminated water and wastewater. PMID:23416994

  1. Small-scale spatial heterogeneity of ecosystem properties, microbial community composition and microbial activities in a temperate mountain forest soil.

    PubMed

    Štursová, Martina; Bárta, Jiří; Šantrůčková, Hana; Baldrian, Petr

    2016-12-01

    Forests are recognised as spatially heterogeneous ecosystems. However, knowledge of the small-scale spatial variation in microbial abundance, community composition and activity is limited. Here, we aimed to describe the heterogeneity of environmental properties, namely vegetation, soil chemical composition, fungal and bacterial abundance and community composition, and enzymatic activity, in the topsoil in a small area (36 m 2 ) of a highly heterogeneous regenerating temperate natural forest, and to explore the relationships among these variables. The results demonstrated a high level of spatial heterogeneity in all properties and revealed differences between litter and soil. Fungal communities had substantially higher beta-diversity than bacterial communities, which were more uniform and less spatially autocorrelated. In litter, fungal communities were affected by vegetation and appeared to be more involved in decomposition. In the soil, chemical composition affected both microbial abundance and the rates of decomposition, whereas the effect of vegetation was small. Importantly, decomposition appeared to be concentrated in hotspots with increased activity of multiple enzymes. Overall, forest topsoil should be considered a spatially heterogeneous environment in which the mean estimates of ecosystem-level processes and microbial community composition may confound the existence of highly specific microenvironments. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Control of aerosol contaminants in indoor air: combining the particle concentration reduction with microbial inactivation.

    PubMed

    Grinshpun, Sergey A; Adhikari, Atin; Honda, Takeshi; Kim, Ki Youn; Toivola, Mika; Rao, K S Ramchander; Reponen, Tiina

    2007-01-15

    An indoor air purification technique, which combines unipolar ion emission and photocatalytic oxidation (promoted by a specially designed RCI cell), was investigated in two test chambers, 2.75 m3 and 24.3 m3, using nonbiological and biological challenge aerosols. The reduction in particle concentration was measured size selectively in real-time, and the Air Cleaning Factor and the Clean Air Delivery Rate (CADR) were determined. While testing with virions and bacteria, bioaerosol samples were collected and analyzed, and the microorganism survival rate was determined as a function of exposure time. We observed that the aerosol concentration decreased approximately 10 to approximately 100 times more rapidly when the purifier operated as compared to the natural decay. The data suggest that the tested portable unit operating in approximately 25 m3 non-ventilated room is capable to provide CADR-values more than twice as great than the conventional closed-loop HVAC system with a rating 8 filter. The particle removal occurred due to unipolar ion emission, while the inactivation of viable airborne microorganisms was associated with photocatalytic oxidation. Approximately 90% of initially viable MS2 viruses were inactivated resulting from 10 to 60 min exposure to the photocatalytic oxidation. Approximately 75% of viable B. subtilis spores were inactivated in 10 min, and about 90% or greater after 30 min. The biological and chemical mechanisms that led to the inactivation of stress-resistant airborne viruses and bacterial spores were reviewed.

  3. Inactivation of Salmonella in tomato stem scars by organic acid wash and chitosan-allyl isothiocyanate coating

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to evaluate inactivation of inoculated Salmonella enterica on tomato stem scars exploiting integrated treatment of organic acid wash (AW) followed by chitosan-allyl isothiocyanate (CT-AIT) coating. The treatment effect on microbial loads and fruit quality during 21 d...

  4. Weed seed inactivation in soil mesocosms via biosolarization with mature compost and tomato processing waste amendments.

    PubMed

    Achmon, Yigal; Fernández-Bayo, Jesús D; Hernandez, Katie; McCurry, Dlinka G; Harrold, Duff R; Su, Joey; Dahlquist-Willard, Ruth M; Stapleton, James J; VanderGheynst, Jean S; Simmons, Christopher W

    2017-05-01

    Biosolarization is a fumigation alternative that combines passive solar heating with amendment-driven soil microbial activity to temporarily create antagonistic soil conditions, such as elevated temperature and acidity, that can inactivate weed seeds and other pest propagules. The aim of this study was to use a mesocosm-based field trial to assess soil heating, pH, volatile fatty acid accumulation and weed seed inactivation during biosolarization. Biosolarization for 8 days using 2% mature green waste compost and 2 or 5% tomato processing residues in the soil resulted in accumulation of volatile fatty acids in the soil, particularly acetic acid, and >95% inactivation of Brassica nigra and Solanum nigrum seeds. Inactivation kinetics data showed that near complete weed seed inactivation in soil was achieved within the first 5 days of biosolarization. This was significantly greater than the inactivation achieved in control soils that were solar heated without amendment or were amended but not solar heated. The composition and concentration of organic matter amendments in soil significantly affected volatile fatty acid accumulation at various soil depths during biosolarization. Combining solar heating with organic matter amendment resulted in accelerated weed seed inactivation compared with either approach alone. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  5. Microbial and physical properties as indicators of sandy soil quality under cropland and grassland

    NASA Astrophysics Data System (ADS)

    Frac, Magdalena; Lipiec, Jerzy; Usowicz, Boguslaw; Oszust, Karolina; Brzezinska, Malgorzata

    2017-04-01

    Land use is one of the key factor driving changes in soil properties influencing on soil health and quality. Microbial diversity and physical properties are sensitive indicators for assessing soil health and quality. The alterations of microbial diversity and physical properties following land use changes have not been sufficiently elucidated, especially for sandy soils. We investigated microbial diversity indicators including fungal communities composition and physical properties of sandy acid soil under cropland and more than 20-yr-old grassland (after cropland) in Trzebieszów, Podlasie Region, Poland (N 51° 59' 24", E 22° 33' 37"). The study included four depths within 0-60 cm. Microbial genetic diversity was assessed by terminal restriction fragment length polymorphism (t-RFLP) analysis, fungal community composition was evaluated by next generation sequencing (NGS) analysis and functional diversity was determined by Biolog EcoPlate method. Overall microbial activity was assessed by soil enzymes (dehydrogenases, β-glucosidase) and respiration test. At the same places soil texture, organic carbon content, pH, bulk density, water holding capacity were determined. Our results showed that grassland soil was characterized by higher activity of soil enzymes than cropland. The average well color development of soil microorganisms, the microbial functional diversity and the number of carbon source utilization were significantly affected by land use type and were differentiated among soil depths. In grassland compared to cropland soil a significant increase of carboxylic acids and decrease of amino acids utilization was observed. The quantitative and qualitative differences were found in community of ammonia oxidizing archaea in cropland and grassland soil. The results of fungal community composition help to explain the soil health of grassland and cropland based on the appearance of phytopathogenic and antagonistic fungi. In general bulk density and field water

  6. Inactivation properties of voltage-gated K+ channels altered by presence of beta-subunit.

    PubMed

    Rettig, J; Heinemann, S H; Wunder, F; Lorra, C; Parcej, D N; Dolly, J O; Pongs, O

    1994-05-26

    Structural and functional diversity of voltage-gated Kv1-type potassium channels in rat brain is enhanced by the association of two different types of subunits, the membrane-bound, poreforming alpha-subunits and a peripheral beta-subunit. We have cloned a beta-subunit (Kv beta 1) that is specifically expressed in the rat nervous system. Association of Kv beta 1 with alpha-subunits confers rapid A-type inactivation on non-inactivating Kv1 channels (delayed rectifiers) in expression systems in vitro. This effect is mediated by an inactivating ball domain in the Kv beta 1 amino terminus.

  7. Identifying possible non-thermal effects of radio frequency energy on inactivating food microorganisms.

    PubMed

    Kou, Xiaoxi; Li, Rui; Hou, Lixia; Zhang, Lihui; Wang, Shaojin

    2018-03-23

    Radio frequency (RF) heating has been successfully used for inactivating microorganisms in agricultural and food products. Athermal (non-thermal) effects of RF energy on microorganisms have been frequently proposed in the literature, resulting in difficulties for developing effective thermal treatment protocols. The purpose of this study was to identify if the athermal inactivation of microorganisms existed during RF treatments. Escherichia coli and Staphylococcus aureus in apple juice and mashed potato were exposed to both RF and conventional thermal energies to compare their inactivation populations. A thermal death time (TDT) heating block system was used as conventional thermal energy source to simulate the same heating treatment conditions, involving heating temperature, heating rate and uniformity, of a RF treatment at a frequency of 27.12 MHz. Results showed that a similar and uniform temperature distribution in tested samples was achieved in both heating systems, so that the central sample temperature could be used as representative one for evaluating thermal inactivation of microorganisms. The survival patterns of two target microorganisms in two food samples were similar both for RF and heating block treatments since their absolute difference of survival populations was <1 log CFU/ml. The statistical analysis indicated no significant difference (P > 0.05) in inactivating bacteria between the RF and the heating block treatments at each set of temperatures. The solid temperature and microbial inactivation data demonstrated that only thermal effect of RF energy at 27.12 MHz was observed on inactivating microorganisms in foods. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Microbial inactivation of paprika by a high-temperature short-X time treatment. Influence on color properties.

    PubMed

    Almela, Luis; Nieto-Sandoval, José M; Fernández López, José A

    2002-03-13

    High-temperature short-time (HTST) treatments have been used to destroy the bioburden of paprika. With this in mind, we have designed a device to treat samples of paprika with a gas whose temperature, pressure, and composition can be selected. Temperatures and treatment times ranged from 130 to 170 degrees C and 4 to 6 s, respectively. The survival of the most commonly found microorganisms in paprika and any alteration in extractable and superficial color were examined. Data showed that the optimum HTST conditions were 145 degrees C, 1.5 kg/cm2 of overpressure, 6 s operation time, and a thermal fluid of saturated steam. No microbial growth was detected during storage after thermal treatment. To minimize the color losses, treated (HTST) paprika samples should be kept under refrigeration.

  9. Inactivation of Caliciviruses

    PubMed Central

    Nims, Raymond; Plavsic, Mark

    2013-01-01

    The Caliciviridae family of viruses contains clinically important human and animal pathogens, as well as vesivirus 2117, a known contaminant of biopharmaceutical manufacturing processes employing Chinese hamster cells. An extensive literature exists for inactivation of various animal caliciviruses, especially feline calicivirus and murine norovirus. The caliciviruses are susceptible to wet heat inactivation at temperatures in excess of 60 °C with contact times of 30 min or greater, to UV-C inactivation at fluence ≥30 mJ/cm2, to high pressure processing >200 MPa for >5 min at 4 °C, and to certain photodynamic inactivation approaches. The enteric caliciviruses (e.g.; noroviruses) display resistance to inactivation by low pH, while the non-enteric species (e.g.; feline calicivirus) are much more susceptible. The caliciviruses are inactivated by a variety of chemicals, including alcohols, oxidizing agents, aldehydes, and β-propiolactone. As with inactivation of viruses in general, inactivation of caliciviruses by the various approaches may be matrix-, temperature-, and/or contact time-dependent. The susceptibilities of the caliciviruses to the various physical and chemical inactivation approaches are generally similar to those displayed by other small, non-enveloped viruses, with the exception that the parvoviruses and circoviruses may require higher temperatures for inactivation, while these families appear to be more susceptible to UV-C inactivation than are the caliciviruses. PMID:24276023

  10. Synthetic microbial ecology and the dynamic interplay between microbial genotypes.

    PubMed

    Dolinšek, Jan; Goldschmidt, Felix; Johnson, David R

    2016-11-01

    Assemblages of microbial genotypes growing together can display surprisingly complex and unexpected dynamics and result in community-level functions and behaviors that are not readily expected from analyzing each genotype in isolation. This complexity has, at least in part, inspired a discipline of synthetic microbial ecology. Synthetic microbial ecology focuses on designing, building and analyzing the dynamic behavior of ‘ecological circuits’ (i.e. a set of interacting microbial genotypes) and understanding how community-level properties emerge as a consequence of those interactions. In this review, we discuss typical objectives of synthetic microbial ecology and the main advantages and rationales of using synthetic microbial assemblages. We then summarize recent findings of current synthetic microbial ecology investigations. In particular, we focus on the causes and consequences of the interplay between different microbial genotypes and illustrate how simple interactions can create complex dynamics and promote unexpected community-level properties. We finally propose that distinguishing between active and passive interactions and accounting for the pervasiveness of competition can improve existing frameworks for designing and predicting the dynamics of microbial assemblages.

  11. Inactivation and pharmacological properties of sqKv1A homotetramers in Xenopus oocytes cannot account for behavior of the squid "delayed rectifier" K(+) conductance.

    PubMed Central

    Jerng, Henry H; Gilly, William F

    2002-01-01

    Considerable published evidence suggests that alpha-subunits of the cloned channel sqKv1A compose the "delayed rectifier" in the squid giant axon system, but discrepancies regarding inactivation properties of cloned versus native channels exist. In this paper we define the mechanism of inactivation for sqKv1A channels in Xenopus oocytes to investigate these and other discrepancies. Inactivation of sqKv1A in Xenopus oocytes was found to be unaffected by genetic truncation of the N-terminus, but highly sensitive to certain amino acid substitutions around the external mouth of the pore. External TEA and K(+) ions slowed inactivation of sqKv1A channels in oocytes, and chloramine T (Chl-T) accelerated inactivation. These features are all consistent with a C-type inactivation mechanism as defined for Shaker B channels. Treatment of native channels in giant fiber lobe neurons with TEA or high K(+) does not slow inactivation, nor does Chl-T accelerate it. Pharmacological differences between the two channel types were also found for 4-aminopyridine (4AP). SqKv1A's affinity for 4AP was poor at rest and increased after activation, whereas 4AP block occurred much more readily at rest with native channels than when they were activated. These results suggest that important structural differences between sqKv1A homotetramers and native squid channels are likely to exist around the external and internal mouths of the pore. PMID:12023225

  12. Dual effectiveness of sodium chlorite for enzymatic browning inhibition and Escherichia coli inactivation on fresh-cut apples

    USDA-ARS?s Scientific Manuscript database

    This study investigated the dual effectiveness of sodium chlorite (SC) for browning inhibition and microbial inactivation on fresh-cut apples. The SC treatment exhibited a strong inhibition on browning reaction of fresh-cut Red Delicious apples during cold storage. Test results from examination of t...

  13. Inactivation of Bacteria in Oil Field Injected Water by a Pulsed Plasma Discharge Process

    NASA Astrophysics Data System (ADS)

    Xin, Qing; Li, Zhongjian; Lei, Lecheng; Yang, Bin

    2016-09-01

    Pulsed plasma discharge was employed to inactivate bacteria in the injection water for an oil field. The effects of water conductivity and initial concentration of bacteria on elimination efficiency were investigated in the batch and continuous flow modes. It was demonstrated that Fe2+ contained in injection water could enhance the elimination efficiency greatly. The addition of reducing agent glutathione (GSH) indicated that active radicals generated by pulsed plasma discharges played an important role in the inactivation of bacteria. Moreover, it was found that the microbial inactivation process for both batch and continuous flow mode well fitted the model based on the Weibull's survival function. supported by Zhejiang Province Welfare Technology Applied Research Project of China (No. 2014C31137), National Natural Science Foundation of China (Nos. 21436007 and U1462201), and the Fundamental Research Funds for the Central Universities of China (No. 2015QNA4032)

  14. Effect of initial microbial density on inactivation of Giardia muris by ozone.

    PubMed

    Haas, Charles N; Kaymak, Baris

    2003-07-01

    Inactivation of microorganisms by disinfectants frequently shows non-linear behavior on a semilogarithmic plot of log survival ratio versus time. A number of models have been developed to depict these deviations from Chick's Law. Some of the models predict that the log survival ratio (at a particular disinfectant dose and contact time, even in absence of demand) would be a function of the initial concentration of microorganisms (N(0)), while other models do not predict such an effect. The effect of N(0) on the survival ratio has not been deliberately tested. This work examined the inactivation of Giardia muris by ozone in batch systems, deliberately varying the disinfectant dose and N(0). It was found that the models predicting a dependency of survival on N(0) gave a better description to the data than models that did not predict such a dependency. Hence there is an apparent decrease in disinfection efficiency of ozone against Giardia muris (at pH 8 and 15 degrees C) as the initial microorganism concentration decreases. This phenomena should be taken into account by both disinfection researchers and by process design engineers.

  15. Infective and inactivated filamentous phage as carriers for immunogenic peptides.

    PubMed

    Samoylova, Tatiana I; Norris, Mandy D; Samoylov, Alexandre M; Cochran, Anna M; Wolfe, Karen G; Petrenko, Valery A; Cox, Nancy R

    2012-07-01

    The focus of this study is on development of vaccines using filamentous phage as a delivery vector for immunogenic peptides. The use of phage as a carrier for immunogenic peptides provides significant benefits such as high immunogenicity, low production costs, and high stability of phage preparations. However, introduction of live recombinant phage into the environment might represent a potential ecological problem. This, for example, may occur when vaccines are used in oral or nasal formulations in field conditions for wild and feral animals. To address this issue, comparative studies of antigenic properties of live and inactivated (non-viable) phage were accomplished. Inactivated phage, if released, will not propagate and will degrade as any other protein. In these experiments, a model phage clone that was previously selected from a phage display library and shown to stimulate production of anti-sperm antibodies with contraceptive properties was used. Multiple methods of phage inactivation were tested, including drying, freezing, autoclaving, heating, and UV irradiation. Under studied conditions, heating at 76°C for 3h, UV irradiation, and autoclaving resulted in complete phage inactivation. Phage samples treated by heat and UV were characterized by spectrophotometry and electron microscopy. To test antigenicity, live and inactivated phage preparations were injected into mice and antibody responses assayed by ELISA. It was found that phage killed by heat causes little to no immune responses, probably due to destruction of phage particles. In contrast, UV-inactivated phage stimulated production of IgG serum antibodies at the levels comparable to live phage. Thus, vaccines formulated to include UV-inactivated filamentous phage might represent environmentally safe alternatives to live phage vaccines. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. Effects of ultraviolet light emitting diodes (LEDs) on microbial and enzyme inactivation of apple juice.

    PubMed

    Akgün, Merve Pelvan; Ünlütürk, Sevcan

    2017-11-02

    In this study, the effects of Ultraviolet light-emitting diodes (UV-LEDs) on the inactivation of E. coli K12 (ATCC 25253), an indicator organism of E. coli O157:H7, and polyphneoloxidase (PPO) in cloudy apple juice (CAJ) were investigated. The clear (AJ) and cloudy apple juice were exposed to UV rays for 40min by using a UV device composed of four UV-LEDs with peak emissions at 254 and 280nm and coupled emissions as follows: 254/365, 254/405, 280/365, 280/405 and 254/280/365/405nm. UV-LEDs at 254nm achieved 1.6±0.1 log 10 CFU/mL inactivation of E. coli K12 at UV dose of 707.2mJ/cm 2 . The highest inactivation of E. coli K12 (2.0±0.1log 10 CFU/mL and 2.0±0.4log 10 CFU/mL) was achieved when the cloudy apple juice was treated with both 280nm and 280/365nm UV-LEDs. For clear apple juice the highest inactivation 4.4log 10 CFU/mL obtained for E. coli K12 was achieved using 4 lamps emitting light at 280nm for 40min exposure time. For the same treatment time, the experiments using a combination of lamps emitting light at 280 and 365nm (2lamp/2lamp) were resulted in 3.9±0.2log 10 CFU/mL reductions. UV-A and UV-C rays in combination showed a better inactivation effect on PPO than UV-C rays used separately. Residual activity of PPO in CAJ was reduced to 32.58% when treated with UV-LED in combination of UV-C (280nm) and UV-A (365nm) rays. Additionally, the total color change (ΔE) of CAJ subjected to combined UV-LED irradiation at 280/365nm was the lowest compared to other studied processing conditions. This study provides key implications for the future application of UV-LEDs to fruit juice pasteurization. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Enhancement of microbial quality and inactivation of pathogenic bacteria by gamma irradiation of ready-to-cook Iranian barbecued chicken

    NASA Astrophysics Data System (ADS)

    Fallah, Aziz A.; Siavash Saei-Dehkordi, S.; Rahnama, Mohammad

    2010-10-01

    Ready-to-cook Iranian barbecued chicken consists of cubed chicken breast, lemon juice, salt, red pepper, onion, saffron and vegetable oil with an overall pH value of about 5.5. This product is sometimes consumed under-cooked, hence it may pose health hazards to consumers when contaminated with food-borne pathogens. In this study, the effect of gamma irradiation (0, 1.5, 3 and 4.5 kGy) on the microbial quality of ready-to-cook (RTC) barbecued chicken samples stored at 4 °C for 15 days was investigated. Moreover, the effectiveness of irradiation for inactivating Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella typhimurium inoculated into the samples was also studied. Irradiation of the samples resulted in dose dependent reduction in counts of aerobic mesophilic bacteria, yeasts and molds, Enterobacteriaceae and lactic acid bacteria. Among the microbial flora, yeasts and molds and Enterobacteriaceae were more sensitive to irradiation and got completely eliminated at dose of 3 kGy. D10 values of L. monocytogenes, E. coli O157:H7 and S. typhimurium inoculated into the samples were 0.680, 0.397 and 0.601 kGy, respectively. An irradiation dose of 3 kGy reduced the counts of E. coli O157:H7 to an undetectable level in RTC barbecued chicken but was ineffective on elimination of L. monocytogenes and S. typhimurium. However, none of the food-borne pathogens were detected in the samples irradiated at 4.5 kGy. This study showed that irradiation had no undesirable effects on the initial sensory attributes of barbecued chicken. At the end of the storage period, irradiated samples were more acceptable compared to non-irradiated ones.

  18. Antimicrobial photodynamic inactivation: a bright new technique to kill resistant microbes

    PubMed Central

    Hamblin, Michael R

    2016-01-01

    Photodynamic therapy (PDT) uses photosensitizers (non-toxic dyes) that are activated by absorption of visible light to form reactive oxygen species (including singlet oxygen) that can oxidize biomolecules and destroy cells. Antimicrobial photodynamic inactivation (aPDI) can treat localized infections. aPDI neither causes any resistance to develop in microbes, nor is affected by existing drug resistance status. We discuss some recent developments in aPDI. New photosensitizers including polycationic conjugates, stable synthetic bacteriochlorins and functionalized fullerenes are described. The microbial killing by aPDI can be synergistically potentiated (several logs) by harmless inorganic salts via photochemistry. Genetically engineered bioluminescent microbial cells allow PDT to treat infections in animal models. Photoantimicrobials have a promising future in the face of the unrelenting increase in antibiotic resistance. PMID:27421070

  19. Inactivation of bacterial biofilms using visible-light-activated unmodified ZnO nanorods

    NASA Astrophysics Data System (ADS)

    Aponiene, Kristina; Serevičius, Tomas; Luksiene, Zivile; Juršėnas, Saulius

    2017-09-01

    Various zinc oxide (ZnO) nanostructures are widely used for photocatalytic antibacterial applications. Since ZnO possesses a wide bandgap, it is believed that only UV light may efficiently assist bacterial inactivation, and diverse crystal lattice modifications should be applied in order to narrow the bandgap for efficient visible-light absorption. In this work we show that even unmodified ZnO nanorods grown by an aqueous chemical growth technique are found to possess intrinsic defects that can be activated by visible light (λ = 405 nm) and successfully applied for total inactivation of various highly resistant bacterial biofilms rather than more sensitive planktonic bacteria. Time-resolved fluorescence analysis has revealed that visible-light excitation creates long-lived charge carriers (τ > 1 μs), which might be crucial for destructive biochemical reactions achieving significant bacterial biofilm inactivation. ZnO nanorods covered with bacterial biofilms of Enterococcus faecalis MSCL 302 after illumination by visible light (λ = 405 nm) were inactivated by 2 log, and Listeria monocytogenes ATCL3C 7644 and Escherichia coli O157:H7 biofilms by 4 log. Heterogenic waste-water microbial biofilms, consisting of a mixed population of mesophilic bacteria after illumination with visible light were also completely destroyed.

  20. Microbial Diversity and Mineralogical-Mechanical Properties of Calcitic Cave Speleothems in Natural and in Vitro Biomineralization Conditions

    PubMed Central

    Dhami, Navdeep K.; Mukherjee, Abhijit; Watkin, Elizabeth L. J.

    2018-01-01

    Natural mineral formations are a window into important processes leading to carbon storage and mineralized carbonate structures formed through abiotic and biotic processes. In the current study, we made an attempt to undertake a comprehensive approach to characterize the mineralogical, mechanical, and microbial properties of different kinds of speleothems from karstic caves; with an aim to understand the bio-geo-chemical processes in speleothem structures and their impact on nanomechanical properties. We also investigated the biomineralization abilities of speleothem surface associated microbial communities in vitro. Mineralogical profiling using techniques such as X-ray powder Diffraction (XRD) and Tescan Integrated Mineral Analyzer (TIMA) demonstrated that calcite was the dominant mineral in the majority of speleothems with Energy Dispersive X-ray Analysis (EDS) indicating a few variations in the elemental components. Differing proportions of polymorphs of calcium carbonate such as aragonite and vaterite were also recorded. Significant variations in trace metal content were recorded through Inductively Coupled Plasma Mass Spectrometer (ICP-MS). Scanning Electron Microscopy (SEM) analysis revealed differences in morphological features of the crystals which varied from triangular prismatic shapes to etched spiky forms. Microbial imprints and associations were seen in a few sections. Analysis of the associated microbial diversity showed significant differences between various speleothems at Phylum level; although Proteobacteria and Actinobacteria were found to be the predominant groups. Genus level microbial associations showed a relationship with the geochemistry, mineralogical composition, and metal content of the speleothems. The assessment of nanomechanical properties measured by Nanoindentation revealed that the speleothems with a dominance of calcite were stronger than the speleothems with mixed calcium carbonate polymorphs and silica content. The in vitro

  1. Kv4 channels underlie A-currents with highly variable inactivation time courses but homogeneous other gating properties in the nucleus tractus solitarii.

    PubMed

    Strube, Caroline; Saliba, Layal; Moubarak, Estelle; Penalba, Virginie; Martin-Eauclaire, Marie-France; Tell, Fabien; Clerc, Nadine

    2015-04-01

    In the nucleus of the tractus solitarii (NTS), a large proportion of neurones express transient A-type potassium currents (I KA) having deep influence on the fidelity of the synaptic transmission of the visceral primary afferent inputs to second-order neurones. Up to now, the strong impact of I KA within the NTS was considered to result exclusively from its variation in amplitude, and its molecular correlate(s) remained unknown. In order to identify which Kv channels underlie I KA in NTS neurones, the gating properties and the pharmacology of this current were determined using whole cell patch clamp recordings in slices. Complementary information was brought by immunohistochemistry. Strikingly, two neurone subpopulations characterized by fast or slow inactivation time courses (respectively about 50 and 200 ms) were discriminated. Both characteristics matched those of the Kv4 channel subfamily. The other gating properties, also matching the Kv4 channel ones, were homogeneous through the NTS. The activation and inactivation occurred at membrane potentials around the threshold for generating action potentials, and the time course of recovery from inactivation was rapid. Pharmacologically, I KA in NTS neurones was found to be resistant to tetraethylammonium (TEA), sea anemone toxin blood-depressing substance (BDS) and dendrotoxin (DTX), whereas Androctonus mauretanicus mauretanicus toxin 3 (AmmTX3), a scorpion toxin of the α-KTX 15 family that has been shown to block all the members of the Kv4 family, inhibited 80 % of I KA irrespectively of its inactivation time course. Finally, immunohistochemistry data suggested that, among the Kv4 channel subfamily, Kv4.3 is the prevalent subunit expressed in the NTS.

  2. Pathogens Inactivated by Low-Energy-Electron Irradiation Maintain Antigenic Properties and Induce Protective Immune Responses

    PubMed Central

    Fertey, Jasmin; Bayer, Lea; Grunwald, Thomas; Pohl, Alexandra; Beckmann, Jana; Gotzmann, Gaby; Casado, Javier Portillo; Schönfelder, Jessy; Rögner, Frank-Holm; Wetzel, Christiane; Thoma, Martin; Bailer, Susanne M.; Hiller, Ekkehard; Rupp, Steffen; Ulbert, Sebastian

    2016-01-01

    Inactivated vaccines are commonly produced by incubating pathogens with chemicals such as formaldehyde or β-propiolactone. This is a time-consuming process, the inactivation efficiency displays high variability and extensive downstream procedures are often required. Moreover, application of chemicals alters the antigenic components of the viruses or bacteria, resulting in reduced antibody specificity and therefore stimulation of a less effective immune response. An alternative method for inactivation of pathogens is ionizing radiation. It acts very fast and predominantly damages nucleic acids, conserving most of the antigenic structures. However, currently used irradiation technologies (mostly gamma-rays and high energy electrons) require large and complex shielding constructions to protect the environment from radioactivity or X-rays generated during the process. This excludes them from direct integration into biological production facilities. Here, low-energy electron irradiation (LEEI) is presented as an alternative inactivation method for pathogens in liquid solutions. LEEI can be used in normal laboratories, including good manufacturing practice (GMP)- or high biosafety level (BSL)-environments, as only minor shielding is necessary. We show that LEEI efficiently inactivates different viruses (influenza A (H3N8), porcine reproductive and respiratory syndrome virus (PRRSV), equine herpesvirus 1 (EHV-1)) and bacteria (Escherichia coli) and maintains their antigenicity. Moreover, LEEI-inactivated influenza A viruses elicit protective immune responses in animals, as analyzed by virus neutralization assays and viral load determination upon challenge. These results have implications for novel ways of developing and manufacturing inactivated vaccines with improved efficacy. PMID:27886076

  3. Mechanism of acetaldehyde-induced deactivation of microbial lipases

    PubMed Central

    2011-01-01

    Background Microbial lipases represent the most important class of biocatalysts used for a wealth of applications in organic synthesis. An often applied reaction is the lipase-catalyzed transesterification of vinyl esters and alcohols resulting in the formation of acetaldehyde which is known to deactivate microbial lipases, presumably by structural changes caused by initial Schiff-base formation at solvent accessible lysine residues. Previous studies showed that several lipases were sensitive toward acetaldehyde deactivation whereas others were insensitive; however, a general explanation of the acetaldehyde-induced inactivation mechanism is missing. Results Based on five microbial lipases from Candida rugosa, Rhizopus oryzae, Pseudomonas fluorescens and Bacillus subtilis we demonstrate that the protonation state of lysine ε-amino groups is decisive for their sensitivity toward acetaldehyde. Analysis of the diverse modification products of Bacillus subtilis lipases in the presence of acetaldehyde revealed several stable products such as α,β-unsaturated polyenals, which result from base and/or amino acid catalyzed aldol condensation of acetaldehyde. Our studies indicate that these products induce the formation of stable Michael-adducts at solvent-accessible amino acids and thus lead to enzyme deactivation. Further, our results indicate Schiff-base formation with acetaldehyde to be involved in crosslinking of lipase molecules. Conclusions Differences in stability observed with various commercially available microbial lipases most probably result from different purification procedures carried out by the respective manufacturers. We observed that the pH of the buffer used prior to lyophilization of the enzyme sample is of utmost importance. The mechanism of acetaldehyde-induced deactivation of microbial lipases involves the generation of α,β-unsaturated polyenals from acetaldehyde which subsequently form stable Michael-adducts with the enzymes. Lyophilization of

  4. Arginine inactivates human herpesvirus 2 and inhibits genital herpesvirus infection.

    PubMed

    Ikeda, Keiko; Yamasaki, Hisashi; Minami, Sawako; Suzuki, Yukiko; Tsujimoto, Kazuko; Sekino, Yoshihisa; Irie, Hiroshi; Arakawa, Tsutomu; Koyama, A Hajime

    2012-12-01

    Arginine, among the amino acids, has demonstrated unique properties, including suppression of protein-protein interactions and virus inactivation. We investigated the effects of arginine on the infectivity of human herpesvirus 2 (HHV-2) and the potential application of arginine as a chemotherapeutic agent against genital herpes. Arginine directly inactivated HHV-2 and characterization of the inactivation demonstrated that 1 M arginine at pH 4.3 inactivated the virus more efficiently compared to 0.1 M citrate or 1 M sodium chloride, indicating that neither acidic pH nor ionic strength alone is sufficient for virus inactivation. The effect of arginine was rapid and concentration-dependent. Although virus inactivation was efficient at an acidic pH, arginine inactivated the virus even at a neutral pH, provided that a higher arginine concentration and prolonged incubation time were used. In addition, arginine suppressed the multiplication of HHV-2 under the conditions at which its effect on cell viability was insignificant. Pilot mouse model studies revealed a marked suppression of death by arginine when the mice were infected with HHV-2 through the vaginal route, followed by an intermittent application of acidic arginine by vaginal instillation.

  5. Inactivation of complement by Loxosceles reclusa spider venom.

    PubMed

    Gebel, H M; Finke, J H; Elgert, K D; Cambell, B J; Barrett, J T

    1979-07-01

    Zymosan depletion of serum complement in guinea pigs rendered them highly resistant to lesion by Loxosceles reclusa spider venom. Guinea pigs deficient in C4 of the complement system are as sensitive to the venom as normal guinea pigs. The injection of 35 micrograms of whole recluse venom intradermally into guinea pigs lowered their complement level by 35.7%. Brown recluse spider venom in concentrations as slight as 0.02 micrograms protein/ml can totally inactivate one CH50 of guinea pig complement in vitro. Bee, scorpion, and other spider venoms had no influence on the hemolytic titer of complement. Fractionation of recluse spider venom by Sephadex G-200 filtration separated the complement-inactivating property of the venom into three major regions which could be distinguished on the basis of heat stability as well as size. None was neutralized by antivenom. Polyacrylamide gel electrophoresis of venom resolved the complement inactivators into five fractions. Complement inactivated by whole venom or the Sephadex fractions could be restored to hemolytic activity by supplements of fresh serum but not by heat-inactivated serum, pure C3, pure C5, or C3 and C5 in combination.

  6. Microbial communities in natural rubber coagula during maturation: impacts on technological properties of dry natural rubber.

    PubMed

    Salomez, M; Subileau, M; Vallaeys, T; Santoni, S; Bonfils, F; Sainte-Beuve, J; Intapun, J; Granet, F; Vaysse, L; Dubreucq, É

    2018-02-01

    To characterize microbial communities present in natural rubber (NR) coagula from Hevea brasiliensis latex during maturation and identify microbial taxa (bacteria and fungi) having an impact on dry NR properties. Microbial community dynamics in NR coagula maturated under controlled conditions were compared and related with the evolution of dry NR properties. The pyrosequencing of 16S (119 837 effective reads) and 18S (131 879 effective reads) rRNA gene regions was performed on 21 samples covering different maturation times and two aeration conditions. Results showed a relatively high bacterial richness (Chao1 estimates of 200-1000) associated with significant bacterial dynamics. Lactic acid bacteria (LAB) were dominant in the first days of maturation. Then, in aerobic conditions, development of Actinobacteria represented by the family Microbacteriaceae was associated with alkalinization of the samples and a higher sensitivity of NR to thermo-oxidation as evaluated by its plasticity retention index (PRI). In anaerobiosis, the reduced development of bacteria, mostly LAB present, was associated with improved NR properties (higher initial plasticity P 0 and PRI). The involvement of micro-organisms in the evolution of dry NR properties during the maturation of NR coagula was confirmed. The importance of the structure and dynamics of microbial communities is specifically highlighted. Natural rubber is a key elastomer for the tyre industry and for a variety of other applications. The majority of raw NR is obtained by natural coagulation of H. brasiliensis latex under the activity of micro-organisms. An improved understanding of the microbial communities involved in the maturation of NR coagula may lead to an improvement in the production process of raw NR to provide a better consistency in NR quality. © 2017 The Society for Applied Microbiology.

  7. Light based technologies for microbial inactivation of liquids, bead surfaces and powdered infant formula.

    PubMed

    Arroyo, Cristina; Dorozko, Anna; Gaston, Edurne; O'Sullivan, Michael; Whyte, Paul; Lyng, James G

    2017-10-01

    This study evaluates the potential of continuous wave Ultraviolet C light (UV-C) and broad-spectrum intense pulsed light (in this study referred to as High Intensity Light Pulses, HILP) for the inactivation of pathogens of public concern in powdered infant formula (PIF) producers. To achieve this goal a sequential set of experiments were performed, firstly in clear liquid media, secondly on the surface of spherical beads under agitation and, finally in PIF. L. innocua was the most sensitive microorganism to both technologies under all conditions studied with reductions exceeding 4 log 10 cycles in PIF. In the clear liquid medium, the maximum tolerance to light was observed for C. sakazakii against UV-C light and for B. subtilis spores against HILP, with a fluence of approximately 17 mJ/cm 2 required for a 1 log 10 cycle inactivation (D value) of each species. In PIF it was possible to inactivate >99% of the vegetative cell populations by HILP with a fluence of 199 mJ/cm 2 and of B. subtilis spores by doubling the fluence. By contrast, for UV-C treatments a fluence of 2853 mJ/cm 2 was needed for 99.9% reduction of C. sakazakii, which was the most light-resistant microorganism to UV-C. Results here obtained clearly show the potential for light-based interventions to improve PIF microbiological safety. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Impact of food model (micro)structure on the microbial inactivation efficacy of cold atmospheric plasma.

    PubMed

    Smet, C; Noriega, E; Rosier, F; Walsh, J L; Valdramidis, V P; Van Impe, J F

    2017-01-02

    The large potential of cold atmospheric plasma (CAP) for food decontamination has recently been recognized. Room-temperature gas plasmas can decontaminate foods without causing undesired changes. This innovative technology is a promising alternative for treating fresh produce. However, more fundamental studies are needed before its application in the food industry. The impact of the food structure on CAP decontamination efficacy of Salmonella Typhimurium and Listeria monocytogenes was studied. Cells were grown planktonically or as surface colonies in/on model systems. Both microorganisms were grown in lab culture media in petri dishes at 20°C until cells reached the stationary phase. Before CAP treatment, cells were deposited in a liquid carrier, on a solid(like) surface or on a filter. A dielectric barrier discharge reactor generated helium-oxygen plasma, which was used to treat samples up to 10min. Although L. monocytogenes is more resistant to CAP treatment, similar trends in inactivation behavior as for S. Typhimurium are observed, with log reductions in the range [1.0-2.9] for S. Typhimurium and [0.2-2.2] for L. monocytogenes. For both microorganisms, cells grown planktonically are easily inactivated, as compared to surface colonies. More stressing growth conditions, due to cell immobilization, result in more resistant cells during CAP treatment. The main difference between the inactivation support systems is the absence or presence of a shoulder phase. For experiments in the liquid carrier, which exhibit a long shoulder, the plasma components need to diffuse and penetrate through the medium. This explains the higher efficacies of CAP treatment on cells deposited on a solid(like) surface or on a filter. This research demonstrates that the food structure influences the cell inactivation behavior and efficacy of CAP, and indicates that food intrinsic factors need to be accounted when designing plasma treatment. Copyright © 2016. Published by Elsevier B.V.

  9. Near-infrared photodynamic inactivation of S. pneumoniae and its interaction with RAW 264.7 macrophages.

    PubMed

    Leite, Ilaiáli S; Geralde, Mariana C; Salina, Ana C G; Medeiros, Alexandra I; Dovigo, Lívia N; Bagnato, Vanderlei S; Inada, Natalia M

    2018-01-01

    Pneumonia is the main cause of children mortality worldwide, and its major treatment obstacle stems from the microorganisms increasing development of resistance to several antibiotics. Photodynamic therapy has been presenting, for the last decades, promising results for some subtypes of cancer and infections. In this work we aimed to develop a safe and efficient in vitro protocol for photodynamic inactivation of Streptococcus pneumoniae, one of the most commonly found bacteria in pneumonia cases, using two near-infrared light sources and indocyanine green, a FDA approved dye. Photodynamic inactivation experiments with bacteria alone allowed to determine the best parameters for microbial inactivation. Cytotoxicity assays with RAW 264.7 macrophages evaluated the safety of the PDI. To determine if the photodynamic inactivation had a positive or negative effect on the natural killing action of macrophages, we selected and tested fewer indocyanine green concentrations and 10 J/cm 2 on macrophage-S. pneumoniae co-cultures. We concluded that ICG has potential as a photosensitizer for near-infrared photodynamic inactivation of S. pneumoniae, producing minimum negative impact on RAW 264.7 macrophages and having a positive interaction with the immune cell's microbicidal action. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Pathogen Inactivating Properties and Increased Sensitivity in Molecular Diagnostics by PAXgene, a Novel Non-Crosslinking Tissue Fixative.

    PubMed

    Loibner, Martina; Buzina, Walter; Viertler, Christian; Groelz, Daniel; Hausleitner, Anja; Siaulyte, Gintare; Kufferath, Iris; Kölli, Bettina; Zatloukal, Kurt

    2016-01-01

    Requirements on tissue fixatives are getting more demanding as molecular analysis becomes increasingly relevant for routine diagnostics. Buffered formaldehyde in pathology laboratories for tissue fixation is known to cause chemical modifications of biomolecules which affect molecular testing. A novel non-crosslinking tissue preservation technology, PAXgene Tissue (PAXgene), was developed to preserve the integrity of nucleic acids in a comparable way to cryopreservation and also to preserve morphological features comparable to those of formalin fixed samples. Because of the excellent preservation of biomolecules by PAXgene we investigated its pathogen inactivation ability and biosafety in comparison to formalin by in-vitro testing of bacteria, human relevant fungi and human cytomegalovirus (CMV). Guidelines for testing disinfectants served as reference for inactivation assays. Furthermore, we tested the properties of PAXgene for detection of pathogens by PCR based assays. All microorganisms tested were similarly inactivated by PAXgene and formalin except Clostridium sporogenes, which remained viable in seven out of ten assays after PAXgene treatment and in three out of ten assays after formalin fixation. The findings suggest that similar biosafety measures can be applied for PAXgene and formalin fixed samples. Detection of pathogens in PCR-based diagnostics using two CMV assays resulted in a reduction of four to ten quantification cycles of PAXgene treated samples which is a remarkable increase of sensitivity. PAXgene fixation might be superior to formalin fixation when molecular diagnostics and highly sensitive detection of pathogens is required in parallel to morphology assessment.

  11. Nondeterministic computational fluid dynamics modeling of Escherichia coli inactivation by peracetic acid in municipal wastewater contact tanks.

    PubMed

    Santoro, Domenico; Crapulli, Ferdinando; Raisee, Mehrdad; Raspa, Giuseppe; Haas, Charles N

    2015-06-16

    Wastewater disinfection processes are typically designed according to heuristics derived from batch experiments in which the interaction among wastewater quality, reactor hydraulics, and inactivation kinetics is often neglected. In this paper, a computational fluid dynamics (CFD) study was conducted in a nondeterministic (ND) modeling framework to predict the Escherichia coli inactivation by peracetic acid (PAA) in municipal contact tanks fed by secondary settled wastewater effluent. The extent and variability associated with the observed inactivation kinetics were both satisfactorily predicted by the stochastic inactivation model at a 95% confidence level. Moreover, it was found that (a) the process variability induced by reactor hydraulics is negligible when compared to the one caused by inactivation kinetics, (b) the PAA dose required for meeting regulations is dictated equally by the fixed limit of the microbial concentration as well as its probability of occurrence, and (c) neglecting the probability of occurrence during process sizing could lead to an underestimation of the PAA dose required by as much as 100%. Finally, the ND-CFD model was used to generate sizing information in the form of probabilistic disinfection curves relating E. coli inactivation and probability of occurrence with the average PAA dose and PAA residual concentration at the outlet of the contact tank.

  12. Inactivation of pathogenic bacteria inoculated onto a Bacto™ agar model surface using TiO2-UVC photocatalysis, UVC and chlorine treatments.

    PubMed

    Yoo, S; Ghafoor, K; Kim, S; Sun, Y W; Kim, J U; Yang, K; Lee, D-U; Shahbaz, H M; Park, J

    2015-09-01

    The aim of this study was to study inactivation of different pathogenic bacteria on agar model surface using TiO2-UV photocatalysis (TUVP). A unified food surface model was simulated using Bacto(™) agar, a routinely used microbial medium. The foodborne pathogenic bacteria Escherichia coli K12 (as a surrogate for E. coli O157:H7), Salmonella Typhimurium, Staphylococcus aureus and Listeria monocytogenes were inoculated onto the agar surface, followed by investigation of TUVP-assisted inactivation and morphological changes in bacterial cells. The TUVP process showed higher bacterial inactivation, particularly for Gram-negative bacteria, than UVC alone and a control (dark reaction). A TUVP treatment of 17·2 mW cm(-2) (30% lower than the UVC light intensity) reduced the microbial load on the agar surface by 4·5-6·0 log CFU cm(-2). UVC treatment of 23·7 mW cm(-2) caused 3·0-5·3 log CFU cm(-2) reduction. The use of agar model surface is effective for investigation of bacterial disinfection and TUVP is a promising nonthermal technique. The results showing effects of photocatalysis and other treatments for inactivation of bacterial pathogens on model surface can be useful for applying such processes for disinfection of fruit, vegetables and other similar surfaces. © 2015 The Society for Applied Microbiology.

  13. High pressure inactivation of Brettanomyces bruxellensis in red wine.

    PubMed

    van Wyk, Sanelle; Silva, Filipa V M

    2017-05-01

    Brettanomyces bruxellensis ("Brett") is a major spoilage concern for the wine industry worldwide, leading to undesirable sensory properties. Sulphur dioxide, is currently the preferred method for wine preservation. However, due to its negative effects on consumers, the use of new alternative non-thermal technologies are increasingly being investigated. The aim of this study was to determine and model the effect of high pressure processing (HPP) conditions and yeast strain on the inactivation of "Brett" in Cabernet Sauvignon wine. Processing at 200 MPa for 3 min resulted in 5.8 log reductions. However higher pressure is recommended to achieve high throughput in the wine industry, for example >6.0 log reductions were achieved after 400 MPa for 5 s. The inactivation of B. bruxellensis is pressure and time dependent, with increased treatment time and pressure leading to increased yeast inactivation. It was also found that yeast strain had a significant effect on HPP inactivation, with AWRI 1499 being the most resistant strain. The Weibull model successfully described the HPP "Brett" inactivation. HPP is a viable alternative for the inactivation of B. bruxellensis in wine, with the potential to reduce the industry's reliance on sulphur dioxide. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Removal of Microbial Contamination from Surface by Plasma

    NASA Astrophysics Data System (ADS)

    Feng, Xinxin; Liu, Hongxia; Shen, Zhenxing; Wang, Taobo

    2018-01-01

    Microbial contamination is closely associated with human and environmental health, they can be tested on food surfaces, medical devices, packing material and so on. In this paper the removal of the microbial contamination from surface using plasma treatment is investigated. The Escherichia coli (E. coli) has been chosen as a bio-indicator enabling to evaluate the effect of plasma assisted microbial inactivation. Oxygen gas was as the working gas. The plasma RF power, plasma exposition time, gas flow and the concentration of organic pollutant were varied in order to see the effect of the plasma treatment on the Gram-negative germ removal. After the treatment, the microbial abatement was evaluated by the standard plate count method. This proved a positive effect of the plasma treatment on Gram-negative germ removal. The kinetics and mathematical model of removal were studied after plasma treatment, and then the removing course of E. coli was analyzed. This work is meaningful for deepening our understanding of the fundamental scientific principles regarding microbial contamination from surface by plasma.

  15. Nanoscale Structural and Mechanical Analysis of Bacillus anthracis Spores Inactivated with Rapid Dry Heating

    PubMed Central

    Felker, Daniel L.; Burggraf, Larry W.

    2014-01-01

    Effective killing of Bacillus anthracis spores is of paramount importance to antibioterrorism, food safety, environmental protection, and the medical device industry. Thus, a deeper understanding of the mechanisms of spore resistance and inactivation is highly desired for developing new strategies or improving the known methods for spore destruction. Previous studies have shown that spore inactivation mechanisms differ considerably depending upon the killing agents, such as heat (wet heat, dry heat), UV, ionizing radiation, and chemicals. It is believed that wet heat kills spores by inactivating critical enzymes, while dry heat kills spores by damaging their DNA. Many studies have focused on the biochemical aspects of spore inactivation by dry heat; few have investigated structural damages and changes in spore mechanical properties. In this study, we have inactivated Bacillus anthracis spores with rapid dry heating and performed nanoscale topographical and mechanical analysis of inactivated spores using atomic force microscopy (AFM). Our results revealed significant changes in spore morphology and nanomechanical properties after heat inactivation. In addition, we also found that these changes were different under different heating conditions that produced similar inactivation probabilities (high temperature for short exposure time versus low temperature for long exposure time). We attributed the differences to the differential thermal and mechanical stresses in the spore. The buildup of internal thermal and mechanical stresses may become prominent only in ultrafast, high-temperature heat inactivation when the experimental timescale is too short for heat-generated vapor to efficiently escape from the spore. Our results thus provide direct, visual evidences of the importance of thermal stresses and heat and mass transfer to spore inactivation by very rapid dry heating. PMID:24375142

  16. Inactivation of possible micromycete food contaminants using the low-temperature plasma and hydrogen peroxide

    NASA Astrophysics Data System (ADS)

    Čeřovský, M.; Khun, J.; Rusová, K.; Scholtz, V.; Soušková, H.

    2013-09-01

    The inhibition effect of hydrogen peroxide aerosol, low-temperature plasma and their combinations has been studied on several micromycetes spores. The low-temperature plasma was generated in corona discharges in the open air apparatus with hydrogen peroxide aerosol. Micromycete spores were inoculated on the surface of agar plates, exposed solely to the hydrogen peroxide aerosol, corona discharge or their combination. After incubation the diameter of inhibition zone was measured. The solely positive corona discharge exhibits no inactivation effect, the solely negative corona discharge and solely hydrogen peroxide aerosol exhibit the inactivation effect, however their combinations exhibit to be much more effective. Low-temperature plasma and hydrogen peroxide aerosol present a possible alternative method of microbial decontamination of food, food packages or other thermolabile materials.

  17. Cationic, anionic and neutral dyes: effects of photosensitizing properties and experimental conditions on the photodynamic inactivation of pathogenic bacteria.

    PubMed

    Sabbahi, Sonia; Ben Ayed, Layla; Boudabbous, Abdellatif

    2013-12-01

    The aim of this study was to evaluate the photobactericidal effect of four photosensitizers (PSs) with different structural and physico-photochemical properties, namely mesotetracationic porphyrin (T4MPyP), dianionic rose Bengal (RB), monocationic methylene blue (MB) and neutral red (NR). Their photokilling activity was tested in vitro on pathogenic bacteria such as Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) suspended in nutrient broth (NB) and in phosphate buffered saline (PBS) through following their influence on the PSs antimicrobial efficacy. Photodynamic inactivation (PDI) experiments were performed using visible light (L) and different PSs concentrations (20-70 μM). The ability of these PSs to mediate bacterial photodynamic inactivation was investigated as a function of type of PS and its concentrations, spectral and physico-chemical properties, bacterial strain, irradiation time and suspending medium. Indeed, they showed antibacterial effects against S. aureus and P. aeruginosa with significant difference in potency. Staphylococcus aureus suspended in NB showed 0.92 log units reduction in viable count in the presence of T4MPyP at 20 μM. Changing the suspending medium from NB to PBS, S. aureus was successfully photoinactivated by T4MPyP (20 μM) when suspended in PBS at least time exposure (10 and 30 min), followed by MB and RB.

  18. Inactivation of Nonpathogenic Escherichia coli, Escherichia coli O157:H7, Salmonella enterica Typhimurium, and Listeria monocytogenes in Ice Using a UVC Light-Emitting Diode.

    PubMed

    Murashita, Suguru; Kawamura, Shuso; Koseki, Shigenobu

    2017-07-01

    Ice, widely used in the food industry, is a potential cause of food poisoning resulting from microbial contamination. Direct microbial inactivation of ice is necessary because microorganisms may have been present in the source water used to make it and/or may have been introduced due to poor hygiene during production or handling of the ice. Nonthermal and nondestructive microbial inactivation technologies are needed to control microorganisms in ice. We evaluated the applicability of a UVC light-emitting diode (UVC-LED) for microbial inactivation in ice. The effects of UV intensity and UV dose of the UVC-LED on Escherichia coli ATCC 25922 and a comparison of UVC-LED with a conventional UV lamp for effective bacterial inactivation in distilled water and ice cubes were investigated to evaluate the performance of the UVC-LED. Finally, we assessed the effects of the UVC-LED on pathogens such as E. coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in ice cubes. The results indicated that UVC-LED effectiveness depended on the UV dose at all UV intensity conditions (0.084, 0.025, 0.013, 0.007, and 0.005 mW/cm 2 ) in ice and that UVC-LED could more efficiently inactivate E. coli ATCC 25922 in distilled water and ice than the UV lamp. At a UV dose of 2.64 mJ/cm 2 , E. coli in distilled water was decreased by 0.90 log CFU/mL (UV lamp) and by more than 7.0 log CFU/mL (UVC-LED). At 15.2 mJ/cm 2 , E. coli in ice was decreased by 3.18 log CFU/mL (UV lamp) and by 4.45 CFU/mL (UVC-LED). Furthermore, UVC-LED irradiation reduced the viable number of pathogens by 6 to 7 log cycles at 160 mJ/cm 2 , although the bactericidal effect was somewhat dependent on the type of bacteria. L. monocytogenes in ice was relatively more sensitive to UVC irradiation than were E. coli O157:H7 and Salmonella Typhimurium. These results demonstrate that UVC-LED irradiation could contribute to the safety of ice in the food industry.

  19. Selective control of cortical axonal spikes by a slowly inactivating K+ current

    PubMed Central

    Shu, Yousheng; Yu, Yuguo; Yang, Jing; McCormick, David A.

    2007-01-01

    Neurons are flexible electrophysiological entities in which the distribution and properties of ionic channels control their behaviors. Through simultaneous somatic and axonal whole-cell recording of layer 5 pyramidal cells, we demonstrate a remarkable differential expression of slowly inactivating K+ currents. Depolarizing the axon, but not the soma, rapidly activated a low-threshold, slowly inactivating, outward current that was potently blocked by low doses of 4-aminopyridine, α-dendrotoxin, and rTityustoxin-Kα. Block of this slowly inactivating current caused a large increase in spike duration in the axon but only a small increase in the soma and could result in distal axons generating repetitive discharge in response to local current injection. Importantly, this current was also responsible for slow changes in the axonal spike duration that are observed after somatic membrane potential change. These data indicate that low-threshold, slowly inactivating K+ currents, containing Kv1.2 α subunits, play a key role in the flexible properties of intracortical axons and may contribute significantly to intracortical processing. PMID:17581873

  20. Soil Microbial Properties and Plant Growth Responses to Carbon and Water Addition in a Temperate Steppe: The Importance of Nutrient Availability

    PubMed Central

    Guo, Chengyuan; Wang, Renzhong; Xiao, Chunwang

    2012-01-01

    Background Global climatic change is generally expected to stimulate net primary production, and consequently increase soil carbon (C) input. The enhanced C input together with potentially increased precipitation may affect soil microbial processes and plant growth. Methodology/Principal Findings To examine the effects of C and water additions on soil microbial properties and plant growth, we conducted an experiment lasting two years in a temperate steppe of northeastern China. We found that soil C and water additions significantly affected microbial properties and stimulated plant growth. Carbon addition significantly increased soil microbial biomass and activity but had a limited effect on microbial community structure. Water addition significantly increased soil microbial activity in the first year but the response to water decreased in the second year. The water-induced changes of microbial activity could be ascribed to decreased soil nitrogen (N) availability and to the shift in soil microbial community structure. However, no water effect on soil microbial activity was visible under C addition during the two years, likely because C addition alleviated nutrient limitation of soil microbes. In addition, C and water additions interacted to affect plant functional group composition. Water addition significantly increased the ratio of grass to forb biomass in C addition plots but showed only minor effects under ambient C levels. Our results suggest that soil microbial activity and plant growth are limited by nutrient (C and N) and water availability, and highlight the importance of nutrient availability in modulating the responses of soil microbes and plants to potentially increased precipitation in the temperate steppe. Conclusions/Significance Increased soil C input and precipitation would show significant effects on soil microbial properties and plant growth in the temperate steppe. These findings will improve our understanding of the responses of soil microbes

  1. DPP10 splice variants are localized in distinct neuronal populations and act to differentially regulate the inactivation properties of Kv4-based ion channels.

    PubMed

    Jerng, Henry H; Lauver, Aaron D; Pfaffinger, Paul J

    2007-08-01

    Dipeptidyl peptidase-like proteins (DPLs) and Kv-channel-interacting proteins (KChIPs) join Kv4 pore-forming subunits to form multi-protein complexes that underlie subthreshold A-type currents (I(SA)) in neuronal somatodendritic compartments. Here, we characterize the functional effects and brain distributions of N-terminal variants belonging to the DPL dipeptidyl peptidase 10 (DPP10). In the Kv4.2+KChIP3+DPP10 channel complex, all DPP10 variants accelerate channel gating kinetics; however, the splice variant DPP10a produces uniquely fast inactivation kinetics that accelerates with increasing depolarization. This DPP10a-specific inactivation dominates in co-expression studies with KChIP4a and other DPP10 isoforms. Real-time qRT-PCR and in situ hybridization analyses reveal differential expression of DPP10 variants in rat brain. DPP10a transcripts are prominently expressed in the cortex, whereas DPP10c and DPP10d mRNAs exhibit more diffuse distributions. Our results suggest that DPP10a underlies rapid inactivation of cortical I(SA), and the regulation of isoform expression may contribute to the variable inactivation properties of I(SA) across different brain regions.

  2. Plant diversity does not buffer drought effects on early-stage litter mass loss rates and microbial properties.

    PubMed

    Vogel, Anja; Eisenhauer, Nico; Weigelt, Alexandra; Scherer-Lorenzen, Michael

    2013-09-01

    Human activities are decreasing biodiversity and changing the climate worldwide. Both global change drivers have been shown to affect ecosystem functioning, but they may also act in concert in a non-additive way. We studied early-stage litter mass loss rates and soil microbial properties (basal respiration and microbial biomass) during the summer season in response to plant species richness and summer drought in a large grassland biodiversity experiment, the Jena Experiment, Germany. In line with our expectations, decreasing plant diversity and summer drought decreased litter mass loss rates and soil microbial properties. In contrast to our hypotheses, however, this was only true for mass loss of standard litter (wheat straw) used in all plots, and not for plant community-specific litter mass loss. We found no interactive effects between global change drivers, that is, drought reduced litter mass loss rates and soil microbial properties irrespective of plant diversity. High mass loss rates of plant community-specific litter and low responsiveness to drought relative to the standard litter indicate that soil microbial communities were adapted to decomposing community-specific plant litter material including lower susceptibility to dry conditions during summer months. Moreover, higher microbial enzymatic diversity at high plant diversity may have caused elevated mass loss of standard litter. Our results indicate that plant diversity loss and summer drought independently impede soil processes. However, soil decomposer communities may be highly adapted to decomposing plant community-specific litter material, even in situations of environmental stress. Results of standard litter mass loss moreover suggest that decomposer communities under diverse plant communities are able to cope with a greater variety of plant inputs possibly making them less responsive to biotic changes. © 2013 John Wiley & Sons Ltd.

  3. High hydrostatic pressure inactivation of Lactobacillus plantarum cells in (O/W)-emulsions is independent from cell surface hydrophobicity and lipid phase parameters

    NASA Astrophysics Data System (ADS)

    Kafka, T. A.; Reitermayer, D.; Lenz, C. A.; Vogel, R. F.

    2017-07-01

    Inactivation efficiency of high hydrostatic pressure (HHP) processing of food is strongly affected by food matrix composition. We investigated effects of fat on HHP inactivation of spoilage-associated Lactobacillus (L.) plantarum strains using defined oil-in-water (O/W)-emulsion model systems. Since fat-mediated effects on HHP inactivation could be dependent on interactions between lipid phase and microbial cells, three major factors possibly influencing such interactions were considered, that is, cell surface hydrophobicity, presence and type of surfactants, and oil droplet size. Pressure tolerance varied noticeably among L. plantarum strains and was independent of cell surface hydrophobicity. We showed that HHP inactivation of all strains tended to be more effective in presence of fat. The observation in both, surfactant-stabilized and surfactant-free (O/W)-emulsion, indicates that cell surface hydrophobicity is no intrinsic pressure resistance factor. In contrast to the presence of fat per se, surfactant type and oil droplet size did not affect inactivation efficiency.

  4. Trends in microbial control techniques for poultry products.

    PubMed

    Silva, Filomena; Domingues, Fernanda C; Nerín, Cristina

    2018-03-04

    Fresh poultry meat and poultry products are highly perishable foods and high potential sources of human infection due to the presence of several foodborne pathogens. Focusing on the microbial control of poultry products, the food industry generally implements numerous preventive measures based on the Hazard Analysis and Critical Control Points (HACCP) food safety management system certification together with technological steps, such as refrigeration coupled to modified atmosphere packaging that are able to control identified potential microbial hazards during food processing. However, in recent years, to meet the demand of consumers for minimally processed, high-quality, and additive-free foods, technologies are emerging associated with nonthermal microbial inactivation, such as high hydrostatic pressure, irradiation, and natural alternatives, such as biopreservation or the incorporation of natural preservatives in packaging materials. These technologies are discussed throughout this article, emphasizing their pros and cons regarding the control of poultry microbiota and their effects on poultry sensory properties. The discussion for each of the preservation techniques mentioned will be provided with as much detail as the data and studies provided in the literature for poultry meat and products allow. These new approaches, on their own, have proved to be effective against a wide range of microorganisms in poultry meat. However, since some of these emergent technologies still do not have full consumer's acceptability and, taking into consideration the hurdle technology concept for poultry processing, it is suggested that they will be used as combined treatments or, more frequently, in combination with modified atmosphere packaging.

  5. Microbial UV fluence-response assessment using a novel UV-LED collimated beam system.

    PubMed

    Bowker, Colleen; Sain, Amanda; Shatalov, Max; Ducoste, Joel

    2011-02-01

    A research study has been performed to determine the ultraviolet (UV) fluence-response of several target non-pathogenic microorganisms to UV light emitting diodes (UV-LEDs) by performing collimated beam tests. UV-LEDs do not contain toxic mercury, offer design flexibility due to their small size, and have a longer operational life than mercury lamps. Comsol Multiphysics was utilized to create an optimal UV-LED collimated beam design based on number and spacing of UV-LEDs and distance of the sample from the light source while minimizing the overall cost. The optimized UV-LED collimated beam apparatus and a low-pressure mercury lamp collimated beam apparatus were used to determine the UV fluence-response of three surrogate microorganisms (Escherichia coli, MS-2, T7) to 255 nm UV-LEDs, 275 nm UV-LEDs, and 254 nm low-pressure mercury lamps. Irradiation by low-pressure mercury lamps produced greater E. coli and MS-2 inactivation than 255 nm and 275 nm UV-LEDs and similar T7 inactivation to irradiation by 275 nm UV-LEDs. The 275 nm UV-LEDs produced more efficient T7 and E. coli inactivation than 255 nm UV-LEDs while both 255 nm and 275 nm UV-LEDs produced comparable microbial inactivation for MS-2. Differences may have been caused by a departure from the time-dose reciprocity law due to microbial repair mechanisms. Copyright © 2010 Elsevier Ltd. All rights reserved.

  6. Skewed X-chromosome inactivation in women affected by Alzheimer's disease.

    PubMed

    Bajic, Vladan; Mandusic, Vesna; Stefanova, Elka; Bozovic, Ana; Davidovic, Radoslav; Zivkovic, Lada; Cabarkapa, Andrea; Spremo-Potparevic, Biljana

    2015-01-01

    X-chromosome instability has been a long established feature in Alzheimer's disease (AD). Premature centromere division and aneuploidy of the X-chromosome has been found in peripheral blood lymphocytes and neuronal tissue in female AD patients. Interestingly, only one chromosome of the X pair has been affected. These results raised a question, "Is the X-chromosome inactivation pattern altered in peripheral blood lymphocytes of women affected by AD?" To address this question, we analyzed the methylation status of androgen receptor promoter which may show us any deviation from the 50 : 50% X inactivation status in peripheral blood lymphocytes of women with AD. Our results showed skewed inactivation patterns (>90%). These findings suggest that an epigenetic alteration on the inactivation centers of the X-chromosome (or skewing) relates not only to aging, by might be a novel property that could account for the higher incidence of AD in women.

  7. Flash Inactivation of Oxygen Evolution

    PubMed Central

    Frasch, Wayne D.; Cheniae, George M.

    1980-01-01

    Brief saturating light flashes were used to probe the mechanism of inactivation of O2 evolution by Tris in chloroplasts. Maximum inactivation with a single flash and an oscillation with period of four on subsequent flashes was observed. Analyses of the oscillations suggested that only the charge-collecting O2-evolving catalyst of photosystem II (S2-state) was a target of inactivation by Tris. This conclusion was supported by the following observations: (a) hydroxylamine preequilibration caused a three-flash delay in the inactivation pattern; (b) the lifetimes of the Tris-inactivable and S2-states were similar; and (c) reagents accelerating S2 deactivation decreased the lifetime of the inactivable state. Inactivation proved to be moderated by F, the precursor of Signal IIs, as shown by a one flash delay with chloroplasts having high abundance of F. Evidence was obtained for cooperativity effects in inactivation and NH3 was shown to be a competitive inhibitor of the Tris-induced inactivation. S2-dependent inactivation was inhibited by glutaraldehyde fixation of chloroplasts, possibly suggesting that inactivation proceeds via conformational changes of the S2-state. PMID:16661270

  8. Inactivation of possible micromycete food contaminants using the low-temperature plasma and hydrogen peroxide

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

    Čeřovský, M., E-mail: scholtz@aldebaran.cz; Khun, J.; Rusová, K.

    2013-09-15

    The inhibition effect of hydrogen peroxide aerosol, low-temperature plasma and their combinations has been studied on several micromycetes spores. The low-temperature plasma was generated in corona discharges in the open air apparatus with hydrogen peroxide aerosol. Micromycete spores were inoculated on the surface of agar plates, exposed solely to the hydrogen peroxide aerosol, corona discharge or their combination. After incubation the diameter of inhibition zone was measured. The solely positive corona discharge exhibits no inactivation effect, the solely negative corona discharge and solely hydrogen peroxide aerosol exhibit the inactivation effect, however their combinations exhibit to be much more effective. Low-temperaturemore » plasma and hydrogen peroxide aerosol present a possible alternative method of microbial decontamination of food, food packages or other thermolabile materials.« less

  9. Inactivation gating of Kv7.1 channels does not involve concerted cooperative subunit interactions.

    PubMed

    Meisel, Eshcar; Tobelaim, William; Dvir, Meidan; Haitin, Yoni; Peretz, Asher; Attali, Bernard

    2018-01-01

    Inactivation is an intrinsic property of numerous voltage-gated K + (Kv) channels and can occur by N-type or/and C-type mechanisms. N-type inactivation is a fast, voltage independent process, coupled to activation, with each inactivation particle of a tetrameric channel acting independently. In N-type inactivation, a single inactivation particle is necessary and sufficient to occlude the pore. C-type inactivation is a slower process, involving the outermost region of the pore and is mediated by a concerted, highly cooperative interaction between all four subunits. Inactivation of Kv7.1 channels does not exhibit the hallmarks of N- and C-type inactivation. Inactivation of WT Kv7.1 channels can be revealed by hooked tail currents that reflects the recovery from a fast and voltage-independent inactivation process. However, several Kv7.1 mutants such as the pore mutant L273F generate an additional voltage-dependent slow inactivation. The subunit interactions during this slow inactivation gating remain unexplored. The goal of the present study was to study the nature of subunit interactions along Kv7.1 inactivation gating, using concatenated tetrameric Kv7.1 channel and introducing sequentially into each of the four subunits the slow inactivating pore mutation L273F. Incorporating an incremental number of inactivating mutant subunits did not affect the inactivation kinetics but slowed down the recovery kinetics from inactivation. Results indicate that Kv7.1 inactivation gating is not compatible with a concerted cooperative process. Instead, adding an inactivating subunit L273F into the Kv7.1 tetramer incrementally stabilizes the inactivated state, which suggests that like for activation gating, Kv7.1 slow inactivation gating is not a concerted process.

  10. Effects of 1-Alkyl-3-Methylimidazolium Nitrate on Soil Physical and Chemical Properties and Microbial Biomass.

    PubMed

    Zhou, Tongtong; Wang, Jun; Ma, Zhiqiang; Du, Zhongkun; Zhang, Cheng; Zhu, Lusheng; Wang, Jinhua

    2018-05-01

    Ionic liquids (ILs), also called room temperature ILs, are widely applied in many fields on the basis of their unique physical and chemical properties. However, numerous ILs may be released into and gradually accumulate in the environment due to their extensive use and absolute solubility. The effects of 1-alkyl-3-methylimidazolium nitrate ([C n mim]NO 3 , n = 4, 6, 8) on soil pH, conductivity, cation exchange capacity, microbial biomass carbon, and microbial biomass nitrogen were examined at the doses of 1, 10, and 100 mg/kg on days 10, 20, 30, and 40. The results demonstrated that the soil pH decreased and the conductivity increased with increasing IL doses. No significant differences were observed in the soil cation-exchange capacity. All three of the tested ILs decreased the soil microbial biomass carbon and nitrogen. Additionally, there were few differences among the ILs with different alkyl chain lengths on the tested indicators except for the microbial biomass nitrogen. The present study addressed a gap in the literature regarding the effects of the aforementioned ILs with different alkyl side chains on the physicochemical properties of soil, and the results could provide the basic data for future studies on their toxicity to soil organisms, such as earthworms and soil microbes.

  11. Photodynamic inactivation of multiresistant bacteria (KPC) using zinc(II)phthalocyanines.

    PubMed

    Miretti, Mariana; Clementi, Romina; Tempesti, Tomas C; Baumgartner, María T

    2017-09-15

    The worldwide increase in antibiotic resistance has led to search of alternatives anti-microbial therapies such as photodynamic inactivation. The aim of this paper was to evaluate the photodynamic activity in vitro of a neutral and two cationic Zn phthalocyanines. Their photokilling activity was tested on Escherichia coli ATCC 25922 and Klebsiella pneumoniae Carbapenemase (KPC)-producing. After treating bacteria with phthalocyanines, the cultures were irradiated with white light. As a result, the bacteria were inactivated in presence of cationic phthalocyanines. The photoinactivation was dependent of the irradiation time and phthalocyanine concentration. The most effective photosensitizer on KPC-producing was Zinc(II)tetramethyltetrapyridino[2,3-b:2',3'-g:2″,3″-l:2‴,3‴-q]porphyrazinium methylsulfate (ZnTM2,3PyPz). After irradiation using the water soluble ZnTM2,3PyPz (3μM) the viability of KPC (30min of irradiation) and E. coli (10min of irradiation) decreased ≈99.995%. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Inactivation of Escherichia coli in milk and concentrated milk using pulsed-light treatment.

    PubMed

    Miller, B M; Sauer, A; Moraru, C I

    2012-10-01

    Pulsed light (PL) treatment has been viewed as an alternative to thermal treatments for the inactivation of pathogenic and spoilage microorganisms in recent years. The objectives of this study were to quantify the effectiveness of PL on inactivating Escherichia coli in cow milk and to evaluate the effect of total solids and fat content on inactivation. Samples of reconstituted milk with variable total solids levels (9.8, 25, and 45%) and commercial cow milk with different fat contents (skim milk, 2% fat, and whole milk) were inoculated with nonpathogenic E. coli ATCC 25922 at a concentration of 10(7)cfu/mL. One milliliter of the inoculated sample was placed in a thin layer in a glass chamber and exposed to PL doses of up to 14.9 J/cm(2), both in static mode and turbulent mode. Survivors were quantified using standard plate counting. All experiments were performed in triplicate. Pulsed light treatment of the concentrated milks of 25 and 45% solids content resulted in reductions of less than 1 log, even in turbulent mode, whereas for the milk with 9.8% solids content, reduction levels of 2.5 log cfu were obtained after treatment with 8.4 J/cm(2) in turbulent mode. In the skim milk, a 3.4 log cfu reduction at 14.9 J/cm(2) was obtained and a plateau of the inactivation curve typical of PL treatment was not achieved. Under the same conditions, both 2% and whole milk attained inactivation levels greater than 2.5 log cfu. These data indicate that PL is effective for the inactivation of E. coli in milk, but has limited effectiveness for microbial inactivation in concentrated milk, due to the absorption of light by the milk solids and shielding of the bacteria in the concentrated substrates. Milk fat also diminishes the effectiveness of PL to some extent, due to light-scattering effects. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  13. [Effect of long-term fertilizing regime on soil microbial diversity and soil property].

    PubMed

    Li, Chenhua; Zhang, Caixia; Tang, Lisong; Xiong, Zhengqin; Wang, Baozhan; Jia, Zhongjun; Li, Yan

    2014-03-04

    To evaluate the effect of long-term fertilization on soil microbial community and soil chemical and physical properties. Using a high-throughput pyrosequencing technique, we studied microbial community in the 0-300 cm soil samples covering a 20-year field-experiment with different fertilization applications including inorganic fertilizer alone (N 300 kg/hm2, P2O5 150 kg/hm2 and K2O 60 kg/hm2) and inorganic fertilizer combined with straw (same application rate of N and P fertilizer combined with 5.4 t straw). Actinobacteria and alpha-proteobacteria were the predominant groups in the topsoil (0-20 cm). As the soil depth increased, the relative abundance of actinobacteria decreased whereas that of proteobacteria, especially gamma-proteobacteria and beta-proteobacteria increased and gradually became the dominant groups in the subsoil (20-300 cm). Long-term fertilizing applications significantly affected soil microbial communities throughout the soil profile, and increased the relative abundance of ammonia-oxidizing archaea at 0-40 cm depth. In addition, agriculture management, e. g. irrigation may be an important driving factor for the distribution of ammonia-oxidizing bacteria in soil profile. Total nitrogen and organic carbon contents were the most influential factors on microbial community in the topsoil and in the subsoil, respectively. Long-term fertilizer applications altered soil nutrient availability within the soil profile, which was likely to result in the different microbial community structure between the fertilizer treatments, especially for the subsoil.

  14. Land-use types and soil chemical properties influence soil microbial communities in the semiarid Loess Plateau region in China

    PubMed Central

    Tian, Qin; Taniguchi, Takeshi; Shi, Wei-Yu; Li, Guoqing; Yamanaka, Norikazu; Du, Sheng

    2017-01-01

    Similar land-use types usually have similar soil properties, and, most likely, similar microbial communities. Here, we assessed whether land-use types or soil chemical properties are the primary drivers of soil microbial community composition, and how changes in one part of the ecosystem affect another. We applied Ion Torrent sequencing to the bacterial and fungal communities of five different land-use (vegetation) types in the Loess Plateau of China. We found that the overall trend of soil quality was natural forest > plantation > bare land. Dominant bacterial phyla consisted of Proteobacteria (42.35%), Actinobacteria (15.61%), Acidobacteria (13.32%), Bacteroidetes (8.43%), and Gemmatimonadetes (6.0%). The dominant fungi phyla were Ascomycota (40.39%), Basidiomycota (38.01%), and Zygomycota (16.86%). The results of Canonical Correspondence Analysis (CCA) and Redundancy Analysis (RDA) based on land-use types displayed groups according to the land-use types. Furthermore, the bacterial communities were mainly organized by soil organic carbon (SOC). The fungal communities were mainly related to available phosphorus (P). The results suggested that the changes of land use type generated changes in soil chemical properties, controlling the composition of microbial community in the semiarid Loess Plateau region. The microbial community could be an indicator for soil quality with respect to ecological restoration. PMID:28349918

  15. Land-use types and soil chemical properties influence soil microbial communities in the semiarid Loess Plateau region in China

    NASA Astrophysics Data System (ADS)

    Tian, Qin; Taniguchi, Takeshi; Shi, Wei-Yu; Li, Guoqing; Yamanaka, Norikazu; Du, Sheng

    2017-03-01

    Similar land-use types usually have similar soil properties, and, most likely, similar microbial communities. Here, we assessed whether land-use types or soil chemical properties are the primary drivers of soil microbial community composition, and how changes in one part of the ecosystem affect another. We applied Ion Torrent sequencing to the bacterial and fungal communities of five different land-use (vegetation) types in the Loess Plateau of China. We found that the overall trend of soil quality was natural forest > plantation > bare land. Dominant bacterial phyla consisted of Proteobacteria (42.35%), Actinobacteria (15.61%), Acidobacteria (13.32%), Bacteroidetes (8.43%), and Gemmatimonadetes (6.0%). The dominant fungi phyla were Ascomycota (40.39%), Basidiomycota (38.01%), and Zygomycota (16.86%). The results of Canonical Correspondence Analysis (CCA) and Redundancy Analysis (RDA) based on land-use types displayed groups according to the land-use types. Furthermore, the bacterial communities were mainly organized by soil organic carbon (SOC). The fungal communities were mainly related to available phosphorus (P). The results suggested that the changes of land use type generated changes in soil chemical properties, controlling the composition of microbial community in the semiarid Loess Plateau region. The microbial community could be an indicator for soil quality with respect to ecological restoration.

  16. Virus inactivation by grapes and wines.

    PubMed Central

    Konowalchuk, J; Speirs, J I

    1976-01-01

    Infusions and extracts of different grapes inactivated poliovirus; agents responsible for this property resided in the skin of the grape. Commercial grape juice at both natural and neutral pH inactivate various enteric viruses and herpes simplex virus; a 1,000-fold reduction in poliovirus infectivity occurred after incubation with grape juice, pH 7.0, for 24 h at 4 degrees C. A variety of wines were antiviral but to a lesser extent than grape juice; red wines were more antiviral than white. Antiviral activity was demonstrable in fractions of grape juice varying in molecular weight from less than 1,000 to greater than 30,000 as determined by membrane filtration. Some restoration of poliovirus infectivity from virus-grape juice complexes was achieved with 1% gelatin, 0.1% Tween 80, 0.5% polyvinyl pyrrolidone, and 0.5% polyethylene glycol. PMID:12719

  17. The effectiveness of radiant catalytic ionization in inactivation of Listeria monocytogenes planktonic and biofilm cells from food and food contact surfaces as a method of food preservation.

    PubMed

    Skowron, K; Grudlewska, K; Krawczyk, A; Gospodarek-Komkowska, E

    2018-06-01

    The aim of the study was to evaluate the microbicidal effectiveness of radiant catalytic ionization (RCI) against Listeria monocytogenes strains in the form of planktonic cells and biofilm on food products and food contact surfaces as a method of food preservation. The study material comprised six strains of L. monocytogenes, isolated from food. Samples of different types of food available by retail (raw carrot, frozen salmon filets, soft cheese) and the fragments of surfaces (stainless steel AISI 304, rubber, milled rock tiles, polypropylene) were used in the experiment. The obtained results showed the effectiveness of RCI in the inactivation of both forms of the tested L. monocytogenes strains on all the surfaces. The effectiveness of RCI for biofilm forms was lower as compared with planktonic forms. The PRR value ranged from 18·19 to 99·97% for planktonic form and from 3·92 to 70·10% for biofilm. The RCI phenomenon induces the inactivation of L. monocytogenes on surfaces of food and materials used in the processing industry to a varying degree, depending on the manner of surface contamination, the properties of the contaminated materials as well as on the origin of the strain and the properties of surrounding dispersive environment in which the micro-organisms were suspended. Searching of new actions aimed at the reduction of the microbial contamination of food and food contact surfaces are extremely important. RCI method has been already described as an effective technique of microbial and abiotic pollution removal from air. However, our studies provide new, additional data related to evaluation the RCI efficacy against microbes on different surfaces, both in planktonic and biofilm form. © 2018 The Society for Applied Microbiology.

  18. Concepts and tools for predictive modeling of microbial dynamics.

    PubMed

    Bernaerts, Kristel; Dens, Els; Vereecken, Karen; Geeraerd, Annemie H; Standaert, Arnout R; Devlieghere, Frank; Debevere, Johan; Van Impe, Jan F

    2004-09-01

    Description of microbial cell (population) behavior as influenced by dynamically changing environmental conditions intrinsically needs dynamic mathematical models. In the past, major effort has been put into the modeling of microbial growth and inactivation within a constant environment (static models). In the early 1990s, differential equation models (dynamic models) were introduced in the field of predictive microbiology. Here, we present a general dynamic model-building concept describing microbial evolution under dynamic conditions. Starting from an elementary model building block, the model structure can be gradually complexified to incorporate increasing numbers of influencing factors. Based on two case studies, the fundamentals of both macroscopic (population) and microscopic (individual) modeling approaches are revisited. These illustrations deal with the modeling of (i) microbial lag under variable temperature conditions and (ii) interspecies microbial interactions mediated by lactic acid production (product inhibition). Current and future research trends should address the need for (i) more specific measurements at the cell and/or population level, (ii) measurements under dynamic conditions, and (iii) more comprehensive (mechanistically inspired) model structures. In the context of quantitative microbial risk assessment, complexity of the mathematical model must be kept under control. An important challenge for the future is determination of a satisfactory trade-off between predictive power and manageability of predictive microbiology models.

  19. Advances in antimicrobial photodynamic inactivation at the nanoscale

    PubMed Central

    Kashef, Nasim; Huang, Ying-Ying; Hamblin, Michael R.

    2017-01-01

    The alarming worldwide increase in antibiotic resistance amongst microbial pathogens necessitates a search for new antimicrobial techniques, which will not be affected by, or indeed cause resistance themselves. Light-mediated photoinactivation is one such technique that takes advantage of the whole spectrum of light to destroy a broad spectrum of pathogens. Many of these photoinactivation techniques rely on the participation of a diverse range of nanoparticles and nanostructures that have dimensions very similar to the wavelength of light. Photodynamic inactivation relies on the photochemical production of singlet oxygen from photosensitizing dyes (type II pathway) that can benefit remarkably from formulation in nanoparticle-based drug delivery vehicles. Fullerenes are a closed-cage carbon allotrope nanoparticle with a high absorption coefficient and triplet yield. Their photochemistry is highly dependent on microenvironment, and can be type II in organic solvents and type I (hydroxyl radicals) in a biological milieu. Titanium dioxide nanoparticles act as a large band-gap semiconductor that can carry out photo-induced electron transfer under ultraviolet A light and can also produce reactive oxygen species that kill microbial cells. We discuss some recent studies in which quite remarkable potentiation of microbial killing (up to six logs) can be obtained by the addition of simple inorganic salts such as the non-toxic sodium/potassium iodide, bromide, nitrite, and even the toxic sodium azide. Interesting mechanistic insights were obtained to explain this increased killing. PMID:29226063

  20. Advances in antimicrobial photodynamic inactivation at the nanoscale

    NASA Astrophysics Data System (ADS)

    Kashef, Nasim; Huang, Ying-Ying; Hamblin, Michael R.

    2017-08-01

    The alarming worldwide increase in antibiotic resistance amongst microbial pathogens necessitates a search for new antimicrobial techniques, which will not be affected by, or indeed cause resistance themselves. Light-mediated photoinactivation is one such technique that takes advantage of the whole spectrum of light to destroy a broad spectrum of pathogens. Many of these photoinactivation techniques rely on the participation of a diverse range of nanoparticles and nanostructures that have dimensions very similar to the wavelength of light. Photodynamic inactivation relies on the photochemical production of singlet oxygen from photosensitizing dyes (type II pathway) that can benefit remarkably from formulation in nanoparticle-based drug delivery vehicles. Fullerenes are a closed-cage carbon allotrope nanoparticle with a high absorption coefficient and triplet yield. Their photochemistry is highly dependent on microenvironment, and can be type II in organic solvents and type I (hydroxyl radicals) in a biological milieu. Titanium dioxide nanoparticles act as a large band-gap semiconductor that can carry out photo-induced electron transfer under ultraviolet A light and can also produce reactive oxygen species that kill microbial cells. We discuss some recent studies in which quite remarkable potentiation of microbial killing (up to six logs) can be obtained by the addition of simple inorganic salts such as the non-toxic sodium/potassium iodide, bromide, nitrite, and even the toxic sodium azide. Interesting mechanistic insights were obtained to explain this increased killing.

  1. Inactivation of Salmonella Typhimurium and Listeria monocytogenes on ham with nonthermal atmospheric pressure plasma

    PubMed Central

    Lis, Karolina Anna; Binder, Sylvia; Li, Yangfang; Kehrenberg, Corinna; Zimmermann, Julia Louise; Ahlfeld, Birte

    2018-01-01

    The application of cold atmospheric pressure plasma (CAP) for decontamination of sliced ready-to-eat (RTE) meat products (in this case, rolled fillets of ham), inoculated with Salmonella (S.) Typhimurium and Listeria (L.) monocytogenes was investigated. Cold atmospheric plasma (CAP) is an ionised gas that includes highly reactive species and ozone, interacting with cell membranes and DNA of bacteria. The mode of action of CAPs includes penetration and disruption of the outer cell membrane or intracellular destruction of DNA located in the cytoplasm. Inoculated ham was treated for 10 and 20 min with CAP generated by a surface-micro-discharge-plasma source using cost-effective ambient air as working gas with different humidity levels of 45–50 and 90%. The chosen plasma modes had a peak-to-peak voltage of 6.4 or 10 kV and a frequency of 2 and 10 kHz. Under the tested conditions, the direct effectiveness of CAP on microbial inactivation was limited. Although all treated samples showed significant reductions in the microbial load subsequent to plasma treatment, the maximum inactivation of S. Typhimurium was 1.14 lg steps after 20 min of CAP-treatment (p<0.05), and L. monocytogenes was reduced by 1.02 lg steps (p<0.05) using high peak-to-peak voltage of 10 kV and a frequency of 2 kHz regardless of moisture content. However, effective inactivation was achieved by a combination of CAP-treatment and cold storage at 8°C ± 0.5°C for 7 and 14 days after packaging under sealed high nitrogen gas flush (70% N2, 30% CO2). Synergistic effects of CAP and cold storage for 14 days led to a clearer decrease in the microbial load of 1.84 lg steps for S. Typhimurium (p<0.05) and 2.55 lg steps for L. monocytogenes (p<0.05). In the case of L. monocytogenes, subsequent to CAP-treatment (10 kV, 2 kHz) and cold storage, microbial counts were predominantly below the detection limit. Measurement showed that after CAP-treatment, surface temperature of ham did not exceed the room temperature

  2. Inactivation of Salmonella Typhimurium and Listeria monocytogenes on ham with nonthermal atmospheric pressure plasma.

    PubMed

    Lis, Karolina Anna; Boulaaba, Annika; Binder, Sylvia; Li, Yangfang; Kehrenberg, Corinna; Zimmermann, Julia Louise; Klein, Günter; Ahlfeld, Birte

    2018-01-01

    The application of cold atmospheric pressure plasma (CAP) for decontamination of sliced ready-to-eat (RTE) meat products (in this case, rolled fillets of ham), inoculated with Salmonella (S.) Typhimurium and Listeria (L.) monocytogenes was investigated. Cold atmospheric plasma (CAP) is an ionised gas that includes highly reactive species and ozone, interacting with cell membranes and DNA of bacteria. The mode of action of CAPs includes penetration and disruption of the outer cell membrane or intracellular destruction of DNA located in the cytoplasm. Inoculated ham was treated for 10 and 20 min with CAP generated by a surface-micro-discharge-plasma source using cost-effective ambient air as working gas with different humidity levels of 45-50 and 90%. The chosen plasma modes had a peak-to-peak voltage of 6.4 or 10 kV and a frequency of 2 and 10 kHz. Under the tested conditions, the direct effectiveness of CAP on microbial inactivation was limited. Although all treated samples showed significant reductions in the microbial load subsequent to plasma treatment, the maximum inactivation of S. Typhimurium was 1.14 lg steps after 20 min of CAP-treatment (p<0.05), and L. monocytogenes was reduced by 1.02 lg steps (p<0.05) using high peak-to-peak voltage of 10 kV and a frequency of 2 kHz regardless of moisture content. However, effective inactivation was achieved by a combination of CAP-treatment and cold storage at 8°C ± 0.5°C for 7 and 14 days after packaging under sealed high nitrogen gas flush (70% N2, 30% CO2). Synergistic effects of CAP and cold storage for 14 days led to a clearer decrease in the microbial load of 1.84 lg steps for S. Typhimurium (p<0.05) and 2.55 lg steps for L. monocytogenes (p<0.05). In the case of L. monocytogenes, subsequent to CAP-treatment (10 kV, 2 kHz) and cold storage, microbial counts were predominantly below the detection limit. Measurement showed that after CAP-treatment, surface temperature of ham did not exceed the room temperature

  3. Inactivation kinetics of foodborne pathogens by UV-C radiation and its subsequent growth in fresh-cut kailan-hybrid broccoli.

    PubMed

    Martínez-Hernández, Ginés Benito; Huertas, Juan-Pablo; Navarro-Rico, Javier; Gómez, Perla A; Artés, Francisco; Palop, Alfredo; Artés-Hernández, Francisco

    2015-04-01

    The inactivation of Escherichia coli, S. Enteritidis and Listeria monocytogenes after UV-C radiation with 0, 2.5, 5, 7.5, 10 and 15 kJ UV-C m(-2) on fresh-cut kailan-hybrid broccoli was explored. Inactivation did not follow linear kinetics. Hence, it was modelled by using the Weibull distribution function, obtaining adjusted R(2) values higher than 94%, indicative of the accuracy of the model to the experimental data. The UV-C doses needed to reduce 1 log cycle the E. coli, S. Enteritidis and L. monocytogenes counts were 1.07, 0.02 and 9.26 kJ m(-2), respectively, being S. Enteritidis the most sensitive microorganism to UV-C radiation while L. monocytogenes was the most resistant. According to experimental data, UV-C doses higher than 2.5 kJ m(-2) did not achieve great microbial reductions. No differences in the growth behaviour of these microorganisms was observed in the treated samples stored under air conditions at 5, 10 and 15 °C, compared to the control. Conclusively, low UV-C doses are effective to reduce E. coli, S. Enteritidis and L. monocytogenes populations in fresh-cut kailan-hybrid broccoli keeping such counts stable during shelf life at 5-10 °C. The current study provides inactivation models for these foodborne pathogens that can be used in microbial risk assessment. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Inactivation of bacterial pathogens in yoba mutandabota, a dairy product fermented with the probiotic Lactobacillus rhamnosus yoba.

    PubMed

    Mpofu, Augustine; Linnemann, Anita R; Nout, Martinus J R; Zwietering, Marcel H; Smid, Eddy J; den Besten, Heidy M W

    2016-01-18

    Mutandabota is a dairy product consumed as a major source of proteins and micronutrients in Southern Africa. In this study the microbial safety of traditional and a variant of mutandabota fermented with the probiotic Lactobacillus rhamnosus yoba (yoba mutandabota) was investigated by challenging the products with five important food pathogens: Listeria monocytogenes, Salmonella spp., Campylobacter jejuni, Escherichia coli O157:H7 and Bacillus cereus. Pasteurized full-fat cow's milk was used for producing traditional and yoba mutandabota, and was inoculated with a cocktail of strains of the pathogens at an inoculum level of 5.5 log cfu/mL. Survival of the pathogens was monitored over a potential consumption time of 24h for traditional mutandabota, and over 24h of fermentation followed by 24h of potential consumption time for yoba mutandabota. In traditional mutandabota (pH3.4 ± 0.1) no viable cells of B. cereus and C. jejuni were detected 3h after inoculation, while L. monocytogenes, E. coli O157:H7 and Salmonella spp. significantly declined (P<0.05), but could still be detected (<3.5 log inactivation) at the end of the potential consumption time. This indicated that consumption of traditional mutandabota exposes consumers to the risk of food-borne microbial infections. In yoba mutandabota, L. rhamnosus yoba grew from 5.5 ± 0.1 log cfu/mL to 9.1 ± 0.4 log cfu/mL in the presence of pathogens. The pH of yoba mutandabota dropped from 4.2 ± 0.1 to 3.3 ± 0.1 after 24h of fermentation, mainly due to organic acids produced during fermentation. Only Salmonella spp. was able to grow in yoba mutandabota during the first 9h of fermentation, but then decreased in viable plate count. None of the tested pathogens were detected (>3.5 log inactivation) after 3h into potential consumption time of yoba mutandabota. Inactivation of pathogens in mutandabota is of public health significance because food-borne pathogens endanger public health upon consumption of contaminated food

  5. Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels.

    PubMed

    Fineberg, Jeffrey D; Ritter, David M; Covarrubias, Manuel

    2012-11-01

    A-type voltage-gated K(+) (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na(+) channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously

  6. Modeling-independent elucidation of inactivation pathways in recombinant and native A-type Kv channels

    PubMed Central

    Fineberg, Jeffrey D.; Ritter, David M.

    2012-01-01

    A-type voltage-gated K+ (Kv) channels self-regulate their activity by inactivating directly from the open state (open-state inactivation [OSI]) or by inactivating before they open (closed-state inactivation [CSI]). To determine the inactivation pathways, it is often necessary to apply several pulse protocols, pore blockers, single-channel recording, and kinetic modeling. However, intrinsic hurdles may preclude the standardized application of these methods. Here, we implemented a simple method inspired by earlier studies of Na+ channels to analyze macroscopic inactivation and conclusively deduce the pathways of inactivation of recombinant and native A-type Kv channels. We investigated two distinct A-type Kv channels expressed heterologously (Kv3.4 and Kv4.2 with accessory subunits) and their native counterparts in dorsal root ganglion and cerebellar granule neurons. This approach applies two conventional pulse protocols to examine inactivation induced by (a) a simple step (single-pulse inactivation) and (b) a conditioning step (double-pulse inactivation). Consistent with OSI, the rate of Kv3.4 inactivation (i.e., the negative first derivative of double-pulse inactivation) precisely superimposes on the profile of the Kv3.4 current evoked by a single pulse because the channels must open to inactivate. In contrast, the rate of Kv4.2 inactivation is asynchronous, already changing at earlier times relative to the profile of the Kv4.2 current evoked by a single pulse. Thus, Kv4.2 inactivation occurs uncoupled from channel opening, indicating CSI. Furthermore, the inactivation time constant versus voltage relation of Kv3.4 decreases monotonically with depolarization and levels off, whereas that of Kv4.2 exhibits a J-shape profile. We also manipulated the inactivation phenotype by changing the subunit composition and show how CSI and CSI combined with OSI might affect spiking properties in a full computational model of the hippocampal CA1 neuron. This work unambiguously

  7. Maple sap predominant microbial contaminants are correlated with the physicochemical and sensorial properties of maple syrup.

    PubMed

    Filteau, Marie; Lagacé, Luc; Lapointe, Gisèle; Roy, Denis

    2012-03-01

    Maple sap processing and microbial contamination are significant aspects that affect maple syrup quality. In this study, two sample sets from 2005 and 2008 were used to assess the maple syrup quality variation and its relationship to microbial populations, with respect to processing, production site and harvesting period. The abundance of maple sap predominant bacteria (Pseudomonas fluorescens group and two subgroups, Rahnella spp., Janthinobacterium spp., Leuconostoc mesenteroides) and yeast (Mrakia spp., Mrakiella spp.,Guehomyces pullulans) was assessed by quantitative PCR. Maple syrup properties were analyzed by physicochemical and sensorial methods. Results indicate that P. fluorescens, Mrakia spp., Mrakiella spp. G. pullulans and Rahnella spp. are stable contaminants of maple sap, as they were found for every production site throughout the flow period. Multiple factor analysis reports a link between the relative abundance of P. fluorescens group and Mrakia spp. in maple sap with maple and vanilla odor as well as flavor of maple syrup. This evidence supports the contribution of these microorganisms or a consortium of predominant microbial contaminants to the characteristic properties of maple syrup. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Stabilization of red fruit-based smoothies by high-pressure processing. Part A. Effects on microbial growth, enzyme activity, antioxidant capacity and physical stability.

    PubMed

    Hurtado, Adriana; Guàrdia, Maria Dolors; Picouet, Pierre; Jofré, Anna; Ros, José María; Bañón, Sancho

    2017-02-01

    Non-thermal pasteurization by high-pressure processing (HPP) is increasingly replacing thermal processing (TP) to maintain the properties of fresh fruit products. However, most of the research on HPP-fruit products only partially addresses fruit-pressure interaction, which limits its practical interest. The objective of this study was to assess the use of a mild HPP treatment to stabilize red fruit-based smoothies (microbial, enzymatic, oxidative and physical stability). HPP (350 MPa/10 °C/5 min) was slightly less effective than TP (85 °C/7 min) in inactivating microbes (mesophilic and psychrophilic bacteria, coliforms, yeasts and moulds) in smoothies kept at 4 °C for up to 28 days. The main limitation of using HPP was its low efficacy in inactivating oxidative (polyphenol oxidase and peroxidase) and hydrolytic (pectin methyl esterase) enzymes. Data on antioxidant status, colour parameters, browning index, transmittance, turbidity and viscosity confirmed that the HPP-smoothies have a greater tendency towards oxidation and clarification, which might lead to undesirable sensory and nutritional changes (see Part B). The microbial quality of smoothies was adequately controlled by mild HPP treatment without affecting their physical-chemical characteristics; however, oxidative and hydrolytic enzymes are highly pressure-resistant, which suggests that additional strategies should be used to stabilize smoothies. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  9. Alpha-momorcharin: a ribosome-inactivating protein from Momordica charantia, possessing DNA cleavage properties.

    PubMed

    Wang, Shuzhen; Zheng, Yinzhen; Yan, Junjie; Zhu, Zhixuan; Wu, Zhihua; Ding, Yi

    2013-11-01

    Ribosome-inactivating proteins (RIPs) function to inhibit protein synthesis through the removal of specific adenine residues from eukaryotic ribosomal RNA and rending the 60S subunit unable to bind elongation factor 2. They have received much attention in biological and biomedical research due to their unique activities toward tumor cells, as well as the important roles in plant defense. Alpha-momorcharin (α-MC), a member of the type I family of RIPs, is rich in the seeds of Momordica charantia L. Previous studies demonstrated that α-MC is an effective antifungal and antibacterial protein. In this study, a detailed analysis of the DNase-like activity of α-MC was conducted. Results showed that the DNase-like activity toward plasmid DNA was time-dependent, temperature-related, and pH-stable. Moreover, a requirement for divalent metal ions in the catalytic domain of α-MC was confirmed. Additionally, Tyr(93) was found to be a critical residue for the DNase-like activity, while Tyr(134), Glu(183), Arg(186), and Trp(215) were activity-related residues. This study on the chemico-physical properties and mechanism of action of α-MC will improve its utilization in scientific research, as well as its potential industrial uses. These results may also assist in the characterization and elucidation of the DNase-like enzymatic properties of other RIPs.

  10. What is microbial community ecology?

    PubMed

    Konopka, Allan

    2009-11-01

    The activities of complex communities of microbes affect biogeochemical transformations in natural, managed and engineered ecosystems. Meaningfully defining what constitutes a community of interacting microbial populations is not trivial, but is important for rigorous progress in the field. Important elements of research in microbial community ecology include the analysis of functional pathways for nutrient resource and energy flows, mechanistic understanding of interactions between microbial populations and their environment, and the emergent properties of the complex community. Some emergent properties mirror those analyzed by community ecologists who study plants and animals: biological diversity, functional redundancy and system stability. However, because microbes possess mechanisms for the horizontal transfer of genetic information, the metagenome may also be considered as a community property.

  11. MS2 inactivation by TiO2 nanoparticles in the presence of quartz sand

    NASA Astrophysics Data System (ADS)

    Syngouna, Vasiliki I.; Chrysikopoulos, Constantinos V.

    2017-04-01

    Virus inactivation by nanoparticles (NPs) is hypothesized to affect virus fate and transport in the subsurface. This study examines the interactions of viruses with titanium dioxide (TiO2) anatase NPs, which is a good disinfectant with unique physiochemical properties, using three different virus concentrations. The bacteriophage MS2 was used as a model virus. A series of batch experiments of MS2 inactivation by TiO2 NPs were conducted at room temperature (25 °C), in the presence of quartz sand, with and without ambient light. The virus inactivation experimental data were satisfactorily fitted with a pseudo-first order expression with a time dependent rate coefficient. Quartz sand was shown to affect MS2 inactivation by TiO2 NPs both in the presence and absence of ambient light, because, under the experimental conditions of this study, the quartz sand offers a protection to the attached MS2 against inactivation. Moreover, in most cases similar inactivation rates were observed in reactor and control tubes (absence of TiO2 NPs) suggesting that low TiO2 concentration (10 mg/L) affects only slightly MS2 inactivation with and without ambient light.

  12. Development of methods to measure virus inactivation in fresh waters.

    PubMed Central

    Ward, R L; Winston, P E

    1985-01-01

    This study concerns the identification and correction of deficiencies in methods used to measure inactivation rates of enteric viruses seeded into environmental waters. It was found that viable microorganisms in an environmental water sample increased greatly after addition of small amounts of nutrients normally present in the unpurified seed virus preparation. This burst of microbial growth was not observed after seeding the water with purified virus. The use of radioactively labeled poliovirus revealed that high percentages of virus particles, sometimes greater than 99%, were lost through adherence to containers, especially in less turbid waters. This effect was partially overcome by the use of polypropylene containers and by the absence of movement during incubation. Adherence to containers clearly demonstrated the need for labeled viruses to monitor losses in this type of study. Loss of viral infectivity in samples found to occur during freezing was avoided by addition of broth. Finally, microbial contamination of the cell cultures during infectivity assays was overcome by the use of gentamicin and increased concentrations of penicillin, streptomycin, and amphotericin B. PMID:3004328

  13. Potential contribution of microbial communities in technical ceramics for the improvement of rheological properties

    NASA Astrophysics Data System (ADS)

    Moreira, Bernardino; Miller, Ana Z.; Santos, Ricardo; Monteiro, Sílvia; Dias, Diamantino; Neves, Orquídia; Dionísio, Amélia; Saiz-Jimenez, Cesareo

    2014-05-01

    Several bacterial and fungal species naturally occurring in ceramic raw materials used in construction, such as Aspergillus, Penicillium and Aureobasidium, are known to produce exopolysaccharides (EPS). These polymers excreted by the cells are of widespread occurrence and may confer unique and potentially interesting properties with potential industrial uses, such as viscosity control, gelation, and flocculation, during ceramic manufacturing. In this study, the microbial communities present in clay raw materials were identified by both cultural methods and DNA-based molecular techniques in order to appraise their potential contribution to enhance the performance of technical ceramics through the use of EPS. Mineralogical identification by X- Ray Diffraction (XRD) and Fourier Transform Infrared (FTIR) spectroscopy of the clay raw materials, as well as characterization of rheological properties of ceramic slips were also performed. Microbial EPS production and its introduction into ceramic slips will be then carried out in order to evaluate their effects on the rheological properties of the ceramic slips, powders and conformed bodies. Some positive aspects related to the use of EPS are: reduction of the environmental impact caused by synthetic organic additives, reduction of production costs, as well as the costs related with operator protection systems, gaseous effluent treatments, complex landfill, among others.

  14. Relationships between waste physicochemical properties, microbial activity and vegetation at coal ash and sludge disposal sites.

    PubMed

    Woch, Marcin W; Radwańska, Magdalena; Stanek, Małgorzata; Łopata, Barbara; Stefanowicz, Anna M

    2018-06-11

    The aim of the study was to assess the relationships between vegetation, physicochemical and microbial properties of substrate at coal ash and sludge disposal sites. The study was performed on 32 plots classified into 7 categories: dried ash sedimentation ponds, dominated by a grass Calamagrostis epigejos (AH-Ce), with the admixture of Pinus sylvestris (AH-CePs) or Robinia pseudoacacia (AH-CeRp), dry ash landfill dominated by Betula pendula and Pinus sylvestris (AD-BpPs) or Salix viminalis (AD-Sv) and coal sludge pond with drier parts dominated by Tussilago farfara (CS-Tf) and the wetter ones by Cyperus flavescens (CS-Cf). Ash sites were covered with soil layer imported as a part of technical reclamation. Ash had relatively high concentrations of some alkali and alkaline earth metals, Mn and pH, while coal sludge had high water and C, S, P and K contents. Concentrations of heavy metals were lower than allowable limits in all substrate types. Microbial biomass and, particularly, enzymatic activity in ash and sludge were generally low. The only exception were CS-Tf plots characterized by the highest microbial biomass, presumably due to large deposits of organic matter that became available for aerobic microbial biomass when water level fell. The properties of ash and sludge adversely affected microbial biomass and enzymatic activity as indicated by significant negative correlations between the content of alkali/alkaline earth metals, heavy metals, and macronutrients with enzymatic activity and/or microbial biomass, as well as positive correlations of these parameters with metabolic quotient (qCO 2 ). Plant species richness and cover were relatively high, which may be partly associated with alleviating influence of soil covering the ash. The effect of the admixture of R. pseudoacacia or P. sylvestris to stands dominated by C. epigejos was smaller than expected. The former species increased NNH 4 , NNO 3 and arylsulfatase activity, while the latter reduced activity of

  15. Development of a microbial population within a hot-drinks vending machine and the microbial load of vended hot chocolate drink.

    PubMed

    Hall, A; Short, K; Saltmarsh, M; Fielding, L; Peters, A

    2007-09-01

    In order to understand the development of the microbial population within a hot-drinks vending machine a new machine was placed in a staff area of a university campus vending only hot chocolate. The machine was cleaned weekly using a detergent based protocol. Samples from the mixing bowl, dispense area, and drink were taken over a 19-wk period and enumerated using plate count agar. Bacillus cereus was identified using biochemical methods. Vended drinks were sampled at 0, 3, 6, and 9 min after vending; the hot chocolate powder was also sampled. Over the 1st 8 wk, a significant increase in the microbial load of the machine components was observed. By the end of the study, levels within the vended drink had also increased significantly. Inactivation of the automatic flush over a subsequent 5-wk period led to a statistically but not operationally significant increase in the microbial load of the dispense area and vended drink. The simple weekly clean had a significant impact on the microbial load of the machine components and the vended drink. This study demonstrated that a weekly, detergent-based cleaning protocol was sufficient to maintain the microbial population of the mixing bowl and dispense point in a quasi-steady state below 3.5 log CFU/cm2 ensuring that the microbial load of the vended drinks was maintained below 3.4 log CFU/mL. The microbial load of the drinks showed no significant changes over 9 min after vending, suggesting only spores are present in the final product.

  16. Microbial biosurfactants with their high-value functional properties

    USDA-ARS?s Scientific Manuscript database

    Microbial world is a rich source for finding valuable industrial chemicals and ingredients. Specifically, many microbial metabolites are surface-active compounds that can be developed into bio-based surfactants, detergents, and emulsifiers. Techno-economic analyses for the production of bio-based ...

  17. Competitive resource allocation to metabolic pathways contributes to overflow metabolisms and emergent properties in cross-feeding microbial consortia.

    PubMed

    Carlson, Ross P; Beck, Ashley E; Phalak, Poonam; Fields, Matthew W; Gedeon, Tomas; Hanley, Luke; Harcombe, William R; Henson, Michael A; Heys, Jeffrey J

    2018-04-17

    Resource scarcity is a common stress in nature and has a major impact on microbial physiology. This review highlights microbial acclimations to resource scarcity, focusing on resource investment strategies for chemoheterotrophs from the molecular level to the pathway level. Competitive resource allocation strategies often lead to a phenotype known as overflow metabolism; the resulting overflow byproducts can stabilize cooperative interactions in microbial communities and can lead to cross-feeding consortia. These consortia can exhibit emergent properties such as enhanced resource usage and biomass productivity. The literature distilled here draws parallels between in silico and laboratory studies and ties them together with ecological theories to better understand microbial stress responses and mutualistic consortia functioning. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  18. Quantifying Variability in Growth and Thermal Inactivation Kinetics of Lactobacillus plantarum.

    PubMed

    Aryani, D C; den Besten, H M W; Zwietering, M H

    2016-08-15

    The presence and growth of spoilage organisms in food might affect the shelf life. In this study, the effects of experimental, reproduction, and strain variabilities were quantified with respect to growth and thermal inactivation using 20 Lactobacillus plantarum strains. Also, the effect of growth history on thermal resistance was quantified. The strain variability in μmax was similar (P > 0.05) to reproduction variability as a function of pH, aw, and temperature, while being around half of the reproduction variability (P < 0.05) as a function of undissociated lactic acid concentration [HLa]. The cardinal growth parameters were estimated for the L. plantarum strains, and the pHmin was between 3.2 and 3.5, the aw,min was between 0.936 and 0.953, the [HLamax], at pH 4.5, was between 29 and 38 mM, and the Tmin was between 3.4 and 8.3°C. The average D values ranged from 0.80 min to 19 min at 55°C, 0.22 to 3.9 min at 58°C, 3.1 to 45 s at 60°C, and 1.8 to 19 s at 63°C. In contrast to growth, the strain variability in thermal resistance was on average six times higher than the reproduction variability and more than ten times higher than the experimental variability. The strain variability was also 1.8 times higher (P < 0.05) than the effect of growth history. The combined effects of strain variability and growth history on D value explained all of the variability as found in the literature, although with bias. Based on an illustrative milk-processing chain, strain variability caused ∼2-log10 differences in growth between the most and least robust strains and >10-log10 differences after thermal treatment. Accurate control and realistic prediction of shelf life is complicated by the natural diversity among microbial strains, and limited information on microbiological variability is available for spoilage microorganisms. Therefore, the objectives of the present study were to quantify strain variability, reproduction (biological) variability, and experimental

  19. Inactivation Methods of Trypsin Inhibitor in Legumes: A Review.

    PubMed

    Avilés-Gaxiola, Sara; Chuck-Hernández, Cristina; Serna Saldívar, Sergio O

    2018-01-01

    Seed legumes have played a major role as a crop worldwide, being cultivated on about 12% to 15% of Earth's arable land; nevertheless, their use is limited by, among other things, the presence of several antinutritional factors (ANFs - naturally occurring metabolites that the plant produces to protect itself from pest attacks.) Trypsin inhibitors (TIs) are one of the most relevant ANFs because they reduce digestion and absorption of dietary proteins. Several methods have been developed in order to inactivate TIs, and of these, thermal treatments are the most commonly used. They cause loss of nutrients, affect functional properties, and require high amounts of energy. Given the above, new processes have emerged to improve the nutritional quality of legumes while trying to solve the problems caused by the use of thermal treatments. This review examines and discusses the methods developed by researchers to inactivate TI present in legumes and their effects over nutritional and functional properties. © 2017 Institute of Food Technologists®.

  20. Sputtered Gum metal thin films showing bacterial inactivation and biocompatibility.

    PubMed

    Achache, S; Alhussein, A; Lamri, S; François, M; Sanchette, F; Pulgarin, C; Kiwi, J; Rtimi, S

    2016-10-01

    Super-elastic Titanium based thin films Ti-23Nb-0.7Ta-2Zr-(O) (TNTZ-O) and Ti-24Nb-(N) (TN-N) (at.%) were deposited by direct current magnetron sputtering (DCMS) in different reactive atmospheres. The effects of oxygen doping (TNTZ-O) and/or nitrogen doping (TN-N) on the microstructure, mechanical properties and biocompatibility of the as-deposited coatings were investigated. Nano-indentation measurements show that, in both cases, 1sccm of reactive gas in the mixture is necessary to reach acceptable values of hardness and Young's modulus. Mechanical properties are considered in relation to the films compactness, the compressive stress and the changes in the grain size. Data on Bacterial inactivation and biocompatibility are reported in this study. The biocompatibility tests showed that O-containing samples led to higher cells proliferation. Bacterial inactivation was concomitant with the observed pH and surface potential changes under light and in the dark. The increased cell fluidity leading to bacterial lysis was followed during the bacterial inactivation time. The increasing cell wall fluidity was attributed to the damage of the bacterial outer cell which losing its capacity to regulate the ions exchange in and out of the bacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Influence of Sedimentary and Seagrass Microbial Communities on Shallow-Water Benthic Optical Properties

    DTIC Science & Technology

    2008-09-30

    grass, Thalassia testudinum) and in Monterey Bay (eel grass, Zostera marina). Assess how the microbial community affects the flux of photons to and...optical properties and photosynthetic potential of the seagrasses Thalassia testudinum and Zostera marina”. The manuscript will be submitted to the...crystals in Thalassia from LSI. We have pursued this serendipitous discovery, and at the Estuarine Research Federation meeting this fall, we will

  2. Ribosome-inactivating proteins

    PubMed Central

    Walsh, Matthew J; Dodd, Jennifer E; Hautbergue, Guillaume M

    2013-01-01

    Ribosome-inactivating proteins (RIPs) were first isolated over a century ago and have been shown to be catalytic toxins that irreversibly inactivate protein synthesis. Elucidation of atomic structures and molecular mechanism has revealed these proteins to be a diverse group subdivided into two classes. RIPs have been shown to exhibit RNA N-glycosidase activity and depurinate the 28S rRNA of the eukaryotic 60S ribosomal subunit. In this review, we compare archetypal RIP family members with other potent toxins that abolish protein synthesis: the fungal ribotoxins which directly cleave the 28S rRNA and the newly discovered Burkholderia lethal factor 1 (BLF1). BLF1 presents additional challenges to the current classification system since, like the ribotoxins, it does not possess RNA N-glycosidase activity but does irreversibly inactivate ribosomes. We further discuss whether the RIP classification should be broadened to include toxins achieving irreversible ribosome inactivation with similar turnovers to RIPs, but through different enzymatic mechanisms. PMID:24071927

  3. Modeling the pulsed light inactivation of microorganisms naturally occurring on vegetable substrates.

    PubMed

    Izquier, Adriana; Gómez-López, Vicente M

    2011-09-01

    Pulsed light (PL) is a fast non-thermal method for microbial inactivation. This research studied the kinetics of PL inactivation of microorganisms naturally occurring in some vegetables. Iceberg lettuce, white cabbage and Julienne-style cut carrots were subjected to increasing PL fluences up to 12J/cm(2) in order to study its effect on aerobic mesophilic bacteria determined by plate count. Also, sample temperature increase was determined by infrared thermometry. Survivors' curves were adjusted to several models. No shoulder but tail was observed. The Weibull model showed good fitting performance of data. Results for lettuce were: goodness-of-fit parameter RMSE=0.2289, fluence for the first decimal reduction δ=0.98±0.80J/cm(2) and concavity parameter p=0.33±0.08. Results for cabbage were: RMSE=0.0725, δ=0.81±0.23J/cm(2) and p=0.30±0.02; and for carrot: RMSE=0.1235, δ=0.39±0.24J/cm(2) and p=0.23±0.03. For lettuce, a log-linear and tail model was also suitable. Validation of the Weibull model produced determination coefficients of 0.88-0.96 and slopes of 0.78-0.99. Heating was too low to contribute to inactivation. A single low-energy pulse was enough to achieve one log reduction, with an ultrafast treatment time of 0.5ms. While PL efficacy was found to be limited to high residual counts, the achievable inactivation level may be considered useful for shelf-life extension. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Microbial fibrinolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo.

    PubMed

    Peng, Yong; Yang, Xiaojuan; Zhang, Yizheng

    2005-11-01

    Accumulation of fibrin in the blood vessels usually results in thrombosis, leading to myocardial infarction and other cardiovascular diseases. For thrombolytic therapy, microbial fibrinolytic enzymes have now attracted much more attention than typical thrombolytic agents because of the expensive prices and the undesirable side effects of the latter. The fibrinolytic enzymes were successively discovered from different microorganisms, the most important among which is the genus Bacillus from traditional fermented foods. The physiochemical properties of these enzymes have been characterized, and their effectiveness in thrombolysis in vivo has been further identified. Therefore, microbial fibrinolytic enzymes, especially those from food-grade microorganisms, have the potential to be developed as functional food additives and drugs to prevent or cure thrombosis and other related diseases.

  5. Microbial dynamics in petroleum oilfields and their relationship with physiological properties of petroleum oil reservoirs.

    PubMed

    Varjani, Sunita J; Gnansounou, Edgard

    2017-12-01

    Petroleum is produced by thermal decay of buried organic material over millions of years. Petroleum oilfield ecosystems represent resource of reduced carbon which favours microbial growth. Therefore, it is obvious that many microorganisms have adapted to harsh environmental conditions of these ecosystems specifically temperature, oxygen availability and pressure. Knowledge of microorganisms present in ecosystems of petroleum oil reservoirs; their physiological and biological properties help in successful exploration of petroleum. Understanding microbiology of petroleum oilfield(s) can be used to enhance oil recovery, as microorganisms in oil reservoirs produce various metabolites viz. gases, acids, solvents, biopolymers and biosurfactants. The aim of this review is to discuss characteristics of petroleum oil reservoirs. This review also provides an updated literature on microbial ecology of these extreme ecosystems including microbial origin as well as various types of microorganisms such as methanogens; iron, nitrate and sulphate reducing bacteria, and fermentative microbes present in petroleum oilfield ecosystems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. X-chromosome inactivation and escape

    PubMed Central

    DISTECHE, CHRISTINE M.; BERLETCH, JOEL B.

    2016-01-01

    X-chromosome inactivation, which was discovered by Mary Lyon in 1961 results in random silencing of one X chromosome in female mammals. This review is dedicated to Mary Lyon, who passed away last year. She predicted many of the features of X inactivation, for e.g., the existence of an X inactivation center, the role of L1 elements in spreading of silencing and the existence of genes that escape X inactivation. Starting from her published work here we summarize advances in the field. PMID:26690513

  7. Impact of Fast Sodium Channel Inactivation on Spike Threshold Dynamics and Synaptic Integration

    PubMed Central

    Platkiewicz, Jonathan; Brette, Romain

    2011-01-01

    Neurons spike when their membrane potential exceeds a threshold value. In central neurons, the spike threshold is not constant but depends on the stimulation. Thus, input-output properties of neurons depend both on the effect of presynaptic spikes on the membrane potential and on the dynamics of the spike threshold. Among the possible mechanisms that may modulate the threshold, one strong candidate is Na channel inactivation, because it specifically impacts spike initiation without affecting the membrane potential. We collected voltage-clamp data from the literature and we found, based on a theoretical criterion, that the properties of Na inactivation could indeed cause substantial threshold variability by itself. By analyzing simple neuron models with fast Na inactivation (one channel subtype), we found that the spike threshold is correlated with the mean membrane potential and negatively correlated with the preceding depolarization slope, consistent with experiments. We then analyzed the impact of threshold dynamics on synaptic integration. The difference between the postsynaptic potential (PSP) and the dynamic threshold in response to a presynaptic spike defines an effective PSP. When the neuron is sufficiently depolarized, this effective PSP is briefer than the PSP. This mechanism regulates the temporal window of synaptic integration in an adaptive way. Finally, we discuss the role of other potential mechanisms. Distal spike initiation, channel noise and Na activation dynamics cannot account for the observed negative slope-threshold relationship, while adaptive conductances (e.g. K+) and Na inactivation can. We conclude that Na inactivation is a metabolically efficient mechanism to control the temporal resolution of synaptic integration. PMID:21573200

  8. Turbidity composition and the relationship with microbial attachment and UV inactivation efficacy.

    PubMed

    Farrell, Charlotte; Hassard, Francis; Jefferson, Bruce; Leziart, Tangui; Nocker, Andreas; Jarvis, Peter

    2018-05-15

    Turbidity in water can be caused by a range of different turbidity causing materials (TCM). Here the characteristics and attachment of bacteria to TCMs was assessed and the resultant impact on UV disinfection determined. TCMs represent potential vehicles for bacterial penetration of water treatment barriers, contamination of potable supplies and impact on subsequent human health. The TCMs under investigation were representative of those that may be present in surface and ground waters, both from the source and formed in the treatment process. The TCMs were chalk, Fe (III) hydroxide precipitate, kaolin clay, manganese dioxide and humic acids, at different turbidity levels representative of source waters (0, 0.1, 0.2, 0.4, 1, 2, and 5 NTU). Escherichia coli and Enterococcus faecalis attachment followed the order of Fe(III)>chalk, with little to no attachment seen for MnO 2 , humic acids and clay. The attachment was postulated to be due to chalk and Fe(III) particles having a more neutral surface charge resulting in elevated aggregation with bacteria compared to other TCMs. The humic acids and Fe(III) were the TCMs which influenced inactivation of E. coli and E. faecalis due to decreasing UV transmittance (UVT) with increasing TCM concentration. The presence of the Fe(III) TCM at 0.2 NTU resulted in the poorest E. coli inactivation, with 2.5 log 10 reduction at UV dose of 10mJcm -2 (k d of -0.23cm 2 mJ -1 ) compared to a 3.9 log 10 reduction in the absence of TCMs. E. faecalis had a greater resistance to UV irradiation than E. coli for all TCMs. Effective disinfection of drinking water is a priority for ensuring high public health standards. Uniform regulations for turbidity levels for waters pre-disinfection by UV light set by regulators may not always be appropriate and efficacy is dependent on the type, as well as the amount, of turbidity present in the water. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  9. Chemical and microbial properties in contaminated soils around a magnesite mine in northeast China

    Treesearch

    D Yang; D-H Zeng; J Zhang; L-J Li; R. Mao

    2012-01-01

    We measured soil chemical and microbial properties at a depth of 0–20 cm among mine tailings, abandoned mined land, contaminated cropland, and uncontaminated cropland around a magnesite mine near Haicheng City, Liaoning Province, China. The objective was to clarify the impact of Mg on the soils. We found that soluble Mg2+ concentration and pH...

  10. Fast inactivation of delayed rectifier K conductance in squid giant axon and its cell bodies.

    PubMed

    Mathes, C; Rosenthal, J J; Armstrong, G M; Gilly, W F

    1997-04-01

    Inactivation of delayed rectifier K conductance (gk) was studied in squid giant axons and in the somata of giant fiber lobe (GFL) neurons. Axon measurements were made with an axial wire voltage clamp by pulsing to VK (approximately -10 mV in 50-70 mM external K) for a variable time and then assaying available gK with a strong, brief test pulse. GFL cells were studied with whole-cell patch clamp using the same prepulse procedure as well as with long depolarizations. Under our experimental conditions (12-18 degrees C, 4 mM internal MgATP) a large fraction of gK inactivates within 250 ms at -10 mV in both cell bodies and axons, although inactivation tends to be more complete in cell bodies. Inactivation in both preparations shows two kinetic components. The faster component is more temperature-sensitive and becomes very prominent above 12 degrees C. Contribution of the fast component to inactivation shows a similar voltage dependence to that of gK, suggesting a strong coupling of this inactivation path to the open state. Omission of internal MgATP or application of internal protease reduces the amount of fast inactivation. High external K decreases the amount of rapidly inactivating IK but does not greatly alter inactivation kinetics. Neither external nor internal tetraethylammonium has a marked effect on inactivation kinetics. Squid delayed rectifier K channels in GFL cell bodies and giant axons thus share complex fast inactivation properties that do not closely resemble those associated with either C-type or N-type inactivation of cloned Kvl channels studied in heterologous expression systems.

  11. Fast Inactivation of Delayed Rectifier K Conductance in Squid Giant Axon and Its Cell Bodies

    PubMed Central

    Mathes, Chris; Rosenthal, Joshua J.C.; Armstrong, Clay M.; Gilly, William F.

    1997-01-01

    Inactivation of delayed rectifier K conductance (gK) was studied in squid giant axons and in the somata of giant fiber lobe (GFL) neurons. Axon measurements were made with an axial wire voltage clamp by pulsing to VK (∼−10 mV in 50–70 mM external K) for a variable time and then assaying available gK with a strong, brief test pulse. GFL cells were studied with whole-cell patch clamp using the same prepulse procedure as well as with long depolarizations. Under our experimental conditions (12–18°C, 4 mM internal MgATP) a large fraction of gK inactivates within 250 ms at −10 mV in both cell bodies and axons, although inactivation tends to be more complete in cell bodies. Inactivation in both preparations shows two kinetic components. The faster component is more temperature-sensitive and becomes very prominent above 12°C. Contribution of the fast component to inactivation shows a similar voltage dependence to that of gK, suggesting a strong coupling of this inactivation path to the open state. Omission of internal MgATP or application of internal protease reduces the amount of fast inactivation. High external K decreases the amount of rapidly inactivating IK but does not greatly alter inactivation kinetics. Neither external nor internal tetraethylammonium has a marked effect on inactivation kinetics. Squid delayed rectifier K channels in GFL cell bodies and giant axons thus share complex fast inactivation properties that do not closely resemble those associated with either C-type or N-type inactivation of cloned Kv1 channels studied in heterologous expression systems. PMID:9101403

  12. Application of manures to mitigate the harmful effects of electrokinetic remediation of heavy metals on soil microbial properties in polluted soils.

    PubMed

    Tahmasbian, Iman; Safari Sinegani, Ali Akbar; Nguyen, Thi Thu Nhan; Che, Rongxiao; Phan, Thuc D; Hosseini Bai, Shahla

    2017-12-01

    Ethylenediaminetetraacetic acid (EDTA) used with electrokinetic (EK) to remediate heavy metal-polluted soils is a toxic chelate for soil microorganisms. Therefore, this study aimed to evaluate the effects of alternative organic chelates to EDTA on improving the microbial properties of a heavy metal-polluted soil subjected to EK. Cow manure extract (CME), poultry manure extract (PME) and EDTA were applied to a lead (Pb) and zinc (Zn)-polluted calcareous soil which were subjected to two electric intensities (1.1 and 3.3 v/cm). Soil carbon pools, microbial activity, microbial abundance (e.g., fungal, actinomycetes and bacterial abundances) and diethylenetriaminepentaacetic acid (DTPA)-extractable Pb and Zn (available forms) were assessed in both cathodic and anodic soils. Applying the EK to soil decreased all the microbial variables in the cathodic and anodic soils in the absence or presence of chelates. Both CME and PME applied with two electric intensities decreased the negative effect of EK on soil microbial variables. The lowest values of soil microbial variables were observed when EK was combined with EDTA. The following order was observed in values of soil microbial variables after treating with EK and chelates: EK + CME or EK + PME > EK > EK + EDTA. The CME and PME could increase the concentrations of available Pb and Zn, although the increase was less than that of EDTA. Overall, despite increasing soil available Pb and Zn, the combination of EK with manures (CME or PME) mitigated the negative effects of using EK on soil microbial properties. This study suggested that the synthetic chelates such as EDTA could be replaced with manures to alleviate the environmental risks of EK application.

  13. Bacterial Imaging and Photodynamic Inactivation Using Zinc(II)-Dipicolylamine BODIPY Conjugates†

    PubMed Central

    Rice, Douglas R.; Gan, Haiying; Smith, Bradley D.

    2015-01-01

    Targeted imaging and antimicrobial photodynamic inactivation (PDI) are emerging methods for detecting and eradicating pathogenic microorganisms. This study describes two structurally related optical probes that are conjugates of a zinc(II)-dipicolylamine targeting unit and a BODIPY chromophore. One probe is a microbial targeted fluorescent imaging agent, mSeek, and the other is an oxygen photosensitizing analogue, mDestroy. The conjugates exhibited high fluorescence quantum yield and singlet oxygen production, respectively. Fluorescence imaging and detection studies examined four bacterial strains: E. coli, S. aureus, K. pneumonia, and B. thuringiensis vegetative cells and purified spores. The fluorescent probe, mSeek, is not phototoxic and enabled detection of all tested bacteria at concentrations of ~100 CFU/mL for B. thuringiensis spores, ~1000 CFU/mL for S. aureus and ~10,000 CFU/mL for E. coli. The photosensitizer analogue, mDestroy, inactivated 99–99.99% of bacterial samples and selectively killed bacterial cells in the presence of mammalian cells. However, mDestroy was ineffective against B. thuringiensis spores. Together, the results demonstrate a new two-probe strategy to optimize PDI of bacterial infection/contamination. PMID:26063101

  14. Distinct CD4+-T-cell responses to live and heat-inactivated Aspergillus fumigatus conidia.

    PubMed

    Rivera, Amariliz; Van Epps, Heather L; Hohl, Tobias M; Rizzuto, Gabrielle; Pamer, Eric G

    2005-11-01

    Aspergillus fumigatus is an important fungal pathogen that causes invasive pulmonary disease in immunocompromised hosts. Respiratory exposure to A. fumigatus spores also causes allergic bronchopulmonary aspergillosis, a Th2 CD4+-T-cell-mediated disease that accompanies asthma. The microbial factors that influence the differentiation of A. fumigatus-specific CD4+ T lymphocytes into Th1 versus Th2 cells remain incompletely defined. We therefore examined CD4+-T-cell responses of immunologically intact mice to intratracheal challenge with live or heat-inactivated A. fumigatus spores. Live but not heat-inactivated fungal spores resulted in recruitment of gamma interferon (IFN-gamma)-producing, fungus-specific CD4+ T cells to lung airways, achieving A. fumigatus-specific frequencies exceeding 5% of total CD4+ T cells. While heat-inactivated spores did not induce detectable levels of IFN-gamma-producing, A. fumigatus-specific CD4+ T cells in the airways, they did prime CD4+ T-cell responses in draining lymph nodes that produced greater amounts of interleukin 4 (IL-4) and IL-13 than T cells responding to live conidia. While immunization with live fungal spores induced antibody responses, we found a marked decrease in isotype-switched, A. fumigatus-specific antibodies in sera of mice following immunization with heat-inactivated spores. Our studies demonstrate that robust Th1 T-cell and humoral responses are restricted to challenge with fungal spores that have the potential to germinate and cause invasive infection. How the adaptive immune system distinguishes between metabolically active and inactive fungal spores remains an important question.

  15. Soil biochemical properties and microbial resilience in agroforestry systems: effects on wheat growth under controlled drought and flooding conditions.

    PubMed

    Rivest, David; Lorente, Miren; Olivier, Alain; Messier, Christian

    2013-10-01

    Agroforestry is increasingly viewed as an effective means of maintaining or even increasing crop and tree productivity under climate change while promoting other ecosystem functions and services. This study focused on soil biochemical properties and resilience following disturbance within agroforestry and conventional agricultural systems and aimed to determine whether soil differences in terms of these biochemical properties and resilience would subsequently affect crop productivity under extreme soil water conditions. Two research sites that had been established on agricultural land were selected for this study. The first site included an 18-year-old windbreak, while the second site consisted in an 8-year-old tree-based intercropping system. In each site, soil samples were used for the determination of soil nutrient availability, microbial dynamics and microbial resilience to different wetting-drying perturbations and for a greenhouse pot experiment with wheat. Drying and flooding were selected as water stress treatments and compared to a control. These treatments were initiated at the beginning of the wheat anthesis period and maintained over 10 days. Trees contributed to increase soil nutrient pools, as evidenced by the higher extractable-P (both sites), and the higher total N and mineralizable N (tree-based intercropping site) found in the agroforestry compared to the conventional agricultural system. Metabolic quotient (qCO2) was lower in the agroforestry than in the conventional agricultural system, suggesting higher microbial substrate use efficiency in agroforestry systems. Microbial resilience was higher in the agroforestry soils compared to soils from the conventional agricultural system (windbreak site only). At the windbreak site, wheat growing in soils from agroforestry system exhibited higher aboveground biomass and number of grains per spike than in conventional agricultural system soils in the three water stress treatments. At the tree

  16. Ammonia Inactivation of Ascaris Ova in Ecological Compost by Using Urine and Ash

    PubMed Central

    Parzen, Rebecca E.; Mercado Guzmán, Álvaro

    2012-01-01

    Viable ova of Ascaris lumbricoides, an indicator organism for pathogens, are frequently found in feces-derived compost produced from ecological toilets, demonstrating that threshold levels of time, temperature, pH, and moisture content for pathogen inactivation are not routinely met. Previous studies have determined that NH3 has ovicidal properties for pathogens, including Ascaris ova. This research attempted to achieve Ascaris inactivation via NH3 under environmental conditions commonly found in ecological toilets and using materials universally available in an ecological sanitation setting, including compost (feces and sawdust), urine, and ash. Compost mixed with stored urine and ash produced the most rapid inactivation, with significant inactivation observed after 2 weeks and with a time to 99% ovum inactivation (T99) of 8 weeks. Compost mixed with fresh urine and ash achieved a T99 of 15 weeks, after a 4-week lag phase. Both matrices had relatively high total-ammonia concentrations and pH values of >9.24 (pKa of ammonia). In compost mixed with ash only, and in compost mixed with fresh urine only, inactivation was observed after an 11-week lag phase. These matrices contained NH3 concentrations of 164 to 173 and 102 to 277 mg/liter, respectively, when inactivation occurred, which was below the previously hypothesized threshold for inactivation (280 mg/liter), suggesting that a lower threshold NH3 concentration may be possible with a longer contact time. Other significant results include the hydrolysis of urea to ammonia between pH values of 10.4 and 11.6, above the literature threshold pH of 10. PMID:22582051

  17. Ammonia inactivation of Ascaris ova in ecological compost by using urine and ash.

    PubMed

    McKinley, James W; Parzen, Rebecca E; Mercado Guzmán, Álvaro

    2012-08-01

    Viable ova of Ascaris lumbricoides, an indicator organism for pathogens, are frequently found in feces-derived compost produced from ecological toilets, demonstrating that threshold levels of time, temperature, pH, and moisture content for pathogen inactivation are not routinely met. Previous studies have determined that NH(3) has ovicidal properties for pathogens, including Ascaris ova. This research attempted to achieve Ascaris inactivation via NH(3) under environmental conditions commonly found in ecological toilets and using materials universally available in an ecological sanitation setting, including compost (feces and sawdust), urine, and ash. Compost mixed with stored urine and ash produced the most rapid inactivation, with significant inactivation observed after 2 weeks and with a time to 99% ovum inactivation (T(99)) of 8 weeks. Compost mixed with fresh urine and ash achieved a T(99) of 15 weeks, after a 4-week lag phase. Both matrices had relatively high total-ammonia concentrations and pH values of >9.24 (pK(a) of ammonia). In compost mixed with ash only, and in compost mixed with fresh urine only, inactivation was observed after an 11-week lag phase. These matrices contained NH(3) concentrations of 164 to 173 and 102 to 277 mg/liter, respectively, when inactivation occurred, which was below the previously hypothesized threshold for inactivation (280 mg/liter), suggesting that a lower threshold NH(3) concentration may be possible with a longer contact time. Other significant results include the hydrolysis of urea to ammonia between pH values of 10.4 and 11.6, above the literature threshold pH of 10.

  18. Effect of gamma irradiation under various atmospheres of packaging on the microbial and physicochemical properties of turmeric powder

    NASA Astrophysics Data System (ADS)

    Esmaeili, Saeideh; Barzegar, Mohsen; Sahari, Mohammad Ali; Berengi-Ardestani, Samira

    2018-07-01

    This study investigated the effect of gamma irradiation (0, 5, 10, and 15 kGy) under various atmospheres of packaging (air, N₂, and vacuum) on the microbial load and physicochemical properties of turmeric powder, including antioxidant activities, total phenolic content (TPC), color parameters, and curcuminoid content. The efficiency of irradiation in reducing microbial contamination in the samples was observed even at the lowest dose. By increasing the irradiation dose, the microbial load was not detectable. Irradiation in the presence of oxygen had synergistic effects on the extraction of curcuminoids and TPC, and increased the antioxidant activity of the methanolic extracts: highest activity was observed at 15 kGy. Generally, gamma irradiation up to the dose of 10 kGy under air atmosphere not only ensured microbial safety and desirability of turmeric powder, but also improved the extraction yield of bioactive compounds and, consequently, antioxidant activities of the samples.

  19. Kombucha tea fermentation: Microbial and biochemical dynamics.

    PubMed

    Chakravorty, Somnath; Bhattacharya, Semantee; Chatzinotas, Antonis; Chakraborty, Writachit; Bhattacharya, Debanjana; Gachhui, Ratan

    2016-03-02

    Kombucha tea, a non-alcoholic beverage, is acquiring significant interest due to its claimed beneficial properties. The microbial community of Kombucha tea consists of bacteria and yeast which thrive in two mutually non-exclusive compartments: the soup or the beverage and the biofilm floating on it. The microbial community and the biochemical properties of the beverage have so far mostly been described in separate studies. This, however, may prevent understanding the causal links between the microbial communities and the beneficial properties of Kombucha tea. Moreover, an extensive study into the microbial and biochemical dynamics has also been missing. In this study, we thus explored the structure and dynamics of the microbial community along with the biochemical properties of Kombucha tea at different time points up to 21 days of fermentation. We hypothesized that several biochemical properties will change during the course of fermentation along with the shifts in the yeast and bacterial communities. The yeast community of the biofilm did not show much variation over time and was dominated by Candida sp. (73.5-83%). The soup however, showed a significant shift in dominance from Candida sp. to Lachancea sp. on the 7th day of fermentation. This is the first report showing Candida as the most dominating yeast genus during Kombucha fermentation. Komagateibacter was identified as the single largest bacterial genus present in both the biofilm and the soup (~50%). The bacterial diversity was higher in the soup than in the biofilm with a peak on the seventh day of fermentation. The biochemical properties changed with the progression of the fermentation, i.e., beneficial properties of the beverage such as the radical scavenging ability increased significantly with a maximum increase at day 7. We further observed a significantly higher D-saccharic acid-1,4-lactone content and caffeine degradation property compared to previously described Kombucha tea fermentations. Our

  20. Properties of two Lymantria dispar nuclear polyhedrosis virus isolates obtained from the microbial pesticide Gypchek

    Treesearch

    James M. Slavicek; John Podgwaite; Carita Lanner-Herrera

    1992-01-01

    Two Lymantria dispar nuclear polyhedrosis virus isolates, 5-6 and A2-1, differing in the phenotypic characteristic of the number of viral occlusions in infected cells, were obtained from a production lot of the microbial pesticide Gypchek and several of their replication properties were investigated and compared. Budded virus titer produced in cell...

  1. Photodynamic inactivation of Paracoccidioides brasiliensis helps the outcome of oral paracoccidiodomycosis.

    PubMed

    Dos Santos, Letícia F M; Melo, Nathália B; de Carli, Marina L; Mendes, Ana Carolina S C; Bani, Giulia Maria A C; Verinaud, Liana M; Burger, Eva; de Oliveira I Moraes, Gabriel; Pereira, Alessandro A C; Brigagão, Maísa R L; Hanemann, João Adolfo C; Sperandio, Felipe F

    2017-05-01

    The antifungal drug therapy often employed to treat paracoccidiodomycosis (PCM), an important neglected fungal systemic infection, leads to offensive adverse effects, besides being very long-lasting. In addition, PCM compromises the oral health of patients by leading to oral lesions that are very painful and disabling. In that way, photodynamic therapy (PDT) arises as a new promising adjuvant treatment for inactivating Paracoccidioides brasiliensis (Pb), the responsible fungus for PCM, and also for helping the patients to deal with such debilitating oral lesions. PDT has been linked to an improved microbial killing, also presenting the advantage of not inducing immediate microbial resistance such as drugs. For the present study, we investigated the generation of reactive oxygen species (ROS) by using the fluorescent probes hydroxyphenyl fluorescein (HPF) and aminophenyl fluorescein (APF) after toluidine blue (TBO-37.5 mg/L)-mediated PDT (660 nm, 40 mW, and 0.04 cm 2 spot area) and the action of TBO-PDT upon Pb cultures grown for 7 or 15 days in semisolid Fava Netto's culture medium; we also targeted oral PCM manifestations by reporting the first clinical cases (three patients) to receive topic PDT for such purpose. We were able to show a significant generation of hydroxyl radicals and hypochlorite after TBO-PDT with doses around 90 J/cm 2 ; such ROS generation was particularly useful to attack and inactivate Pb colonies at 7 and 15 days. All three patients reported herein related an immediate relief when it came to pain, mouth opening, and also the ability to chew and swallow. As extracted from our clinical results, which are in fact based on in vitro outcomes, TBO-PDT is a very safe, inexpensive, and promising therapy for the oral manifestations of PCM.

  2. Determination of Time Dependent Virus Inactivation Rates

    NASA Astrophysics Data System (ADS)

    Chrysikopoulos, C. V.; Vogler, E. T.

    2003-12-01

    A methodology is developed for estimating temporally variable virus inactivation rate coefficients from experimental virus inactivation data. The methodology consists of a technique for slope estimation of normalized virus inactivation data in conjunction with a resampling parameter estimation procedure. The slope estimation technique is based on a relatively flexible geostatistical method known as universal kriging. Drift coefficients are obtained by nonlinear fitting of bootstrap samples and the corresponding confidence intervals are obtained by bootstrap percentiles. The proposed methodology yields more accurate time dependent virus inactivation rate coefficients than those estimated by fitting virus inactivation data to a first-order inactivation model. The methodology is successfully applied to a set of poliovirus batch inactivation data. Furthermore, the importance of accurate inactivation rate coefficient determination on virus transport in water saturated porous media is demonstrated with model simulations.

  3. The immunomodulatory properties of probiotic microorganisms beyond their viability (ghost probiotics: proposal of paraprobiotic concept).

    PubMed

    Taverniti, Valentina; Guglielmetti, Simone

    2011-08-01

    The probiotic approach represents a potentially effective and mild alternative strategy for the prevention and treatment of either inflammatory or allergic diseases. Several studies have shown that different bacterial strains can exert their probiotic abilities by influencing the host's immune system, thereby modulating immune responses. However, the emerging concern regarding safety problems arising from the extensive use of live microbial cells is enhancing the interest in non-viable microorganisms or microbial cell extracts, as they could eliminate shelf-life problems and reduce the risks of microbial translocation and infection. The purpose of this review is to provide an overview of the scientific literature concerning studies in which dead microbial cells or crude microbial cell fractions have been used as health-promoting agents. Particular attention will be given to the modulation of host immune responses. Possible mechanisms determining the effect on the immune system will also be discussed. Finally, in the light of the FAO/WHO definition of probiotics, indicating that the word 'probiotic' should be restricted to products that contain live microorganisms, and considering the scientific evidence indicating that inactivated microbes can positively affect human health, we propose the new term 'paraprobiotic' to indicate the use of inactivated microbial cells or cell fractions to confer a health benefit to the consumer.

  4. Urease from Helicobacter pylori is inactivated by sulforaphane and other isothiocyanates.

    PubMed

    Fahey, Jed W; Stephenson, Katherine K; Wade, Kristina L; Talalay, Paul

    2013-05-24

    Infections by Helicobacter pylori are very common, causing gastroduodenal inflammation including peptic ulcers, and increasing the risk of gastric neoplasia. The isothiocyanate (ITC) sulforaphane [SF; 1-isothiocyanato-4-(methylsulfinyl)butane] derived from edible crucifers such as broccoli is potently bactericidal against Helicobacter, including antibiotic-resistant strains, suggesting a possible dietary therapy. Gastric H. pylori infections express high urease activity which generates ammonia, neutralizes gastric acidity, and promotes inflammation. The finding that SF inhibits (inactivates) urease (jack bean and Helicobacter) raised the issue of whether these properties might be functionally related. The rates of inactivation of urease activity depend on enzyme and SF concentrations and show first order kinetics. Treatment with SF results in time-dependent increases in the ultraviolet absorption of partially purified Helicobacter urease in the 260-320 nm region. This provides direct spectroscopic evidence for the formation of dithiocarbamates between the ITC group of SF and cysteine thiols of urease. The potencies of inactivation of Helicobacter urease by isothiocyanates structurally related to SF were surprisingly variable. Natural isothiocyanates closely related to SF, previously shown to be bactericidal (berteroin, hirsutin, phenethyl isothiocyanate, alyssin, and erucin), did not inactivate urease activity. Furthermore, SF is bactericidal against both urease positive and negative H. pylori strains. In contrast, some isothiocyanates such as benzoyl-ITC, are very potent urease inactivators, but are not bactericidal. The bactericidal effects of SF and other ITC against Helicobacter are therefore not obligatorily linked to urease inactivation, but may reduce the inflammatory component of Helicobacter infections. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Urease from Helicobacter pylori is inactivated by sulforaphane and other isothiocyanates

    PubMed Central

    Fahey, Jed W.; Stephenson, Katherine K.; Wade, Kristina L.; Talalay, Paul

    2013-01-01

    Infections by Helicobacter pylori are very common, causing gastroduodenal inflammation including peptic ulcers, and increasing the risk of gastric neoplasia. The isothiocyanate (ITC) sulforaphane [SF; 1-isothiocyanato-4-(methylsulfinyl)butane] derived from edible crucifers such as broccoli is potently bactericidal against Helicobacter, including antibiotic-resistant strains, suggesting a possible dietary therapy. Gastric H. pylori infections express high urease activity which generates ammonia, neutralizes gastric acidity, and promotes inflammation. The finding that SF inhibits (inactivates) urease (jack bean and Helicobacter) raised the issue of whether these properties might be functionally related. The rates of inactivation of urease activity depend on enzyme and SF concentrations and show first order kinetics. Treatment with SF results in time-dependent increases in the ultraviolet absorption of partially purified Helicobacter urease in the 280–340 nm region. This provides direct spectroscopic evidence for the formation of dithiocarbamates between the ITC group of SF and cysteine thiols of urease. The potencies of inactivation of Helicobacter urease by isothiocyanates structurally related to SF were surprisingly variable. Natural isothiocyanates closely related to SF, previously shown to be bactericidal (berteroin, hirsutin, phenethyl isothiocyanate, alyssin, and erucin), did not inactivate urease activity. Furthermore, SF is bactericidal against both urease positive and negative H. pylori strains. In contrast, some isothiocyanates such as benzoyl-ITC, are very potent urease inactivators, but are not bactericidal. The bactericidal effects of SF and other ITC against Helicobacter are therefore not obligatorily linked to urease inactivation, but may reduce the inflammatory component of Helicobacter infections. PMID:23583386

  6. A Conducting State with Properties of a Slow Inactivated State in a Shaker K+ Channel Mutant

    PubMed Central

    Olcese, Riccardo; Sigg, Daniel; Latorre, Ramon; Bezanilla, Francisco; Stefani, Enrico

    2001-01-01

    In Shaker K+ channel, the amino terminus deletion Δ6-46 removes fast inactivation (N-type) unmasking a slow inactivation process. In Shaker Δ6-46 (Sh-IR) background, two additional mutations (T449V-I470C) remove slow inactivation, producing a noninactivating channel. However, despite the fact that Sh-IR-T449V-I470C mutant channels remain conductive, prolonged depolarizations (1 min, 0 mV) produce a shift of the QV curve by about −30 mV, suggesting that the channels still undergo the conformational changes typical of slow inactivation. For depolarizations longer than 50 ms, the tail currents measured during repolarization to −90 mV display a slow component that increases in amplitude as the duration of the depolarizing pulse increases. We found that the slow development of the QV shift had a counterpart in the amplitude of the slow component of the ionic tail current that is not present in Sh-IR. During long depolarizations, the time course of both the increase in the slow component of the tail current and the change in voltage dependence of the charge movement could be well fitted by exponential functions with identical time constant of 459 ms. Single channel recordings revealed that after prolonged depolarizations, the channels remain conductive for long periods after membrane repolarization. Nonstationary autocovariance analysis performed on macroscopic current in the T449V-I470C mutant confirmed that a novel open state appears with increasing prepulse depolarization time. These observations suggest that in the mutant studied, a new open state becomes progressively populated during long depolarizations (>50 ms). An appealing interpretation of these results is that the new open state of the mutant channel corresponds to a slow inactivated state of Sh-IR that became conductive. PMID:11158167

  7. Influenza Virus Inactivation for Studies of Antigenicity and Phenotypic Neuraminidase Inhibitor Resistance Profiling ▿

    PubMed Central

    Jonges, Marcel; Liu, Wai Ming; van der Vries, Erhard; Jacobi, Ronald; Pronk, Inge; Boog, Claire; Koopmans, Marion; Meijer, Adam; Soethout, Ernst

    2010-01-01

    Introduction of a new influenza virus in humans urges quick analysis of its virological and immunological characteristics to determine the impact on public health and to develop protective measures for the human population. At present, however, the necessity of executing pandemic influenza virus research under biosafety level 3 (BSL-3) high-containment conditions severely hampers timely characterization of such viruses. We tested heat, formalin, Triton X-100, and β-propiolactone treatments for their potencies in inactivating human influenza A(H3N2) and avian A(H7N3) viruses, as well as seasonal and pandemic A(H1N1) virus isolates, while allowing the specimens to retain their virological and immunological properties. Successful heat inactivation coincided with the loss of hemagglutinin (HA) and neuraminidase (NA) characteristics, and β-propiolactone inactivation reduced the hemagglutination titer and NA activity of the human influenza virus 10-fold or more. Although Triton X-100 treatment resulted in inconsistent HA activity, the NA activities in culture supernatants were enhanced consistently. Nonetheless, formalin treatment permitted the best retention of HA and NA properties. Triton X-100 treatment proved to be the easiest-to-use influenza virus inactivation protocol for application in combination with phenotypic NA inhibitor susceptibility assays, while formalin treatment preserved B-cell and T-cell epitope antigenicity, allowing the detection of both humoral and cellular immune responses. In conclusion, we demonstrated successful influenza virus characterization using formalin- and Triton X-100-inactivated virus samples. Application of these inactivation protocols limits work under BSL-3 conditions to virus culture, thus enabling more timely determination of public health impact and development of protective measures when a new influenza virus, e.g., pandemic A(H1N1)v virus, is introduced in humans. PMID:20089763

  8. Infectivity of RNA from inactivated poliovirus.

    PubMed

    Nuanualsuwan, Suphachai; Cliver, Dean O

    2003-03-01

    During inactivation of poliovirus type 1 (PV-1) by exposure to UV, hypochlorite, and heat (72 degrees C), the infectivity of the virus was compared with that of its RNA. DEAE-dextran (1-mg/ml concentration in Dulbecco's modified Eagle medium buffered with 0.05 M Tris, pH 7.4) was used to facilitate transfecting PV-1 RNA into FRhK-4 host cells. After interaction of PV-1 RNA with cell monolayer at room temperature (21 to 22 degrees C) for 20 min, the monolayers were washed with 5 ml of Hanks balanced salt solution. The remainder of the procedure was the same as that for the conventional plaque technique, which was also used for quantifying the PV-1 whole-particle infectivity. Plaque formation by extracted RNA was approximately 100,000-fold less efficient than that by whole virions. The slopes of best-fit regression lines of inactivation curves for virion infectivity and RNA infectivity were compared to determine the target of inactivation. For UV and hypochlorite inactivation the slopes of inactivation curves of virion infectivity and RNA infectivity were not statistically different. However, the difference of slopes of inactivation curves of virion infectivity and RNA infectivity was statistically significant for thermal inactivation. The results of these experiments indicate that viral RNA is a primary target of UV and hypochlorite inactivations but that the sole target of thermal inactivation is the viral capsid.

  9. Infectivity of RNA from Inactivated Poliovirus

    PubMed Central

    Nuanualsuwan, Suphachai; Cliver, Dean O.

    2003-01-01

    During inactivation of poliovirus type 1 (PV-1) by exposure to UV, hypochlorite, and heat (72°C), the infectivity of the virus was compared with that of its RNA. DEAE-dextran (1-mg/ml concentration in Dulbecco's modified Eagle medium buffered with 0.05 M Tris, pH 7.4) was used to facilitate transfecting PV-1 RNA into FRhK-4 host cells. After interaction of PV-1 RNA with cell monolayer at room temperature (21 to 22°C) for 20 min, the monolayers were washed with 5 ml of Hanks balanced salt solution. The remainder of the procedure was the same as that for the conventional plaque technique, which was also used for quantifying the PV-1 whole-particle infectivity. Plaque formation by extracted RNA was approximately 100,000-fold less efficient than that by whole virions. The slopes of best-fit regression lines of inactivation curves for virion infectivity and RNA infectivity were compared to determine the target of inactivation. For UV and hypochlorite inactivation the slopes of inactivation curves of virion infectivity and RNA infectivity were not statistically different. However, the difference of slopes of inactivation curves of virion infectivity and RNA infectivity was statistically significant for thermal inactivation. The results of these experiments indicate that viral RNA is a primary target of UV and hypochlorite inactivations but that the sole target of thermal inactivation is the viral capsid. PMID:12620852

  10. Effects of Microbial Transglutaminase on Physicochemical, Microbial and Sensorial Properties of Kefir Produced by Using Mixture Cow's and Soymilk.

    PubMed

    Temiz, Hasan; Dağyıldız, Kübra

    2017-01-01

    The objective of this research was to investigate the effects microbial transglutaminase (m-TGs) on the physicochemical, microbial and sensory properties of kefir produced by using mix cow and soymilk. Kefir batches were prepared using 0, 0.5, 1 and 1.5 Units m-TGs for per g of milk protein. Adding m-TGs to milk caused an increase in the pH and viscosity and caused a decrease in titratable acidity and syneresis in the kefir samples. Total bacteria, lactobacilli and streptococci counts decreased, while yeast counts increased in all the samples during storage. Alcohols and acids compounds have increased in all the samples except in the control samples, while carbonyl compounds have decreased in all the samples during storage (1-30 d). The differences in the percentage of alcohols, carbonyl compounds and acids in total volatiles on the 1st and the 30th d of storage were observed at 8.47-23.52%, 6.94-25.46% and 59.64-63.69%, respectively. The consumer evaluation of the kefir samples showed that greater levels of acceptability were found for samples which had been added 1.5 U m-TGs for per g of milk protein.

  11. Effect of gamma irradiation on microbial load, physicochemical and sensory characteristics of soybeans (Glycine max L. Merrill)

    USDA-ARS?s Scientific Manuscript database

    Gamma irradiation is highly effective in inactivating microorganisms in various foods and offers a safe alternative method of food decontamination. In the present study, soybeans (Glycine max L. Merrill) were treated with 0, 1.0, 3.0, 5.0 and 10.0 KGy of gamma irradiation. Microbial populations on s...

  12. Ultraviolet-C irradiation for inactivation of viruses in foetal bovine serum.

    PubMed

    Vaidya, Vivek; Dhere, Rajeev; Agnihotri, Snehal; Muley, Ravindra; Patil, Sanjay; Pawar, Amit

    2018-07-05

    Foetal Bovine Serum (FBS) and porcine trypsin are one of the essential raw materials used in the manufacturing of cell culture based viral vaccines. Being from animal origin, these raw materials can potentially contaminate the final product by known or unknown adventitious agents. The issue is more serious in case of live attenuated viral vaccines, where there is no inactivation step which can take care of such adventitious agents. It is essential to design production processes which can offer maximum viral clearance potential for animal origin products. Ultraviolet-C irradiation is known to inactivate various adventitious viral agents; however there are limited studies on ultraviolet inactivation of viruses in liquid media. We obtained a recently developed UVivatec ultraviolet-C (UV-C) irradiation based viral clearance system for evaluating its efficacy to inactivate selected model viruses. This system has a unique design with spiral path of liquid allowing maximum exposure to UV-C light of a short wavelength of 254 nm. Five live attenuated vaccine viruses and four other model viruses were spiked in tissue culture media and exposed to UV-C irradiation. The pre and post UV-C irradiation samples were analyzed for virus content to find out the extent of inactivation of various viruses. These experiments showed substantial log reduction for the majority of the viruses with few exceptions based on the characteristics of these viruses. Having known the effect of UV irradiation on protein structure, we also evaluated the post irradiation samples of culture media for growth promoting properties using one of the most fastidious human diploid cells (MRC-5). UV-C exposure did not show any notable impact on the nutritional properties of culture media. The use of an UV-C irradiation based system is considered to be promising approach to mitigate the risk of adventitious agents in cell culture media arising through animal derived products. Copyright © 2018 Elsevier Ltd. All

  13. Microbial Safety and Shelf Life of UV-C Treated Freshly Squeezed White Grape Juice.

    PubMed

    Unluturk, Sevcan; Atilgan, Mehmet R

    2015-08-01

    The effects of UV-C irradiation on the inactivation of Escherichia coli K-12 (ATCC 25253), a surrogate of E. coli O157:H7, and on the shelf life of freshly squeezed turbid white grape juice (FSWGJ) were investigated. FSWGJ samples were processed at 0.90 mL/s for 32 min by circulating 8 times in an annular flow UV system. The UV exposure time was 244 s per cycle. The population of E. coli K-12 was reduced by 5.34 log cycles after exposure to a total UV dosage of 9.92 J/cm(2) (1.24 J/cm(2) per cycle) at 0.90 mL/s flow rate. The microbial shelf life of UV-C treated FSWGJ was extended up to 14 d at 4 °C. UV exposure was not found to alter pH, total soluble solid, and titratable acidity of juice. There was a significant effect (P < 0.05) on turbidity, absorbance coefficient, color, and ascorbic acid content. Furthermore, all physicochemical properties were altered during refrigerated storage. The microbial shelf life of FSWGJ was doubled after UV-C treatment, whereas the quality of juice was adversely affected similarly observed in the control samples. © 2015 Institute of Food Technologists®

  14. Rust preventive oil additives based on microbial fats

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

    Salenko, V.I.; Fedorov, V.V.; Kazantsev, Yu.E.

    1983-03-01

    This article investigates the composition and lubricating properties of microbial fats obtained from microorganisms grown on various hydrocarbon substrates (n-paraffins, alcohols, natural gas, petroleum distillates, etc.). Focuses on the protective functions of the 4 main fractions (unsaponifiables, free fatty acids, glycerides, and phospholipids) which comprise the microbial fat from a yeast grown on purified liquid n-paraffins. Concludes that neutralized microbial fats can be used as preservative additives; that the principal components of the microbial fats have the properties necessary for oil-soluble corrosion inhibitors; that the phospholipids of the microbial fat can fulfill the functions of not only preservative additives, butmore » also highly effective operational/ preservative additives; and that fats of microbial origin can be used in the development of multipurpose polyfunctional additives.« less

  15. Inactivation of coliphage Q beta by potassium ferrate.

    PubMed

    Kazama, F

    1994-05-15

    The kinetics of inactivation of a bacteriophage by potassium ferrate were studied with the F-specific RNA-coliphage Q beta. Inactivation in phosphate buffer (pH 6, 7 and 8) containing ferrate could be described by Hom's model. The inactivation rate depended on the pH. However, the relative effects of ferrate concentration and exposure time on inactivation were not affected by a change in pH from 6 to 8. In a study of the mechanism by which ferrate inactivated the virus, the efficiency of viral inactivation after ferrate decomposed in buffer was assayed. Inactivation was still effective and still followed Hom's equation after the complete decomposition of ferrate ion; however, the efficiency of that inactivation disappeared when sodium thiosulfate was added, suggesting that long-lived oxidative intermediates capable of viral inactivation were generated during the decomposition of ferrate ions.

  16. Treatment alternatives of slaughterhouse wastes, and their effect on the inactivation of different pathogens: a review.

    PubMed

    Franke-Whittle, Ingrid H; Insam, Heribert

    2013-05-01

    Slaughterhouse wastes are a potential reservoir of bacterial, viral, prion and parasitic pathogens, capable of infecting both animals and humans. A quick, cost effective and safe disposal method is thus essential in order to reduce the risk of disease following animal slaughter. Different methods for the disposal of such wastes exist, including composting, anaerobic digestion (AD), alkaline hydrolysis (AH), rendering, incineration and burning. Composting is a disposal method that allows a recycling of the slaughterhouse waste nutrients back into the earth. The high fat and protein content of slaughterhouse wastes mean however, that such wastes are an excellent substrate for AD processes, resulting in both the disposal of wastes, a recycling of nutrients (soil amendment with sludge), and in methane production. Concerns exist as to whether AD and composting processes can inactivate pathogens. In contrast, AH is capable of the inactivation of almost all known microorganisms. This review was conducted in order to compare three different methods of slaughterhouse waste disposal, as regards to their ability to inactivate various microbial pathogens. The intention was to investigate whether AD could be used for waste disposal (either alone, or in combination with another process) such that both energy can be obtained and potentially hazardous materials be disposed of.

  17. Treatment alternatives of slaughterhouse wastes, and their effect on the inactivation of different pathogens: A review

    PubMed Central

    2013-01-01

    Slaughterhouse wastes are a potential reservoir of bacterial, viral, prion and parasitic pathogens, capable of infecting both animals and humans. A quick, cost effective and safe disposal method is thus essential in order to reduce the risk of disease following animal slaughter. Different methods for the disposal of such wastes exist, including composting, anaerobic digestion (AD), alkaline hydrolysis (AH), rendering, incineration and burning. Composting is a disposal method that allows a recycling of the slaughterhouse waste nutrients back into the earth. The high fat and protein content of slaughterhouse wastes mean however, that such wastes are an excellent substrate for AD processes, resulting in both the disposal of wastes, a recycling of nutrients (soil amendment with sludge), and in methane production. Concerns exist as to whether AD and composting processes can inactivate pathogens. In contrast, AH is capable of the inactivation of almost all known microorganisms. This review was conducted in order to compare three different methods of slaughterhouse waste disposal, as regards to their ability to inactivate various microbial pathogens. The intention was to investigate whether AD could be used for waste disposal (either alone, or in combination with another process) such that both energy can be obtained and potentially hazardous materials be disposed of. PMID:22694189

  18. Soil microbial properties after 5 years of consecutive amendment with composted tannery sludge.

    PubMed

    Araujo, Ademir Sérgio Ferreira; Miranda, Ana Roberta Lima; Oliveira, Mara Lucia Jacinto; Santos, Vilma Maria; Nunes, Luís Alfredo Pinheiro Leal; Melo, Wanderley José

    2015-01-01

    Composting has been recognised an alternative method to tannery sludge recycling and afterwards to be used in agriculture. As the tannery sludge contains salts and chromium, the application of composted tannery sludge (CTS) should be performed carefully to minimise negative effects on soil microbial properties. Therefore, this study evaluated the effects of 5-year repeated CTS amendment on soil microbial biomass (SMB) and enzyme activities in a tropical soil. CTS was applied during 5 years at 0, 2.5, 5, 10 and 20 Mg ha(-1), and at the fifth year, the microbial biomass C (MBC) and N (MBN), basal and substrate-induced respiration (SIR), metabolic quotient (qCO₂) and dehydrogenase (DHA) and fluorescein diacetate (FDA) hydrolysis were determined in the soil samples. Soil MBC and MBN showed the highest values with the amendment of 5 Mg ha(-1) CTS. Soil respiration increased with the increase in CTS rates, while SIR showed the highest values with the amendment of 0, 2.5 and 5 Mg ha(-1) CTS. DHA activity showed the highest values with the amendment up to 2.5 Mg ha(-1), while FDA hydrolysis increased up to the rate of 5 Mg ha(-1) CTS. The results show that after 5 years of permanent amendment of CTS, soils amended with 2.5 Mg ha(-1) have SMB and enzymatic activities similar to those in unamended soil.

  19. Peracetic Acid Treatment Generates Potent Inactivated Oral Vaccines from a Broad Range of Culturable Bacterial Species

    PubMed Central

    Moor, Kathrin; Wotzka, Sandra Y.; Toska, Albulena; Diard, Médéric; Hapfelmeier, Siegfried; Slack, Emma

    2016-01-01

    Our mucosal surfaces are the main sites of non-vector-borne pathogen entry, as well as the main interface with our commensal microbiota. We are still only beginning to understand how mucosal adaptive immunity interacts with commensal and pathogenic microbes to influence factors such as infectivity, phenotypic diversity, and within-host evolution. This is in part due to difficulties in generating specific mucosal adaptive immune responses without disrupting the mucosal microbial ecosystem itself. Here, we present a very simple tool to generate inactivated mucosal vaccines from a broad range of culturable bacteria. Oral gavage of 1010 peracetic acid-inactivated bacteria induces high-titer-specific intestinal IgA in the absence of any measurable inflammation or species invasion. As a proof of principle, we demonstrate that this technique is sufficient to provide fully protective immunity in the murine model of invasive non-typhoidal Salmonellosis, even in the face of severe innate immune deficiency. PMID:26904024

  20. Bacterial inactivation of the anticancer drug doxorubicin.

    PubMed

    Westman, Erin L; Canova, Marc J; Radhi, Inas J; Koteva, Kalinka; Kireeva, Inga; Waglechner, Nicholas; Wright, Gerard D

    2012-10-26

    Microbes are exposed to compounds produced by members of their ecological niche, including molecules with antibiotic or antineoplastic activities. As a result, even bacteria that do not produce such compounds can harbor the genetic machinery to inactivate or degrade these molecules. Here, we investigated environmental actinomycetes for their ability to inactivate doxorubicin, an aminoglycosylated anthracycline anticancer drug. One strain, Streptomyces WAC04685, inactivates doxorubicin via a deglycosylation mechanism. Activity-based purification of the enzymes responsible for drug inactivation identified the NADH dehydrogenase component of respiratory electron transport complex I, which was confirmed by gene inactivation studies. A mechanism where reduction of the quinone ring of the anthracycline by NADH dehydrogenase leads to deglycosylation is proposed. This work adds anticancer drug inactivation to the enzymatic inactivation portfolio of actinomycetes and offers possibilities for novel applications in drug detoxification. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Thin-film fixed-bed reactor for solar photocatalytic inactivation of Aeromonas hydrophila: influence of water quality

    PubMed Central

    2012-01-01

    Background Controlling fish disease is one of the major concerns in contemporary aquaculture. The use of antibiotics or chemical disinfection cannot provide a healthy aquaculture system without residual effects. Water quality is also important in determining the success or failure of fish production. Several solar photocatalytic reactors have been used to treat drinking water or waste water without leaving chemical residues. This study has investigated the impact of several key aspects of water quality on the inactivation of the pathogenic bacterium Aeromonas hydrophila using a pilot-scale thin-film fixed-bed reactor (TFFBR) system. Results The level of inactivation of Aeromonas hydrophila ATCC 35654 was determined using a TFFBR with a photocatalytic area of 0.47 m2 under the influence of various water quality variables (pH, conductivity, turbidity and colour) under high solar irradiance conditions (980–1100 W m-2), at a flow rate of 4.8 L h-1 through the reactor. Bacterial enumeration were obtained through conventional plate count using trypticase soy agar media, cultured in conventional aerobic conditions to detect healthy cells and under ROS-neutralised conditions to detect both healthy and sub-lethally injured (oxygen-sensitive) cells. The results showed that turbidity has a major influence on solar photocatalytic inactivation of A. hydrophila. Humic acids appear to decrease TiO2 effectiveness under full sunlight and reduce microbial inactivation. pH in the range 7–9 and salinity both have no major effect on the extent of photoinactivation or sub-lethal injury. Conclusions This study demonstrates the effectiveness of the TFFBR in the inactivation of Aeromonas hydrophila under the influence of several water quality variables at high solar irradiance, providing an opportunity for the application of solar photocatalysis in aquaculture systems, as long as turbidity remains low. PMID:23194331

  2. Effective Chemical Inactivation of Ebola Virus

    PubMed Central

    Haddock, Elaine; Feldmann, Friederike

    2016-01-01

    Reliable inactivation of specimens before removal from high-level biocontainment is crucial for safe operation. To evaluate efficacy of methods of chemical inactivation, we compared in vitro and in vivo approaches using Ebola virus as a surrogate pathogen. Consequently, we have established parameters and protocols leading to reliable and effective inactivation. PMID:27070504

  3. Efficacy of atmospheric pressure dielectric barrier discharge for inactivating airborne pathogens

    DOE PAGES

    Romero-Mangado, Jaione; Dey, Avishek; Diaz-Cartagena, Diana C.; ...

    2017-07-05

    Atmospheric pressure plasmas have gained attention in recent years for several environmental applications. This technology could potentially be used to deactivate airborne microorganisms, surface-bound microorganisms, and biofilms. Here, the authors explore the efficacy of the atmospheric pressure dielectric barrier discharge (DBD) to inactivate airborne Staphylococcus epidermidis and Aspergillus niger that are opportunistic pathogens associated with nosocomial infections. This technology uses air as the source of gas and does not require any process gas such as helium, argon, nitrogen, or hydrogen. Moreover, the effect of DBD was studied on aerosolized S. epidermidis and aerosolized A. niger spores via scanning electron microscopymore » (SEM). The morphology observed on the SEM micrographs showed deformations in the cellular structure of both microorganisms. Cell structure damage upon interaction with the DBD suggests leakage of vital cellular materials, which is a key mechanism for microbial inactivation. The chemical structure of the cell surface of S. epidermidis was also analyzed by near edge x-ray absorption fine structure spectroscopy before and after DBD exposure. Our results from surface analysis revealed that reactive oxygen species from the DBD discharge contributed to alterations on the chemistry of the cell membrane/cell wall of S. epidermidis.« less

  4. Efficacy of atmospheric pressure dielectric barrier discharge for inactivating airborne pathogens

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

    Romero-Mangado, Jaione; Dey, Avishek; Diaz-Cartagena, Diana C.

    Atmospheric pressure plasmas have gained attention in recent years for several environmental applications. This technology could potentially be used to deactivate airborne microorganisms, surface-bound microorganisms, and biofilms. Here, the authors explore the efficacy of the atmospheric pressure dielectric barrier discharge (DBD) to inactivate airborne Staphylococcus epidermidis and Aspergillus niger that are opportunistic pathogens associated with nosocomial infections. This technology uses air as the source of gas and does not require any process gas such as helium, argon, nitrogen, or hydrogen. Moreover, the effect of DBD was studied on aerosolized S. epidermidis and aerosolized A. niger spores via scanning electron microscopymore » (SEM). The morphology observed on the SEM micrographs showed deformations in the cellular structure of both microorganisms. Cell structure damage upon interaction with the DBD suggests leakage of vital cellular materials, which is a key mechanism for microbial inactivation. The chemical structure of the cell surface of S. epidermidis was also analyzed by near edge x-ray absorption fine structure spectroscopy before and after DBD exposure. Our results from surface analysis revealed that reactive oxygen species from the DBD discharge contributed to alterations on the chemistry of the cell membrane/cell wall of S. epidermidis.« less

  5. Long-Term Coffee Monoculture Alters Soil Chemical Properties and Microbial Communities.

    PubMed

    Zhao, Qingyun; Xiong, Wu; Xing, Yizhang; Sun, Yan; Lin, Xingjun; Dong, Yunping

    2018-04-17

    Long-term monoculture severely inhibits coffee plant growth, decreases its yield and results in serious economic losses in China. Here, we selected four replanted coffee fields with 4, 18, 26 and 57 years of monoculture history in Hainan China to investigate the influence of continuous cropping on soil chemical properties and microbial communities. Results showed long-term monoculture decreased soil pH and organic matter content and increased soil EC. Soil bacterial and fungal richness decreased with continuous coffee cropping. Principal coordinate analysis suggested monoculture time was a major determinant of bacterial and fungal community structures. Relative abundances of bacterial Proteobacteria, Bacteroidetes and Nitrospira and fungal Ascomycota phyla decreased over time. At genus level, potentially beneficial microbes such as Nitrospira and Trichoderma, significantly declined over time and showed positive relationships with coffee plant growth in pots. In conclusion, continuous coffee cropping decreased soil pH, organic matter content, potentially beneficial microbes and increased soil EC, which might lead to the poor growth of coffee plants in pots and decline of coffee yields in fields. Thus, developing sustainable agriculture to improve soil pH, organic matter content, microbial activity and reduce the salt stress under continuous cropping system is important for coffee production in China.

  6. The Protein/Peptide Direct Virus Inactivation During Chromatographic Process: Developing Approaches.

    PubMed

    Volkov, Georgii L; Havryliuk, Sergiy P; Krasnobryzha, Ievgenia M; Havryliuk, Olena S

    2017-01-01

    Virus clearance is required for pharmaceutical preparations derived from animal or human sources such as blood products, vaccines, recombinant proteins produced in mammalian cell lines, etc. High cost and substantial protein losses during virus inactivation are significant problems for protein/peptide manufacturing. The goal of this project was to develop a method to perform virus inactivation in a course of protein chromatographic purification. Another goal was to show that the chromatographic adsorbent can serve as reliable "sieva" for mechanical washing away of infecting viruses. Using chromatographic, photometric, IFA, and RT-PCR approaches, it was discovered that high temperature-depending dynamic capacity of adsorbent allowed to perform a virus inactivation directly in a chromatographic column by solvent/detergent treatment. The peptide/protein biological activity was completely preserved. Using this new approach enveloped and nonenveloped viruses were effectively removed protein preparation. In addition, it was shown that RT-PCR method demonstrates more precise and reproducible results and robust properties for assessment of virus reduction than virus titer followed by infectivity studies. Presented method allowed to obtain the factor of virus concentration decrease (FVD) values that were higher than those provided by known technologies and was sufficient for a full inactivation of viruses. The method is recommended to use in pharmaceutical industry.

  7. Spatial heterogeneity of physicochemical properties explains differences in microbial composition in arid soils from Cuatro Cienegas, Mexico.

    PubMed

    Pajares, Silvia; Escalante, Ana E; Noguez, Ana M; García-Oliva, Felipe; Martínez-Piedragil, Celeste; Cram, Silke S; Eguiarte, Luis Enrique; Souza, Valeria

    2016-01-01

    Arid ecosystems are characterized by high spatial heterogeneity, and the variation among vegetation patches is a clear example. Soil biotic and abiotic factors associated with these patches have also been well documented as highly heterogeneous in space. Given the low vegetation cover and little precipitation in arid ecosystems, soil microorganisms are the main drivers of nutrient cycling. Nonetheless, little is known about the spatial distribution of microorganisms and the relationship that their diversity holds with nutrients and other physicochemical gradients in arid soils. In this study, we evaluated the spatial variability of soil microbial diversity and chemical parameters (nutrients and ion content) at local scale (meters) occurring in a gypsum-based desert soil, to gain knowledge on what soil abiotic factors control the distribution of microbes in arid ecosystems. We analyzed 32 soil samples within a 64 m(2) plot and: (a) characterized microbial diversity using T-RFLPs of the bacterial 16S rRNA gene, (b) determined soil chemical parameters, and (c) identified relationships between microbial diversity and chemical properties. Overall, we found a strong correlation between microbial composition heterogeneity and spatial variation of cations (Ca(2), K(+)) and anions (HCO[Formula: see text], Cl(-), SO[Formula: see text]) content in this small plot. Our results could be attributable to spatial differences of soil saline content, favoring the patchy emergence of salt and soil microbial communities.

  8. Microbial utilization of low molecular weight organics in soil depends on the substances properties

    NASA Astrophysics Data System (ADS)

    Gunina, Anna

    2016-04-01

    Utilization of low molecular weight organic substances (LMWOS) in soil is regulated by microbial uptake from solution and following incorporation of into specific cell cycles. Various chemical properties of LMWOS, namely oxidation state, number of carbon (C) atoms, number of carboxylic (-COOH) groups, can affect their uptake from soil solution and further microbial utilization. The aim of the study was to trace the initial fate (including the uptake from soil solution and utilization by microorganisms) of three main classes of LMWOS, having contrast properties - sugars, carboxylic and amino acids. Top 10 cm of mineral soil were collected under Silver birch stands within the Bangor DIVERSE experiment, UK. Soil solution was extracted by centrifugation at 4000 rpm during 15 min. Soil was spiked with 14C glucose or fructose; malic, succinic or formic acids; alanine or glycine. No additional non-labeled LMWOS were added. 14C was traced in the dissolved organic matter (DOM), CO2, cytosol and soil organic matter (SOM) during one day. To estimate half-life times (T1 /2)of LMWOS in soil solution and in SOM pools, the single and double first order kinetic equations were fitted to the uptake and mineralization dynamics, respectively. The LMWOS T1 /2in DOM pool varied between 0.6-5 min, with the highest T1 /2for sugars (3.7 min) and the lowest for carboxylic acids (0.6-1.4 min). Thus, initial uptake of LMWOS is not a limiting step of microbial utilization. The T1 /2 of carboxylic and amino acids in DOM were closely related with oxidation state, showing that reduced substances remain in soil solution longer, than oxidized. The initial T1 /2 of LMWOS in SOM ranged between 30-80 min, with the longest T1 /2 for amino acids (50-80 min) and the shortest for carboxylic acids (30-48 min). These T1 /2values were in one-two orders of magnitude higher than LMWOS T1 /2 in soil solution, pointing that LMWOS mineralization occur with a delay after the uptake. Absence of correlations between

  9. Long-Term Application of Bioorganic Fertilizers Improved Soil Biochemical Properties and Microbial Communities of an Apple Orchard Soil

    PubMed Central

    Wang, Lei; Yang, Fang; E, Yaoyao; Yuan, Jun; Raza, Waseem; Huang, Qiwei; Shen, Qirong

    2016-01-01

    Soil biochemical properties and microbial communities are usually considered as important indicators of soil health because of their association with plant nutrition. In this study, we investigated the impact of long-term application of bioorganic fertilizer (BOF) on soil biochemical properties and microbial communities in the apple orchard soil of the Loess Plateau. The experiment included three treatments: (1) control without fertilization (CK); (2) chemical fertilizer application (CF); and (3) bioorganic fertilizer application (BOF). The high throughput sequencing was used to examine the bacterial and fungal communities in apple orchard soil. The results showed that the BOF treatment significantly increased the apple yield during the experimental time (2009–2015). The application of BOF significantly increased the activities of catalase and invertase compared to those in CK and CF treatments. The high throughput sequencing data showed that the application of BOF changed the microbial community composition of all soil depths considered (0–20 cm, 20–40 cm, and 40–60 cm), e.g., the relative abundance of bio-control bacteria (Xanthomonadales, Lysobacter, Pseudomonas, and Bacillus), Proteobacteria, Bacteroidetes, Ohtaekwangia, Ilyonectria, and Lecanicillium was increased while that of Acidobacteria, Chloroflexi, Gp4, Gp6 and Sphaerobacter was decreased. The increase in apple yield after the application of BOF might be due to increase in organic matter, total nitrogen and catalase and invertase activities of soil and change in the bacterial community composition by enriching Bacillus, Pseudomonas, Lysobacter, and Ohtaekwangia. These results further enhance the understanding on how BOFs alter soil microbial community composition to stimulate soil productivity. PMID:27965631

  10. Long-Term Application of Bioorganic Fertilizers Improved Soil Biochemical Properties and Microbial Communities of an Apple Orchard Soil.

    PubMed

    Wang, Lei; Yang, Fang; E, Yaoyao; Yuan, Jun; Raza, Waseem; Huang, Qiwei; Shen, Qirong

    2016-01-01

    Soil biochemical properties and microbial communities are usually considered as important indicators of soil health because of their association with plant nutrition. In this study, we investigated the impact of long-term application of bioorganic fertilizer (BOF) on soil biochemical properties and microbial communities in the apple orchard soil of the Loess Plateau. The experiment included three treatments: (1) control without fertilization (CK); (2) chemical fertilizer application (CF); and (3) bioorganic fertilizer application (BOF). The high throughput sequencing was used to examine the bacterial and fungal communities in apple orchard soil. The results showed that the BOF treatment significantly increased the apple yield during the experimental time (2009-2015). The application of BOF significantly increased the activities of catalase and invertase compared to those in CK and CF treatments. The high throughput sequencing data showed that the application of BOF changed the microbial community composition of all soil depths considered (0-20 cm, 20-40 cm, and 40-60 cm), e.g., the relative abundance of bio-control bacteria ( Xanthomonadales, Lysobacter, Pseudomonas , and Bacillus ), Proteobacteria, Bacteroidetes, Ohtaekwangia, Ilyonectria , and Lecanicillium was increased while that of Acidobacteria, Chloroflexi, Gp4, Gp6 and Sphaerobacter was decreased. The increase in apple yield after the application of BOF might be due to increase in organic matter, total nitrogen and catalase and invertase activities of soil and change in the bacterial community composition by enriching Bacillus, Pseudomonas, Lysobacter , and Ohtaekwangia . These results further enhance the understanding on how BOFs alter soil microbial community composition to stimulate soil productivity.

  11. Long-term reactive nitrogen loading alters soil carbon and microbial community properties in a subalpine forest ecosystem

    USGS Publications Warehouse

    Boot, Claudia M.; Hall, Ed K.; Denef, Karolien; Baron, Jill S.

    2016-01-01

    Elevated nitrogen (N) deposition due to increased fossil fuel combustion and agricultural practices has altered global carbon (C) cycling. Additions of reactive N to N-limited environments are typically accompanied by increases in plant biomass. Soil C dynamics, however, have shown a range of different responses to the addition of reactive N that seem to be ecosystem dependent. We evaluated the effect of N amendments on biogeochemical characteristics and microbial responses of subalpine forest organic soils in order to develop a mechanistic understanding of how soils are affected by N amendments in subalpine ecosystems. We measured a suite of responses across three years (2011–2013) during two seasons (spring and fall). Following 17 years of N amendments, fertilized soils were more acidic (control mean 5.09, fertilized mean 4.68), and had lower %C (control mean 33.7% C, fertilized mean 29.8% C) and microbial biomass C by 22% relative to control plots. Shifts in biogeochemical properties in fertilized plots were associated with an altered microbial community driven by reduced arbuscular mycorrhizal (control mean 3.2 mol%, fertilized mean 2.5 mol%) and saprotrophic fungal groups (control mean 17.0 mol%, fertilized mean 15.2 mol%), as well as a decrease in N degrading microbial enzyme activity. Our results suggest that decreases in soil C in subalpine forests were in part driven by increased microbial degradation of soil organic matter and reduced inputs to soil organic matter in the form of microbial biomass.

  12. Different efficiency of ozonated water washing to inactivate Salmonella enterica typhimurium on green onions, grape tomatoes, and green leaf lettuces.

    PubMed

    Xu, Wenqing; Wu, Changqing

    2014-03-01

    Ozonated water washing is one of the emerging techniques to inactivate foodborne pathogens on produce, and limited information is available to optimize processing parameters (treatment time, temperature, and pH) to improve ozone efficacy on Salmonella inactivation for different produce. The efficacy of ozonated water washing for inactivation of Salmonella enterica Typhimurium on green onions, grape tomatoes and green leaf lettuces were studied in our research. Surface inoculated fresh produce were washed by ozonated water for 1, 5, or 10 min at room temperature and pH 5.60 ± 0.03. Then efficacy of ozonated water washing at mild heated (50 °C) and refrigerated (4 °C) temperature for 5 min with pH 5.60 ± 0.03 was investigated. Salmonella inactivation efficacy under pH 5.60 ± 0.03 and 2.64 ± 0.02 with 5 min washing at room temperature were also compared. Our results showed that Salmonella inactivation by ozonated water was time-dependent for 3 fresh produce. Mild heated temperature (50 °C) and pH 2.64 ± 0.02 improved efficacy of ozonated water to inactivate Salmonella on tomatoes and lettuces, but not on green onions. It is suggested that different surface structures of fresh produce significantly impact the antimicrobial efficacy of ozonated water washing operated under various parameters (time, temperature, and pH). Washing is the essential step for green onions and lettuces in the packinghouse and grape tomatoes in the restaurants and grocery stores having salad bars. Ozonated water can be used as disinfectant to reduce microbial contamination (FDA). The effectiveness of this disinfectant depends on the type of product and treatment conditions, such as water temperature, acidity, contact time. Our study showed that Salmonella inactivation by ozonated water washing was time-dependent. Mild heat and low pH improved inactivation efficacy on tomatoes and lettuces, but not on green onions. Processors should consider adjustments that are most appropriate for

  13. Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components. [examined with a scanning electron microscope

    NASA Technical Reports Server (NTRS)

    Campbell, J. E.

    1974-01-01

    The uses of scanning electron microscopy in assessing changes that occur in spores exposed to wet and dry heat cycles at elevated temperatures were examined. Several species of Bacillus and other nonspore-forming species of organisms were used for the experiment. Surface morphology of viable and nonviable organisms was clearly detectable by this method, making it a potentially useful technique for investigating microbial inactivation on space vehicle surfaces and components. Micrographs of the spores and bacterial cells are provided.

  14. Optimization of a nanotechnology based antimicrobial platform for food safety applications using Engineered Water Nanostructures (EWNS)

    PubMed Central

    Pyrgiotakis, Georgios; Vedantam, Pallavi; Cirenza, Caroline; McDevitt, James; Eleftheriadou, Mary; Leonard, Stephen S.; Demokritou, Philip

    2016-01-01

    A chemical free, nanotechnology-based, antimicrobial platform using Engineered Water Nanostructures (EWNS) was recently developed. EWNS have high surface charge, are loaded with reactive oxygen species (ROS), and can interact-with, and inactivate an array of microorganisms, including foodborne pathogens. Here, it was demonstrated that their properties during synthesis can be fine tuned and optimized to further enhance their antimicrobial potential. A lab based EWNS platform was developed to enable fine-tuning of EWNS properties by modifying synthesis parameters. Characterization of EWNS properties (charge, size and ROS content) was performed using state-of-the art analytical methods. Further their microbial inactivation potential was evaluated with food related microorganisms such as Escherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium parafortuitum, and Saccharomyces cerevisiae inoculated onto the surface of organic grape tomatoes. The results presented here indicate that EWNS properties can be fine-tuned during synthesis resulting in a multifold increase of the inactivation efficacy. More specifically, the surface charge quadrupled and the ROS content increased. Microbial removal rates were microorganism dependent and ranged between 1.0 to 3.8 logs after 45 mins of exposure to an EWNS aerosol dose of 40,000 #/cm3. PMID:26875817

  15. Optimization of a nanotechnology based antimicrobial platform for food safety applications using Engineered Water Nanostructures (EWNS)

    NASA Astrophysics Data System (ADS)

    Pyrgiotakis, Georgios; Vedantam, Pallavi; Cirenza, Caroline; McDevitt, James; Eleftheriadou, Mary; Leonard, Stephen S.; Demokritou, Philip

    2016-02-01

    A chemical free, nanotechnology-based, antimicrobial platform using Engineered Water Nanostructures (EWNS) was recently developed. EWNS have high surface charge, are loaded with reactive oxygen species (ROS), and can interact-with, and inactivate an array of microorganisms, including foodborne pathogens. Here, it was demonstrated that their properties during synthesis can be fine tuned and optimized to further enhance their antimicrobial potential. A lab based EWNS platform was developed to enable fine-tuning of EWNS properties by modifying synthesis parameters. Characterization of EWNS properties (charge, size and ROS content) was performed using state-of-the art analytical methods. Further their microbial inactivation potential was evaluated with food related microorganisms such as Escherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium parafortuitum, and Saccharomyces cerevisiae inoculated onto the surface of organic grape tomatoes. The results presented here indicate that EWNS properties can be fine-tuned during synthesis resulting in a multifold increase of the inactivation efficacy. More specifically, the surface charge quadrupled and the ROS content increased. Microbial removal rates were microorganism dependent and ranged between 1.0 to 3.8 logs after 45 mins of exposure to an EWNS aerosol dose of 40,000 #/cm3.

  16. Optimization of a nanotechnology based antimicrobial platform for food safety applications using Engineered Water Nanostructures (EWNS).

    PubMed

    Pyrgiotakis, Georgios; Vedantam, Pallavi; Cirenza, Caroline; McDevitt, James; Eleftheriadou, Mary; Leonard, Stephen S; Demokritou, Philip

    2016-02-15

    A chemical free, nanotechnology-based, antimicrobial platform using Engineered Water Nanostructures (EWNS) was recently developed. EWNS have high surface charge, are loaded with reactive oxygen species (ROS), and can interact-with, and inactivate an array of microorganisms, including foodborne pathogens. Here, it was demonstrated that their properties during synthesis can be fine tuned and optimized to further enhance their antimicrobial potential. A lab based EWNS platform was developed to enable fine-tuning of EWNS properties by modifying synthesis parameters. Characterization of EWNS properties (charge, size and ROS content) was performed using state-of-the art analytical methods. Further their microbial inactivation potential was evaluated with food related microorganisms such as Escherichia coli, Salmonella enterica, Listeria innocua, Mycobacterium parafortuitum, and Saccharomyces cerevisiae inoculated onto the surface of organic grape tomatoes. The results presented here indicate that EWNS properties can be fine-tuned during synthesis resulting in a multifold increase of the inactivation efficacy. More specifically, the surface charge quadrupled and the ROS content increased. Microbial removal rates were microorganism dependent and ranged between 1.0 to 3.8 logs after 45 mins of exposure to an EWNS aerosol dose of 40,000 #/cm(3).

  17. Modulation of slow inactivation in human cardiac Kv1.5 channels by extra- and intracellular permeant cations

    PubMed Central

    Fedida, David; Maruoka, Neil D; Lin, Shunping

    1999-01-01

    The properties and regulation of slow inactivation by intracellular and extracellular cations in the human heart K+ channel hKv1.5 have been investigated. Extensive NH2- and COOH-terminal deletions outside the central core of transmembrane domains did not affect the degree of inactivation. The voltage dependence of steady-state inactivation curves of hKv1.5 channels was unchanged in Rb+ and Cs+, compared with K+, but biexponential inactivation over 10 s was reduced from ∼100% of peak current in Na+ to ∼65% in K+, ∼50% in Rb+ and ∼30% in Cs+. This occurred as a result of a decrease in both fast and slow components of inactivation, with little change in inactivation time constants. Changes in extracellular cation species and concentration (5-300 mM) had only small effects on the rates of inactivation and recovery from inactivation (τrecovery∼1 s). Mutation of residues at a putative regulatory site at R487 in the outer pore mouth did not affect slow inactivation or recovery from inactivation of hKv1.5, although sensitivity to extracellular TEA was conferred. Symmetrical reduction of both intra- and extracellular cation concentrations accelerated and augmented both components of inactivation of K+ (Kd = 34.7 mM) and Cs+ (Kd = 20.5 mM) currents. These effects could be quantitatively accounted for by unilateral reduction of intracellular K+ (Ki+) (Kd = 43.4 mM) or Csi+ with constant 135 mM external ion concentrations. We conclude that inactivation and recovery from inactivation in hKv1.5 were not typically C-type in nature. However, the ion species dependence of inactivation was still closely coupled to ion permeation through the pore. Intracellular ion modulatory actions were more potent than extracellular actions, although still of relatively low affinity. These results suggest the presence of ion binding sites capable of regulating inactivation located on both intracellular and extracellular sides of the pore selectivity filter. PMID:10050000

  18. Chlorine inactivation of Tubifex tubifex in drinking water and the synergistic effect of sequential inactivation with UV irradiation and chlorine.

    PubMed

    Nie, Xiao-Bao; Li, Zhi-Hong; Long, Yuan-Nan; He, Pan-Pan; Xu, Chao

    2017-06-01

    The inactivation of Tubifex tubifex is important to prevent contamination of drinking water. Chlorine is a widely-used disinfectant and the key factor in the inactivation of T. tubifex. This study investigated the inactivation kinetics of chlorine on T. tubifex and the synergistic effect of the sequential use of chlorine and UV irradiation. The experimental results indicated that the Ct (concentration × time reaction ) concept could be used to evaluate the inactivation kinetics of T. tubifex with chlorine, thus allowing for the use of a simpler Ct approach for the assessment of T. tubifex chlorine inactivation requirements. The inactivation kinetics of T. tubifex by chlorine was found to be well-fitted to a delayed pseudo first-order Chick-Watson expression. Sequential experiments revealed that UV irradiation and chlorine worked synergistically to effectively inactivate T. tubifex as a result of the decreased activation energy, E a , induced by primary UV irradiation. Furthermore, the inactivation effectiveness of T. tubifex by chlorine was found to be affected by several drinking water quality parameters including pH, turbidity, and chemical oxygen demand with potassium permanganate (COD Mn ) concentration. High pH exhibited pronounced inactivation effectiveness and the decrease in turbidity and COD Mn concentrations contributed to the inactivation of T. tubifex. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Microbial risk assessment in heterogeneous aquifers: 1. Pathogen transport

    NASA Astrophysics Data System (ADS)

    Molin, S.; Cvetkovic, V.

    2010-05-01

    Pathogen transport in heterogeneous aquifers is investigated for microbial risk assessment. A point source with time-dependent input of pathogens is assumed, exemplified as a simple on-site sanitation installation, intermingled with water supply wells. Any pathogen transmission pathway (realization) to the receptor from a postulated infection hazard is viewed as a random event, with the hydraulic conductivity varying spatially. For aquifers where VAR[lnK] < 1 and the integral scale is finite, we provide relatively simple semianalytical expressions for pathogen transport that incorporate the colloid filtration theory. We test a wide range of Damkohler numbers in order to assess the significance of rate limitations on the aquifer barrier function. Even slow immobile inactivation may notably affect the retention of pathogens. Analytical estimators for microbial peak discharge are evaluated and are shown to be applicable using parameters representative of rotavirus and Hepatitis A with input of 10-20 days duration.

  20. Microbial mat of the thermal springs Kuchiger Republic of Buryatia: species composition, biochemical properties and electrogenic activity in biofuel cell

    NASA Astrophysics Data System (ADS)

    Aleksandrovich Yuriev, Denis; Viktorovna Zaitseva, Svetlana; Olegovna Zhdanova, Galina; Yurievich Tolstoy, Mikhail; Dondokovna Barkhutova, Darima; Feodorovna Vyatchina, Olga; Yuryevna Konovalova, Elena; Iosifovich Stom, Devard

    2018-02-01

    Electrogenic, molecular and some other properties of a microbial mat isolated from the Kuchiger hot spring (Kurumkansky District, Republic of Buryatia) were studied. Molecular analysis showed that representatives of Proteobacteria (85.5 % of the number of classified bacterial sequences) prevailed in the microbial mat of the Kuchiger springs, among which sulfur bacteria of the genus Thiothrix were the most numerous. In the microbial mat there were bacteria from the families Rhodocyclaceae, Comamonadaceae and Flavobacteriaceae. Phylum Bacteroidetes, Cyanobacteria/Chloroplast, Fusobacteria, Fibrobacteres, Acidobacteria, Chlorobi, Spirochaetes, Verrucomicrobia, Firmicutes, Deinococcus-Thermus, Chloroflexi and Actinobacteria are also noted in the composition of the microbial mat. Under the experimental conditions using Kuchiger-mat 16 as bioagents, glucose and peptone as substrates, the power of BFC was 240 and 221 mW / m2, respectively. When replacing the substrate with sodium acetate, the efficiency of the BFC was reduced by a factor of 10 (20 mW / m2). The prospects of using a microbial mat “Kuchiger-16” as an electrogen in BFC when utilizing alkaline waste water components to generate electricity are discussed.

  1. Adsorption, sedimentation, and inactivation of E. coli within wastewater treatment wetlands.

    PubMed

    Boutilier, L; Jamieson, R; Gordon, R; Lake, C; Hart, W

    2009-09-01

    Bacteria fate and transport within constructed wetlands must be understood if engineered wetlands are to become a reliable form of wastewater treatment. This study investigated the relative importance of microbial treatment mechanisms in constructed wetlands treating both domestic and agricultural wastewater. Escherichia coli (E. coli) inactivation, adsorption, and settling rates were measured in the lab within two types of wastewater (dairy wastewater lagoon effluent and domestic septic tank effluent). In situ E. coli inactivation was also measured within a domestic wastewater treatment wetland and the adsorption of E. coli was also measured within the wetland effluent. Inactivation of E. coli appears to be the most significant contributor to E. coli removal within the wastewaters and wetland environments examined in this study. E. coli survived longer within the dairy wastewater (DW) compared to the domestic wastewater treatment wetland water (WW). First order rate constants for E. coli inactivation within the WW in the lab ranged from 0.09 day(-1) (d(-1)) at 7.6 degrees C to 0.18d(-1) at 22.8 degrees C. The average in situ rate constant observed within the domestic wetland ranged from 0.02 d(-1) to 0.03 d(-1) at an average water temperature of 17 degrees C. First order rate constants for E. coli inactivation within the DW ranged from 0.01 d(-1) at 7.7 degrees C to 0.04 d(-1) at 24.6 degrees C. Calculated distribution coefficients (K(d)) were 19,000 mL g(-1), 324,000 mL g(-1), and 293 mL g(-1) for E. coli with domestic septic tank effluent (STE), treated wetland effluent (WLE), and DW, respectively. Approximately 50%, 20%, and 90% of E. coli were "free floating" or associated with particles <5 microm in size within the STE, WLE, and DW respectively. Although 10-50% of E. coli were found to associate with particles >5 microm within both the STE and DW, settling did not appear to contribute to E. coli removal within sedimentation experiments, indicating that the

  2. Photochemical and microbial alterations of DOM spectroscopic properties in the estuarine system Ria de Aveiro.

    PubMed

    Santos, L; Santos, E B H; Dias, J M; Cunha, A; Almeida, A

    2014-08-01

    The influence of photochemical transformations of chromophoric dissolved organic matter (CDOM) on microbial communities was evaluated in the estuarine system Ria de Aveiro. Two sites, representative of the marine and brackish water zones of the estuary, were surveyed regularly in order to determine seasonal and vertical profiles of variation of CDOM properties. Optical parameters of CDOM indicative of aromaticity and molecular weight were used to establish CDOM sources, and microbial abundance and activity was characterized. Additionally, microcosm experiments were performed in order to simulate photochemical reactions of CDOM and to evaluate microbial responses to light-induced changes in CDOM composition. The CDOM of the two estuarine zones showed different spectral characteristics, with significantly higher values of the specific ultra-violet absorbance at 254 nm (SUVA254) (5.5 times) and of the absorption coefficient at 350 nm (a350) (12 times) and lower SR (S275-295/S350-400) ratio at brackish water compared with the marine zone, reflecting the different amounts and prevailing sources of organic matter, as well as distinct riverine and oceanic influences. At the marine zone, the abundance of bacteria and the activity of Leu-AMPase correlated with a350 and a254, suggesting a microbial contribution to the HMW CDOM pool. The irradiation of DOM resulted in a decrease of the values of a254 and a350 and an increase of the slope S275-295 and of the ratios E2 : E3 (a250/a365) and SR, which in turn increase its bioavailability. However, the extent of photoinduced transformations and microbial responses was dependent on the initial optical characteristics of CDOM. In Ria de Aveiro both photochemical and microbial processes yielded optical changes in CDOM and the overall results of these combined processes determine the fate of CDOM in the estuarine system and have an influence on local productivity and in adjacent coastal areas.

  3. Inactivation of phosphorylase b by potassium ferrate, a new reactive analogue of the phosphate group.

    PubMed

    Lee, Y M; Benisek, W F

    1976-03-25

    Rabbit muscle phosphorylase b reacts with the phosphate-like reagent potassium ferrate, K2FeO4, a potent oxidizing agent. The reaction results in inactivation of the enzyme and abolition of the ability of the enzyme to bind 5'-AMP. Activating and nonactivating nucleotides which bind at the 5'-AMP binding site such as 5'-AMP, 2'-AMP, 3'-AMP, and 5'-IMP substantially protect the enzyme from inactivation by ferrate. One to two residues of tyrosine and approximately 1 residue of cysteine are modified by ferrate under the conditions employed. Tyrosine is protected by 5-AMP, whereas cysteine is not. The tyrosine modification is suggested as the inactivating chemical reaction. The location of the inactivating reaction is suggested to be in or near the 5'-AMP binding site. The structural and chemical properties of ferrate ion are discussed and compared to those of phosphate. Ferrate ion may be a reagent useful for phosphate group binding site-directed modification of proteins.

  4. Effect of surface roughness on inactivation of Escherichia coli O157:H7 87-23 by new organic acid-surfactant combinations on alfalfa, broccoli, and radish seeds.

    PubMed

    Fransisca, Lilia; Feng, Hao

    2012-02-01

    Surface roughness has been reported as one of the factors affecting microbial attachment and removal. Seed surfaces are complex, and different seed varieties have different surface topographies. As a result, a sanitizer effective in eliminating pathogenic bacteria on one seed may not be as effective when applied to another seed. The objectives of this research were (i) to investigate the efficacy of malic acid and thiamine dilaurylsulfate (TDS) combined treatments for inactivation of E. coli O157:H7 strain 87-23 on alfalfa, broccoli, and radish seeds, (ii) to quantify surface roughness of the seeds, and (iii) to determine the correlation between microbial removal and surface roughness. The surface roughness of each seed type was measured by confocal laser scanning microscopy (CLSM) and surface profilometry. Surface roughness (Ra) values of the seeds were then calculated from CLSM data. Seeds inoculated with E. coli O157:H7 87-23 were washed for 20 min in malic acid and TDS solutions and rinsed for 10 min in tap water. Radish seeds had the highest Ra values, followed by broccoli and alfalfa seeds. A combination of 10% malic acid and 1% TDS was more effective than 20,000 ppm of Ca(OCl)(2) for inactivation of E. coli O157:H7 87-23 on broccoli seeds, while the inactivation on radish and alfalfa seeds was not significantly different compared with the 20,000-ppm Ca(OCl)(2) wash. Overall, a negative correlation existed between the seeds' Ra values and microbial removal. Different seeds had different surface roughness, contributing to discrepancies in the ability of the sanitizers to eliminate E. coli O157:H7 87-23 on the seeds. Therefore, the effectiveness of one sanitizer on one seed type should not be translated to all seed varieties.

  5. A novel N-terminal motif of dipeptidyl peptidase-like proteins produces rapid inactivation of KV4.2 channels by a pore-blocking mechanism.

    PubMed

    Jerng, Henry H; Dougherty, Kevin; Covarrubias, Manuel; Pfaffinger, Paul J

    2009-11-01

    The somatodendritic subthreshold A-type K(+) current in neurons (I(SA)) depends on its kinetic and voltage-dependent properties to regulate membrane excitability, action potential repetitive firing, and signal integration. Key functional properties of the K(V)4 channel complex underlying I(SA) are determined by dipeptidyl peptidase-like proteins known as dipeptidyl peptidase 6 (DPP6) and dipeptidyl peptidase 10 (DPP10). Among the multiple known DPP10 isoforms with alternative N-terminal sequences, DPP10a confers exceptionally fast inactivation to K(V)4.2 channels. To elucidate the molecular basis of this fast inactivation, we investigated the structure-function relationship of the DPP10a N-terminal region and its interaction with the K(V)4.2 channel. Here, we show that DPP10a shares a conserved N-terminal sequence (MNQTA) with DPP6a (aka DPP6-E), which also induces fast inactivation. Deletion of the NQTA sequence in DPP10a eliminates this dramatic fast inactivation, and perfusion of MNQTA peptide to the cytoplasmic face of inside-out patches inhibits the K(V)4.2 current. DPP10a-induced fast inactivation exhibits competitive interactions with internally applied tetraethylammonium (TEA), and elevating the external K(+) concentration accelerates recovery from DPP10a-mediated fast inactivation. These results suggest that fast inactivation induced by DPP10a or DPP6a is mediated by a common N-terminal inactivation motif via a pore-blocking mechanism. This mechanism may offer an attractive target for novel pharmacological interventions directed at impairing I(SA) inactivation and reducing neuronal excitability.

  6. X inactivation in Rett syndrome: A preliminary study showing partial preferential inactivation of paternal X with the M27{beta} probe

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

    Camus, P.; Abbadi, N.; Gilgenkrantz, S.

    1994-04-15

    Rett syndrome (RS) is a severe progressive neurological disorder occurring exclusively in females. Most cases are sporadic. The few familial cases (less than 1%) cannot be explained by a simple mode of inheritance. Several hypotheses have been proposed: X-linked male lethal mutation, maternal uniparental disomy, fresh mutation on the X chromosome, involvement of mitochondrial DNA and differential inactivation with metabolic interference of X-borne alleles. The authors have examined the pattern of X inactivation in 10 affected girls who were selected according to the clinical criteria previously described and accepted by the French Rett Scientific Committee. The X inactivation pattern wasmore » studied by analysis of methylation at the hypervariable locus DXS255 with the M27{beta} probe. The results show a more-or-less skewed inactivation of paternal X in 8 Rett females, and 2 cases of symmetrical inactivation. In control girls, inactivation was symmetrical cases and the maternal X has been preferentially inactivated in the other 2 cases. In no case was a total skewed inactivation observed. Though there was clear evidence for a preferential paternal X inactivation that was statistically significant further studies are necessary to establish a relationship between X inactivation pattern and Rett syndrome.« less

  7. Evaluation of the factors controlling the time-dependent inactivation rate coefficients of bacteriophage MS2 and PRD1

    USGS Publications Warehouse

    Anders, R.; Chrysikopoulos, C.V.

    2006-01-01

    Static and dynamic batch experiments were conducted to study the effects of temperature and the presence of sand on the inactivation of bacteriophage MS2 and PRD1. The experimental data suggested that the inactivation process can be satisfactorily represented by a pseudo-first-order expression with time-dependent rate coefficients. The time-dependent rate coefficients were used to determine pertinent thermodynamic properties required for the analysis of the molecular processes involved in the inactivation of each bacteriophage. A combination of high temperature and the presence of sand appears to produce the greatest disruption to the surrounding protein coat of MS2. However, the lower activation energies for PRD1 indicate a weaker dependence of the inactivation rate on temperature. Instead, the presence of air-liquid and air-solid interfaces appears to produce the greatest damage to specific viral components that are related to infection. These results indicate the importance of using thermodynamic parameters based on the time-dependent inactivation model to better predict the inactivation of viruses in groundwater. ?? 2006 American Chemical Society.

  8. Screening, identification, and characterization of a GH43 family β-xylosidase/α-arabinofuranosidase from a compost microbial metagenome.

    PubMed

    Matsuzawa, Tomohiko; Kaneko, Satoshi; Yaoi, Katsuro

    2015-11-01

    A putative glycoside hydrolase family 43 β-xylosidase/α-arabinofuranosidase (CoXyl43) that promotes plant biomass saccharification was isolated via functional screening of a compost microbial metagenomic library and characterized. CoXyl43 promoted the saccharification of plant biomasses, including xylans (xylan and arabinoxylan), rice straw, and Erianthus, by degrading xylooligosaccharide residues to monosaccharide residues. The recombinant CoXyl43 protein exhibited both β-xylosidase and α-arabinofuranosidase activities for chromogenic substrates, with optimal activity at pH 7.5 and 55 °C. Both of these activities were inactivated by ethanol, dimethylsulfoxide, and zinc and copper ions but were activated by manganese ions. Only the β-xylosidase activity of recombinant CoXyl43 was enhanced in the presence of calcium ions. These results indicate that CoXyl43 exhibits unique enzymatic properties useful for biomass saccharification.

  9. Physicochemical, microbial, and sensory properties of nanopowdered eggshell-supplemented yogurt during storage.

    PubMed

    Al Mijan, Mohammad; Choi, Kyung-Hoon; Kwak, Hae-Soo

    2014-01-01

    This study was carried out to investigate the possibility of adding nanopowdered eggshell (NPES) into yogurt to improve the functionality of yogurt and the effects of adding NPES on the physicochemical, microbial, and sensory properties of the products during storage. The pH and mean lactic acid bacteria counts of NPES-added (0.15-0.45%, wt/vol) yogurt ranged from 4.31 to 4.66 and from 6.56 × 10(8) to 8.56 × 10(8)cfu/mL, respectively, whereas these values ranged from 4.13 to 4.44 and 8.46 × 10(8) to 1.39 × 10(9), respectively, for the control samples during storage at 5 °C for 16d, which indicates a prolonged shelf-life with NPES-supplemented yogurt. Color analysis showed that the lightness (L*) and position between red and green (a*) values were not significantly influenced by the addition of NPES. However, the position between yellow and blue (b*) value significantly increased with the addition of the concentration (0.45%, wt/vol) of NPES at d 16 of storage. Sensory evaluation revealed that NPES-added yogurts showed a notably less sourness score and a higher astringency score than the control. An earthy flavor was higher in 0.45% NPES-supplemented yogurt compared with the control. Based on the results obtained from the current study, the concentration (0.15 to 0.30%, wt/vol) of NPES can be used to formulate NPES-supplemented yogurt without any significant adverse effects on the physicochemical, microbial, and sensory properties. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  10. Inactivation of the Lactobacillus leichmannii ribonucleoside triphosphate reductase by 2'-chloro-2'-deoxyuridine 5'-triphosphate: stoichiometry of inactivation, site of inactivation, and mechanism of the protein chromophore formation

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

    Ashley, G.W.; Harris, G.; Stubbe, J.A.

    1988-06-14

    The ribonucleoside triphosphate reductase (RTPR) of Lactobacillus leichmannii is inactivated by the substrate analogue 2'-chloro-2'-deoxyuridine 5'-triphosphate (ClUTP). Inactivation is due to alkylation by 2-methylene-3(2H)-furanone, a decomposition product of the enzymic product 3'-keto-2'-deoxyuridine triphosphate. The former has been unambiguously identified as 2-((ethylthio)methyl)-3(2H)-furanone, an ethanethiol trapped adduct, which is identical by /sup 1/H NMR spectroscopy with material synthesized chemically. Subsequent to rapid inactivation, a slow process occurs that results in formation of a new protein-associated chromophore absorbing maximally near 320 nm. The terminal stages of the inactivation have now been investigated in detail. The alkylation and inactivation stoichiometries were studied as amore » function of the ratio of ClUTP to enzyme. The amount of labeling of RTPR increased with increasing ClUTP concentration up to the maximum of approximately 4 labels/RTPR, yet the degree of inactivation did not increase proportionally. This suggests that (1) RTPR may be inactivated by alkylation of a single site and (2) decomposition of 3'-keto-dUTP is not necessarily enzyme catalyzed. The formation of the new protein chromophore was also monitored during inactivation and found to reach its full extent upon the first alkylation . Thus, out of four alkylation sites, only one appears capable of undergoing the subsequent reaction to form the new chromophore. Model studies suggest that the new chromophore is due to addition of an amino group to the 5-position of enzyme-bound furanone, followed by ring opening and tautomerization to give a ..beta..-aminoenone structure.« less

  11. Mechanisms of Inactivation of Dry Escherichia coli by High-Pressure Carbon Dioxide

    PubMed Central

    Chen, Yuan Yao; Temelli, Feral

    2017-01-01

    ABSTRACT High-pressure carbon dioxide processing is a promising technology for nonthermal food preservation. However, few studies have determined the lethality of high-pressure CO2 on dry bacterial cells, and the mechanism of inactivation remains unknown. This study explored the mechanisms of inactivation by using Escherichia coli AW1.7 and mutant strains differing in heat and acid resistance, in membrane composition based on disruption of the locus of heat resistance, and in genes coding for glutamate decarboxylases and cyclopropane fatty acid synthase. The levels of lethality of treatments with liquid, gaseous, and supercritical CO2 were compared. The cell counts of E. coli AW1.7 and mutants with a water activity (aW) of 1.0 were reduced by more than 3 log10 (CFU/ml) after supercritical CO2 treatment at 35°C for 15 min; increasing the pressure generally enhanced inactivation, except for E. coli AW1.7 ΔgadAB. E. coli AW1.7 Δcfa was more susceptible than E. coli AW1.7 after treatment at 10 and 40 MPa; other mutations did not affect survival. Dry cells of E. coli were resistant to treatments with supercritical and liquid CO2 at any temperature. Treatments with gaseous CO2 at 65°C were more bactericidal than those with supercritical CO2 or treatments at 65°C only. Remarkably, E. coli AW1.7 was more susceptible than E. coli AW1.7 Δcfa when subjected to the gaseous CO2 treatment. This study identified CO2-induced membrane fluidization and permeabilization as causes of supercritical mediated microbial inactivation, and diffusivity was a dominant factor for gaseous CO2. IMPORTANCE The safety of dry foods is of increasing concern for public health. Desiccated microorganisms, including pathogens, remain viable over long periods of storage and generally tolerate environmental insults that are lethal to the same organisms at high water activity. This study explored the use of high-pressure carbon dioxide to determine its lethality for dried Escherichia coli and to

  12. Mechanisms of Inactivation of Dry Escherichia coli by High-Pressure Carbon Dioxide.

    PubMed

    Chen, Yuan Yao; Temelli, Feral; Gänzle, Michael G

    2017-05-15

    High-pressure carbon dioxide processing is a promising technology for nonthermal food preservation. However, few studies have determined the lethality of high-pressure CO 2 on dry bacterial cells, and the mechanism of inactivation remains unknown. This study explored the mechanisms of inactivation by using Escherichia coli AW1.7 and mutant strains differing in heat and acid resistance, in membrane composition based on disruption of the locus of heat resistance, and in genes coding for glutamate decarboxylases and cyclopropane fatty acid synthase. The levels of lethality of treatments with liquid, gaseous, and supercritical CO 2 were compared. The cell counts of E. coli AW1.7 and mutants with a water activity (a W ) of 1.0 were reduced by more than 3 log 10 (CFU/ml) after supercritical CO 2 treatment at 35°C for 15 min; increasing the pressure generally enhanced inactivation, except for E. coli AW1.7 Δ gadAB E. coli AW1.7 Δ cfa was more susceptible than E. coli AW1.7 after treatment at 10 and 40 MPa; other mutations did not affect survival. Dry cells of E. coli were resistant to treatments with supercritical and liquid CO 2 at any temperature. Treatments with gaseous CO 2 at 65°C were more bactericidal than those with supercritical CO 2 or treatments at 65°C only. Remarkably, E. coli AW1.7 was more susceptible than E. coli AW1.7 Δ cfa when subjected to the gaseous CO 2 treatment. This study identified CO 2 -induced membrane fluidization and permeabilization as causes of supercritical mediated microbial inactivation, and diffusivity was a dominant factor for gaseous CO 2 IMPORTANCE The safety of dry foods is of increasing concern for public health. Desiccated microorganisms, including pathogens, remain viable over long periods of storage and generally tolerate environmental insults that are lethal to the same organisms at high water activity. This study explored the use of high-pressure carbon dioxide to determine its lethality for dried Escherichia coli and to

  13. Thin-film fixed-bed reactor (TFFBR) for solar photocatalytic inactivation of aquaculture pathogen Aeromonas hydrophila

    PubMed Central

    2012-01-01

    Background Outbreaks of infectious diseases by microbial pathogens can cause substantial losses of stock in aquaculture systems. There are several ways to eliminate these pathogens including the use of antibiotics, biocides and conventional disinfectants, but these leave undesirable chemical residues. Conversely, using sunlight for disinfection has the advantage of leaving no chemical residue and is particularly suited to countries with sunny climates. Titanium dioxide (TiO2) is a photocatalyst that increases the effectiveness of solar disinfection. In recent years, several different types of solar photocatalytic reactors coated with TiO2 have been developed for waste water and drinking water treatment. In this study a thin-film fixed-bed reactor (TFFBR), designed as a sloping flat plate reactor coated with P25 DEGUSSA TiO2, was used. Results The level of inactivation of the aquaculture pathogen Aeromonas hydrophila ATCC 35654 was determined after travelling across the TFFBR under various natural sunlight conditions (300-1200 W m-2), at 3 different flow rates (4.8, 8.4 and 16.8 L h-1). Bacterial numbers were determined by conventional plate counting using selective agar media, cultured (i) under conventional aerobic conditions to detect healthy cells and (ii) under conditions designed to neutralise reactive oxygen species (agar medium supplemented with the peroxide scavenger sodium pyruvate at 0.05% w/v, incubated under anaerobic conditions), to detect both healthy and sub-lethally injured (oxygen-sensitive) cells. The results clearly demonstrate that high sunlight intensities (≥ 600 W m-2) and low flow rates (4.8 L h-1) provided optimum conditions for inactivation of A. hydrophila ATCC 3564, with greater overall inactivation and fewer sub-lethally injured cells than at low sunlight intensities or high flow rates. Low sunlight intensities resulted in reduced overall inactivation and greater sub-lethal injury at all flow rates. Conclusions This is the first

  14. Can microbial cells develop resistance to oxidative stress in antimicrobial photodynamic inactivation?

    PubMed

    Kashef, Nasim; Hamblin, Michael R

    2017-03-01

    Infections have been a major cause of disease throughout the history of humans on earth. With the introduction of antibiotics, it was thought that infections had been conquered. However, bacteria have been able to develop resistance to antibiotics at an exponentially increasing rate. The growing threat from multi-drug resistant organisms calls for intensive action to prevent the emergence of totally resistant and untreatable infections. Novel, non-invasive, non-antibiotic strategies are needed that act more efficiently and faster than current antibiotics. One promising alternative is antimicrobial photodynamic inactivation (APDI), an approach that produces reactive oxygen species when dyes and light are combined. So far, it has been questionable if bacteria can develop resistance against APDI. This review paper gives an overview of recent studies concerning the susceptibility of bacteria towards oxidative stress, and suggests possible mechanisms of the development of APDI-resistance that should at least be addressed. Some ways to potentiate APDI and also to overcome future resistance are suggested. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Microbial-Induced Heterogeneity in the Acoustic Properties of Porous Media

    EPA Science Inventory

    Acoustic wave data were acquired over a two-dimensional region of a microbial-stimulated sand column and an unstimulated sand column to assess the spatiotemporal changes in a porous medium caused by microbial growth and biofilm formation. The acoustic signals from the unstimulate...

  16. Pressure- and heat-induced inactivation of butyrylcholinesterase: evidence for multiple intermediates and the remnant inactivation process.

    PubMed Central

    Weingand-Ziade, A; Ribes, F; Renault, F; Masson, P

    2001-01-01

    The inactivation process of native (N) human butyrylcholinesterase (BuChE) by pressure and/or heat was found to be multi-step. It led to irreversible formation of an active intermediate (I) state and a denatured state. This series-inactivation process was described by expanding the Lumry-Eyring [Lumry, R. and Eyring, H. (1954) J. Phys. Chem. 58, 110-120] model. The intermediate state (I) was found to have a K(m) identical with that of the native state and a turnover rate (k(cat)) twofold higher than that of the native state with butyrylthiocholine as the substrate. The increased catalytic efficiency (k(cat)/K(m)) of I can be explained by a conformational change in the active-site gorge and/or restructuring of the water-molecule network in the active-site pocket, making the catalytic steps faster. However, a pressure/heat-induced covalent modification of native BuChE, affecting the catalytic machinery, cannot be ruled out. The inactivation process of BuChE induced by the combined action of pressure and heat was found to continue after interruption of pressure/temperature treatment. This secondary inactivation process was termed 'remnant inactivation'. We hypothesized that N and I were in equilibrium with populated metastable N' and I' states. The N' and I' states can either return to the active forms, N and I, or develop into inactive forms, N(')(in) and I(')(in). Both active N' and I' intermediate states displayed different rates of remnant inactivation depending on the pressure and temperature pretreatments and on the storage temperature. A first-order deactivation model describing the kinetics of the remnant inactivation of BuChE is proposed. PMID:11368776

  17. Enzyme reactor design under thermal inactivation.

    PubMed

    Illanes, Andrés; Wilson, Lorena

    2003-01-01

    Temperature is a very relevant variable for any bioprocess. Temperature optimization of bioreactor operation is a key aspect for process economics. This is especially true for enzyme-catalyzed processes, because enzymes are complex, unstable catalysts whose technological potential relies on their operational stability. Enzyme reactor design is presented with a special emphasis on the effect of thermal inactivation. Enzyme thermal inactivation is a very complex process from a mechanistic point of view. However, for the purpose of enzyme reactor design, it has been oversimplified frequently, considering one-stage first-order kinetics of inactivation and data gathered under nonreactive conditions that poorly represent the actual conditions within the reactor. More complex mechanisms are frequent, especially in the case of immobilized enzymes, and most important is the effect of catalytic modulators (substrates and products) on enzyme stability under operation conditions. This review focuses primarily on reactor design and operation under modulated thermal inactivation. It also presents a scheme for bioreactor temperature optimization, based on validated temperature-explicit functions for all the kinetic and inactivation parameters involved. More conventional enzyme reactor design is presented merely as a background for the purpose of highlighting the need for a deeper insight into enzyme inactivation for proper bioreactor design.

  18. Phospholipids fatty acids of drinking water reservoir sedimentary microbial community: Structure and function responses to hydrostatic pressure and other physico-chemical properties.

    PubMed

    Chai, Bei-Bei; Huang, Ting-Lin; Zhao, Xiao-Guang; Li, Ya-Jiao

    2015-07-01

    Microbial communities in three drinking water reservoirs, with different depth in Xi'an city, were quantified by phospholipids fatty acids analysis and multivariate statistical analysis was employed to interpret their response to different hydrostatic pressure and other physico-chemical properties of sediment and overlying water. Principle component analyses of sediment characteristics parameters showed that hydrostatic pressure was the most important effect factor to differentiate the overlying water quality from three drinking water reservoirs from each other. NH4+ content in overlying water was positive by related to hydrostatic pressure, while DO in water-sediment interface and sediment OC in sediment were negative by related with it. Three drinking water reservoir sediments were characterized by microbial communities dominated by common and facultative anaerobic Gram-positive bacteria, as well as, by sulfur oxidizing bacteria. Hydrostatic pressure and physico-chemical properties of sediments (such as sediment OC, sediment TN and sediment TP) were important effect factors to microbial community structure, especially hydrostatic pressure. It is also suggested that high hydrostatic pressure and low dissolved oxygen concentration stimulated Gram-positive and sulfate-reducing bacteria (SRB) bacterial population in drinking water reservoir sediment. This research supplied a successful application of phospholipids fatty acids and multivariate analysis to investigate microbial community composition response to different environmental factors. Thus, few physico-chemical factors can be used to estimate composition microbial of community as reflected by phospholipids fatty acids, which is difficult to detect.

  19. Effects of Microbial Transglutaminase on Physicochemical, Microbial and Sensorial Properties of Kefir Produced by Using Mixture Cow’s and Soymilk

    PubMed Central

    2017-01-01

    The objective of this research was to investigate the effects microbial transglutaminase (m-TGs) on the physicochemical, microbial and sensory properties of kefir produced by using mix cow and soymilk. Kefir batches were prepared using 0, 0.5, 1 and 1.5 Units m-TGs for per g of milk protein. Adding m-TGs to milk caused an increase in the pH and viscosity and caused a decrease in titratable acidity and syneresis in the kefir samples. Total bacteria, lactobacilli and streptococci counts decreased, while yeast counts increased in all the samples during storage. Alcohols and acids compounds have increased in all the samples except in the control samples, while carbonyl compounds have decreased in all the samples during storage (1-30 d). The differences in the percentage of alcohols, carbonyl compounds and acids in total volatiles on the 1st and the 30th d of storage were observed at 8.47-23.52%, 6.94-25.46% and 59.64-63.69%, respectively. The consumer evaluation of the kefir samples showed that greater levels of acceptability were found for samples which had been added 1.5 U m-TGs for per g of milk protein. PMID:28943774

  20. Phylogenetic Structure and Metabolic Properties of Microbial Communities in Arsenic-Rich Waters of Geothermal Origin

    PubMed Central

    Crognale, Simona; Zecchin, Sarah; Amalfitano, Stefano; Fazi, Stefano; Casentini, Barbara; Corsini, Anna; Cavalca, Lucia; Rossetti, Simona

    2017-01-01

    Arsenic (As) is a toxic element released in aquatic environments by geogenic processes or anthropic activities. To counteract its toxicity, several microorganisms have developed mechanisms to tolerate and utilize it for respiratory metabolism. However, still little is known about identity and physiological properties of microorganisms exposed to natural high levels of As and the role they play in As transformation and mobilization processes. This work aims to explore the phylogenetic composition and functional properties of aquatic microbial communities in As-rich freshwater environments of geothermal origin and to elucidate the key microbial functional groups that directly or indirectly may influence As-transformations across a natural range of geogenic arsenic contamination. Distinct bacterial communities in terms of composition and metabolisms were found. Members of Proteobacteria, affiliated to Alpha- and Betaproteobacteria were mainly retrieved in groundwaters and surface waters, whereas Gammaproteobacteria were the main component in thermal waters. Most of the OTUs from thermal waters were only distantly related to 16S rRNA gene sequences of known taxa, indicating the occurrence of bacterial biodiversity so far unexplored. Nitrate and sulfate reduction and heterotrophic As(III)-oxidization were found as main metabolic traits of the microbial cultivable fraction in such environments. No growth of autotrophic As(III)-oxidizers, autotrophic and heterotrophic As(V)-reducers, Fe-reducers and oxidizers, Mn-reducers and sulfide oxidizers was observed. The ars genes, involved in As(V) detoxifying reduction, were found in all samples whereas aioA [As(III) oxidase] and arrA genes [As(V) respiratory reductase] were not found. Overall, we found that As detoxification processes prevailed over As metabolic processes, concomitantly with the intriguing occurrence of novel thermophiles able to tolerate high levels of As. PMID:29312179

  1. Phylogenetic Structure and Metabolic Properties of Microbial Communities in Arsenic-Rich Waters of Geothermal Origin.

    PubMed

    Crognale, Simona; Zecchin, Sarah; Amalfitano, Stefano; Fazi, Stefano; Casentini, Barbara; Corsini, Anna; Cavalca, Lucia; Rossetti, Simona

    2017-01-01

    Arsenic (As) is a toxic element released in aquatic environments by geogenic processes or anthropic activities. To counteract its toxicity, several microorganisms have developed mechanisms to tolerate and utilize it for respiratory metabolism. However, still little is known about identity and physiological properties of microorganisms exposed to natural high levels of As and the role they play in As transformation and mobilization processes. This work aims to explore the phylogenetic composition and functional properties of aquatic microbial communities in As-rich freshwater environments of geothermal origin and to elucidate the key microbial functional groups that directly or indirectly may influence As-transformations across a natural range of geogenic arsenic contamination. Distinct bacterial communities in terms of composition and metabolisms were found. Members of Proteobacteria , affiliated to Alpha - and Betaproteobacteria were mainly retrieved in groundwaters and surface waters, whereas Gammaproteobacteria were the main component in thermal waters. Most of the OTUs from thermal waters were only distantly related to 16S rRNA gene sequences of known taxa, indicating the occurrence of bacterial biodiversity so far unexplored. Nitrate and sulfate reduction and heterotrophic As(III)-oxidization were found as main metabolic traits of the microbial cultivable fraction in such environments. No growth of autotrophic As(III)-oxidizers, autotrophic and heterotrophic As(V)-reducers, Fe-reducers and oxidizers, Mn-reducers and sulfide oxidizers was observed. The ars genes, involved in As(V) detoxifying reduction, were found in all samples whereas aioA [As(III) oxidase] and arrA genes [As(V) respiratory reductase] were not found. Overall, we found that As detoxification processes prevailed over As metabolic processes, concomitantly with the intriguing occurrence of novel thermophiles able to tolerate high levels of As.

  2. Microbial properties explain temporal variation in soil respiration in a grassland subjected to nitrogen addition

    PubMed Central

    Li, Yue; Liu, Yinghui; Wu, Shanmei; Niu, Lei; Tian, Yuqiang

    2015-01-01

    The role of soil microbial variables in shaping the temporal variability of soil respiration has been well acknowledged but is poorly understood, particularly under elevated nitrogen (N) deposition conditions. We measured soil respiration along with soil microbial properties during the early, middle, and late growing seasons in temperate grassland plots that had been treated with N additions of 0, 2, 4, 8, 16, or 32 g N m−2 yr−1 for 10 years. Representing the averages over three observation periods, total (Rs) and heterotrophic (Rh) respiration were highest with 4 g N m−2 yr−1, but autotrophic respiration (Ra) was highest with 8 to 16 g N m−2 yr−1. Also, the responses of Rh and Ra were unsynchronized considering the periods separately. N addition had no significant impact on the temperature sensitivity (Q10) for Rs but inhibited the Q10 for Rh. Significant interactions between observation period and N level occurred in soil respiration components, and the temporal variations in soil respiration components were mostly associated with changes in microbial biomass carbon (MBC) and phospholipid fatty acids (PLFAs). Further observation on soil organic carbon and root biomass is needed to reveal the long-term effect of N deposition on soil C sequestration. PMID:26678303

  3. INACTIVATION AND REACTIVATION OF B. MEGATHERIUM PHAGE

    PubMed Central

    Northrop, John H.

    1955-01-01

    Preparation of Reversibly Inactivated (R.I.) Phage.— If B. megatherium phage (of any type, or in any stage of purification) is suspended in dilute salt solutions at pH 5–6, it is completely inactivated; i.e., it does not form plaques, or give rise to more phage when mixed with a sensitive organism (Northrop, 1954). The inactivation occurs when the phage is added to the dilute salt solution. If a suspension of the inactive phage in pH 7 peptone is titrated to pH 5 and allowed to stand, the activity gradually returns. The inactivation is therefore reversible. Properties of R.I. Phage.— The R.I. phage is adsorbed by sensitive cells at about the same rate as the active phage. It kills the cells, but no active phage is produced. The R.I. phage therefore has the properties of phage "ghosts" (Herriott, 1951) or of colicines (Gratia, 1925), or phage inactivated by ultraviolet light (Luria, 1947). The R.I. phage is sedimented in the centrifuge at the same rate as active phage. It is therefore about the same size as the active phage. The R.I. phage is most stable in pH 7, 5 per cent peptone, and may be kept in this solution for weeks at 0°C. The rate of digestion of R.I. phage by trypsin, chymotrypsin, or desoxyribonuclease is about the same as that of active phage (Northrop, 1955 a). Effect of Various Substances on the Formation of R.I. Phage.— There is an equilibrium between R.I. phage and active phage. The R.I. form is the stable one in dilute salt solution, pH 5 to 6.5 and at low temperature (<20°C.). At pH >6.5, in dilute salt solution, the R.I. phage changes to the active form. The cycle, active ⇌ inactive phage, may be repeated many times at 0°C. by changing the pH of the solution back and forth between pH 7 and pH 6. Irreversible inactivation is caused by distilled water, some heavy metals, concentrated urea or quanidine solutions, and by l-arginine. Reversible inactivation is prevented by all salts tested (except those causing irreversible inactivation

  4. Ammonia produced by bacterial colonies promotes growth of ampicillin-sensitive Serratia sp. by means of antibiotic inactivation.

    PubMed

    Cepl, Jaroslav; Blahůšková, Anna; Cvrčková, Fatima; Markoš, Anton

    2014-05-01

    Volatiles produced by bacterial cultures are known to induce regulatory and metabolic alterations in nearby con-specific or heterospecific bacteria, resulting in phenotypic changes including acquisition of antibiotic resistance. We observed unhindered growth of ampicillin-sensitive Serratia rubidaea and S. marcescens on ampicillin-containing media, when exposed to volatiles produced by dense bacterial growth. However, this phenomenon appeared to result from pH increase in the medium caused by bacterial volatiles rather than alterations in the properties of the bacterial cultures, as alkalization of ampicillin-containing culture media to pH 8.5 by ammonia or Tris exhibited the same effects, while pretreatment of bacterial cultures under the same conditions prior to antibiotic exposure did not increase ampicillin resistance. Ampicillin was readily inactivated at pH 8.5, suggesting that observed bacterial growth results from metabolic alteration of the medium, rather than an active change in the target bacterial population (i.e. induction of resistance or tolerance). However, even such seemingly simple mechanism may provide a biologically meaningful basis for protection against antibiotics in microbial communities growing on semi-solid media. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  5. The voltage sensor of excitation–contraction coupling in mammals: Inactivation and interaction with Ca2+

    PubMed Central

    2017-01-01

    In skeletal muscle, the four-helix voltage-sensing modules (VSMs) of CaV1.1 calcium channels simultaneously gate two Ca2+ pathways: the CaV1.1 pore itself and the RyR1 calcium release channel in the sarcoplasmic reticulum. Here, to gain insight into the mechanism by which VSMs gate RyR1, we quantify intramembrane charge movement associated with VSM activation (sensing current) and gated Ca2+ release flux in single muscle cells of mice and rats. As found for most four-helix VSMs, upon sustained depolarization, rodent VSMs lose the ability to activate Ca2+ release channels opening; their properties change from a functionally capable mode, in which the mobile sensor charge is called charge 1, to an inactivated mode, charge 2, with a voltage dependence shifted toward more negative voltages. We find that charge 2 is promoted and Ca2+ release inactivated when resting, well-polarized muscle cells are exposed to low extracellular [Ca2+] and that the opposite occurs in high [Ca2+]. It follows that murine VSMs are partly inactivated at rest, which establishes the reduced availability of voltage sensing as a pathogenic mechanism in disorders of calcemia. We additionally find that the degree of resting inactivation is significantly different in two mouse strains, which underscores the variability of voltage sensor properties and their vulnerability to environmental conditions. Our studies reveal that the resting and activated states of VSMs are equally favored by extracellular Ca2+. Promotion by an extracellular species of two states of the VSM that differ in the conformation of the activation gate requires the existence of a second gate, inactivation, topologically extracellular and therefore accessible from outside regardless of the activation state. PMID:29021148

  6. Thermal Inactivation of Viruses

    DTIC Science & Technology

    1977-10-01

    thermal inactivation . 12 Bibliography 20 Table 3. Thermal inactivation of viruses in foods 25 Bibliography 31 Table 4, Agents modifying...the presence of protective agents that reduce the lethal effect of heat on the viruses at temperatures below 60 C In addition, whtn solid foods...systems but were observed j when animal inoculation was utilized. It is possible that free virus nucl’lc < acid may have been the causative agent in

  7. Photodynamic-induced inactivation of Propionibacterium acnes

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Teschke, M.; Eick, Stephen G.; Pfister, W.; Meyer, Herbert; Halbhuber, Karl-Juergen

    1998-05-01

    We report on photodynamically induced inactivation of the skin bacterium Propionibacterium acnes (P. acnes) using endogenous as well as exogenous photosensitizers and red light sources. P. acnes is involved in the pathogenesis of the skin disease acne vulgaris. The skin bacterium is able to synthesize the metal-free fluorescent porphyrins protoporphyrin IX (PP) and coproporphyrin (CP) as shown by in situ spectrally-resolved detection of natural autofluorescence of human skin and bacteria colonies. These naturally occurring intracellular porphyrins act as efficient endogenous photosensitizers. Inactivation of P. acnes suspensions was achieved by irradiation with He-Ne laser light in the red spectral region (632.8 nm). We monitored the photodynamically-induced death of single bacteria using a fluorescent viability kit in combination with confocal laser scanning microscopy. In addition, the photo-induced inactivation was calculated by CFU (colony forming units) determination. We found 633 nm-induced inactivation (60 mW, 0.12 cm2 exposure area, 1 hour irradiation) of 72% in the case of non-incubated bacteria based on the destructive effect of singlet oxygen produced by red light excited endogenous porphyrins and subsequent energy transfer to molecular oxygen. In order to achieve a nearly complete inactivation within one exposure procedure, the exogenous photosensitizer Methylene Blue (Mb) was added. Far red exposure of Mb-labeled bacteria using a krypton ion laser at 647 nm and 676 nm resulted in 99% inactivation.

  8. Slowly inactivating component of Na+ current in peri-somatic region of hippocampal CA1 pyramidal neurons

    PubMed Central

    Park, Yul Young; Johnston, Daniel

    2013-01-01

    The properties of voltage-gated ion channels on the neuronal membrane shape electrical activity such as generation and backpropagation of action potentials, initiation of dendritic spikes, and integration of synaptic inputs. Subthreshold currents mediated by sodium channels are of interest because of their activation near rest, slow inactivation kinetics, and consequent effects on excitability. Modulation of these currents can also perturb physiological responses of a neuron that might underlie pathological states such as epilepsy. Using nucleated patches from the peri-somatic region of hippocampal CA1 neurons, we recorded a slowly inactivating component of the macroscopic Na+ current (which we have called INaS) that shared many biophysical properties with the persistent Na+ current, INaP, but showed distinctively faster inactivating kinetics. Ramp voltage commands with a velocity of 400 mV/s were found to elicit this component of Na+ current reliably. INaS also showed a more hyperpolarized I-V relationship and slower inactivation than those of the fast transient Na+ current (INaT) recorded in the same patches. The peak amplitude of INaS was proportional to the peak amplitude of INaT but was much smaller in amplitude. Hexanol, riluzole, and ranolazine, known Na+ channel blockers, were tested to compare their effects on both INaS and INaT. The peak conductance of INaS was preferentially blocked by hexanol and riluzole, but the shift of half-inactivation voltage (V1/2) was only observed in the presence of riluzole. Current-clamp measurements with hexanol suggested that INaS was involved in generation of an action potential and in upregulation of neuronal excitability. PMID:23236005

  9. High-Pressure Inactivation of Rotaviruses: Role of Treatment Temperature and Strain Diversity in Virus Inactivation

    PubMed Central

    Araud, Elbashir; DiCaprio, Erin; Yang, Zhihong; Li, Xinhui; Lou, Fangfei; Hughes, John H.; Chen, Haiqiang

    2015-01-01

    Rotavirus (RV) is the major etiological agent of acute gastroenteritis in infants worldwide. Although high-pressure processing (HPP) is a popular method to inactivate enteric pathogens in food, the sensitivity of different virus strains within same species and serotype to HPP is variable. This study aimed to compare the barosensitivities of seven RV strains derived from four serotypes (serotype G1, strains Wa, Ku, and K8; serotype G2, strain S2; serotype G3, strains SA-11 and YO; and serotype G4, strain ST3) following high-pressure treatment. RV strains showed various responses to HPP based on the initial temperature and had different inactivation profiles. Ku, K8, S2, SA-11, YO, and ST3 showed enhanced inactivation at 4°C compared to 20°C. In contrast, strain Wa was not significantly impacted by the initial treatment temperature. Within serotype G1, strain Wa was significantly (P < 0.05) more resistant to HPP than strains Ku and K8. Overall, the resistance of the human RV strains to HPP at 4°C can be ranked as Wa > Ku = K8 > S2 > YO > ST3, and in terms of serotype the ranking is G1 > G2 > G3 > G4. In addition, pressure treatment of 400 MPa for 2 min was sufficient to eliminate the Wa strain, the most pressure-resistant RV, from oyster tissues. HPP disrupted virion structure but did not degrade viral protein or RNA, providing insight into the mechanism of viral inactivation by HPP. In conclusion, HPP is capable of inactivating RV at commercially acceptable pressures, and the efficacy of inactivation is strain dependent. PMID:26187961

  10. Inactivation of conserved genes induces microbial aversion, drug detoxification, and innate immunity in C.elegans

    PubMed Central

    Melo, Justine A.; Ruvkun, Gary

    2012-01-01

    Summary The nematode C. elegans consumes benign bacteria such as E. coli and is repelled by pathogens and toxins. Here we show that RNAi and toxin-mediated disruption of core cellular activities, including translation, respiration, and protein turnover, stimulates behavioral avoidance of attractive E. coli. RNAi of such essential processes also induces expression of detoxification and innate immune response genes in the absence of toxins or pathogens. Disruption of core processes in non-neuronal tissues can stimulate aversion behavior, revealing a neuroendocrine axis of control. Microbial avoidance requires serotonergic and Jnk kinase signaling. We propose that surveillance pathways oversee critical cellular activities to detect pathogens, many of which deploy toxins and virulence factors to disrupt these same host pathways. Variation in cellular surveillance and endocrine pathways controlling behavior, detoxification and immunity selected by past toxin or microbial interactions could underlie aberrant responses to foods, medicines, and microbes. PMID:22500807

  11. Capsid functions of inactivated human picornaviruses and feline calicivirus.

    PubMed

    Nuanualsuwan, Suphachai; Cliver, Dean O

    2003-01-01

    The exceptional stability of enteric viruses probably resides in their capsids. The capsid functions of inactivated human picornaviruses and feline calicivirus (FCV) were determined. Viruses were inactivated by UV, hypochlorite, high temperature (72 degrees C), and physiological temperature (37 degrees C), all of which are pertinent to transmission via food and water. Poliovirus (PV) and hepatitis A virus (HAV) are transmissible via water and food, and FCV is the best available surrogate for the Norwalk-like viruses, which are leading causes of food-borne and waterborne disease in the United States. The capsids of all 37 degrees C-inactivated viruses still protected the viral RNA against RNase, even in the presence of proteinase K, which contrasted with findings with viruses inactivated at 72 degrees C. The loss of ability of the virus to attach to homologous cell receptors was universal, regardless of virus type and inactivation method, except for UV-inactivated HAV, and so virus inactivation was almost always accompanied by the loss of virus attachment. Inactivated HAV and FCV were captured by homologous antibodies. However, inactivated PV type 1 (PV-1) was not captured by homologous antibody and 37 degrees C-inactivated PV-1 was only partially captured. The epitopes on the capsids of HAV and FCV are evidently discrete from the receptor attachment sites, unlike those of PV-1. These findings indicate that the primary target of UV, hypochlorite, and 72 degrees C inactivation is the capsid and that the target of thermal inactivation (37 degrees C versus 72 degrees C) is temperature dependent.

  12. Lactose digestion by yogurt beta-galactosidase: influence of pH and microbial cell integrity.

    PubMed

    Martini, M C; Bollweg, G L; Levitt, M D; Savaiano, D A

    1987-02-01

    Lactase-deficient subjects more effectively digest lactose in yogurt than lactose in other dairy products, apparently due to yogurt microbial beta-galactosidase (beta-gal) which is active in the GI tract. We evaluated the effects of buffering capacity of yogurt, gastric pH, and microbial cell disruption on beta-gal activity and lactose digestion. Three times more acid was required to acidify yogurt than to acidify milk. Yogurt beta-gal was stable at pH 4.0 but inactivated at lower pH. When yogurt was sonicated to disrupt microbial cell structure, only 20% activity remained after incubation at pH 4.0 for 60 min. In vivo gastric pH remained greater than 2.7 for 3 h after ingestion of yogurt. Acidified milk alone or with disrupted yogurt microorganisms caused twice as much lactose malabsorption as did acidified milk containing intact yogurt microorganisms. The results provide a possible explanation for the survival of beta-gal activity from yogurt in the GI tract.

  13. Vertical variation of a black soil's properties in response to freeze-thaw cycles and its links to shift of microbial community structure.

    PubMed

    Han, Ziming; Deng, Mingwen; Yuan, Anqi; Wang, Jiahui; Li, Hao; Ma, Jincai

    2018-06-01

    Soil freeze-thaw cycles (FTCs) change soil physical, chemical, and biological properties, however information regarding their vertical variations in response to FTCs is limited. In this work, black soil (silty loam) packed soil columns were exposed to 8 FTCs, and soil properties were determined for each of vertical layer of soil columns. The results revealed that after FTCs treatment, moisture and electrical conductivity (EC) salinity tended to increase in upper soil layers. Increments of ammonium nitrogen (NH 4 + -N) and nitrate nitrogen (NO 3 - -N) in top layers (0-10cm) were greater than those in other layers, and increments of water soluble organic carbon (WSOC) and decrease of microbial biomass carbon (MBC) in middle layers (10-20cm) were greater than those in both ends. Overall, microbial community structure was mainly influenced by soil physical properties (moisture and EC) and chemical properties (pH and WSOC). For bacterial (archaeal) and fungal communities, soil physical properties, chemical properties and their interaction explained 79.73% and 82.66% of total variation, respectively. Our results provided insights into the vertical variation of soil properties caused by FTCs, and such variation had a major impact on the change of structure and composition of soil bacterial and fungal communities. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Extracting survival parameters from isothermal, isobaric, and "iso-concentration" inactivation experiments by the "3 end points method".

    PubMed

    Corradini, M G; Normand, M D; Newcomer, C; Schaffner, D W; Peleg, M

    2009-01-01

    Theoretically, if an organism's resistance can be characterized by 3 survival parameters, they can be found by solving 3 simultaneous equations that relate the final survival ratio to the lethal agent's intensity. (For 2 resistance parameters, 2 equations will suffice.) In practice, the inevitable experimental scatter would distort the results of such a calculation or render the method unworkable. Averaging the results obtained with more than 3 final survival ratio triplet combinations, determined in four or more treatments, can remove this impediment. This can be confirmed by the ability of a kinetic inactivation model derived from the averaged parameters to predict survival patterns under conditions not employed in their determination, as demonstrated with published isothermal survival data of Clostridium botulinum spores, isobaric data of Escherichia coli under HPP, and Pseudomonas exposed to hydrogen peroxide. Both the method and the underlying assumption that the inactivation followed a Weibull-Log logistic (WeLL) kinetics were confirmed in this way, indicating that when an appropriate survival model is available, it is possible to predict the entire inactivation curves from several experimental final survival ratios alone. Where applicable, the method could simplify the experimental procedure and lower the cost of microbial resistance determinations. In principle, the methodology can be extended to deteriorative chemical reactions if they too can be characterized by 2 or 3 kinetic parameters.

  15. Photodynamic Inactivation of Mammalian Viruses and Bacteriophages

    PubMed Central

    Costa, Liliana; Faustino, Maria Amparo F.; Neves, Maria Graça P. M. S.; Cunha, Ângela; Almeida, Adelaide

    2012-01-01

    Photodynamic inactivation (PDI) has been used to inactivate microorganisms through the use of photosensitizers. The inactivation of mammalian viruses and bacteriophages by photosensitization has been applied with success since the first decades of the last century. Due to the fact that mammalian viruses are known to pose a threat to public health and that bacteriophages are frequently used as models of mammalian viruses, it is important to know and understand the mechanisms and photodynamic procedures involved in their photoinactivation. The aim of this review is to (i) summarize the main approaches developed until now for the photodynamic inactivation of bacteriophages and mammalian viruses and, (ii) discuss and compare the present state of the art of mammalian viruses PDI with phage photoinactivation, with special focus on the most relevant mechanisms, molecular targets and factors affecting the viral inactivation process. PMID:22852040

  16. Non-linear pressure/temperature-dependence of high pressure thermal inactivation of proteolytic Clostridium botulinum type B in foods.

    PubMed

    Maier, Maximilian B; Lenz, Christian A; Vogel, Rudi F

    2017-01-01

    The effect of high pressure thermal (HPT) processing on the inactivation of spores of proteolytic type B Clostridium botulinum TMW 2.357 in four differently composed low-acid foods (green peas with ham, steamed sole, vegetable soup, braised veal) was studied in an industrially feasible pressure range and temperatures between 100 and 120°C. Inactivation curves exhibited rapid inactivation during compression and decompression followed by strong tailing effects. The highest inactivation (approx. 6-log cycle reduction) was obtained in braised veal at 600 MPa and 110°C after 300 s pressure-holding time. In general, inactivation curves exhibited similar negative exponential shapes, but maximum achievable inactivation levels were lower in foods with higher fat contents. At high treatment temperatures, spore inactivation was more effective at lower pressure levels (300 vs. 600 MPa), which indicates a non-linear pressure/temperature-dependence of the HPT spore inactivation efficiency. A comparison of spore inactivation levels achievable using HPT treatments versus a conventional heat sterilization treatment (121.1°C, 3 min) illustrates the potential of combining high pressures and temperatures to replace conventional retorting with the possibility to reduce the process temperature or shorten the processing time. Finally, experiments using varying spore inoculation levels suggested the presence of a resistant fraction comprising approximately 0.01% of a spore population as reason for the pronounced tailing effects in survivor curves. The loss of the high resistance properties upon cultivation indicates that those differences develop during sporulation and are not linked to permanent modifications at the genetic level.

  17. Microbial production of polyhydroxybutyrate with tailor-made properties: an integrated modelling approach and experimental validation.

    PubMed

    Penloglou, Giannis; Chatzidoukas, Christos; Kiparissides, Costas

    2012-01-01

    The microbial production of polyhydroxybutyrate (PHB) is a complex process in which the final quantity and quality of the PHB depend on a large number of process operating variables. Consequently, the design and optimal dynamic operation of a microbial process for the efficient production of PHB with tailor-made molecular properties is an extremely interesting problem. The present study investigates how key process operating variables (i.e., nutritional and aeration conditions) affect the biomass production rate and the PHB accumulation in the cells and its associated molecular weight distribution. A combined metabolic/polymerization/macroscopic modelling approach, relating the process performance and product quality with the process variables, was developed and validated using an extensive series of experiments and measurements. The model predicts the dynamic evolution of the biomass growth, the polymer accumulation, the consumption of carbon and nitrogen sources and the average molecular weights of the PHB in a bioreactor, under batch and fed-batch operating conditions. The proposed integrated model was used for the model-based optimization of the production of PHB with tailor-made molecular properties in Azohydromonas lata bacteria. The process optimization led to a high intracellular PHB accumulation (up to 95% g of PHB per g of DCW) and the production of different grades (i.e., different molecular weight distributions) of PHB. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Human platelet gel supernatant inactivates opportunistic wound pathogens on skin.

    PubMed

    Edelblute, Chelsea M; Donate, Amy L; Hargrave, Barbara Y; Heller, Loree C

    2015-01-01

    Activation of human platelets produces a gel-like substance referred to as platelet rich plasma or platelet gel. Platelet gel is used clinically to promote wound healing; it also exhibits antimicrobial properties that may aid in the healing of infected wounds. The purpose of this study was to quantify the efficacy of human platelet gel against the opportunistic bacterial wound pathogens Acinetobacter baumannii, Pseudomonas aeruginosa, and Staphylococcus aureus on skin. These opportunistic pathogens may exhibit extensive antibiotic resistance, necessitating the development of alternative treatment options. The antimicrobial efficacy of platelet gel supernatants was quantified using an in vitro broth dilution assay, an ex vivo inoculated skin assay, and in an in vivo skin decontamination assay. Human platelet gel supernatants were highly bactericidal against A. baumannii and moderately but significantly bactericidal against S. aureus in vitro and in the ex vivo skin model. P. aeruginosa was not inactivated in vitro; a low but significant inactivation level was observed ex vivo. These supernatants were quite effective at inactivating a model organism on skin in vivo. These results suggest application of platelet gel has potential clinical applicability, not only in the acceleration of wound healing, but also against relevant bacteria causing wound infections.

  19. Microbial Communities as Experimental Units

    PubMed Central

    DAY, MITCH D.; BECK, DANIEL; FOSTER, JAMES A.

    2011-01-01

    Artificial ecosystem selection is an experimental technique that treats microbial communities as though they were discrete units by applying selection on community-level properties. Highly diverse microbial communities associated with humans and other organisms can have significant impacts on the health of the host. It is difficult to find correlations between microbial community composition and community-associated diseases, in part because it may be impossible to define a universal and robust species concept for microbes. Microbial communities are composed of potentially thousands of unique populations that evolved in intimate contact, so it is appropriate in many situations to view the community as the unit of analysis. This perspective is supported by recent discoveries using metagenomics and pangenomics. Artificial ecosystem selection experiments can be costly, but they bring the logical rigor of biological model systems to the emerging field of microbial community analysis. PMID:21731083

  20. Anti-microbial properties of histone H2A from skin secretions of rainbow trout, Oncorhynchus mykiss.

    PubMed Central

    Fernandes, Jorge M O; Kemp, Graham D; Molle, M Gerard; Smith, Valerie J

    2002-01-01

    Skin exudates of rainbow trout contain a potent 13.6 kDa anti-microbial protein which, from partial internal amino acid sequencing, peptide mass fingerprinting, matrix-associated laser desorption/ionization MS and amino acid analysis, seems to be histone H2A, acetylated at the N-terminus. The protein, purified to homogeneity by ion-exchange and reversed-phase chromatography, exhibits powerful anti-bacterial activity against Gram-positive bacteria, with minimal inhibitory concentrations in the submicromolar range. Kinetic analysis revealed that at a concentration of 0.3 microM all test bacteria lose viability after 30 min incubation. Weaker activity is also displayed against the yeast Saccharomyces cerevisiae. The protein is salt-sensitive and has no haemolytic activity towards trout erythrocytes at concentrations below 0.3 microM. Reconstitution of the protein in a planar lipid bilayer strongly disturbs the membrane but does not form stable ion channels, indicating that its anti-bacterial activity is probably not due to pore-forming properties. This is the first report to show that, in addition to its classical function in the cell, histone H2A has extremely strong anti-microbial properties and could therefore help contribute to protection against bacterial invasion. PMID:12164782

  1. Quasi-specific access of the potassium channel inactivation gate

    PubMed Central

    Venkataraman, Gaurav; Srikumar, Deepa; Holmgren, Miguel

    2014-01-01

    Many voltage-gated potassium channels open in response to membrane depolarization and then inactivate within milliseconds. Neurons use these channels to tune their excitability. In Shaker K+ channels, inactivation is caused by the cytoplasmic amino terminus, termed the inactivation gate. Despite having four such gates, inactivation is caused by the movement of a single gate into a position that occludes ion permeation. The pathway that this single inactivation gate takes into its inactivating position remains unknown. Here we show that a single gate threads through the intracellular entryway of its own subunit, but the tip of the gate has sufficient freedom to interact with all four subunits deep in the pore, and does so with equal probability. This pathway demonstrates that flexibility afforded by the inactivation peptide segment at the tip of the N-terminus is used to mediate function. PMID:24909510

  2. Effects of root pruning on the physicochemical properties and microbial activities of poplar rhizosphere soil.

    PubMed

    Jing, Da-Wei; Liu, Fang-Chun; Wang, Ming-You; Ma, Hai-Lin; Du, Zhen-Yu; Ma, Bing-Yao; Dong, Yu-Feng

    2017-01-01

    This study aimed to determine the effects of root pruning on the physicochemical characteristics and microbial activities of poplar rhizosphere soil. The root systems of 5-year-old poplar (Populus×euramericana cv. 'Neva') trees were manually pruned at 6, 8, or 10 times diameter at breast height (DBH) from the trunk (severe, moderate, and light, respectively) along both inter-row sides. Moderate root pruning significantly increased the concentrations of amino acids, organic acids, and total sugars in the root exudates and decreased the pH of rhizosphere soil. This treatment also increased the contents of available nitrogen, phosphorus, potassium, and total organic carbon as well as high-, medium-, and low-activity organic carbon in rhizosphere soil. Moreover, moderate pruning increased the contents of microbial biomass carbon and nitrogen, and enhanced basal respiration, in addition to decreasing the metabolic quotients in rhizosphere soil by 8.9%, 5.0%, and 11.4% compared with control, light, and severe root pruning treatments, respectively. Moderate pruning increased the growth rates of DBH, tree height, and volume to the highest levels. Furthermore, these indices were not significantly different between the light root pruning and control groups, but varied significantly between severe and moderate root-pruning treatments. Thus, root pruning, depending on the distance from the trunk, significantly influences the physicochemical properties and microbial activities in poplar rhizosphere soil.

  3. Effects of root pruning on the physicochemical properties and microbial activities of poplar rhizosphere soil

    PubMed Central

    Jing, Da-Wei; Liu, Fang-Chun; Wang, Ming-You; Ma, Hai-Lin; Du, Zhen-Yu; Ma, Bing-Yao; Dong, Yu-Feng

    2017-01-01

    This study aimed to determine the effects of root pruning on the physicochemical characteristics and microbial activities of poplar rhizosphere soil. The root systems of 5-year-old poplar (Populus×euramericana cv. ‘Neva’) trees were manually pruned at 6, 8, or 10 times diameter at breast height (DBH) from the trunk (severe, moderate, and light, respectively) along both inter-row sides. Moderate root pruning significantly increased the concentrations of amino acids, organic acids, and total sugars in the root exudates and decreased the pH of rhizosphere soil. This treatment also increased the contents of available nitrogen, phosphorus, potassium, and total organic carbon as well as high-, medium-, and low-activity organic carbon in rhizosphere soil. Moreover, moderate pruning increased the contents of microbial biomass carbon and nitrogen, and enhanced basal respiration, in addition to decreasing the metabolic quotients in rhizosphere soil by 8.9%, 5.0%, and 11.4% compared with control, light, and severe root pruning treatments, respectively. Moderate pruning increased the growth rates of DBH, tree height, and volume to the highest levels. Furthermore, these indices were not significantly different between the light root pruning and control groups, but varied significantly between severe and moderate root-pruning treatments. Thus, root pruning, depending on the distance from the trunk, significantly influences the physicochemical properties and microbial activities in poplar rhizosphere soil. PMID:29117215

  4. Engineering of Microbial Cell Factories for the Production of Plant Polyphenols with Health-Beneficial Properties.

    PubMed

    Dudnik, Alexey; Gaspar, Paula; Neves, Ana Rute; Forster, Jochen

    2018-05-15

    Polyphenols form a group of important natural bioactive compounds with numerous ascribed health-beneficial attributes (e.g. antioxidant, anti-inflammatory, anti-microbial and tumor-suppressing properties). Some polyphenols can also be used as natural dyes or plastic precursors. Notwithstanding their relevance, production of most of these compounds still relies on extraction from plant material, which for most of it is a costly and an inefficient procedure. The use of microbial cell factories for this purpose is an emerging alternative that could allow a more efficient and sustainable production. The most recent advances in molecular biology and genetic engineering, combined with the ever-growing understanding of microbial physiology have led to multiple success stories. Production of multiple polyphenolic compounds or their direct precursors has been achieved not only in the common production hosts, such as Escherichia coli and Saccharomyces cerevisiae but also in Corynebacterium glutamicum and Lactococcus lactis. However, boosting production of native compounds or introduction of heterologous biosynthetic pathways also brings certain challenges, such as the need to express, balance and maintain efficient precursor supply. This review will discuss the most recent advances in the field of metabolic engineering of microorganisms for polyphenol biosynthesis and its future perspectives, as well as outlines their potential health benefits and current production methods. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  5. Mathematical quantification of the induced stress resistance of microbial populations during non-isothermal stresses.

    PubMed

    Garre, Alberto; Huertas, Juan Pablo; González-Tejedor, Gerardo A; Fernández, Pablo S; Egea, Jose A; Palop, Alfredo; Esnoz, Arturo

    2018-02-02

    This contribution presents a mathematical model to describe non-isothermal microbial inactivation processes taking into account the acclimation of the microbial cell to thermal stress. The model extends the log-linear inactivation model including a variable and model parameters quantifying the induced thermal resistance. The model has been tested on cells of Escherichia coli against two families of non-isothermal profiles with different constant heating rates. One of the families was composed of monophasic profiles, consisting of a non-isothermal heating stage from 35 to 70°C; the other family was composed of biphasic profiles, consisting of a non-isothermal heating stage followed by a holding period at constant temperature of 57.5°C. Lower heating rates resulted in a higher thermal resistance of the bacterial population. This was reflected in a higher D-value. The parameter estimation was performed in two steps. Firstly, the D and z-values were estimated from the isothermal experiments. Next, the parameters describing the acclimation were estimated using one of the biphasic profiles. This set of parameters was able to describe the remaining experimental data. Finally, a methodology for the construction of diagrams illustrating the magnitude of the induced thermal resistance is presented. The methodology has been illustrated by building it for a biphasic temperature profile with a linear heating phase and a holding phase. This diagram provides a visualization of how the shape of the temperature profile (heating rate and holding temperature) affects the acclimation of the cell to the thermal stress. This diagram can be used for the design of inactivation treatments by industry taking into account the acclimation of the cell to the thermal stress. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Non-thermal plasma for inactivated-vaccine preparation.

    PubMed

    Wang, Guomin; Zhu, Ruihao; Yang, Licong; Wang, Kaile; Zhang, Qian; Su, Xia; Yang, Bing; Zhang, Jue; Fang, Jing

    2016-02-17

    Vaccines are of great importance in controlling the spread of infectious diseases in poultry farming. The safety and efficacy of vaccines are also essential. To explore the feasibility of a novel technology (non-thermal plasma) in inactivated vaccine preparation, an alternating current atmospheric pressure non-thermal plasma (NTP) jet with Ar/O2/N2 as the operating gas was used to inactivate a Newcastle disease virus (NDV, LaSota) strain and H9N2 avian influenza virus (AIV, A/Chicken/Hebei/WD/98) for vaccine preparation. The results showed that complete inactivation could be achieved with 2 min of NTP treatment for both NDV and AIV. Moreover, a proper NTP treatment time is needed for inactivation of a virus without destruction of the antigenic determinants. Compared to traditional formaldehyde-inactivated vaccine, the vaccine made from NDV treated by NTP for 2 min (NTP-2 min-NDV-vaccine) could induce a higher NDV-specific antibody titer in specific pathogen-free (SPF) chickens, and the results of a chicken challenge experiment showed that NTP-2 min-NDV-vaccine could protect SPF chickens from a lethal NDV challenge. Vaccines made from AIV treated by NTP for 2 min (NTP-2 min-AIV-vaccine) also showed a similar AIV-specific antibody titer compared with traditional AIV vaccines prepared using formaldehyde inactivation. Studies of the morphological changes of the virus, chemical analysis of NDV allantoic fluid and optical emission spectrum analysis of NTP suggested that reactive oxygen species and reactive nitrogen species produced by NTP played an important role in the virus inactivation process. All of these results demonstrated that it could be feasible to use non-thermal NTP as an alternative strategy to prepare inactivated vaccines for Newcastle disease and avian influenza. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Impact of Organic and Conventional Systems of Coffee Farming on Soil Properties and Culturable Microbial Diversity

    PubMed Central

    2016-01-01

    A study was undertaken with an objective of evaluating the long-term impacts of organic (ORG) and conventional (CON) methods of coffee farming on soil physical, chemical, biological, and microbial diversity. Electrical conductivity and bulk density were found to increase by 34% and 21%, respectively, in CON compared to ORG system, while water holding capacity was found decreased in both the systems. Significant increase in organic carbon was observed in ORG system. Major nutrients, nitrogen and potassium, levels showed inclination in both ORG and CON system, but the trend was much more pronounced in CON system. Phosphorus was found to increase in both ORG and CON system, but its availability was found to be more with CON system. In biological attributes, higher soil respiration and fluorescein diacetate activity were recorded in ORG system compared to CON system. Higher soil urease activity was observed in CON system, while dehydrogenase activity does not show significant differences between ORG and CON systems. ORG system was found to have higher macrofauna (31.4%), microbial population (34%), and microbial diversity indices compared to CON system. From the present study, it is accomplished that coffee soil under long-term ORG system has better soil properties compared to CON system. PMID:27042378

  8. Impact of Organic and Conventional Systems of Coffee Farming on Soil Properties and Culturable Microbial Diversity.

    PubMed

    Velmourougane, Kulandaivelu

    2016-01-01

    A study was undertaken with an objective of evaluating the long-term impacts of organic (ORG) and conventional (CON) methods of coffee farming on soil physical, chemical, biological, and microbial diversity. Electrical conductivity and bulk density were found to increase by 34% and 21%, respectively, in CON compared to ORG system, while water holding capacity was found decreased in both the systems. Significant increase in organic carbon was observed in ORG system. Major nutrients, nitrogen and potassium, levels showed inclination in both ORG and CON system, but the trend was much more pronounced in CON system. Phosphorus was found to increase in both ORG and CON system, but its availability was found to be more with CON system. In biological attributes, higher soil respiration and fluorescein diacetate activity were recorded in ORG system compared to CON system. Higher soil urease activity was observed in CON system, while dehydrogenase activity does not show significant differences between ORG and CON systems. ORG system was found to have higher macrofauna (31.4%), microbial population (34%), and microbial diversity indices compared to CON system. From the present study, it is accomplished that coffee soil under long-term ORG system has better soil properties compared to CON system.

  9. Pulsed electric fields for pasteurization: defining processing conditions

    USDA-ARS?s Scientific Manuscript database

    Application of pulsed electric fields (PEF) technology in food pasteurization has been extensively studied. Optimal PEF treatment conditions for maximum microbial inactivation depend on multiple factors including PEF processing conditions, production parameters and product properties. In order for...

  10. Inactivation of murine norovirus by chemical biocides on stainless steel

    PubMed Central

    2009-01-01

    Background Human norovirus (NoV) causes more than 80% of nonbacterial gastroenteritis in Europe and the United States. NoV transmission via contaminated surfaces may be significant for the spread of viruses. Therefore, measures for prevention and control, such as surface disinfection, are necessary to interrupt the dissemination of human NoV. Murine norovirus (MNV) as a surrogate for human NoV was used to study the efficacy of active ingredients of chemical disinfectants for virus inactivation on inanimate surfaces. Methods The inactivating properties of different chemical biocides were tested in a quantitative carrier test with stainless steel discs without mechanical action. Vacuum-dried MNV was exposed to different concentrations of alcohols, peracetic acid (PAA) or glutaraldehyde (GDA) for 5 minutes exposure time. Detection of residual virus was determined by endpoint-titration on RAW 264.7 cells. Results PAA [1000 ppm], GDA [2500 ppm], ethanol [50% (v/v)] and 1-propanol [30% (v/v)] were able to inactivate MNV under clean conditions (0.03% BSA) on the carriers by ≥ 4 log10 within 5 minutes exposure time, whereas 2-propanol showed a reduced effectiveness even at 60% (v/v). Furthermore, there were no significant differences in virus reduction whatever interfering substances were used. When testing with ethanol, 1- and 2-propanol, results under clean conditions were nearly the same as in the presence of dirty conditions (0.3% BSA plus 0.3% erythrocytes). Conclusion Products based upon PAA, GDA, ethanol and 1-propanol should be used for NoV inactivation on inanimate surfaces. Our data provide valuable information for the development of strategies to control NoV transmission via surfaces. PMID:19583832

  11. Biophysical characterization and functional consequences of a slowly inactivating potassium current in neostriatal neurons.

    PubMed

    Gabel, L A; Nisenbaum, E S

    1998-04-01

    Neostriatal spiny projection neurons can display a pronounced delay in their transition to action potential discharge that is mediated by a slowly developing ramp depolarization. The possible contribution of a slowly inactivating A-type K+ current (IAs) to this delayed excitation was investigated by studying the biophysical and functional properties of IAs using whole cell voltage- and current-clamp recording from acutely isolated neostriatal neurons. Isolation of IAs from other voltage-gated, calcium-independent K+ currents was achieved through selective blockade of IAs with low concentrations (10 microM) of the benzazepine derivative, 6-chloro-7,8-dihydroxy-3-allyl- 1-phenyl-2,3,4,5-tetra-hydro-1H-3-benzazepine (APB; SKF82958) and subsequent current subtraction. Examination of the voltage dependence of activation showed that IAs began to flow at approximately -60 mV in response to depolarization. The voltage dependence of inactivation revealed that approximately 50% of IAs channels were available at the normal resting potential (-80 mV) of these cells, but that only 20% of the channels were available at membrane potentials corresponding to spike threshold (about -40 mV). At these depolarized membrane potentials, the rate of activation was moderately rapid (tau approximately 60 ms), whereas the rate of inactivation was slow (tau approximately 1.5 s). The time course of removal of inactivation of IAs at -80 mV also was relatively slow (tau approximately 1.0 s). The subthreshold availability of IAs combined with its rapid activation and slow inactivation rates suggested that this current should be capable of dampening the onset of prolonged depolarizing responses, but over time its efficacy should diminish, slowly permitting the membrane to depolarize toward spike threshold. Voltage recording experiments confirmed this hypothesis by demonstrating that application of APB at a concentration (10 microM) that selectively blocks IAs substantially decreased the latency to

  12. Effect of gamma irradiation on microbial load, physicochemical and sensory characteristics of soybeans (Glycine max L. Merrill)

    NASA Astrophysics Data System (ADS)

    Yun, Juan; Li, Xihong; Fan, Xuetong; Tang, Yao; Xiao, Yao; Wan, Sen

    2012-08-01

    Gamma irradiation is highly effective in inactivating microorganisms in various foods and offers a safe alternative method of food decontamination. In the present study, soybeans (Glycine max L. Merrill) were treated with 0, 1.0, 3.0, 5.0 and 10.0 KGy of gamma irradiation. Microbial populations on soybeans, isoflavone, tocopherol contents, raffinose family oligosaccharides, color and sensory properties were evaluated as a function of irradiation dose. The results indicated that gamma irradiation reduced aerobic bacterial and fungal load. Irradiation at the doses applied did not cause any significant change (p>0.05) in the contents of isoflavone of soybeans, but decreased tocopherol contents. The content of key flatulence-producing raffinose family oligosaccharides in irradiated soybeans (10.0 kGy) decreased by 82.1% compared to the control. Sensory analysis showed that the odor of the soybeans was organoleptically acceptable at doses up to 5.0 kGy and no significant differences were observed between irradiated and nonirradiated samples in flavor, texture and color after irradiation.

  13. Inactivation of Penicillins by Thiol Broth

    PubMed Central

    Murray, Patrick R.; Niles, Ann C.

    1982-01-01

    Thiol broth with sodium polyanetholesulfonate inactivated penicillin G, carbenicillin, nafcillin, oxacillin, and gentamicin, but had no effect on cephalothin, cefoxitin, clindamycin, chloramphenicol, erythromycin, and tetracycline. Only Thiol broth was capable of this inactivation, which was not influenced by the presence of blood. PMID:7153352

  14. Deep Diversity: Novel Approach to Overcoming the PCR Bias Encountered During Environmental Analysis of Microbial Populations for Alpha-Diversity

    NASA Technical Reports Server (NTRS)

    Ramirez, Gustavo A; Vaishampayan, Parag A.

    2011-01-01

    Alpha-diversity studies are of crucial importance to environmental microbiologists. The polymerase chain reaction (PCR) method has been paramount for studies interrogating microbial environmental samples for taxon richness. Phylogenetic studies using this technique are based on the amplification and comparison of the 16S rRNA coding regions. PCR, due disproportionate distribution of microbial species in the environment, increasingly favors the amplification of the most predominant phylotypes with every subsequent reaction cycle. The genetic and chemical complexity of environmental samples are intrinsic factors that exacerbate an inherit bias in PCR-based quantitative and qualitative studies of microbial communities. We report that treatment of a genetically complex total genomic environmental DNA extract with Propidium Monoazide (PMA), a DNA intercalating molecule capable of forming a covalent cross-linkage to organic moieties upon light exposure, disproportionally inactivates predominant phylotypes and results in the exponential amplification of previously shadowed microbial ?-diversity quantified as a 19.5% increase in OUTs reported via phylogenetic screening using PhyloChip.

  15. Voltage Sensor Inactivation in Potassium Channels

    PubMed Central

    Bähring, Robert; Barghaan, Jan; Westermeier, Regina; Wollberg, Jessica

    2012-01-01

    In voltage-gated potassium (Kv) channels membrane depolarization causes movement of a voltage sensor domain. This conformational change of the protein is transmitted to the pore domain and eventually leads to pore opening. However, the voltage sensor domain may interact with two distinct gates in the pore domain: the activation gate (A-gate), involving the cytoplasmic S6 bundle crossing, and the pore gate (P-gate), located externally in the selectivity filter. How the voltage sensor moves and how tightly it interacts with these two gates on its way to adopt a relaxed conformation when the membrane is depolarized may critically determine the mode of Kv channel inactivation. In certain Kv channels, voltage sensor movement leads to a tight interaction with the P-gate, which may cause conformational changes that render the selectivity filter non-conductive (“P/C-type inactivation”). Other Kv channels may preferably undergo inactivation from pre-open closed-states during voltage sensor movement, because the voltage sensor temporarily uncouples from the A-gate. For this behavior, known as “preferential” closed-state inactivation, we introduce the term “A/C-type inactivation”. Mechanistically, P/C- and A/C-type inactivation represent two forms of “voltage sensor inactivation.” PMID:22654758

  16. Soil microbial properties after long-term swine slurry application to conventional and no-tillage systems in Brazil.

    PubMed

    Balota, Elcio L; Machineski, Oswaldo; Hamid, Karima I A; Yada, Ines F U; Barbosa, Graziela M C; Nakatani, Andre S; Coyne, Mark S

    2014-08-15

    Swine waste can be used as an agricultural fertilizer, but large amounts may accumulate excess nutrients in soil or contaminate the surrounding environment. This study evaluated long-term soil amendment (15 years) with different levels of swine slurry to conventional (plow) tillage (CT) and no tillage (NT) soils. Long-term swine slurry application did not affect soil organic carbon. Some chemical properties, such as calcium, base saturation, and aluminum saturation were significantly different within and between tillages for various application rates. Available P and microbial parameters were significantly affected by slurry addition. Depending on tillage, soil microbial biomass and enzyme activity increased up to 120 m(3) ha(-1) year(-1) in all application rates. The NT system had higher microbial biomass and activity than CT at all application levels. There was an inverse relationship between the metabolic quotient (qCO2) and MBC, and the qCO2 was 53% lower in NT than CT. Swine slurry increased overall acid phosphatase activity, but the phosphatase produced per unit of microbial biomass decreased. A comparison of data obtained in the 3rd and 15th years of swine slurry application indicated that despite slurry application the CT system degraded with time while the NT system had improved values of soil quality indicators. For these Brazilian oxisols, swine slurry amendment was insufficient to maintain soil quality parameters in annual crop production without additional changes in tillage management. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Selective removal and inactivation of bacteria by nanoparticle composites prepared by surface modification of montmorillonite with quaternary ammonium compounds.

    PubMed

    Khalil, Rowaida K S

    2013-10-01

    The purpose of the present study was to prepare new nanocomposites with antibacterial activities by surface modification of montmorillonite using quaternary ammonium compounds that are widely applied as disinfectants and antiseptics in food-processing environments. The intercalation of four quaternary ammonium compounds namely benzalkonium chloride, cetylpyridinium chloride monohydrate, hexadecyltrimethylammonium bromide, tetraethylammonium chloride hydrate into montmorillonite layers was confirmed by X-ray diffraction. The antibacterial influences of the modified clay variants against important foodborne pathogens differed based on modifiers quantities, microbial cell densities, and length of contact. Elution experiments through 0.1 g of the studied montmorillonite variants indicated that Staphylococcus aureus, Pseudomonas aeroginosa, and Listeria monocytogenes were the most sensitive strains. 1 g of hexadecyltrimethylammonium bromide intercalated montmorillonites demonstrated maximum inactivation of L. monocytogenes populations, with 4.5 log c.f.u./ml units of reduction. In adsorption experiments, 0.1 g of tetraethylammonium chloride hydrate montmorillonite variants significantly reduced the growth of Escherichia coli O157:H7, L. monocytogenes, and S. aureus populations by 5.77, 6.33, and 7.38 log units respectively. Growth of wide variety of microorganisms was strongly inhibited to undetectable levels (microbial populations adsorbed to the new nanocomposites was substantially different from that in elution experiments, where interactions of nanocomposites with bacteria were specific and more complex than simple ability to inactivate. Treatment columns packed with modified variants maintained their inactivation capacity to the growth of Salmonella Tennessee and S. aureus populations after 48 h of incubation

  18. Lack of X inactivation associated with maternal X isodisomy: evidence for a counting mechanism prior to X inactivation during human embryogenesis.

    PubMed

    Migeon, B R; Jeppesen, P; Torchia, B S; Fu, S; Dunn, M A; Axelman, J; Schmeckpeper, B J; Fantes, J; Zori, R T; Driscoll, D J

    1996-01-01

    We have previously reported functional disomy for X-linked genes in females with tiny ring X chromosomes and a phenotype significantly more abnormal than Turner syndrome. In such cases the disomy results from failure of these X chromosomes to inactivate because they lack DNA sequences essential for cis X inactivation. Here we describe a novel molecular mechanism for functional X disomy that is associated with maternal isodisomy. In this case, the severe mental retardation and multiple congenital abnormalities in a female with a mosaic 45,X/ 46,X,del(X)(q21.3-qter)/ 46X,r(X) karyotype are associated with overexpression of the genes within Xpter to Xq21.31 in many of her cells. Her normal X, ring X, and deleted linear X chromosomes originate from the same maternal X chromosome, and all are transcriptionally active. None expresses X inactive specific transcript (XIST), although the locus and region of the putative X inactivation center (XIC) are present on both normal and linear deleted X chromosomes. To our knowledge, this is the first report of a functional maternal X isodisomy, and the largest X chromosome to escape inactivation. In addition, these results (1) show that cis inactivation does not invariably occur in human females with two X chromosomes, even when the XIC region is present on both of them; (2) provide evidence for a critical time prior to the visible onset of X inactivation in the embryo when decisions about X inactivation are made; and (3) support the hypothesis that the X chromosome counting mechanism involves chromosomal imprinting, occurs prior to the onset of random inactivation, and is required for subsequent inactivation of the chromosome.

  19. Inactivation of virus in solution by cold atmospheric pressure plasma: identification of chemical inactivation pathways

    NASA Astrophysics Data System (ADS)

    Aboubakr, Hamada A.; Gangal, Urvashi; Youssef, Mohammed M.; Goyal, Sagar M.; Bruggeman, Peter J.

    2016-05-01

    Cold atmospheric pressure plasma (CAP) inactivates bacteria and virus through in situ production of reactive oxygen and nitrogen species (RONS). While the bactericidal and virucidal efficiency of plasmas is well established, there is limited knowledge about the chemistry leading to the pathogen inactivation. This article describes a chemical analysis of the CAP reactive chemistry involved in the inactivation of feline calicivirus. We used a remote radio frequency CAP produced in varying gas mixtures leading to different plasma-induced chemistries. A study of the effects of selected scavengers complemented with positive control measurements of relevant RONS reveal two distinctive pathways based on singlet oxygen and peroxynitrous acid. The first mechanism is favored in the presence of oxygen and the second in the presence of air when a significant pH reduction is induced in the solution by the plasma. Additionally, smaller effects of the H2O2, O3 and \\text{NO}2- produced were also found. Identification of singlet oxygen-mediated 2-imidazolone/2-oxo-His (His  +14 Da)—an oxidative modification of His 262 comprising the capsid protein of feline calicivirus links the plasma induced singlet oxygen chemistry to viral inactivation.

  20. Microbial inactivation and MLF performances of Tempranillo Rioja wines treated with PEF after alcoholic fermentation.

    PubMed

    González-Arenzana, Lucía; López-Alfaro, Isabel; Garde-Cerdán, Teresa; Portu, Javier; López, Rosa; Santamaría, Pilar

    2018-03-23

    This study was performed with the aim of reducing the microbial communities of wines after alcoholic fermentation to improve the establishment of commercial Oenococcus oeni inoculum for developing the malolactic fermentation. Microbial community reduction was accomplished by applying Pulsed Electric Field (PEF) technology to four different wines. Overall, significant reductions in yeast population were observed. To a lesser extent, lactic acid bacteria were reduced while acetic acid bacteria were completely eliminated after the PEF treatment. In three out of the four tested wines, a decrease in the competitive pressure between microorganisms due to the detected reduction led to a general but slight shortening of the malolactic fermentation duration. In the wine with the most adverse conditions to commercial starter establishment, the shortest malolactic fermentation was reached after PEF treatment. Finally, the sensorial quality of three out of the four treated wines was considered better after the PEF treatment. Therefore, PEF technology meant an important tool for improving the malolactic fermentation performance. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. GEOELECTRICAL EVIDENCE OF MICROBIAL DEGRADATION OF DIESEL CONTAMINATED SEDIMENTS

    EPA Science Inventory

    The alteration of physical properties by microbial activity in petroleum contaminated sediments was investigated using geophysical techniques in laboratory column experiments. Microbial population growth was determined by the Most Probable Number technique (MPN), community dynami...

  2. Multi-Layered TiO2 Films towards Enhancement of Escherichia coli Inactivation

    PubMed Central

    Yoriya, Sorachon; Chumphu, Angkana; Pookmanee, Pusit; Laithong, Wreerat; Thepa, Sirichai; Songprakorp, Roongrojana

    2016-01-01

    Crystalline TiO2 has shown its great photocatalytic properties in bacterial inactivation. This work presents a design fabrication of low-cost, layered TiO2 films assembled reactors and a study of their performance for a better understanding to elucidate the photocatalytic effect on inactivation of E. coli in water. The ability to reduce the number of bacteria in water samples for the layered TiO2 composing reactors has been investigated as a function of time, while varying the parameters of light sources, initial concentration of bacteria, and ratios of TiO2 film area and volume of water. Herein, the layered TiO2 films have been fabricated on the glass plates by thermal spray coating prior to screen printing, allowing a good adhesion of the films. Surface topology and crystallographic phase of TiO2 for the screen-printed active layer have been characterized, resulting in the ratio of anatase:rutile being 80:20. Under exposure to sunlight and a given condition employed in this study, the optimized film area:water volume of 1:2.62 has shown a significant ability to reduce the E. coli cells in water samples. The ratio of surface area of photocatalytic active base to volume of water medium is believed to play a predominant role facilitating the cells inactivation. The kinetic rate of inactivation and its behavior are also described in terms of adsorption of reaction species at different contact times. PMID:28773930

  3. Randomized Trials Comparing Inactivated Vaccine After Medium- or High-titer Measles Vaccine With Standard Titer Measles Vaccine After Inactivated Vaccine: A Meta-analysis.

    PubMed

    Aaby, Peter; Ravn, Henrik; Benn, Christine S; Rodrigues, Amabelia; Samb, Badara; Ibrahim, Salah A; Libman, Michael D; Whittle, Hilton C

    2016-11-01

    Observational studies have suggested that girls have higher mortality if their most recent immunization is an inactivated vaccine rather than a live vaccine. We therefore reanalyzed 5 randomized trials of early measles vaccine (MV) in which it was possible to compare an inactivated vaccines [after medium-titer MV (MTMV) or high-titer MV (HTMV)] and a live standard titer MV (after an initial inactivated vaccine). The trials were conducted in Sudan, Senegal, The Gambia and Guinea-Bissau. The intervention group received live MTMV or HTMV from 4 to 5 months and then an inactivated vaccine from 9 to 10 months of age; the control children received inactivated vaccine/placebo from 4 to 5 months and standard titer MV from 9 to 10 months of age. We compared mortality from 9 months until end of study at 3 to 5 years of age for children who received inactivated vaccine (after MTMV or HTMV) and standard titer MV (after inactivated vaccine), respectively. The original datasets were analyzed using a Cox proportional hazards model stratified by trial. The mortality rate ratio (MRR) was 1.38 (95% confidence interval: 1.05-1.83) after an inactivated vaccine (after MTMV or HTMV) compared with a standard titer MV (after inactivated vaccine). Girls had a MRR of 1.89 (1.27-2.80), whereas there was no effect for boys, the sex-differential effect being significant (P = 0.02). Excluding measles cases did not alter these conclusions, the MRR after inactivated vaccines (after MTMV or HTMV) being 1.40 (1.06-1.86) higher overall and 1.92 (1.29-2.86) for girls. Control for variations in national immunization schedules for other vaccines did not modify these results. After 9 months of age, all children had been immunized against measles, and mortality in girls was higher when they had received inactivated vaccines (after MTMV or HTMV) rather than live standard titer MV (after an inactivated vaccine).

  4. Effect of a residue after evaporation from industrial vitamin C fermentation on chemical and microbial properties of alkali-saline soil.

    PubMed

    Kong, Tao; Xu, Hui; Wang, Zhenyu; Sun, Hao; Wang, Lihua

    2014-07-01

    Residue after evaporation (RAE) from industrial vitamin C fermentation is emitted as a waste product at an amount of 60,000 tons per year in China. The disposal of RAE is difficult because of its high chemical oxygen demand (1.17×10(6) mg/l) and low pH (0.27). We hypothesized that RAE could be used as an ameliorant for alkali-saline soils, and tried to verify it by carrying out a pot experiment of pakchoi cultivation and to explore its effect on soil chemical and microbial properties. The results showed that pakchoi yield was increased by 28.13% and pakchoi quality was also enhanced under RAE treatment. The improved chemical and microbial properties of treated soil were also observed: soil pH was decreased from 9.19 to 9.03; total organic carbon, available phosphorus and available potassium were increased by 49.15%, 34.91% and 42.02%, respectively; number of culturable bacteria, actinomycetes and fungi, microbial biomass carbon and enzyme activity number were improved by 52.97%, 104.05%, 79.09%, 57.82% and 31.16%, respectively. These results suggested the residue application led to an improved soil quality and subsequently a higher yield and quality of pakchoi. This study provided a strong evidence for the feasibility of RAE as an ameliorant for alkali-saline soil.

  5. A review on the application of inorganic nano-structured materials in the modification of textiles: focus on anti-microbial properties.

    PubMed

    Dastjerdi, Roya; Montazer, Majid

    2010-08-01

    Textiles can provide a suitable substrate to grow micro-organisms especially at appropriate humidity and temperature in contact to human body. Recently, increasing public concern about hygiene has been driving many investigations for anti-microbial modification of textiles. However, using many anti-microbial agents has been avoided because of their possible harmful or toxic effects. Application of inorganic nano-particles and their nano-composites would be a good alternative. This review paper has focused on the properties and applications of inorganic nano-structured materials with good anti-microbial activity potential for textile modification. The discussed nano-structured anti-microbial agents include TiO(2) nano-particles, metallic and non-metallic TiO(2) nano-composites, titania nanotubes (TNTs), silver nano-particles, silver-based nano-structured materials, gold nano-particles, zinc oxide nano-particles and nano-rods, copper nano-particles, carbon nanotubes (CNTs), nano-clay and its modified forms, gallium, liposomes loaded nano-particles, metallic and inorganic dendrimers nano-composite, nano-capsules and cyclodextrins containing nano-particles. This review is also concerned with the application methods for the modification of textiles using nano-structured materials. Copyright 2010 Elsevier B.V. All rights reserved.

  6. Spatial heterogeneity of physicochemical properties explains differences in microbial composition in arid soils from Cuatro Cienegas, Mexico

    PubMed Central

    Pajares, Silvia; Noguez, Ana M.; García-Oliva, Felipe; Martínez-Piedragil, Celeste; Cram, Silke S.; Eguiarte, Luis Enrique; Souza, Valeria

    2016-01-01

    Arid ecosystems are characterized by high spatial heterogeneity, and the variation among vegetation patches is a clear example. Soil biotic and abiotic factors associated with these patches have also been well documented as highly heterogeneous in space. Given the low vegetation cover and little precipitation in arid ecosystems, soil microorganisms are the main drivers of nutrient cycling. Nonetheless, little is known about the spatial distribution of microorganisms and the relationship that their diversity holds with nutrients and other physicochemical gradients in arid soils. In this study, we evaluated the spatial variability of soil microbial diversity and chemical parameters (nutrients and ion content) at local scale (meters) occurring in a gypsum-based desert soil, to gain knowledge on what soil abiotic factors control the distribution of microbes in arid ecosystems. We analyzed 32 soil samples within a 64 m2 plot and: (a) characterized microbial diversity using T-RFLPs of the bacterial 16S rRNA gene, (b) determined soil chemical parameters, and (c) identified relationships between microbial diversity and chemical properties. Overall, we found a strong correlation between microbial composition heterogeneity and spatial variation of cations (Ca2, K+) and anions (HCO\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{upgreek} \\usepackage{mathrsfs} \\setlength{\\oddsidemargin}{-69pt} \\begin{document} }{}${}_{3}^{-}$\\end{document}3−, Cl−, SO\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{upgreek} \\usepackage{mathrsfs} \\setlength{\\oddsidemargin}{-69pt} \\begin{document} }{}${}_{4}^{2-}$\\end{document}42−) content in this small plot. Our results could be attributable to spatial differences of soil saline content, favoring the patchy emergence of

  7. Investigation of E. coli bacteria inactivation by photocatalytic activity of TiO2 coated expanded polystyrene foam

    NASA Astrophysics Data System (ADS)

    Varnagiris, S.; Sakalauskaite, S.; Tuckute, S.; Lelis, M.; Daugelavicius, R.; Milcius, D.

    2017-03-01

    Photocatalytic properties of anatase and other TiO2 polymorphs are widely researched and applied in practical application. In current study TiO2 films on the plasma pre-treated expanded polystyrene (EPS) foam were deposited using magnetron sputtering technique. Main properties of the films were characterised using combination of XRD, XPS and SEM techniques. Photocatalytic properties of the observed crystalline anatase phase were tested by investigating bleaching of the methylene blue (MB) aqueous solution and by testing Escherichia coli (E. coli) viability after incubation under UV-B irradiation. E. coli viability experiments indicated that there are two mechanisms of E. coli bacteria inactivation. UV irradiation alone causes rapid damage to the outer membrane of E. coli bacteria. The second mechanism of E. coli inactivation is invoked only with synergistic combination of TiO2 and UV. Acting as photocatalyst TiO2 generates active radicals who initiate the chain peroxidation of organic molecules and within 45 min reduce E. coli bacteria viability by nearly 90%.

  8. Optimization of microbial detoxification for an aquatic mercury-contaminated environment.

    PubMed

    Figueiredo, Neusa L; Canário, João; Serralheiro, Maria Luísa; Carvalho, Cristina

    2017-01-01

    Mercury (Hg) reduction performed by microorganisms is well recognized as a biological means for remediation of contaminated environment. Recently, studies demonstrated that Hg-resistant microorganisms of Tagus Estuary are involved in metal reduction processes. In the present study, aerobic microbial community isolated from a highly Hg-contaminated area of Tagus Estuary was used to determine the optimization of the reduction process in conditions such as the contaminated ecosystem. Factorial design methodology was employed to examine the influence of glucose, sulfate, iron, and chloride on Hg reduction. In the presence of several concentrations of these elements, microbial community reduced Hg in a range of 37-61% of the initial 0.1 mg/ml Hg 2+ levels. The response prediction through central composite design showed that the increase of sulfate concentration led to an optimal response in Hg reduction by microbial community, while the rise in chloride levels markedly decreased metal reduction. Iron may exert antagonistic effects depending upon the media composition. These results are useful in understanding the persistence of Hg contamination in Tagus Estuary after inactivation of critical industrial units, as well as data might also be beneficial for development of new bioremediation strategies either in Tagus Estuary and/or in other Hg-contaminated aquatic environments.

  9. Visualization of physico-chemical properties and microbial distribution in soil and root microenvironments

    NASA Astrophysics Data System (ADS)

    Eickhorst, Thilo; Schmidt, Hannes

    2016-04-01

    Plant root development is influenced by soil properties and environmental factors. In turn plant roots can also change the physico-chemical conditions in soil resulting in gradients between roots and the root-free bulk soil. By releasing a variety of substances roots facilitate microbial activities in their direct vicinity, the rhizosphere. The related microorganisms are relevant for various ecosystem functions in the root-soil interface such as nutrient cycling. It is therefore important to study the impact and dynamics of microorganisms associated to different compartments in root-soil interfaces on a biologically meaningful micro-scale. The analysis of microorganisms in their habitats requires microscopic observations of the respective microenvironment. This can be obtained by preserving the complex soil structure including the root system by resin impregnation resulting in high quality thin sections. The observation of such sections via fluorescence microscopy, SEM-EDS, and Nano-SIMS will be highlighted in this presentation. In addition, we will discuss the combination of this methodological approach with other imaging techniques such as planar optodes or non-invasive 3D X-ray CT to reveal the entire spatial structure and arrangement of soil particles and roots. When combining the preservation of soil structure via resin impregnation with 16S rRNA targeted fluorescence in situ hybridization (FISH) single microbial cells can be visualized, localized, and quantified in the undisturbed soil matrix including the root-soil interfaces. The simultaneous use of multiple oligonucleotide probes thereby provides information on the spatial distribution of microorganisms belonging to different phylogenetic groups. Results will be shown for paddy soils, where management induced physico-chemical dynamics (flooding and drying) as well as resulting microbial dynamics were visualized via correlative microscopy in resin impregnated samples.

  10. Evaluation of the Efficiency of the Sample Inactivation Reagent in the Abbott RealTime MTB Assay for Inactivation of Mycobacterium tuberculosis

    PubMed Central

    Wallis, Carole; Pahalawatta, Vihanga; Frank, Andrea; Ramdin, Neeshan; Viana, Raquel; Abravaya, Klara; Leckie, Gregor; Tang, Ning

    2015-01-01

    The Abbott RealTime MTB assay is a nucleic acid amplification test (NAAT) for the detection of Mycobacterium tuberculosis complex DNA. The sample inactivation procedure used in the assay, consisting of one part sample treated with 3 parts inactivation reagent for 60 min, effectively reduced viscosity and inactivated M. tuberculosis in clinical specimens. PMID:26085611

  11. Microbial decontamination of cosmetic raw materials and personal care products by irradiation

    NASA Astrophysics Data System (ADS)

    Katušin-Ražem, Branka; Mihaljević, Branka; Ražem, Dušan

    2003-03-01

    Typical levels of sporadically occurring (dynamic) microbial contamination of cosmetic raw materials: pigments, abrasives and liposomes, as well as of final products for personal care: toothpaste, crayons, shampoos, cleansers and creams, were evaluated. In most cases the contamination was dominated by a single population of microorganisms, either Gram-negative bacteria or molds. The feasibility of microbial decontamination by irradiation was studied by determining the resistance to gamma radiation of contaminating microflora in situ. It was expressed as a dose required for the first 90% reduction, D first 90% red . The values in the range 1-2 kGy for molds and 0.1-0.6 kGy for Gram-negative bacteria were obtained. This relatively high susceptibility to irradiation allowed inactivation factors close to 6 to be achieved with doses generally not exceeding 3 kGy, and yielding endpoint contamination less than 10/g.

  12. Interactions of the H5 pore region and hydroxylamine with N-type inactivation in the Shaker K+ channel.

    PubMed Central

    Yool, A J; Schwarz, T L

    1995-01-01

    Mutations at sites in the H5 region of the Shaker B K+ channel were used to analyze the influence of the pore on N-type inactivation. Single-channel and two-electrode voltage clamp analyses showed that mutations at residues T441 and T442, which are thought to lie at the internal mouth of the pore, produced opposite effects on inactivation: the inactivated state is stabilized by T441S and destabilized by T442S. In addition, an ammonium derivative, hydroxylamine (OH-(NH3)+), appears to bind in the pore region of T441S and further decreases the rate of recovery from N-type inactivation. This effect relies on the presence of the amino-terminal. The effect of hydroxylamine on the T441S mutation of this K+ channel shows several properties analogous to those of local anesthetics on the Na+ channel. These results can be interpreted to suggest that part of the H5 region contributes to the receptor for the inactivation particle and that a hydroxylamine ion trapped near that site can stabilize their interaction. Images FIGURE 8 PMID:7696498

  13. Opposing effects of two osmolytes--trehalose and glycerol--on thermal inactivation of rabbit muscle 6-phosphofructo-1-kinase.

    PubMed

    Faber-Barata, Joana; Sola-Penna, Mauro

    2005-01-01

    Trehalose and glycerol are known as good stabilizers of function and structure of several macromolecules against stress conditions. We previously reported that they have comparable effectiveness on protecting two yeast cytosolic enzymes against thermal inactivation. However, enzyme protection has always been associated to a decrease in catalytic activity at the stabilizing conditions i.e., the presence of the protective molecule. In the present study we tested trehalose and glycerol on thermal protection of the mammalian cytosolic enzyme phosphofructokinase. Here we found that trehalose was able to protect phosphofructokinase against thermal inactivation as well as to promote an activation of its catalytic activity. The enzyme incubated in the presence of 1 M trehalose did not present any significant inactivation within 2 h of incubation at 50 degrees C, contrasting to control experiments where the enzyme was fully inactivated during the same period exhibiting a t0.5 for thermal inactivation of 56+/-5 min. On the other hand, enzyme incubated in the presence of 37.5% (v/v) glycerol was not protected against incubation at 50 degrees C. Indeed, when phosphofructokinase was incubated for 45 min at 50 degrees C in the presence of lower concentrations of glycerol (7.5-25%, v/v), the remaining activity was 2-4 times lower than control. These data show that the compatibility of effects previously shown for trehalose and glycerol with some yeast cytosolic enzymes can not be extended to all globular enzyme system. In the case of phosphofructokinase, we believe that its property of shifting between several different complex oligomers configurations can be influenced by the physicochemical properties of the stabilizing molecules.

  14. Fabrication of magnetic Fe@ZnO0.6S0.4 nanocomposite for visible-light-driven photocatalytic inactivation of Escherichia coli

    NASA Astrophysics Data System (ADS)

    Peng, Ziling; Wu, Dan; Wang, Wei; Tan, Fatang; Ng, Tsz Wai; Chen, Jianguo; Qiao, Xueliang; Wong, Po Keung

    2017-02-01

    Bacterial inactivation by magnetic photocatalysts has now received growing interests due to the easy separation for recycle and reuse of photocatalysts. In this study, magnetic Fe@ZnO0.6S0.4 photocatalyst was prepared by a facile two-step precipitation method. Multiple techniques such as X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), UV-vis diffused reflectance spectra (UV-vis DRS) and vibrating sample magnetometer (VSM) were employed to characterize the structure, morphology and physicochemical properties of the photocatalyst. The as-obtained Fe@ZnO0.6S0.4 possessing magnetic property was easily collected from the reaction system by a magnet. Under white light-emitting-diode (LED) lamp irradiation, Fe@ZnO0.6S0.4 nanocomposite could completely inactivate 7-log of Escherichia coli K-12 within 5 h. More importantly, almost no decrease of photocatalytic efficiency in bacterial inactivation was observed even after five consecutive cycles, demonstrating Fe@ZnO0.6S0.4 exhibited good stability for reuse. The low released rate of Fe2+/Fe3+ and Zn2+ from Fe@ZnO0.6S0.4 composite further indicated the photocatalyst showed low cytotoxicity to bacterium and high stability under LED lamp irradiation. Facile preparation, high photocatalytic efficiency, good stability and reusability, and magnetic recovery property endow Fe@ZnO0.6S0.4 nanocomposite to be a promising photocatalytic material for bacterial inactivation.

  15. Knockout of the BK β2 subunit abolishes inactivation of BK currents in mouse adrenal chromaffin cells and results in slow-wave burst activity

    PubMed Central

    Martinez-Espinosa, Pedro L.; Yang, Chengtao; Gonzalez-Perez, Vivian; Xia, Xiao-Ming

    2014-01-01

    Rat and mouse adrenal medullary chromaffin cells (CCs) express an inactivating BK current. This inactivation is thought to arise from the assembly of up to four β2 auxiliary subunits (encoded by the kcnmb2 gene) with a tetramer of pore-forming Slo1 α subunits. Although the physiological consequences of inactivation remain unclear, differences in depolarization-evoked firing among CCs have been proposed to arise from the ability of β2 subunits to shift the range of BK channel activation. To investigate the role of BK channels containing β2 subunits, we generated mice in which the gene encoding β2 was deleted (β2 knockout [KO]). Comparison of proteins from wild-type (WT) and β2 KO mice allowed unambiguous demonstration of the presence of β2 subunit in various tissues and its coassembly with the Slo1 α subunit. We compared current properties and cell firing properties of WT and β2 KO CCs in slices and found that β2 KO abolished inactivation, slowed action potential (AP) repolarization, and, during constant current injection, decreased AP firing. These results support the idea that the β2-mediated shift of the BK channel activation range affects repetitive firing and AP properties. Unexpectedly, CCs from β2 KO mice show an increased tendency toward spontaneous burst firing, suggesting that the particular properties of BK channels in the absence of β2 subunits may predispose to burst firing. PMID:25267913

  16. Evaluation of the Efficiency of the Sample Inactivation Reagent in the Abbott RealTime MTB Assay for Inactivation of Mycobacterium tuberculosis.

    PubMed

    Qi, Chao; Wallis, Carole; Pahalawatta, Vihanga; Frank, Andrea; Ramdin, Neeshan; Viana, Raquel; Abravaya, Klara; Leckie, Gregor; Tang, Ning

    2015-09-01

    The Abbott RealTime MTB assay is a nucleic acid amplification test (NAAT) for the detection of Mycobacterium tuberculosis complex DNA. The sample inactivation procedure used in the assay, consisting of one part sample treated with 3 parts inactivation reagent for 60 min, effectively reduced viscosity and inactivated M. tuberculosis in clinical specimens. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Microbial factories for recombinant pharmaceuticals

    PubMed Central

    Ferrer-Miralles, Neus; Domingo-Espín, Joan; Corchero, José Luis; Vázquez, Esther; Villaverde, Antonio

    2009-01-01

    Most of the hosts used to produce the 151 recombinant pharmaceuticals so far approved for human use by the Food and Drug Administration (FDA) and/or by the European Medicines Agency (EMEA) are microbial cells, either bacteria or yeast. This fact indicates that despite the diverse bottlenecks and obstacles that microbial systems pose to the efficient production of functional mammalian proteins, namely lack or unconventional post-translational modifications, proteolytic instability, poor solubility and activation of cell stress responses, among others, they represent convenient and powerful tools for recombinant protein production. The entering into the market of a progressively increasing number of protein drugs produced in non-microbial systems has not impaired the development of products obtained in microbial cells, proving the robustness of the microbial set of cellular systems (so far Escherichia coli and Saccharomyces cerevisae) developed for protein drug production. We summarize here the nature, properties and applications of all those pharmaceuticals and the relevant features of the current and potential producing hosts, in a comparative way. PMID:19317892

  18. Study of phytochemical, anti-microbial, anti-oxidant, and anti-cancer properties of Allium wallichii.

    PubMed

    Bhandari, Jaya; Muhammad, BushraTaj; Thapa, Pratiksha; Shrestha, Bhupal Govinda

    2017-02-08

    There is growing interest in the use of plants for the treatment and prevention of cancer. Medicinal plants are currently being evaluated as source of promising anticancer agents. In this paper, we have investigated the anticancer potential of plant Allium wallichii, a plant native to Nepal and growing at elevations of 2300-4800 m. This is the first study of its kind for the plant mentioned. The dried plant was extracted in aqueous ethanol. Phytochemical screening, anti-microbial assay, anti-oxidant assay, cytotoxicity assay and the flow-cytometric analysis were done for analyzing different phytochemicals present, anti-microbial activity, anti-oxidant activity and anti-cancer properties of Allium wallichii. We observed the presence of steroids, terpenoids, flavonoids, reducing sugars and glycosides in the plant extract and the plant showed moderate anti-microbial and anti-oxidant activity. The IC 50 values of Allium wallichii in different cancer cell lines are 69.69 μg/ml for Prostate cancer (PC3) cell line, 55.29 μg/ml for Breast Cancer (MCF-7) cell line and 46.51 μg/ml for cervical cancer (HeLa) cell line as compared to Doxorubicin (0.85 μg/ml). The cell viability assay using FACS showed that the IC 50 value of Allium wallichii for Burkitt's lymphoma (B-Lymphoma) cell line was 3.817 ± 1.99 mg/ml. Allium wallichii can be an important candidate to be used as an anticancer agent. Separation of pure compounds with bioassay guided extraction, spectrometric analysis and subsequent cytotoxicity assay of the pure bioactive compounds from Allium wallichii is highly recommended as the crude extract itself showed promising cytotoxicity.

  19. Gamma irradiation of sorghum flour: Effects on microbial inactivation, amylase activity, fermentability, viscosity and starch granule structure

    NASA Astrophysics Data System (ADS)

    Mukisa, Ivan M.; Muyanja, Charles M. B. K.; Byaruhanga, Yusuf B.; Schüller, Reidar B.; Langsrud, Thor; Narvhus, Judith A.

    2012-03-01

    Malted and un-malted sorghum ( Sorghum bicolor (L.) Moench) flour was gamma irradiated with a dose of 10 kGy and then re-irradiated with 25 kGy. The effects of irradiation on microbial decontamination, amylase activity, fermentability (using an amylolytic L. plantarum MNC 21 strain), starch granule structure and viscosity were determined. Standard methods were used during determinations. The 10 kGy dose had no effect on microbial load of un-malted flour but reduced that of malted flour by 3 log cycles. Re-irradiation resulted in complete decontamination. Irradiation of malt caused a significant ( p<0.05) reduction in alpha and beta amylase activity (22% and 32%, respectively). Irradiation of un-malted flour increased the rates of utilization of glucose and maltose by 53% and 100%, respectively, during fermentation. However, microbial growth, rate of lactic acid production, final lactic acid concentration and pH were not affected. Starch granules appeared normal externally even after re-irradiation, however, granules ruptured and dissolved easily after hydration and gelatinization. Production of high dry matter density porridge (200 g dry matter/L) with a viscosity of 3500 cP was achieved by irradiation of un-malted flout at 10 kGy. Gamma irradiation can be used to decontaminate flours and could be utilized to produce weaning porridge from sorghum.

  20. Random X inactivation in the mule and horse placenta.

    PubMed

    Wang, Xu; Miller, Donald C; Clark, Andrew G; Antczak, Douglas F

    2012-10-01

    In eutherian mammals, dosage compensation of X-linked genes is achieved by X chromosome inactivation. X inactivation is random in embryonic and adult tissues, but imprinted X inactivation (paternal X silencing) has been identified in the extra-embryonic membranes of the mouse, rat, and cow. Few other species have been studied for this trait, and the data from studies of the human placenta have been discordant or inconclusive. Here, we quantify X inactivation using RNA sequencing of placental tissue from reciprocal hybrids of horse and donkey (mule and hinny). In placental tissue from the equid hybrids and the horse parent, the allelic expression pattern was consistent with random X inactivation, and imprinted X inactivation can clearly be excluded. We characterized horse and donkey XIST gene and demonstrated that XIST allelic expression in female hybrid placental and fetal tissues is negatively correlated with the other X-linked genes chromosome-wide, which is consistent with the XIST-mediated mechanism of X inactivation discovered previously in mice. As the most structurally and morphologically diverse organ in mammals, the placenta also appears to show diverse mechanisms for dosage compensation that may result in differences in conceptus development across species.

  1. Impact of pulsed-electric field and high-voltage electrical discharges on red wine microbial stabilization and quality characteristics.

    PubMed

    Delsart, C; Grimi, N; Boussetta, N; Miot Sertier, C; Ghidossi, R; Vorobiev, E; Mietton Peuchot, M

    2016-01-01

    In this study, pulsed-electric fields (PEF) and high-voltage electrical discharges (HVED) are proposed as new techniques for the microbial stabilization of red wines before bottling. The efficiency of the treatment was then evaluated. PEF and HVED-treatments have been applied to wine for the inactivation of Oenococcus oeni CRBO 9304, O. oeni CRBO 0608, Pediococcus parvulus CRBO 2.6 and Brettanomyces bruxellensis CB28. Different treatment times (1, 2, 4, 6, 8 and 10 ms) were used at 20 kV cm(-1) for the PEF treatments and at 40 kV for the HVED treatments, which correspond to applied energies from 80 to 800 kJ l(-1) . The effects of the treatments on the microbial inactivation rate and on various characteristics of red wines (phenolic composition, chromatic characteristics and physico-chemical parameters) were measured. The application of PEF or HVED treatments on red wine allowed the inactivation of alteration yeasts (B. bruxellensis CB28) and bacteria (O. oeni CRBO 9304, O. oeni CRBO 0608 and P. parvulus CRBO 2.6). The electric discharges at 40 kV were less effective than the PEF even after 10 ms of treatments. Indeed, 4 ms of PEF treatment at 20 kV cm(-1) were sufficient to inactivate all micro-organisms present in the wines. Also, the use of PEF had no negative impact on the composition of wines compared to the HVED treatments. Contrary to PEF, the phenolics compounds were degraded after the HVED treatment and the physico-chemical composition of wine were modified with HVED. PEF technology seems to be an interesting alternative to stabilize microbiologically wines before bottling and without modifying their composition. This process offers many advantages for winemakers: no chemical inputs, low energy consumption (320 kJ l(-1) ), fast (treatment time of 4 ms) and athermal (ΔT ≈ 10°C). © 2015 The Society for Applied Microbiology.

  2. High pressure thermal inactivation of Clostridium botulinum type E endospores – kinetic modeling and mechanistic insights

    PubMed Central

    Lenz, Christian A.; Reineke, Kai; Knorr, Dietrich; Vogel, Rudi F.

    2015-01-01

    Cold-tolerant, neurotoxigenic, endospore forming Clostridium (C.) botulinum type E belongs to the non-proteolytic physiological C. botulinum group II, is primarily associated with aquatic environments, and presents a safety risk for seafood. High pressure thermal (HPT) processing exploiting the synergistic effect of pressure and temperature can be used to inactivate bacterial endospores. We investigated the inactivation of C. botulinum type E spores by (near) isothermal HPT treatments at 300–1200 MPa at 30–75°C for 1 s to 10 min. The occurrence of heat and lysozyme susceptible spore fractions after such treatments was determined. The experimental data were modeled to obtain kinetic parameters and represented graphically by isoeffect lines. In contrast to findings for spores of other species and within the range of treatment parameters applied, zones of spore stabilization (lower inactivation than heat treatments alone), large heat susceptible (HPT-induced germinated) or lysozyme-dependently germinable (damaged coat layer) spore fractions were not detected. Inactivation followed first order kinetics. Dipicolinic acid release kinetics allowed for insights into possible inactivation mechanisms suggesting a (poorly effective) physiologic-like (similar to nutrient-induced) germination at ≤450 MPa/≤45°C and non-physiological germination at >500 MPa/>60–70°C. Results of this study support the existence of some commonalities in the HPT inactivation mechanism of C. botulinum type E spores and Bacillus spores although both organisms have significantly different HPT resistance properties. The information presented here contributes to closing the gap in knowledge regarding the HPT inactivation of spore formers relevant to food safety and may help industrial implementation of HPT processing. The markedly lower HPT resistance of C. botulinum type E spores compared with the resistance of spores from other C. botulinum types could allow for the implementation of

  3. INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE

    EPA Science Inventory

    Ozone inactivation rates for Cryptosporidium parvum (C. parvum) oocysts were determined with an in-vitro excystation method based on excysted sporozoite counts. Results were consistent with published animal infectivity data for the same C. parvum strain. The inactivation kinetics...

  4. Biogeochemical Processes in Microbial Ecosystems

    NASA Technical Reports Server (NTRS)

    DesMarais, David J.

    2001-01-01

    The hierarchical organization of microbial ecosystems determines process rates that shape Earth's environment, create the biomarker sedimentary and atmospheric signatures of life, and define the stage upon which major evolutionary events occurred. In order to understand how microorganisms have shaped the global environment of Earth and, potentially, other worlds, we must develop an experimental paradigm that links biogeochemical processes with ever-changing temporal and spatial distributions of microbial populations and their metabolic properties. Additional information is contained in the original extended abstract.

  5. Enzymatic Production of Biodiesel: Strategies to Overcome Methanol Inactivation.

    PubMed

    Lotti, Marina; Pleiss, Jürgen; Valero, Francisco; Ferrer, Pau

    2018-05-01

    Lipase-catalyzed transesterification of triglycerides and alcohols to obtain biodiesel is an environmentally friendly and sustainable route for fuels production since, besides proceeding in mild reaction conditions, it allows for the use of low-cost feedstocks that contain water and free fatty acids, for example non-edible oils and waste oils. This review article reports recent advances in the field and focus in particular on a major issue in the enzymatic process, the inactivation of most lipases caused by methanol, the preferred acyl acceptor used for alcoholysis. The recent results about immobilization of enzymes on nano-materials and the use of whole-cell biocatalysts, as well as the use of cell-surface display technologies and metabolic engineering strategies for microbial production of biodiesel are described. It is discussed also insight into the effects of methanol on lipases obtained by modeling approaches and report on studies aimed at mining novel alcohol stable enzymes or at improving robustness in existing ones by protein engineering. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Phage Therapy and Photodynamic Therapy: Low Environmental Impact Approaches to Inactivate Microorganisms in Fish Farming Plants

    PubMed Central

    Almeida, Adelaide; Cunha, Ângela; Gomes, Newton C.M.; Alves, Eliana; Costa, Liliana; Faustino, Maria A.F.

    2009-01-01

    Owing to the increasing importance of aquaculture to compensate for the progressive worldwide reduction of natural fish and to the fact that several fish farming plants often suffer from heavy financial losses due to the development of infections caused by microbial pathogens, including multidrug resistant bacteria, more environmentally-friendly strategies to control fish infections are urgently needed to make the aquaculture industry more sustainable. The aim of this review is to briefly present the typical fish farming diseases and their threats and discuss the present state of chemotherapy to inactivate microorganisms in fish farming plants as well as to examine the new environmentally friendly approaches to control fish infection namely phage therapy and photodynamic antimicrobial therapy. PMID:19841715

  7. Systems Biology of Microbial Exopolysaccharides Production.

    PubMed

    Ates, Ozlem

    2015-01-01

    Exopolysaccharides (EPSs) produced by diverse group of microbial systems are rapidly emerging as new and industrially important biomaterials. Due to their unique and complex chemical structures and many interesting physicochemical and rheological properties with novel functionality, the microbial EPSs find wide range of commercial applications in various fields of the economy such as food, feed, packaging, chemical, textile, cosmetics and pharmaceutical industry, agriculture, and medicine. EPSs are mainly associated with high-value applications, and they have received considerable research attention over recent decades with their biocompatibility, biodegradability, and both environmental and human compatibility. However, only a few microbial EPSs have achieved to be used commercially due to their high production costs. The emerging need to overcome economic hurdles and the increasing significance of microbial EPSs in industrial and medical biotechnology call for the elucidation of the interrelations between metabolic pathways and EPS biosynthesis mechanism in order to control and hence enhance its microbial productivity. Moreover, a better understanding of biosynthesis mechanism is a significant issue for improvement of product quality and properties and also for the design of novel strains. Therefore, a systems-based approach constitutes an important step toward understanding the interplay between metabolism and EPS biosynthesis and further enhances its metabolic performance for industrial application. In this review, primarily the microbial EPSs, their biosynthesis mechanism, and important factors for their production will be discussed. After this brief introduction, recent literature on the application of omics technologies and systems biology tools for the improvement of production yields will be critically evaluated. Special focus will be given to EPSs with high market value such as xanthan, levan, pullulan, and dextran.

  8. Systems Biology of Microbial Exopolysaccharides Production

    PubMed Central

    Ates, Ozlem

    2015-01-01

    Exopolysaccharides (EPSs) produced by diverse group of microbial systems are rapidly emerging as new and industrially important biomaterials. Due to their unique and complex chemical structures and many interesting physicochemical and rheological properties with novel functionality, the microbial EPSs find wide range of commercial applications in various fields of the economy such as food, feed, packaging, chemical, textile, cosmetics and pharmaceutical industry, agriculture, and medicine. EPSs are mainly associated with high-value applications, and they have received considerable research attention over recent decades with their biocompatibility, biodegradability, and both environmental and human compatibility. However, only a few microbial EPSs have achieved to be used commercially due to their high production costs. The emerging need to overcome economic hurdles and the increasing significance of microbial EPSs in industrial and medical biotechnology call for the elucidation of the interrelations between metabolic pathways and EPS biosynthesis mechanism in order to control and hence enhance its microbial productivity. Moreover, a better understanding of biosynthesis mechanism is a significant issue for improvement of product quality and properties and also for the design of novel strains. Therefore, a systems-based approach constitutes an important step toward understanding the interplay between metabolism and EPS biosynthesis and further enhances its metabolic performance for industrial application. In this review, primarily the microbial EPSs, their biosynthesis mechanism, and important factors for their production will be discussed. After this brief introduction, recent literature on the application of omics technologies and systems biology tools for the improvement of production yields will be critically evaluated. Special focus will be given to EPSs with high market value such as xanthan, levan, pullulan, and dextran. PMID:26734603

  9. Low temperature MS2 (ATCC15597-B1) virus inactivation using a hot bubble column evaporator (HBCE).

    PubMed

    Garrido, A; Pashley, R M; Ninham, B W

    2017-03-01

    In the treatment of household wastewater viruses are hard to eliminate. A new technique is described which tackles this major problem. The MS2 (ATCC15597-B1) virus was used as a surrogate to estimate the inactivation rates for enteric viruses by a hot (150°C) air bubble column evaporator (HBCE) system Its surface charging properties obtained by dynamic light scattering, have been studied in a range of aqueous salt solutions and secondary treated synthetic sewage water. A combination of MS2 virus surface charge properties with thermal inactivation rates, and an improved double layer plaque assay technique, allows an assessment of the efficiency of the HBCE process for virus removal in water. The system is a new energy efficient treatment for water reuse applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Inactivation of B. subtilis spores by low pressure plasma—influence of optical filters and photon/particle fluxes on the inactivation efficiency

    NASA Astrophysics Data System (ADS)

    Fiebrandt, Marcel; Hillebrand, Bastian; Lackmann, Jan-Wilm; Raguse, Marina; Moeller, Ralf; Awakowicz, Peter; Stapelmann, Katharina

    2018-01-01

    Inactivation experiments were performed with Bacillus subtilis spores in a low pressure double inductively coupled plasma (DICP) system. Argon, nitrogen and oxygen at 5 Pa were used as feed gas to change the emission spectrum in the range of 100 nm to 400 nm, as well as between radical and metastable densities. Optical filters were applied, to block particles and selected wavelengths from the spores. By determining absolute photon fluxes, the sporicidal efficiency of various wavelength ranges was evaluated. The results showed good agreement with other plasma experiments, as well as with monochromatic light inactivation experiments from a synchrotron. The findings indicated that the inactivation rate constants of broadband plasma emission and monochromatic light were identical, and that no synergistic effect exists. Furthermore, the influence of radicals, ions and metastables on the inactivation efficiency was of minor importance in the set-up used, and radiation was the main reason for spore inactivation.

  11. Slow Inactivation in Shaker K Channels Is Delayed by Intracellular Tetraethylammonium

    PubMed Central

    González-Pérez, Vivian; Neely, Alan; Tapia, Christian; González-Gutiérrez, Giovanni; Contreras, Gustavo; Orio, Patricio; Lagos, Verónica; Rojas, Guillermo; Estévez, Tania; Stack, Katherine; Naranjo, David

    2008-01-01

    After removal of the fast N-type inactivation gate, voltage-sensitive Shaker (Shaker IR) K channels are still able to inactivate, albeit slowly, upon sustained depolarization. The classical mechanism proposed for the slow inactivation observed in cell-free membrane patches—the so called C inactivation—is a constriction of the external mouth of the channel pore that prevents K+ ion conduction. This constriction is antagonized by the external application of the pore blocker tetraethylammonium (TEA). In contrast to C inactivation, here we show that, when recorded in whole Xenopus oocytes, slow inactivation kinetics in Shaker IR K channels is poorly dependent on external TEA but severely delayed by internal TEA. Based on the antagonism with internally or externally added TEA, we used a two-pulse protocol to show that half of the channels inactivate by way of a gate sensitive to internal TEA. Such gate had a recovery time course in the tens of milliseconds range when the interpulse voltage was −90 mV, whereas C-inactivated channels took several seconds to recover. Internal TEA also reduced gating charge conversion associated to slow inactivation, suggesting that the closing of the internal TEA-sensitive inactivation gate could be associated with a significant amount of charge exchange of this type. We interpreted our data assuming that binding of internal TEA antagonized with U-type inactivation (Klemic, K.G., G.E. Kirsch, and S.W. Jones. 2001. Biophys. J. 81:814–826). Our results are consistent with a direct steric interference of internal TEA with an internally located slow inactivation gate as a “foot in the door” mechanism, implying a significant functional overlap between the gate of the internal TEA-sensitive slow inactivation and the primary activation gate. But, because U-type inactivation is reduced by channel opening, trapping the channel in the open conformation by TEA would also yield to an allosteric delay of slow inactivation. These results

  12. Hemostatic properties and protein expression profile of therapeutic apheresis plasma treated with amotosalen and ultraviolet A for pathogen inactivation.

    PubMed

    Ohlmann, Philippe; Hechler, Béatrice; Chafey, Philippe; Ravanat, Catherine; Isola, Hervé; Wiesel, Marie-Louise; Cazenave, Jean-Pierre; Gachet, Christian

    2016-09-01

    The INTERCEPT Blood System (IBS) using amotosalen-HCl and ultraviolet (UV)A inactivates a large spectrum of microbial pathogens and white blood cells in therapeutic plasma. Our aim was to evaluate to what extent IBS modifies the capacity of plasma to generate thrombin and induces qualitative or quantitative modifications of plasma proteins. Plasma units from four donors were collected by apheresis. Samples were taken before (control [CTRL]) and after IBS treatment and stored at -80°C until use. The activities of plasma coagulation factors and inhibitors and the thrombin generation potential were determined using assays measuring clotting times and the calibrated automated thrombogram (CAT), respectively. The proteomic profile of plasma proteins was examined using a two-dimensional differential in-gel electrophoresis (2D-DIGE) method. Nearly all of the procoagulant and antithrombotic factors tested retained at least 78% of their initial pre-IBS activity. Only FVII and FVIII displayed a lower level of conservation (67%), which nevertheless remained within the reference range for conventional plasma coagulation factors. The thrombin generation profile of plasma was conserved after IBS treatment. Among the 1331 protein spots revealed by 2D-DIGE analysis, only four were differentially expressed in IBS plasma compared to CTRL plasma and two were identified by mass spectrometric analysis as transthyretin and apolipoprotein A1. The IBS technique for plasma moderately decreases the activities of plasma coagulation factors and antithrombotic proteins, with no impact on the thrombin generation potential of plasma and very limited modifications of the proteomic profile. © 2016 AABB.

  13. Qualitation and Quantitation on Microplasma Jet for Bacteria Inactivation.

    PubMed

    Du, ChangMing; Liu, Ya; Huang, YaNi; Li, ZiMing; Men, Rui; Men, Yue; Tang, Jun

    2016-01-06

    In this work, a self-made microplasma jet system was used to conduct the qualitation and quantitation of inactivation with Escherichia coli as the target bacteria. The logarithmic concentration and the size of antimicrobial rings served as the evaluation parameters, respectively. The effect of various parameters on inactivation effect was studied. The results showed that the majority of bacteria had been inactivated in 30 s. The inactivation effect enhanced and then weakened with the increase of air flow rate, and receded as the extension of treatment distance. The effect with different carrier gases showed as follows: oxygen > air > nitrogen > argon. Meanwhile, the effect of different components of microplasma was studied in the optimum conditions (The flow rate was 5 L/min; inactivation distance was 2 cm). The results showed that electrically neutral active species was the main factor of inactivation rather than heating effect, ultraviolet radiation and charged particles. Finally the experiments of thallus change proved that microplasma jet had etching effect on cell membrane. It also found that microplasma could degrade organic material like protein. Furthermore, the images of scanning electron microscope (SEM) revealed the change of cell morphology step by step in the whole process of inactivation.

  14. Qualitation and Quantitation on Microplasma Jet for Bacteria Inactivation

    NASA Astrophysics Data System (ADS)

    Du, Changming; Liu, Ya; Huang, Yani; Li, Ziming; Men, Rui; Men, Yue; Tang, Jun

    2016-01-01

    In this work, a self-made microplasma jet system was used to conduct the qualitation and quantitation of inactivation with Escherichia coli as the target bacteria. The logarithmic concentration and the size of antimicrobial rings served as the evaluation parameters, respectively. The effect of various parameters on inactivation effect was studied. The results showed that the majority of bacteria had been inactivated in 30 s. The inactivation effect enhanced and then weakened with the increase of air flow rate, and receded as the extension of treatment distance. The effect with different carrier gases showed as follows: oxygen > air > nitrogen > argon. Meanwhile, the effect of different components of microplasma was studied in the optimum conditions (The flow rate was 5 L/min inactivation distance was 2 cm). The results showed that electrically neutral active species was the main factor of inactivation rather than heating effect, ultraviolet radiation and charged particles. Finally the experiments of thallus change proved that microplasma jet had etching effect on cell membrane. It also found that microplasma could degrade organic material like protein. Furthermore, the images of scanning electron microscope (SEM) revealed the change of cell morphology step by step in the whole process of inactivation.

  15. Qualitation and Quantitation on Microplasma Jet for Bacteria Inactivation

    PubMed Central

    Du, ChangMing; Liu, Ya; Huang, YaNi; Li, ZiMing; Men, Rui; Men, Yue; Tang, Jun

    2016-01-01

    In this work, a self-made microplasma jet system was used to conduct the qualitation and quantitation of inactivation with Escherichia coli as the target bacteria. The logarithmic concentration and the size of antimicrobial rings served as the evaluation parameters, respectively. The effect of various parameters on inactivation effect was studied. The results showed that the majority of bacteria had been inactivated in 30 s. The inactivation effect enhanced and then weakened with the increase of air flow rate, and receded as the extension of treatment distance. The effect with different carrier gases showed as follows: oxygen > air > nitrogen > argon. Meanwhile, the effect of different components of microplasma was studied in the optimum conditions (The flow rate was 5 L/min; inactivation distance was 2 cm). The results showed that electrically neutral active species was the main factor of inactivation rather than heating effect, ultraviolet radiation and charged particles. Finally the experiments of thallus change proved that microplasma jet had etching effect on cell membrane. It also found that microplasma could degrade organic material like protein. Furthermore, the images of scanning electron microscope (SEM) revealed the change of cell morphology step by step in the whole process of inactivation. PMID:26732987

  16. FAST TRACK COMMUNICATION: Selective inactivation of micro-organisms with near-infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Tsen, K. T.; Tsen, Shaw-Wei D.; Sankey, Otto F.; Kiang, Juliann G.

    2007-11-01

    We demonstrate an unconventional and revolutionary method for selective inactivation of micro-organisms by using near-infrared femtosecond laser pulses. We show that if the wavelength and pulse width of the excitation femtosecond laser are appropriately selected, there exists a window in power density that enables us to achieve selective inactivation of target viruses and bacteria without causing cytotoxicity in mammalian cells. This strategy targets the mechanical (vibrational) properties of micro-organisms, and thus its antimicrobial efficacy is likely unaffected by genetic mutation in the micro-organisms. Such a method may be effective against a wide variety of drug resistant micro-organisms and has broad implications in disinfection as well as in the development of novel treatments for viral and bacterial pathogens.

  17. Relationship between Sublethal Injury and Microbial Inactivation by the Combination of High Hydrostatic Pressure and Citral or tert-Butyl Hydroquinone ▿

    PubMed Central

    Somolinos, Maria; García, Diego; Pagán, Rafael; Mackey, Bernard

    2008-01-01

    The aim was to investigate (i) the occurrence of sublethal injury in Listeria monocytogenes, Escherichia coli, and Saccharomyces cerevisiae after high hydrostatic pressure (HHP) treatment as a function of the treatment medium pH and composition and (ii) the relationship between the occurrence of sublethal injury and the inactivating effect of a combination of HHP and two antimicrobial compounds, tert-butyl hydroquinone (TBHQ) and citral. The three microorganisms showed a high proportion of sublethally injured cells (up to 99.99% of the surviving population) after HHP. In E. coli and L. monocytogenes, the extent of inactivation and sublethal injury depended on the pH and the composition of the treatment medium, whereas in S. cerevisiae, inactivation and sublethal injury were independent of medium pH or composition under the conditions tested. TBHQ alone was not lethal to E. coli or L. monocytogenes but acted synergistically with HHP and 24-h refrigeration, resulting in a viability decrease of >5 log10 cycles of both organisms. The antimicrobial effect of citral depended on the microorganism and the treatment medium pH. Acting alone for 24 h under refrigeration, 1,000 ppm of citral caused a reduction of 5 log10 cycles of E. coli at pH 7.0 and almost 3 log10 cycles of L. monocytogenes at pH 4.0. The combination of citral and HHP also showed a synergistic effect. Our results have confirmed that the detection of sublethal injury after HHP may contribute to the identification of those treatment conditions under which HHP may act synergistically with other preserving processes. PMID:18952869

  18. 21 CFR 610.11a - Inactivated influenza vaccine, general safety test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Inactivated influenza vaccine, general safety test... Inactivated influenza vaccine, general safety test. For inactivated influenza vaccine, the general safety test... subcutaneous or intraperitoneal injection of 5.0 milliliters of inactivated influenza vaccine into each guinea...

  19. 21 CFR 610.11a - Inactivated influenza vaccine, general safety test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Inactivated influenza vaccine, general safety test... Inactivated influenza vaccine, general safety test. For inactivated influenza vaccine, the general safety test... subcutaneous or intraperitoneal injection of 5.0 milliliters of inactivated influenza vaccine into each guinea...

  20. 21 CFR 610.11a - Inactivated influenza vaccine, general safety test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Inactivated influenza vaccine, general safety test... Inactivated influenza vaccine, general safety test. For inactivated influenza vaccine, the general safety test... subcutaneous or intraperitoneal injection of 5.0 milliliters of inactivated influenza vaccine into each guinea...

  1. 21 CFR 610.11a - Inactivated influenza vaccine, general safety test.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Inactivated influenza vaccine, general safety test... Inactivated influenza vaccine, general safety test. For inactivated influenza vaccine, the general safety test... subcutaneous or intraperitoneal injection of 5.0 milliliters of inactivated influenza vaccine into each guinea...

  2. Coupling among Microbial Communities, Biogeochemistry, and Mineralogy across Biogeochemical Facies

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

    Stegen, James C.; Konopka, Allan; McKinely, Jim

    Physical properties of sediments are commonly used to define subsurface lithofacies and these same physical properties influence subsurface microbial communities. This suggests an (unexploited) opportunity to use the spatial distribution of facies to predict spatial variation in biogeochemically relevant microbial attributes. Here, we characterize three biogeochemical facies—oxidized, reduced, and transition—within one lithofacies and elucidate relationships among facies features and microbial community biomass, diversity, and community composition. Consistent with previous observations of biogeochemical hotspots at environmental transition zones, we find elevated biomass within a biogeochemical facies that occurred at the transition between oxidized and reduced biogeochemical facies. Microbial diversity—the number ofmore » microbial taxa—was lower within the reduced facies and was well-explained by a combination of pH and mineralogy. Null modeling revealed that microbial community composition was influenced by ecological selection imposed by redox state and mineralogy, possibly due to effects on nutrient availability or transport. As an illustrative case, we predict microbial biomass concentration across a three-dimensional spatial domain by coupling the spatial distribution of subsurface biogeochemical facies with biomass-facies relationships revealed here. We expect that merging such an approach with hydro-biogeochemical models will provide important constraints on simulated dynamics, thereby reducing uncertainty in model predictions.« less

  3. Open- and closed-state fast inactivation in sodium channels

    PubMed Central

    Lehmann-Horn, Frank; Holzherr, Boris D

    2011-01-01

    The role of sodium channel closed-state fast inactivation in membrane excitability is not well understood. We compared open- and closed-state fast inactivation, and the gating charge immobilized during these transitions, in skeletal muscle channel hNaV1.4. A significant fraction of total charge movement and its immobilization occurred in the absence of channel opening. Simulated action potentials in skeletal muscle fibers were attenuated when pre-conditioned by subthreshold depolarization. Anthopleurin A, a site-3 toxin that inhibits gating charge associated with the movement of DIVS4, was used to assess the role of this voltage sensor in closed-state fast inactivation. Anthopleurin elicited opposing effects on the gating mode, kinetics and charge immobilized during open- versus closed-state fast inactivation. This same toxin produced identical effects on recovery of channel availability and remobilization of gating charge, irrespective of route of entry into fast inactivation. Our findings suggest that depolarization promoting entry into fast inactivation from open versus closed states provides access to the IFMT receptor via different rate-limiting conformational translocations of DIVS4. PMID:21099342

  4. Guiding bioprocess design by microbial ecology.

    PubMed

    Volmer, Jan; Schmid, Andreas; Bühler, Bruno

    2015-06-01

    Industrial bioprocess development is driven by profitability and eco-efficiency. It profits from an early stage definition of process and biocatalyst design objectives. Microbial bioprocess environments can be considered as synthetic technical microbial ecosystems. Natural systems follow Darwinian evolution principles aiming at survival and reproduction. Technical systems objectives are eco-efficiency, productivity, and profitable production. Deciphering technical microbial ecology reveals differences and similarities of natural and technical systems objectives, which are discussed in this review in view of biocatalyst and process design and engineering strategies. Strategies for handling opposing objectives of natural and technical systems and for exploiting and engineering natural properties of microorganisms for technical systems are reviewed based on examples. This illustrates the relevance of considering microbial ecology for bioprocess design and the potential for exploitation by synthetic biology strategies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Inactivation of Lassa, Marburg, and Ebola viruses by gamma irradiation

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

    Elliott, L.H.; McCormick, J.B.; Johnson, K.M.

    1982-10-01

    Because of the cumbersome conditions experienced in a maximum containment laboratory, methods for inactivating highly pathogenic viruses were investigated. The infectivity of Lassa, Marburg, and Ebola viruses was inactivated without altering the immunological activity after radiation with /sup 60/Co gamma rays. At 4 degrees C, Lassa virus was the most difficult to inactivate with a rate of 5.3 X 10(-6) log 50% tissue culture infective dose per rad of /sup 60/Co radiation, as compared with 6.8 X 10(-6) log 50% tissue culture infective dose per rad for Ebola virus and 8.4 X 10(-6) log 50% tissue culture infective dose permore » rad for Marburg virus. Experimental inactivation curves, as well as curves giving the total radiation needed to inactivate a given concentration of any of the three viruses, are presented. We found this method of inactivation to be superior to UV light or beta-propiolactone inactivation and now routinely use it for preparation of material for protein-chemistry studies or for preparation of immunological reagents.« less

  6. Inactivation of Lassa, Marburg, and Ebola viruses by gamma irradiation

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

    Elliott, L.H.; McCormick, J.B.; Johnson, K.M.

    1982-10-01

    Because of the cumbersome conditions experienced in a maximum containment laboratory, methods for inactivating highly pathogenic viruses were investigated. The infectivity of Lassa, Marburg, and Ebola viruses was inactivated without altering the immunological activity after radiation with /sup 60/CO gamma rays. At 4 degrees C, Lassa virus was the most difficult to inactivate with a rate of 5.3 X 10(-6) log 50% tissue culture infective dose per rad of /sup 60/CO radiation, as compared with 6.8 X 10(-6) log 50% tissue culture infective dose per rad for Ebola virus and 8.4 X 10(-6) log 50% tissue culture infective dose permore » rad for Marburg virus. Experimental inactivation curves, as well as curves giving the total radiation needed to inactivate a given concentration of any of the three viruses, are presented. The authors found this method of inactivation to be superior to UV light or beta-propiolactone inactivation and now routinely use it for preparation of material for protein-chemistry studies or for preparation of immunological reagents.« less

  7. Structure of suicide-inactivated. beta. -hydroxydecanoyl-thioester dehydrase

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

    Schwab, J.M.; Ho, C.K.; Li, W.B.

    ..beta..-Hydroxydecanoylthioester dehydrase, the key enzyme in biosynthesis of unsaturated fatty acids under anaerobic conditions, equilibrates thioesters of (R)-3-hydroxydecanoic acid, E-2-decenoic acid, and Z-3-decenoic acid. Dehydrase is irreversibly inactivated by the N-acetylcysteamine thioester of 3-decynoic acid (3-decynoyl-NAC), via dehydrase-catalyzed isomerization to 2,3-decadienoyl-NAC. To probe the relationship between normal catalysis and suicide inactivation, the structure of the inactivated enzyme has been studied. 3-(2-/sup 13/C)Decynoyl-NAC was synthesized and incubated with dehydrase. /sup 13/C NMR showed that attack of 2,3-decadienoyl-NAC by the active site histidine gives 3-histidinyl-3-decenoyl-NAC, which slowly rearranges to the more stable ..delta../sup 2/ isomer. Model histidine-allene adducts have been made andmore » characterized. Analysis of NMR data show that the C=C configuration of the decenoyl moiety of enzyme-bound inactivator is E. The suggestion that the mechanism of dehydrase inactivation parallels its normal mechanism of action is supported these findings.« less

  8. Photodynamic inactivation of pathogenic species Pseudomonas aeruginosa and Candida albicans with lutetium (III) acetate phthalocyanines and specific light irradiation.

    PubMed

    Mantareva, Vanya; Kussovski, Vesselin; Durmuş, Mahmut; Borisova, Ekaterina; Angelov, Ivan

    2016-11-01

    Photodynamic inactivation (PDI) is a light-associated therapeutic approach suitable for treatment of local acute infections. The method is based on specific light-activated compound which by specific irradiation and in the presence of molecular oxygen produced molecular singlet oxygen and other reactive oxygen species, all toxic for pathogenic microbial cells. The study presents photodynamic impact of two recently synthesized water-soluble cationic lutetium (III) acetate phthalocyanines (LuPc-5 and LuPc-6) towards two pathogenic strains, namely, the Gram-negative bacterium Pseudomonas aeruginosa and a fungus Candida albicans. The photodynamic effect was evaluated for the cells in suspensions and organized in 48-h developed biofilms. The relatively high levels of uptakes of LuPc-5 and LuPc-6 were determined for fungal cells compared to bacterial cells. The penetration depths and distribution of both LuPcs into microbial biofilms were investigated by means of confocal fluorescence microscopy. The photoinactivation efficiency was studied for a wide concentration range (0.85-30 μM) of LuPc-5 and LuPc-6 at a light dose of 50 J cm -2 from red light-emitting diode (LED; 665 nm). The PDI study on microbial biofilms showed incomplete photoinactivation (<3 logs) for the used gentle drug-light protocol.

  9. Thermal inactivation of Listeria innocua in salmon (Oncorhynchus keta) caviar using conventional glass and novel aluminum thermal-death-time tubes.

    PubMed

    Al-Holy, M; Quinde, Z; Guan, D; Tang, J; Rasco, B

    2004-02-01

    Differences in the come-up times and thermal inactivation parameters of Listeria innocua in salmon (Oncorhynchus keta) caviar containing 2.5% salt using conventional thermal-death-time (TDT) glass tubes and a novel aluminum tube were tested and compared. Generally, the come-up times and decimal reduction times (D-values) were shorter and the change in temperature required to change the D-value (z-value) was longer in the aluminum than in the glass tubes. The D-values at 60, 63, and 65 degrees C for the aluminum TDT tubes were 2.97, 0.77, and 0.40 min, respectively, and for the glass TDT tubes, these values were 3.55, 0.84, and 0.41 min. The z-values were 5.7 degrees C in the aluminum and 5.3 degrees C in the glass. Because of the shorter come-up time, the aluminum TDT tubes may provide a more precise measurement of microbial thermal inactivation than the glass TDT tubes, particularly for viscous materials, solid foods, and foods containing particulate matter.

  10. Photodynamic inactivation of foodborne bacteria by eosin Y.

    PubMed

    Bonin, E; Dos Santos, A R; Fiori da Silva, A; Ribeiro, L H; Favero, M E; Campanerut-Sá, P A Z; de Freitas, C F; Caetano, W; Hioka, N; Mikcha, J M G

    2018-06-01

    The aim of this study was evaluate the effect of photodynamic inactivation mediated by eosin Y in Salmonella enterica serotype Typhimurium ATCC 14028, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Bacillus cereus ATCC 11778. Bacteria (10 7 CFU per ml) were incubated with eosin Y at concentrations ranging from 0·1 to 10 μmol l -1 , irradiated by green LED (λ max 490-570 nm) for 5, 10 and 15 min and the cellular viability was determined. Pseudomonas aeruginosa was completely inactivated when treated with 10 μmol l -1 eosin Y for 10 min. Treatments reduced B. cereus and Salm. Typhimurium counts to 2·7 log CFU per ml and 1·7 log CFU per ml, respectively. Escherichia coli counts were slightly reduced. Staphylococcus aureus presented the highest sensitivity, being completely inactivated by eosin Y at 5 μmol l -1 and 5 min of illumination. The reduction of cellular viability of photoinactivated Staph. aureus was also demonstrated by flow cytometry and morphological changes were observed by scanning electron microscopy. Eosin Y in combination with LED produced bacterial inactivation, being a potential candidate for photodynamic inactivation. This study evidenced the efficacy of photodynamic inactivation as a novel and promising alternative to bacterial control. © 2018 The Society for Applied Microbiology.

  11. [Effects of biochar on microbial ecology in agriculture soil: a review].

    PubMed

    Ding, Yan-Li; Liu, Jie; Wang, Ying-Ying

    2013-11-01

    Biochar, as a new type of soil amendment, has been obtained considerable attention in the research field of environmental sciences worldwide. The studies on the effects of biochar in improving soil physical and chemical properties started quite earlier, and already covered the field of soil microbial ecology. However, most of the studies considered the soil physical and chemical properties and the microbial ecology separately, with less consideration of their interactions. This paper summarized and analyzed the interrelationships between the changes of soil physical and chemical properties and of soil microbial community after the addition of biochar. Biochar can not only improve soil pH value, strengthen soil water-holding capacity, increase soil organic matter content, but also affect soil microbial community structure, and alter the abundance of soil bacteria and fungi. After the addition of biochar, the soil environment and soil microorganisms are interacted each other, and promote the improvement of soil microbial ecological system together. This review was to provide a novel perspective for the in-depth studies of the effects of biochar on soil microbial ecology, and to promote the researches on the beneficial effects of biochar to the environment from ecological aspect. The methods to improve the effectiveness of biochar application were discussed, and the potential applications of biochar in soil bioremediation were further analyzed.

  12. Comparison of two radio-frequency plasma sterilization processes using microspot evaluation of microbial inactivation.

    PubMed

    Lassen, Klaus S; Johansen, Jens E; Grün, Reinar

    2006-07-01

    In this study, we evaluated gas plasma surface sterilization methods in a specific sterilizer. We have introduced a new monitoring method using 0.4 microm pore size membranes, which in this study gave the information corresponding to 3000 exposed biological indicators per treatment cycle. This enabled us to compare the fraction of inoculates that showed no growth after exposure for 30 different locations in the chamber, and hereby identify weak and strong spots in the chamber with regard to sporicidal effect. Membranes were also used to expose a broad spectrum of soil bacteria for plasma treatment at four different conditions. The organisms were identified using PCR and sequencing. The test showed that Bacillus stearothermophilus spores were inactivated at the slowest rate among the tested microorganisms. Further alpha-proteobacteria (Gram negative) seemed more sensitive than the rest of the tested organisms. The microspot evaluation approach has been a most useful tool in the assessment of sterilization performance in sterilizers that do not have clear measurable parameters related to the sterilization.

  13. Factor V activation and inactivation by venom proteases.

    PubMed

    Rosing, J; Govers-Riemslag, J W; Yukelson, L; Tans, G

    2001-01-01

    Blood coagulation factor V is a single-chain glycoprotein with M(r) = 330,000 which plays an important role in the procoagulant and anticoagulant pathways. Thrombin activates factor V into factor Va, a two-chain molecule which is composed of a heavy (M(r) = 105,000) and a light chain (M(r) = 71,000/74,000). Factor Va accelerates factor Xa-catalysed prothrombin activation more than 1,000-fold and under physiological conditions the cofactor activity of factor Va in prothrombin activation is down-regulated by activated protein C. Factor V can also be activated by a wide variety of snake venoms (e.g. from Vipera species, Naja naja oxiana, Bothrops atrox) and by proteases present in the bristles of a South American caterpillar (Lonomia achelous). Some venoms, notably of Vipera lebetina turanica and Lonomia achelous, contain proteases that are able to inactivate factor V or factor Va. Venom factor V activators are excellent tools in studying the structure-function relationship of factor V(a) and they are also used in diagnostic tests for quantification of plasma factor V levels and for the screening of defects in the protein C pathway. In this review, the structural and functional properties of animal venom factor V activators and inactivators is described. Copyright 2002 S. Karger AG, Basel

  14. Physico-chemical parameters, bioactive compounds and microbial quality of thermo-sonicated carrot juice during storage.

    PubMed

    Martínez-Flores, Héctor E; Garnica-Romo, Ma Guadalupe; Bermúdez-Aguirre, Daniela; Pokhrel, Prashant Raj; Barbosa-Cánovas, Gustavo V

    2015-04-01

    Thermosonication has been successfully tested in food for microbial inactivation; however, changes in bioactive compounds and shelf-life of treated products have not been thoroughly investigated. Carrot juice was thermo-sonicated (24 kHz, 120 μm amplitude) at 50 °C, 54 °C and 58 °C for 10 min (acoustic power 2204.40, 2155.72, 2181.68 mW/mL, respectively). Quality parameters and microbial growth were evaluated after processing and during storage at 4 °C. Control and sonicated treatments at 50 °C and 54 °C had 10, 12 and 14 d of shelf-life, respectively. Samples sonicated at 58 °C had the best quality; microbial growth remained low at around 3-log for mesophiles, 4.5-log for yeasts and molds and 2-log for enterobacteria after 20 d of storage. Furthermore, thermo-sonicated juice at 58 °C retained >98% of carotenoids and 100% of ascorbic acid. Phenolic compounds increased in all stored, treated juices. Thermo-sonication is therefore a promising technology for preserving the quality of carrot juice by minimising the physicochemical changes during storage, retarding microbial growth and retaining the bioactive compounds. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Microbial mito-pathogens: fact or fiction?

    PubMed

    Bongaerts, Ger P A; van den Heuvel, Lambert P

    2008-01-01

    Mitochondria are bacteria-like semi-autonomous intracellular organelles that function as the powerhouses of eukaryotic cells. Inactivation or destruction of these organelles may have far-reaching consequences regarding the viability of the cells and thus of tissues, organs and finally even the body. Since mitochondria resemble (degenerated) bacteria, we have extrapolated from both cytological and microbiological facts the existence of various (kinds of) mitochondrion-specific microbial pathogens, i.e., pathogenic micro-organisms that may damage or destroy the mitochondria from within. These mito-pathogens may include mitoviruses, mitoviroids and mitobacteria. Although these mito-pathogens have not yet been demonstrated in humans, their theoretical degenerative effect regarding energy production from energy-rich substrates, such as carbohydrates and fats, might explain diseases that have not yet been understood, such as prion diseases and post-traumatic muscle dystrophy. Therefore, these kinds of micro-organisms should be kept in mind.

  16. Inactivation of Viruses by Benzalkonium Chloride

    PubMed Central

    Armstrong, J. A.; Froelich, E. J.

    1964-01-01

    Benzalkonium chloride (as Roccal or Zephiran) was found to inactivate influenza, measles, canine distemper, rabies, fowl laryngotracheitis, vaccinia, Semliki Forest, feline pneumonitis, meningopneumonitis, and herpes simplex viruses after 10 min of exposure at 30 C or at room temperature. Poliovirus and encephalomyocarditis virus were not inactivated under the same conditions. It was concluded that all viruses tested were sensitive except members of the picorna group. The literature was reviewed. PMID:4288740

  17. Viral capsid mobility: a dynamic conduit for inactivation.

    PubMed

    Broo, K; Wei, J; Marshall, D; Brown, F; Smith, T J; Johnson, J E; Schneemann, A; Siuzdak, G

    2001-02-27

    Mass spectrometry and fluorescent probes have provided direct evidence that alkylating agents permeate the protein capsid of naked viruses and chemically inactivate the nucleic acid. N-acetyl-aziridine and a fluorescent alkylating agent, dansyl sulfonate aziridine, inactivated three different viruses, flock house virus, human rhinovirus-14, and foot and mouth disease virus. Mass spectral studies as well as fluorescent probes showed that alkylation of the genome was the mechanism of inactivation. Because particle integrity was not affected by selective alkylation (as shown by electron microscopy and sucrose gradient experiments), it was reasoned that the dynamic nature of the viral capsid acts as a conduit to the interior of the particle. Potential applications include fluorescent labeling for imaging viral genomes in living cells, the sterilization of blood products, vaccine development, and viral inactivation in vivo.

  18. Controlled Microbial Cenoses in Closed Spaces

    NASA Astrophysics Data System (ADS)

    Somova, Lydia; Mikheeva, Galina

    Controlled microbial cenoses have good prospects in closed spaces: for air treatment in LSS and cellars industrial premises; for sewage treatment in LSS; for increase of productivity and protect of plants from infections in LSS. Possible methods of formation of microbiocenoses are: selection, autoselection, artificial formation taking into account their biochemical properties and metabolic interactions. Experimental microbiocenoses, has been produced on the basis of natural association of microorganisms by long cultivation on specially developed medium. Dominating groups are bacteria of genera: Lactobacillus, Streptococcus, Leuconostoc, Bidobac-terium, Rhodopseudomonas and yeast of genera: Kluyveromyces, Saccharomyces and Torulop-sis. Microbiocenoses do not contain pathogenic and conditionally pathogenic microorganisms, they possess opposing and probiotic properties. Different examples of microbial cenoses actions are to be presented in the paper.

  19. Heme impairs the ball-and-chain inactivation of potassium channels.

    PubMed

    Sahoo, Nirakar; Goradia, Nishit; Ohlenschläger, Oliver; Schönherr, Roland; Friedrich, Manfred; Plass, Winfried; Kappl, Reinhard; Hoshi, Toshinori; Heinemann, Stefan H

    2013-10-15

    Fine-tuned regulation of K(+) channel inactivation enables excitable cells to adjust action potential firing. Fast inactivation present in some K(+) channels is mediated by the distal N-terminal structure (ball) occluding the ion permeation pathway. Here we show that Kv1.4 K(+) channels are potently regulated by intracellular free heme; heme binds to the N-terminal inactivation domain and thereby impairs the inactivation process, thus enhancing the K(+) current with an apparent EC50 value of ∼20 nM. Functional studies on channel mutants and structural investigations on recombinant inactivation ball domain peptides encompassing the first 61 residues of Kv1.4 revealed a heme-responsive binding motif involving Cys13:His16 and a secondary histidine at position 35. Heme binding to the N-terminal inactivation domain induces a conformational constraint that prevents it from reaching its receptor site at the vestibule of the channel pore.

  20. Microbial Enzymes: Tools for Biotechnological Processes

    PubMed Central

    Adrio, Jose L.; Demain, Arnold L.

    2014-01-01

    Microbial enzymes are of great importance in the development of industrial bioprocesses. Current applications are focused on many different markets including pulp and paper, leather, detergents and textiles, pharmaceuticals, chemical, food and beverages, biofuels, animal feed and personal care, among others. Today there is a need for new, improved or/and more versatile enzymes in order to develop more novel, sustainable and economically competitive production processes. Microbial diversity and modern molecular techniques, such as metagenomics and genomics, are being used to discover new microbial enzymes whose catalytic properties can be improved/modified by different strategies based on rational, semi-rational and random directed evolution. Most industrial enzymes are recombinant forms produced in bacteria and fungi. PMID:24970208

  1. Evaluation of the treatment of both sides of raw chicken breasts with an atmospheric pressure plasma jet for the inactivation of Escherichia coli.

    PubMed

    Yong, Hae In; Kim, Hyun-Joo; Park, Sanghoo; Choe, Wonho; Oh, Mi Wha; Jo, Cheorun

    2014-08-01

    Atmospheric pressure plasma (APP) is an emerging nonthermal microbial inactivation technique. In this study, agar and raw chicken breast were inoculated with Escherichia coli and treated with an APP jet based on cold arc plasma. The aim of this study was to investigate the optimum conditions for the plasma treatment of an APP jet in order to maximize the efficiency of E. coli inactivation. The combination of N2+O2 (10 standard cubic centimeters per minute) and a longer treatment time (10 min) resulted in the highest inactivation of E. coli on agar plates with an optimum treatment distance of 20 mm. The samples in dry and wet conditions showed similar reductions in E. coli count when one side of the samples was treated at a given treatment time. Treating both sides-2.5 min on each side-resulted in a higher growth inhibition of E. coli than treatment of a single side only for 5 min. However, there was no significant difference between one-side treated samples (10 min) and both-sides treated samples (5+5 min). When the concentration of E. coli in the chicken breast sample was 10(4) colony-forming units (CFU)/g, the reduction rate of the E. coli was the highest, followed by 10(5), 10(6), and 10(7) CFU/g; however, no difference was found between 10(3) and 10(4) CFU/g. In conclusion, various treatment conditions may affect the inactivation efficiency of E. coli. In the present study, the optimum condition was determined as the treatment distance of 20 mm and longer treatment time (10 min) with the addition of oxygen to the nitrogen gas flow. Furthermore, the cell concentration of sample was an important parameter for the efficacy of the inactivation process.

  2. Inactivation of Mycobacterium avium with free chlorine.

    PubMed

    Luh, Jeanne; Mariñas, Benito J

    2007-07-15

    The inactivation kinetics of Mycobacterium avium with free chlorine was characterized by two stages: an initial phase at a relatively fast rate followed by a slower second stage of pseudo first-order kinetics. The inactivation rate of each stage was approximately the same for all experiments performed at a certain condition of pH and temperature; however, variability was observed for the disinfectant exposure at which the transition between the two stages occurred. This variability was not a function of the initial disinfectant concentration, the initial bacterial density, or the bacterial stock. However, the transition to the second stage varied more significantly at high temperatures (30 degrees C), while lower variability was observed at lower temperatures (5 and 20 degrees C). Experiments conducted at pH values in the range of 6-9 revealed that the inactivation of M. avium was primarily due to hypochlorous acid, with little contribution from hypochlorite ion within this pH range. The inactivation kinetics was represented with a two-population model. The activation energies for the resulting pseudo first-order rate constants for the populations with fast and slow kinetics were 100.3 and 96.5 kJ/mol, respectively. The magnitude of these values suggested that for waters of relatively high pH and low temperatures, little inactivation of M. avium would be achieved within treatment plants, providing a seeding source for distribution systems.

  3. Effect of different baculovirus inactivation procedures on the integrity and immunogenicity of porcine parvovirus-like particles.

    PubMed

    Rueda, P; Fominaya, J; Langeveld, J P; Bruschke, C; Vela, C; Casal, J I

    2000-11-22

    We have demonstrated earlier the usefulness of recombinant porcine parvovirus (PPV) virus-like particles (VLPs) as an efficient recombinant vaccine for PPV. Here, we have demonstrated that preparations of PPV VLPs could be contaminated by recombinant baculoviruses. Since these baculoviruses can be a problem for the registration and safety requirements of the recombinant vaccine, we have tested different baculovirus inactivation strategies, studying simultaneously the integrity and immunogenicity of the VLPs. These methods were pasteurization, treatment with detergents and alkylation with binary ethylenimine (BEI). The structural and functional integrity of the PPV VLPs after the inactivation treatments were analyzed by electron microscopy, hemagglutination, double antibody sandwich (DAS)-ELISA and immunogenicity studies. Binary ethylenimine and Triton X-100 inactivated particles maintained all the original structural and antigenic properties. In addition, PPV VLPs were subjected to size-exclusion chromatography to analyze the presence of VP2 monomers or any other contaminant. The resulting highly purified material was used as the standard of reference to quantify PPV VLPs in order to determine the dose of vaccine by DAS-ELISA. After immunization experiments in guinea pigs, the antibody titers obtained with all the inactivation procedures were very similar. Triton X-100 treatment was selected for further testing in animals because of the speed, simplicity and safety of the overall procedure.

  4. Mechanism-based inactivation of CYP2C9 by linderane.

    PubMed

    Wang, Hui; Wang, Kai; Mao, Xu; Zhang, Qingqing; Yao, Tong; Peng, Ying; Zheng, Jiang

    2015-01-01

    1. Linderane (LDR), a furan-containing sesquiterpenoid, is found in Lindera aggregata (Sims) Kosterm, a common traditional Chinese herbal medicine. We thoroughly studied the irreversible inhibitory effect of LDR on cytochrome P450 2C9 (CYP2C9). 2. LDR caused a time- and concentration-dependent inactivation of CYP2C9. In addition, the inactivation of CYP2C9 by LDR was NADPH-dependent and irreversible. More than 50% of CYP2C9 activity was lost after its incubation with LDR at the concentration of 10 μM for 15 min at 30 °C. The maximal rate constant for inactivation (kinact) was found to be 0.0419 min(-1), and the concentration required for half-maximal inactivation (KI) was 1.26 μM, respectively. Glutathione (GSH), catalase, and superoxide dismutase (SOD) failed to protect CYP2C9 against inactivation by LDR. Diclofenac, a substrate of CYP2C9, prevented the enzyme from inactivation produced by LDR. The estimated partition ratio of the inactivation was approximately 227. 3. Two reactive intermediates, including furanoepoxide and γ-ketoenal, might be responsible for the observed enzyme inactivation. The formation of the intermediates was verified by chemical synthesis. Multiple P450 enzymes, including CYPs 1A2, 2B6, 2C9, 2C19, 2D6, 3A4, and 3A5, were found to be involved in the metabolic activation of LDR. In conclusion, LDR was characterized as a mechanism-based inactivator of CYP2C9.

  5. Inactivation of MS2 bacteriophage by streamer corona discharge in water.

    PubMed

    Lee, Changha; Kim, Jaeeun; Yoon, Jeyong

    2011-02-01

    Electrical discharge processes are emerging as water treatment technologies applicable to both the degradation of organic contaminants as well as inactivation of pathogens. Particularly as a disinfection technology, electrical discharge processes do not produce toxic byproducts, and effectively inactivate a wide spectrum of microorganisms by multiple lethal actions generated by the formation of plasma channels. This study demonstrates the inactivation of a virus using the streamer corona discharge process (SCDP) with MS2 phage as a surrogate. A rapid inactivation of MS2 phage (i.e., approximately 4 log inactivation in 5 min) was observed in all experimental runs conducted. Discharge conditions such as applied voltage and storage capacitance significantly affected the inactivation efficiency of MS2 phage, whereas the influence of water quality parameters was minor. In order to elucidate the mechanism of MS2 phage inactivation, potentially lethal factors that can be generated by the SCDP were selected, and their roles in the inactivation of MS2 phage were examined. As a result, effects of UV radiation, chemical oxidants, and pulsed electric fields were found to be insignificant. The shockwave generated upon plasma channel formation appears to be the most important factor responsible for MS2 phage inactivation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  6. Long Non-coding RNAs in the X-inactivation Center

    PubMed Central

    Kalantry, Sundeep

    2014-01-01

    The X-inactivation center is a hotbed of functional long non-coding RNAs in eutherian mammals. These RNAs are thought to help orchestrate the epigenetic transcriptional states of the two X-chromosomes in females as well as of the single X-chromosome in males. To balance X-linked gene expression between the sexes, females undergo transcriptional silencing of most genes on one of the two X-chromosomes in a process termed X-chromosome inactivation. While one X-chromosome is inactivated, the other X-chromosome remains active. Moreover, with a few notable exceptions, the originally established epigenetic transcriptional profiles of the two is maintained as such through many rounds of cell division, essentially for the life of the organism. The stable divergent transcriptional fates of the two X-chromosomes, despite residing in a shared nucleoplasm, make X-inactivation a paradigm of epigenetic transcriptional regulation. Originally proposed in 1961 by Mary Lyon, the X-inactivation hypothesis has been validated through much experimentation over the last fifty years. In the last 25 years, the discovery and functional characterization has firmly established X-linked long non-coding RNAs as key players in choreographing X-chromosome inactivation. PMID:24297756

  7. Inactivation gating determines nicotine blockade of human HERG channels.

    PubMed

    Wang, H Z; Shi, H; Liao, S J; Wang, Z

    1999-09-01

    We have previously found that nicotine blocked multiple K+ currents, including the rapid component of delayed rectifier K+ currents (IKr), by interacting directly with the channels. To shed some light on the mechanisms of interaction between nicotine and channels, we performed detailed analysis on the human ether-à-go-go-related gene (HERG) channels, which are believed to be equivalent to the native I(Kr) when expressed in Xenopus oocytes. Nicotine suppressed the HERG channels in a concentration-dependent manner with greater potency with voltage protocols, which favor channel inactivation. Nicotine caused dramatic shifts of the voltage-dependent inactivation curve to more negative potentials and accelerated the inactivation process. Conversely, maneuvers that weakened the channel inactivation gating considerably relieved the blockade. Elevating the extracellular K+ concentration from 5 to 20 mM increased the nicotine concentration (by approximately 100-fold) needed to achieve the same degree of inhibition. Moreover, nicotine lost its ability to block the HERG channels when a single mutation was introduced to a residue located after transmembrane domain 6 (S631A) to remove the rapid channel inactivation. Our data suggest that the inactivation gating determines nicotine blockade of the HERG channels.

  8. Free chlorine inactivation of fungi in drinking water sources.

    PubMed

    Pereira, V J; Marques, R; Marques, M; Benoliel, M J; Barreto Crespo, M T

    2013-02-01

    The effectiveness of free chlorine for the inactivation of fungi present in settled surface water was tested. In addition, free chlorine inactivation rate constants of Cladosporium tenuissimum, Cladosporium cladosporioides, Phoma glomerata, Aspergillus terreus, Aspergillus fumigatus, Penicillium griseofulvum, and Penicillium citrinum that were found to occur in different source waters were determined in different water matrices (laboratory grade water and settled water). The effect of using different disinfectant concentrations (1 and 3 mg/l), temperatures (21 and 4 °C), and pH levels (6 and 7) was addressed. The sensitivity degree of different fungi isolates to chlorine disinfection varied among different genera with some species showing a higher resistance to disinfection and others expected to be more prone to protection from inactivation by the water matrix components. When the disinfection efficiency measured in terms of the chlorine concentration and contact time (Ct) values needed to achieve 99% inactivation were compared with the Ct values reported as being able to achieve the same degree of inactivation of other microorganisms, fungi were found to be more resistant to chlorine inactivation than bacteria and viruses and less resistant than Cryptosporidium oocysts. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. INACTIVATION OF SOME SEMISYNTHETIC PENICILLINS BY GRAM-NEGATIVE BACILLI

    PubMed Central

    Sabath, Leon; Finland, Maxwell

    1963-01-01

    Sabath, Leon (Boston City Hospital, Boston, Mass.) and Maxwell Finland. Inactivation of some semisynthetic penicillins by gram-negative bacilli. J. Bacteriol. 85:314–321. 1963.—An agar diffusion method was used to test 55 strains of gram-negative bacilli for their ability to inactivate penicillin G, methicillin, biphenylpenicillin, oxacillin, and ampicillin; 26 strains inactivated one or more of them. All strains of Klebsiella-Aerobacter, nearly all of Escherichia coli, and some of Pseudomonas aeruginosa, but not those of Proteus or Salmonella, were active by this method. Penicillin G was inactivated by the largest number of strains, biphenylpenicillin and ampicillin by somewhat fewer, and oxacillin and methicillin by about half as many. When the five penicillins were incubated with four strains of different bacteria in broth at 37 C, all were inactivated to a considerable extent by all the strains, each penicillin to a different degree, but to about the same extent by all the strains. Adsorption alone did not account for the loss of activity. The results suggest that there are qualitative, as well as quantitative, differences among species or even strains of gram-negative bacilli in their ability to inactivate the various penicillins. Images PMID:13975857

  10. A general approach for chemical labeling and rapid, spatially controlled protein inactivation

    PubMed Central

    Marks, Kevin M.; Braun, Patrick D.; Nolan, Garry P.

    2004-01-01

    Chemical labeling of proteins inside of living cells can enable studies of the location, movement, and function of proteins in vivo. Here we demonstrate an approach for chemical labeling of proteins that uses the high-affinity interaction between an FKBP12 mutant (F36V) and a synthetic, engineered ligand (SLF′). A fluorescein conjugate to the engineered ligand (FL-SLF′) retained binding to FKBP12(F36V) and possessed similar fluorescence properties as parental fluorescein. FL-SLF′ labeled FKBP12(F36V) fusion proteins in live mammalian cells, and was used to monitor the subcellular localization of a membrane targeted FKBP12(F36V) construct. Chemical labeling of FKBP12(F36V) fusion proteins with FL-SLF′ was readily detectable at low expression levels of the FKBP12(F36V) fusion, and the level of fluorescent staining with FL-SLF′ was proportional to the FKBP12(F36V) expression level. This FL-SLF′-FKBP12(F36V) labeling technique was tested in fluorophore assisted laser inactivation (FALI), a light-mediated technique to rapidly inactivate fluorophore-labeled target proteins. FL-SLF′ mediated FALI of a β-galactosidase-FKBP12(F36V) fusion protein, causing rapid inactivation of >90% of enzyme activity upon irradiation in vitro. FL-SLF′ also mediated FALI of a β-galactosidase fusion expressed in living NIH 3T3 cells, where β-galactosidase activity was reduced in 15 s. Thus, FL-SLF′ can be used to monitor proteins in vivo and to target rapid, spatially and temporally defined inactivation of target proteins in living cells in a process that we call FK-FALI. PMID:15218100

  11. SURFACE INACTIVATION OF BACTERIAL VIRUSES AND OF PROTEINS

    PubMed Central

    Adams, Mark H.

    1948-01-01

    1. The seven bacterial viruses of the T group active against E. coli, are rapidly inactivated at gas-liquid interfaces. 2. The kinetics of this inactivation whether brought about by shaking or by bubbling with nitrogen are those of a first order reaction. 3. This inactivation may be prevented by the addition of enough protein to maintain the gas-liquid interface in a saturated condition. 4. The analogy between this phenomenon and the surface denaturation of proteins is pointed out and discussed. PMID:18917025

  12. Thermal inactivation and chaperonin-mediated renaturation of mitochondrial aspartate aminotransferase.

    PubMed Central

    Lawton, J M; Doonan, S

    1998-01-01

    Mitochondrial aspartate aminotransferase is inactivated irreversibly on heating. The inactivated protein aggregates, but aggregation is prevented by the presence of the chaperonin 60 from Escherichia coli (GroEL). The chaperonin increases the rate of thermal inactivation in the temperature range 55-65 degrees C but not at lower temperatures. It has previously been shown [Twomey and Doonan (1997) Biochim. Biophys. Acta 1342, 37-44] that the enzyme switches to a modified, but catalytically active, conformation at approx. 55-60 degrees C and the present results show that this conformation is recognized by and binds to GroEL. The thermally inactivated protein can be released from GroEL in an active form by the addition of chaperonin 10 from E. coli (GroES)/ATP, showing that inactivation is not the result of irreversible chemical changes. These results suggest that the irreversibility of thermal inactivation is due to the formation of an altered conformation with a high kinetic barrier to refolding rather than to any covalent changes. In the absence of chaperonin the unfolded molecules aggregate but this is a consequence, rather than the cause, of irreversible inactivation. PMID:9693123

  13. Prevention of microbial biofilms - the contribution of micro and nanostructured materials.

    PubMed

    Grumezescu, Alexandru Mihai; Chifiriuc, Carmen Mariana

    2014-01-01

    Microbial biofilms are associated with drastically enhanced resistance to most of the antimicrobial agents and with frequent treatment failures, generating the search for novel strategies which can eradicate infections by preventing the persistent colonization of the hospital environment, medical devices or human tissues. Some of the current approaches for fighting biofilms are represented by the development of novel biomaterials with increased resistance to microbial colonization and by the improvement of the current therapeutic solutions with the aid of nano (bio)technology. This special issues includes papers describing the applications of nanotechnology and biomaterials science for the development of improved drug delivery systems and nanostructured surfaces for the prevention and treatment of medical biofilms. Nanomaterials display unique and well-defined physical and chemical properties making them useful for biomedical applications, such as: very high surface area to volume ratio, biocompatibility, biodegradation, safety for human ingestion, capacity to support surface modification and therefore, to be combined with other bioactive molecules or substrata and more importantly being seemingly not attracting antimicrobial resistance. The use of biomaterials is significantly contributing to the reduction of the excessive use of antibiotics, and consequently to the decrease of the emergence rate of resistant microorganisms, as well as of the associated toxic effects. Various biomaterials with intrinsic antimicrobial activity (inorganic nanoparticles, polymers, composites), medical devices for drug delivery, as well as factors influencing their antimicrobial properties are presented. One of the presented papers reviews the recent literature on the use of magnetic nanoparticles (MNP)-based nanomaterials in antimicrobial applications for biomedicine, focusing on the growth inhibition and killing of bacteria and fungi, and, on viral inactivation. The anti

  14. Low pH inactivation for xenotropic gamma retrovirus in recombinant human TNF-α receptor immunoglobulin G and mechanism of inactivation.

    PubMed

    Ma, Rong; Cui, Xiaolan

    2014-01-01

    CHO-derived recombinant proteins for human therapeutic are used commonly. There are noninfectious endogenous retroviruses in CHO cells. Validation study for inactivation process is required. Murine xenotropic gamma retrovirus (X-MulV) is a model virus in validation study. In our previous study, optimum conditions for X-MulV inactivation were sifted. In this study, we performed a further research on low pH inactivation for evaluation of X-MulV clearance in manufacturing of recombinant human TNF-α receptor immunoglobulin G fusion proteins (rhTNF-α) for injection. Cell-based infectivity assay was used for the evaluation of X-MulV clearance. RhTNF-α were spiked with X-MulV and were inactivated at pH 3.60 ∼ 3.90, 25 ± 2 °C, and 0 ∼ 240 min, respectively. Samples incubated at the conditions for 15 ∼ 180 min were not inactivated effectively. For 4 h incubation, log10 reductions were achieved 5.0 log10. Biological activity of rhTNF-α incubated at pH 3.60, 25 °C for 4 h, which was assayed on murine L929 fibroblasts cells, was not affected by low pH. Env gene of X-MulV, which was detected by conventional PCR method for the first time, was not detected after incubation at pH 3.60, and it may be the mechanism of low pH inactivation. Copyright © 2013. Published by Elsevier Ltd.

  15. Initial steps of inactivation at the K+ channel selectivity filter

    PubMed Central

    Thomson, Andrew S.; Heer, Florian T.; Smith, Frank J.; Hendron, Eunan; Bernèche, Simon; Rothberg, Brad S.

    2014-01-01

    K+ efflux through K+ channels can be controlled by C-type inactivation, which is thought to arise from a conformational change near the channel’s selectivity filter. Inactivation is modulated by ion binding near the selectivity filter; however, the molecular forces that initiate inactivation remain unclear. We probe these driving forces by electrophysiology and molecular simulation of MthK, a prototypical K+ channel. Either Mg2+ or Ca2+ can reduce K+ efflux through MthK channels. However, Ca2+, but not Mg2+, can enhance entry to the inactivated state. Molecular simulations illustrate that, in the MthK pore, Ca2+ ions can partially dehydrate, enabling selective accessibility of Ca2+ to a site at the entry to the selectivity filter. Ca2+ binding at the site interacts with K+ ions in the selectivity filter, facilitating a conformational change within the filter and subsequent inactivation. These results support an ionic mechanism that precedes changes in channel conformation to initiate inactivation. PMID:24733889

  16. Susceptibility of representative dental pathogens to inactivation by the PDT with water-soluble photosensitizers

    NASA Astrophysics Data System (ADS)

    Angelov, Ivan; Mantareva, Vanya; Kussovski, Veselin; Worle, Diter; Kisov, Hristo; Belcheva, Marieta; Georgieva, Tzvetelina; Dimitrov, Slavcho

    2011-02-01

    In the recent decade the applications of photodynamic therapy (PDT) rapidly increase in several topics and one of areas where the PDT in the future will be play significant role is dentistry. The different photosensitizing complexes with a good water solubility and with absorption with an intensive maximum in the red region (630-690 nm), which makes them suitable for photodynamic treatments, were investigated. The photochemical properties of complexes for singlet oxygen generation were investigated and were shown relations between uptake levels and light intensity to achieve increase in photodynamic efficacy. Photodynamic efficacy against fungi Candida albicans and bacteria's E. faecalis, MRSA and S. Mutans in planktonic media was evaluated. The high photodynamic efficacy was shown for SiPc at very low concentrations (0.9 μM) and light doses of 50 J cm-2 by intensity of light 60 mW cm-2. The photodynamic response for E. faecalis, MRSA and S. Mutans, after treatments with different photosensitizers show strong dependence on concentrations of photsensitzers and micro organisms. The level of inactivation of the pathogen bacteria's from 1-2 degree of initial concentration up to full inactivation was observed. The studied complexes were compared to the recently studied Methylene blue, Haematoporphyrine and tetra-methylpirydiloxy Zn(II)- phthalocyanines and experimental results show that some of them have a good potential for inactivation of representative pathogenic bacterial strains. Experimental results also indicate that photodynamic therapy appears an effective method for inactivation of oral pathogenic bacterias and fungi.

  17. Significance of Inactivated Genes in Leukemia: Pathogenesis and Prognosis

    PubMed Central

    Heidari, Nazanin; Abroun, Saeid; Bertacchini, Jessika; Vosoughi, Tina; Rahim, Fakher; Saki, Najmaldin

    2017-01-01

    Epigenetic and genetic alterations are two mechanisms participating in leukemia, which can inactivate genes involved in leukemia pathogenesis or progression. The purpose of this review was to introduce various inactivated genes and evaluate their possible role in leukemia pathogenesis and prognosis. By searching the mesh words “Gene, Silencing AND Leukemia” in PubMed website, relevant English articles dealt with human subjects as of 2000 were included in this study. Gene inactivation in leukemia is largely mediated by promoter’s hypermethylation of gene involving in cellular functions such as cell cycle, apoptosis, and gene transcription. Inactivated genes, such as ASPP1, TP53, IKZF1 and P15, may correlate with poor prognosis in acute lymphoid leukemia (ALL), chronic lymphoid leukemia (CLL), chronic myelogenous leukemia (CML) and acute myeloid leukemia (AML), respectively. Gene inactivation may play a considerable role in leukemia pathogenesis and prognosis, which can be considered as complementary diagnostic tests to differentiate different leukemia types, determine leukemia prognosis, and also detect response to therapy. In general, this review showed some genes inactivated only in leukemia (with differences between B-ALL, T-ALL, CLL, AML and CML). These differences could be of interest as an additional tool to better categorize leukemia types. Furthermore; based on inactivated genes, a diverse classification of Leukemias could represent a powerful method to address a targeted therapy of the patients, in order to minimize side effects of conventional therapies and to enhance new drug strategies. PMID:28580304

  18. Inactivation of Bacillus atrophaeus by OH radicals

    NASA Astrophysics Data System (ADS)

    Ono, Ryo; Yonetamari, Kenta; Tokumitsu, Yusuke; Yonemori, Seiya; Yasuda, Hachiro; Mizuno, Akira

    2016-08-01

    The inactivation of Bacillus atrophaeus by OH radicals is measured. This study aims to evaluate the bactericidal effects of OH radicals produced by atmospheric-pressure nonthermal plasma widely used for plasma medicine; however, in this study, OH radicals are produced by vacuum ultraviolet (VUV) photolysis of water vapor instead of plasma to allow the production of OH radicals with almost no other reactive species. A 172 nm VUV light from a Xe2 excimer lamp irradiates a He-H2O mixture flowing in a quartz tube to photodissociate H2O to produce OH, H, O, HO2, H2O2, and O3. The produced reactive oxygen species (ROS) flow out of the quartz tube nozzle to the bacteria on an agar plate and cause inactivation. The inactivation by OH radicals among the six ROS is observed by properly setting the experimental conditions with the help of simulations calculating the ROS densities. A 30 s treatment with approximately 0.1 ppm OH radicals causes visible inactivation.

  19. Quantitative assessment of the risk of microbial spoilage in foods. Prediction of non-stability at 55 °C caused by Geobacillus stearothermophilus in canned green beans.

    PubMed

    Rigaux, Clémence; André, Stéphane; Albert, Isabelle; Carlin, Frédéric

    2014-02-03

    Microbial spoilage of canned foods by thermophilic and highly heat-resistant spore-forming bacteria, such as Geobacillus stearothermophilus, is a persistent problem in the food industry. An incubation test at 55 °C for 7 days, then validation of biological stability, is used as an indicator of compliance with good manufacturing practices. We propose a microbial risk assessment model predicting the percentage of non-stability due to G. stearothermophilus in canned green beans manufactured by a French company. The model accounts for initial microbial contaminations of fresh unprocessed green beans with G. stearothermophilus, cross-contaminations in the processing chain, inactivation processes and probability of survival and growth. The sterilization process is modeled by an equivalent heating time depending on sterilization value F₀ and on G. stearothermophilus resistance parameter z(T). Following the recommendations of international organizations, second order Monte-Carlo simulations are used, separately propagating uncertainty and variability on parameters. As a result of the model, the mean predicted non-stability rate is of 0.5%, with a 95% uncertainty interval of [0.1%; 1.2%], which is highly similar to data communicated by the French industry. A sensitivity analysis based on Sobol indices and some scenario tests underline the importance of cross-contamination at the blanching step, in addition to inactivation due to the sterilization process. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Germination and Inactivation of Alicyclobacillus acidoterrestris Spores Induced by Moderate Hydrostatic Pressure.

    PubMed

    Sokołowska, Barbara; Skapska, Sylwia; Fonberg-Broczek, Monika; Niezgoda, Jolanta; Porebska, Izabela; Dekowska, Agnieszka; Rzoska, Sylwester J

    2015-01-01

    Given the importance of spoilage caused by Alicyclobacillus acidoterrestris for the fruit juice industry, the objective of this work was to study the germination and inactivation of A. acidoterrestris spores induced by moderate hydrostatic pressure. Hydrostatic pressure treatment can induce the germination and inactivation of A. acidoterrestris spores. At low pH, spore germination of up to 3.59-3.75 log and inactivation of 1.85-2.04 log was observed in a low pressure window (200-300 MPa) applied at 50 degrees C for 20 min. Neutral pH suppressed inactivation, the number of spores inactivated at pH 7.0 was only 0.24-1.06 log. The pressurization temperature significantly affected spore germination and inactivation. The degree of germination in apple juice after pressurization for 30 min with 200 MPa at 20 degrees C was 2.04 log, with only 0.61 log of spores being inactivated, while at 70 degrees C spore germination was 5.94 log and inactivation 4.72 log. This temperature strongly stimulated germination and inactivation under higher (500 MPa) than lower (200 MPa) pressure. When the oscillatory mode was used, the degree of germination and inactivation was slightly higher than at continuous mode. The degree of germination and inactivation was inversely proportional to the soluble solids content and was lowest in concentrated apple juice.

  1. Inactivation of human and simian rotaviruses by ozone

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

    Vaughn, J.M.; Chen, Y.S.; Lindburg, K.

    1987-09-01

    The inactivation of simian rotavirus Sa-11 and human rotavirus type 2 (Wa) by ozone was compared at 4/sup 0/C by using single-particle virus stocks. Although the human strain was clearly more sensitive, both virus types were rapidly inactivated by ozone concentrations of 0.25 mg/liter or greater at all pH levels tested. Comparison of the virucidal activity of ozone with that of chlorine in identical experiments indicated little significant difference in rotavirus-inactivating efficiencies when the disinfectants were used at concentrations of 0.25 mg/liter or greater.

  2. Inactivation of RNA Viruses by Gamma Irradiation: A Study on Mitigating Factors

    PubMed Central

    Hume, Adam J.; Ames, Joshua; Rennick, Linda J.; Duprex, W. Paul; Marzi, Andrea; Tonkiss, John; Mühlberger, Elke

    2016-01-01

    Effective inactivation of biosafety level 4 (BSL-4) pathogens is vital in order to study these agents safely. Gamma irradiation is a commonly used method for the inactivation of BSL-4 viruses, which among other advantages, facilitates the study of inactivated yet morphologically intact virions. The reported values for susceptibility of viruses to inactivation by gamma irradiation are sometimes inconsistent, likely due to differences in experimental protocols. We analyzed the effects of common sample attributes on the inactivation of a recombinant vesicular stomatitis virus expressing the Zaire ebolavirus glycoprotein and green fluorescent protein. Using this surrogate virus, we found that sample volume and protein content of the sample modulated viral inactivation by gamma irradiation but that air volume within the sample container and the addition of external disinfectant surrounding the sample did not. These data identify several factors which alter viral susceptibility to inactivation and highlight the usefulness of lower biosafety level surrogate viruses for such studies. Our results underscore the need to validate inactivation protocols of BSL-4 pathogens using “worst-case scenario” procedures to ensure complete sample inactivation. PMID:27455307

  3. Effects of Plant Diversity, Functional Group Composition, and Fertilization on Soil Microbial Properties in Experimental Grassland

    PubMed Central

    Strecker, Tanja; Barnard, Romain L.; Niklaus, Pascal A.; Scherer-Lorenzen, Michael; Weigelt, Alexandra; Scheu, Stefan; Eisenhauer, Nico

    2015-01-01

    Background Loss of biodiversity and increased nutrient inputs are two of the most crucial anthropogenic factors driving ecosystem change. Although both received considerable attention in previous studies, information on their interactive effects on ecosystem functioning is scarce. In particular, little is known on how soil biota and their functions are affected by combined changes in plant diversity and fertilization. Methodology/Principal Findings We investigated the effects of plant diversity, functional community composition, and fertilization on the biomass and respiration of soil microbial communities in a long-term biodiversity experiment in semi-natural grassland (Jena Experiment). Plant species richness enhanced microbial basal respiration and microbial biomass, but did not significantly affect microbial specific respiration. In contrast, the presence of legumes and fertilization significantly decreased microbial specific respiration, without altering microbial biomass. The effect of legumes was superimposed by fertilization as indicated by a significant interaction between the presence of legumes and fertilization. Further, changes in microbial stoichiometry (C-to-N ratio) and specific respiration suggest the presence of legumes to reduce N limitation of soil microorganisms and to modify microbial C use efficiency. Conclusions/Significance Our study highlights the role of plant species and functional group diversity as well as interactions between plant community composition and fertilizer application for soil microbial functions. Our results suggest soil microbial stoichiometry to be a powerful indicator of microbial functioning under N limited conditions. Although our results support the notion that plant diversity and fertilizer application independently affect microbial functioning, legume effects on microbial N limitation were superimposed by fertilization, indicating significant interactions between the functional composition of plant communities and

  4. [Effect of various factors on ozone inactivating Giardia in water].

    PubMed

    Ran, Zhi-Lin; Li, Shao-Feng; Huang, Jun-Li; Yuan, Yi-Xing; Cui, Chong-Wei

    2010-06-01

    In order to study the effect of O3 inactivating Giardia in water, different factors (CT value, pH, temperature, turbidity, organic content and inorganic ions) which might influence the inactivation were investigated by using fluorescence staining method. The results indicated that the whole process of O3 inactivating Giardia could be divided into two periods, the inactivated rate in log phase was significantly faster than it in the slow phase [k1 = (5.64 +/- 0.023) x 10(-1) mg x min, k2 = (2.72 +/- 0.002) x 10(-2) mg x min, k1 > k2]. When the turbidity was 0.1 to 20. 0NTU, temperature was 5 to 35 degrees C, pH was 6.0 to 9.0, HA content was 0.5 to 10.0 mg/L, the turbidity was lower, the higher inactivating ratio could be received. With the increasing of temperature, the inactivating effect was decreased. The ability of O3 inactivating Giardia was stronger under acidic condition than it was in alkali circumstance. When the reaction system contained higher concentration of organics, the competition reaction might take place between Giardia and organics with O3, which might reduce inactivation ratio. The sequence of affecting disinfectant ability of O3 was NO3- > None > SO4(2-) > HCO3-, while inorganic cations (Ca2+, Mg2+ and Cu2+) promoted the inactive reaction to a certain extent. If the CT value of O3 was more than 15.0 min x mg/L, the ratio of inactivation could exceed 99.0% during disinfecting drinking water.

  5. Deep subsurface microbial processes

    USGS Publications Warehouse

    Lovley, D.R.; Chapelle, F.H.

    1995-01-01

    Information on the microbiology of the deep subsurface is necessary in order to understand the factors controlling the rate and extent of the microbially catalyzed redox reactions that influence the geophysical properties of these environments. Furthermore, there is an increasing threat that deep aquifers, an important drinking water resource, may be contaminated by man's activities, and there is a need to predict the extent to which microbial activity may remediate such contamination. Metabolically active microorganisms can be recovered from a diversity of deep subsurface environments. The available evidence suggests that these microorganisms are responsible for catalyzing the oxidation of organic matter coupled to a variety of electron acceptors just as microorganisms do in surface sediments, but at much slower rates. The technical difficulties in aseptically sampling deep subsurface sediments and the fact that microbial processes in laboratory incubations of deep subsurface material often do not mimic in situ processes frequently necessitate that microbial activity in the deep subsurface be inferred through nonmicrobiological analyses of ground water. These approaches include measurements of dissolved H2, which can predict the predominant microbially catalyzed redox reactions in aquifers, as well as geochemical and groundwater flow modeling, which can be used to estimate the rates of microbial processes. Microorganisms recovered from the deep subsurface have the potential to affect the fate of toxic organics and inorganic contaminants in groundwater. Microbial activity also greatly influences 1 the chemistry of many pristine groundwaters and contributes to such phenomena as porosity development in carbonate aquifers, accumulation of undesirably high concentrations of dissolved iron, and production of methane and hydrogen sulfide. Although the last decade has seen a dramatic increase in interest in deep subsurface microbiology, in comparison with the study of

  6. Nutrient addition dramatically accelerates microbial community succession.

    PubMed

    Knelman, Joseph E; Schmidt, Steven K; Lynch, Ryan C; Darcy, John L; Castle, Sarah C; Cleveland, Cory C; Nemergut, Diana R

    2014-01-01

    The ecological mechanisms driving community succession are widely debated, particularly for microorganisms. While successional soil microbial communities are known to undergo predictable changes in structure concomitant with shifts in a variety of edaphic properties, the causal mechanisms underlying these patterns are poorly understood. Thus, to specifically isolate how nutrients--important drivers of plant succession--affect soil microbial succession, we established a full factorial nitrogen (N) and phosphorus (P) fertilization plot experiment in recently deglaciated (∼3 years since exposure), unvegetated soils of the Puca Glacier forefield in Southeastern Peru. We evaluated soil properties and examined bacterial community composition in plots before and one year after fertilization. Fertilized soils were then compared to samples from three reference successional transects representing advancing stages of soil development ranging from 5 years to 85 years since exposure. We found that a single application of +NP fertilizer caused the soil bacterial community structure of the three-year old soils to most resemble the 85-year old soils after one year. Despite differences in a variety of soil edaphic properties between fertilizer plots and late successional soils, bacterial community composition of +NP plots converged with late successional communities. Thus, our work suggests a mechanism for microbial succession whereby changes in resource availability drive shifts in community composition, supporting a role for nutrient colimitation in primary succession. These results suggest that nutrients alone, independent of other edaphic factors that change with succession, act as an important control over soil microbial community development, greatly accelerating the rate of succession.

  7. Nutrient Addition Dramatically Accelerates Microbial Community Succession

    PubMed Central

    Knelman, Joseph E.; Schmidt, Steven K.; Lynch, Ryan C.; Darcy, John L.; Castle, Sarah C.; Cleveland, Cory C.; Nemergut, Diana R.

    2014-01-01

    The ecological mechanisms driving community succession are widely debated, particularly for microorganisms. While successional soil microbial communities are known to undergo predictable changes in structure concomitant with shifts in a variety of edaphic properties, the causal mechanisms underlying these patterns are poorly understood. Thus, to specifically isolate how nutrients – important drivers of plant succession – affect soil microbial succession, we established a full factorial nitrogen (N) and phosphorus (P) fertilization plot experiment in recently deglaciated (∼3 years since exposure), unvegetated soils of the Puca Glacier forefield in Southeastern Peru. We evaluated soil properties and examined bacterial community composition in plots before and one year after fertilization. Fertilized soils were then compared to samples from three reference successional transects representing advancing stages of soil development ranging from 5 years to 85 years since exposure. We found that a single application of +NP fertilizer caused the soil bacterial community structure of the three-year old soils to most resemble the 85-year old soils after one year. Despite differences in a variety of soil edaphic properties between fertilizer plots and late successional soils, bacterial community composition of +NP plots converged with late successional communities. Thus, our work suggests a mechanism for microbial succession whereby changes in resource availability drive shifts in community composition, supporting a role for nutrient colimitation in primary succession. These results suggest that nutrients alone, independent of other edaphic factors that change with succession, act as an important control over soil microbial community development, greatly accelerating the rate of succession. PMID:25050551

  8. Film forming microbial biopolymers for commercial applications--a review.

    PubMed

    Vijayendra, S V N; Shamala, T R

    2014-12-01

    Microorganisms synthesize intracellular, structural and extracellular polymers also referred to as biopolymers for their function and survival. These biopolymers play specific roles as energy reserve materials, protective agents, aid in cell functioning, the establishment of symbiosis, osmotic adaptation and support the microbial genera to function, adapt, multiply and survive efficiently under changing environmental conditions. Viscosifying, gelling and film forming properties of these have been exploited for specific significant applications in food and allied industries. Intensive research activities and recent achievements in relevant and important research fields of global interest regarding film forming microbial biopolymers is the subject of this review. Microbial polymers such as pullulan, kefiran, bacterial cellulose (BC), gellan and levan are placed under the category of exopolysaccharides (EPS) and have several other functional properties including film formation, which can be used for various applications in food and allied industries. In addition to EPS, innumerable bacterial genera are found to synthesis carbon energy reserves in their cells known as polyhydroxyalkanoates (PHAs), microbial polyesters, which can be extruded into films with excellent moisture and oxygen barrier properties. Blow moldable biopolymers like PHA along with polylactic acid (PLA) synthesized chemically in vitro using lactic acid (LA), which is produced by LA bacteria through fermentation, are projected as biodegradable polymers of the future for packaging applications. Designing and creating of new property based on requirements through controlled synthesis can lead to improvement in properties of existing polysaccharides and create novel biopolymers of great commercial interest and value for wider applications. Incorporation of antimicrobials such as bacteriocins or silver and copper nanoparticles can enhance the functionality of polymer films especially in food packaging

  9. Design and Mechanism of Tetrahydrothiophene-based GABA Aminotransferase Inactivators

    PubMed Central

    Le, Hoang V.; Hawker, Dustin D.; Wu, Rui; Doud, Emma; Widom, Julia; Sanishvili, Ruslan; Liu, Dali; Kelleher, Neil L.; Silverman, Richard B.

    2015-01-01

    Low levels of γ-aminobutyric acid (GABA), one of two major neurotransmitters that regulate brain neuronal activity, are associated with many neurological disorders, such as epilepsy, Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, and cocaine addiction. One of the main methods to raise the GABA level in human brain is to use small molecules that cross the blood-brain barrier and inhibit the activity of γ-aminobutyric acid aminotransferase (GABA-AT), the enzyme that degrades GABA. We have designed a series of conformationally-restricted, tetrahydrothiophene-based GABA analogs with a properly-positioned leaving group that could facilitate a ring-opening mechanism, leading to inactivation of GABA-AT. One compound in the series is eight times more efficient an inactivator of GABA-AT than vigabatrin, the only FDA-approved inactivator of GABA-AT. Our mechanistic studies show that the compound inactivates GABA-AT by a new mechanism. The metabolite resulting from inactivation does not covalently bind to amino acid residues of GABA-AT but stays in the active site via H-bond interactions with Arg-192, a π-π interaction with Phe-189, and a weak nonbonded S···O=C interaction with Glu-270, thereby inactivating the enzyme. PMID:25781189

  10. Thermal Inactivation of Feline Calicivirus in Pet Food Processing.

    PubMed

    Haines, J; Patel, M; Knight, A I; Corley, D; Gibson, G; Schaaf, J; Moulin, J; Zuber, S

    2015-12-01

    Extrusion is the most common manufacturing process used to produce heat-treated dry dog and cat food (pet food) for domestic use and international trade. Due to reoccurring outbreaks of notifiable terrestrial animal diseases and their impact on international trade, experiments were undertaken to demonstrate the effectiveness of heat-treated extruded pet food on virus inactivation. The impact of extrusion processing in a pet food matrix on virus inactivation has not been previously reported and very few inactivation studies have examined the thermal inactivation of viruses in complex food matrices. The feline calicivirus vaccine strain FCV F-9 was used as a surrogate model RNA virus pathogen. Small-scale heat inactivation experiments using animal-derived pet food raw materials showed that a > 4 log10 reduction (log10 R) in infectivity occurred at 70 °C prior to reaching the minimum extrusion manufacturing operating temperature of 100 °C. As anticipated, small-scale pressure studies at extrusion pressure (1.6 MPa) showed no apparent effect on FCV F-9 inactivation. Additionally, FCV F-9 was shown not to survive the acidic conditions used to produce pet food palatants of animal origin that are typically used as a coating after the extrusion process.

  11. Use of PRD1 bacteriophage in groundwater viral transport, inactivation, and attachment studies

    USGS Publications Warehouse

    Harvey, Ronald W.; Ryan, Joseph N.

    2004-01-01

    PRD1, an icosahedra-shaped, 62 nm (diameter), double-stranded DNA bacteriophage with an internal membrane, has emerged as an important model virus for studying the manner in which microorganisms are transported through a variety of groundwater environments. The popularity of this phage for use in transport studies involving geologic media is due, in part, to its relative stability over a range of temperatures and low degree of attachment in aquifer sediments. Laboratory and field investigations employing PRD1 are leading to a better understanding of viral attachment and transport behaviors in saturated geologic media and to improved methods for describing mathematically subsurface microbial transport at environmentally significant field scales. Radioisotopic labeling of PRD1 is facilitating additional information about the nature of viral interactions with solid surfaces in geologic media, the importance of iron oxide surfaces, and allowing differentiation between inactivation and attachment in field-scale tracer tests.

  12. Conjugated Polymers/DNA Hybrid Materials for Protein Inactivation.

    PubMed

    Zhao, Likun; Zhang, Jiangyan; Xu, Huiming; Geng, Hao; Cheng, Yongqiang

    2016-09-07

    Chromophore-assisted light inactivation (CALI) is a powerful tool for analyzing protein functions due to the high degree of spatial and temporal resolution. In this work, we demonstrate a CALI approach based on conjugated polymers (CPs)/DNA hybrid material for protein inactivation. The target protein is conjugated with single-stranded DNA in advance. Single-stranded DNA can form CPs/DNA hybrid material with cationic CPs via electrostatic and hydrophobic interactions. Through the formation of CPs/DNA hybrid material, the target protein that is conjugated with DNA is brought into close proximity to CPs. Under irradiation, CPs harvest light and generate reactive oxygen species (ROS), resulting in the inactivation of the adjacent target protein. This approach can efficiently inactivate any target protein which is conjugated with DNA and has good specificity and universality, providing a new strategy for studies of protein function and adjustment of protein activity.

  13. In vitro and in vivo anti-microbial activity evaluation of inactivated cells of Lactobacillus salivarius CECT 5713 against Streptococcus mutans.

    PubMed

    Sañudo, Ana I; Luque, Roberto; Díaz-Ropero, Mª Paz; Fonollá, Juristo; Bañuelos, Óscar

    2017-12-01

    Defining the etiology of dental caries is a complex problem. The microbiological approach has included Streptococcus mutans as one of the bacterial species involved in this disease. This research investigates the inhibitory effects of heat-inactivated Lactobacillus salivarius CECT 5713 against S. mutans using in vitro and in vivo assays. On the one hand, the effect of non-viable L. salivarius CECT 5713 on the in vitro adhesion of S. mutans to hydroxyapatite discs was evaluated. On the other hand, levels of Streptococcus mutans, amount of salivary flow and salivary pH before and after taking the rinse with the non-viable L. salivarius CECT 5713 in healthy volunteers were assessed (self-controlled open-label pilot study). The levels of S. mutans seemed to decrease in the in vitro and in vivo assays (p<0.05). The in vitro effect of non-viable L. salivarius was maintained until 36 months of storage. In addition, the reduction of S. mutans salivary concentration in the volunteers was statistically significant from the third day until two weeks of treatment. Heat-inactivated L. salivarius CECT 5713 prevents S. mutans adhesion to hydroxyapatite and could be used as a strategy to reduce the salivary concentration of this oral pathogen. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Validating the Inactivation Effectiveness of Chemicals on Ebola Virus.

    PubMed

    Haddock, Elaine; Feldmann, Friederike

    2017-01-01

    While viruses such as Ebola virus must be handled in high-containment laboratories, there remains the need to process virus-infected samples for downstream research testing. This processing often includes removal to lower containment areas and therefore requires assurance of complete viral inactivation within the sample before removal from high-containment. Here we describe methods for the removal of chemical reagents used in inactivation procedures, allowing for validation of the effectiveness of various inactivation protocols.

  15. Selective Destruction of Protein Function by Chromophore-Assisted Laser Inactivation

    NASA Astrophysics Data System (ADS)

    Jay, Daniel G.

    1988-08-01

    Chromophore-assisted laser inactivation of protein function has been achieved. After a protein binds a specific ligand or antibody conjugated with malachite green (C.I. 42000), it is selectively inactivated by laser irradiation at a wavelength of light absorbed by the dye but not significantly absorbed by cellular components. Ligand-bound proteins in solution and on the surfaces of cells can be denatured without other proteins in the same samples being affected. Chromophore-assisted laser inactivation can be used to study cell surface phenomena by inactivating the functions of single proteins on living cells, a molecular extension of cellular laser ablation. It has an advantage over genetics and the use of specific inhibitors in that the protein function of a single cell within the organism can be inactivated by focusing the laser beam.

  16. Contribution of microbial carbon to soil fractions: significance of diverse microbial group biochemistry

    NASA Astrophysics Data System (ADS)

    Throckmorton, H.; Bird, J. A.; Dane, L.; Firestone, M. K.; Horwath, W. R.

    2011-12-01

    point for both sites. Microbial C inputs were more stable in the OF and MF than the LF throughout the course of the study at both sites. There were no differences in 13C recovery among microbial isolates in any fractions in PR, despite minor differences in overall turnover rates. In CA, there were some differences among microbial isolates in 13C recovery in the LF and OF, which related to 13C recoveries in whole soils. In the CA MF, microbial 13C recoveries did not significantly differ, and in generally low variability among treatments was observed. Results support increased protection of microbial C via association with the MF; however, differential sorption of various microbial group isolates over others was not observed. Overall results suggest that inherent properties of microbial residues may be more important to determining its stability in CA soils when it is 1) unassociated with the mineral matrix (LF); or 2) physically occluded within aggregates; compared with that intimately associated with mineral surfaces (MF). Compound-specific recovery of microbial isolates with Py-GC-MS-IRMS will be discussed.

  17. In Silico Docking and Electrophysiological Characterization of Lacosamide Binding Sites on Collapsin Response Mediator Protein-2 Identifies a Pocket Important in Modulating Sodium Channel Slow Inactivation*

    PubMed Central

    Wang, Yuying; Brittain, Joel M.; Jarecki, Brian W.; Park, Ki Duk; Wilson, Sarah M.; Wang, Bo; Hale, Rachel; Meroueh, Samy O.; Cummins, Theodore R.; Khanna, Rajesh

    2010-01-01

    The anti-epileptic drug (R)-lacosamide ((2R)-2-(acetylamino)-N-benzyl-3-methoxypropanamide (LCM)) modulates voltage-gated sodium channels (VGSCs) by preferentially interacting with slow inactivated sodium channels, but the observation that LCM binds to collapsin response mediator protein 2 (CRMP-2) suggests additional mechanisms of action for LCM. We postulated that CRMP-2 levels affects the actions of LCM on VGSCs. CRMP-2 labeling by LCM analogs was competitively displaced by excess LCM in rat brain lysates. Manipulation of CRMP-2 levels in the neuronal model system CAD cells affected slow inactivation of VGSCs without any effects on other voltage-dependent properties. In silico docking was performed to identify putative binding sites in CRMP-2 that may modulate the effects of LCM on VGSCs. These studies identified five cavities in CRMP-2 that can accommodate LCM. CRMP-2 alanine mutants of key residues within these cavities were functionally similar to wild-type CRMP-2 as assessed by similar levels of enhancement in dendritic complexity of cortical neurons. Next, we examined the effects of expression of wild-type and mutant CRMP-2 constructs on voltage-sensitive properties of VGSCs in CAD cells: 1) steady-state voltage-dependent activation and fast-inactivation properties were not affected by LCM, 2) CRMP-2 single alanine mutants reduced the LCM-mediated effects on the ability of endogenous Na+ channels to transition to a slow inactivated state, and 3) a quintuplicate CRMP-2 alanine mutant further decreased this slow inactivated fraction. Collectively, these results identify key CRMP-2 residues that can coordinate LCM binding thus making it more effective on its primary clinical target. PMID:20538611

  18. Influence of organic waste and residue mud additions on chemical, physical and microbial properties of bauxite residue sand.

    PubMed

    Jones, Benjamin E H; Haynes, Richard J; Phillips, Ian R

    2011-02-01

    In an alumina refinery, bauxite ore is treated with sodium hydroxide at high temperatures and pressures and for every tone of alumina produced, about 2 tones of alkaline, saline bauxite processing waste is also produced. At Alcoa, a dry stacking system of disposal is used, and it is the sand fraction of the processing waste that is rehabilitated. There is little information available regarding the most appropriate amendments to add to the processing sand to aid in revegetation. The purpose of this study was to investigate how the addition of organic wastes (biosolids and poultry manure), in the presence or absence of added residue mud, would affect the properties of the residue sand and its suitability for revegetation. Samples of freshly deposited residue sand were collected from Alcoa's Kwinana refinery. Samples were treated with phosphogypsum (2% v/v), incubated, and leached. A laboratory experiment was then set up in which the two organic wastes were applied at 0 or the equivalent to 60 tones ha(-1) in combination with residue mud added at rates of 0%, 10% and 20% v/v. Samples were incubated for 8 weeks, after which, key chemical, physical and microbial properties of the residue sand were measured along with seed germination. Additions of residue mud increased exchangeable Na(+), ESP and the pH, and HCO (3) (-) and Na(+) concentrations in saturation paste extracts. Additions of biosolids and poultry manure increased concentrations of extractable P, NH (4) (+) , K, Mg, Cu, Zn, Mn and Fe. Addition of residue mud, in combination with organic wastes, caused a marked decrease in macroporosity and a concomitant increase in mesoporosity, available water holding capacity and the quantity of water held at field capacity. With increasing residue mud additions, the percentage of sample present as sand particles (<1 mm diameter) decreased, and the percentage present in aggregated form (>2 mm diameter) increased; greatest aggregation occurred where a combination of residue

  19. The Comprehensive Microbial Resource.

    PubMed

    Peterson, J D; Umayam, L A; Dickinson, T; Hickey, E K; White, O

    2001-01-01

    One challenge presented by large-scale genome sequencing efforts is effective display of uniform information to the scientific community. The Comprehensive Microbial Resource (CMR) contains robust annotation of all complete microbial genomes and allows for a wide variety of data retrievals. The bacterial information has been placed on the Web at http://www.tigr.org/CMR for retrieval using standard web browsing technology. Retrievals can be based on protein properties such as molecular weight or hydrophobicity, GC-content, functional role assignments and taxonomy. The CMR also has special web-based tools to allow data mining using pre-run homology searches, whole genome dot-plots, batch downloading and traversal across genomes using a variety of datatypes.

  20. The Comprehensive Microbial Resource

    PubMed Central

    Peterson, Jeremy D.; Umayam, Lowell A.; Dickinson, Tanja; Hickey, Erin K.; White, Owen

    2001-01-01

    One challenge presented by large-scale genome sequencing efforts is effective display of uniform information to the scientific community. The Comprehensive Microbial Resource (CMR) contains robust annotation of all complete microbial genomes and allows for a wide variety of data retrievals. The bacterial information has been placed on the Web at http://www.tigr.org/CMR for retrieval using standard web browsing technology. Retrievals can be based on protein properties such as molecular weight or hydrophobicity, GC-content, functional role assignments and taxonomy. The CMR also has special web-based tools to allow data mining using pre-run homology searches, whole genome dot-plots, batch downloading and traversal across genomes using a variety of datatypes. PMID:11125067

  1. Nano-Mechanical Properties of Heat Inactivated Bacillus anthracis and Bacillus thuringiensis Spores

    DTIC Science & Technology

    2008-03-01

    Scanner Laser Mirror Cantilever Sample Probe Tip 16 cereus strain 569, and Bacillus globigii var. niger . Zolock determined that there wer...been used to measure the surface elasticities of a variety of microbial organisms including Pseudomonas putida, Bacillus subtilis, Aspergillus ...66:307-311 (2005). Zhao, Liming, David Schaefer, and Mark R. Marten. “Assessment of Elasticity and Topography of Aspergillus nidulans Spores via

  2. Development and Testing of a Method for Validating Chemical Inactivation of Ebola Virus.

    PubMed

    Alfson, Kendra J; Griffiths, Anthony

    2018-03-13

    Complete inactivation of infectious Ebola virus (EBOV) is required before a sample may be removed from a Biosafety Level 4 laboratory. The United States Federal Select Agent Program regulations require that procedures used to demonstrate chemical inactivation must be validated in-house to confirm complete inactivation. The objective of this study was to develop a method for validating chemical inactivation of EBOV and then demonstrate the effectiveness of several commonly-used inactivation methods. Samples containing infectious EBOV ( Zaire ebolavirus ) in different matrices were treated, and the sample was diluted to limit the cytopathic effect of the inactivant. The presence of infectious virus was determined by assessing the cytopathic effect in Vero E6 cells. Crucially, this method did not result in a loss of infectivity in control samples, and we were able to detect less than five infectious units of EBOV ( Zaire ebolavirus ). We found that TRIzol LS reagent and RNA-Bee inactivated EBOV in serum; TRIzol LS reagent inactivated EBOV in clarified cell culture media; TRIzol reagent inactivated EBOV in tissue and infected Vero E6 cells; 10% neutral buffered formalin inactivated EBOV in tissue; and osmium tetroxide vapors inactivated EBOV on transmission electron microscopy grids. The methods described herein are easily performed and can be adapted to validate inactivation of viruses in various matrices and by various chemical methods.

  3. Sunlight inactivation of somatic coliphage in the presence of natural organic matter.

    PubMed

    Sun, Chen-Xi; Kitajima, Masaaki; Gin, Karina Yew-Hoong

    2016-01-15

    Long wavelengths of sunlight spectrum (UVA and visible light), as well as natural organic matter (NOM) are important environmental factors affecting survival of viruses in aquatic environment through direct and indirect inactivation. In order to understand the virus inactivation kinetics under such conditions, this study investigated the effects of Suwannee River natural organic matter (NOM) on the inactivation of a somatic coliphage, phiX174, by UVA and visible light. Experiments were carried out to examine the virucidal effects of UVA/visible light, assess the influence of SRNOM at different concentrations, and identify the effective ROS in virus inactivation. The results from this study showed that the presence of NOM could either enhance virus inactivation or reduce virus inactivation depending on the concentration, where the inactivation rate followed a parabolic relationship against NOM concentration. The results indicated that moderate levels of NOM (11 ppm) had the strongest antiviral activity, while very low or very high NOM concentrations prolonged virus survival. The results also showed that OH▪ was the primary ROS in causing phiX174 (ssDNA virus) inactivation, unlike previous findings where (1)O2 was the primary ROS causing MS2 (ssRNA virus) inactivation. The phiX174 inactivation by OH∙ could be described as k=3.7 ✕ 10(13)[OH∙]+1.404 (R(2)=0.8527). Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Hydrazine vapor inactivates Bacillus spores

    NASA Astrophysics Data System (ADS)

    Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

    2016-05-01

    NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

  5. Spartina alterniflora invasion alters soil microbial community composition and microbial respiration following invasion chronosequence in a coastal wetland of China

    PubMed Central

    Yang, Wen; Jeelani, Nasreen; Leng, Xin; Cheng, Xiaoli; An, Shuqing

    2016-01-01

    The role of exotic plants in regulating soil microbial community structure and activity following invasion chronosequence remains unclear. We investigated soil microbial community structure and microbial respiration following Spartina alterniflora invasion in a chronosequence of 6-, 10-, 17-, and 20-year-old by comparing with bare flat in a coastal wetland of China. S. alterniflora invasion significantly increased soil moisture and salinity, the concentrations of soil water-soluble organic carbon and microbial biomass carbon (MBC), the quantities of total and various types of phospholipid fatty acids (PLFAs), the fungal:bacterial PLFAs ratio and cumulative microbial respiration compared with bare flat. The highest MBC, gram-negative bacterial and saturated straight-chain PLFAs were found in 10-year-old S. alterniflora soil, while the greatest total PLFAs, bacterial and gram-positive bacterial PLFAs were found in 10- and 17-year-old S. alterniflora soils. The monounsaturated:branched PLFAs ratio declined, and cumulative microbial respiration on a per-unit-PLFAs increased following S. alterniflora invasion in the chronosequence. Our results suggest that S. alterniflora invasion significantly increased the biomass of soil various microbial groups and microbial respiration compared to bare flat soil by increasing soil available substrate, and modifying soil physiochemical properties. Soil microbial community reached the most enriched condition in the 10-year-old S. alterniflora community. PMID:27241173

  6. Ecological effects of soil properties and metal concentrations on the composition and diversity of microbial communities associated with land use patterns in an electronic waste recycling region.

    PubMed

    Wu, Wencheng; Dong, Changxun; Wu, Jiahui; Liu, Xiaowen; Wu, Yingxin; Chen, Xianbin; Yu, Shixiao

    2017-12-01

    Soil microbes play vital roles in ecosystem functions, and soil microbial communities may be strongly structured by land use patterns associated with electronic waste (e-waste) recycling activities, which can increase the heavy metal concentration in soils. In this study, a suite of soils from five land use types (paddy field, vegetable field, dry field, forest field, and e-waste recycling site) were collected in Longtang Town, Guangdong Province, South China. Soil physicochemical properties and heavy metal concentrations were measured, and the indigenous microbial assemblages were profiled using 16S rRNA high-throughput sequencing and clone library analyses. The results showed that mercury concentration was positively correlated with both Faith's PD and Chao1 estimates, suggesting that the soil microbial alpha diversity was predominantly regulated by mercury. In addition, redundancy analysis indicated that available phosphorus, soil moisture, and mercury were the three major drivers affecting the microbial assemblages. Overall, the microbial composition was determined primarily by land use patterns, and this study provides a novel insight on the composition and diversity of microbial communities in soils associated with e-waste recycling activities. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Effects of gamma irradiation on physicochemical properties, antioxidant and microbial activities of sour cherry juice

    NASA Astrophysics Data System (ADS)

    Arjeh, Edris; Barzegar, Mohsen; Ali Sahari, Mohammad

    2015-09-01

    Recently, due to the beneficial effects of bioactive compounds, demand for minimally processed fruits and fruit juices has increased rapidly in the world. In this study, sour cherry juice (SCJ) was exposed to gamma irradiation at 0.0, 0.5, 1.5, 3.0, 4.5, and 6.0 kGy and then stored at 4 °C for 60 days. Total soluble solids (TSS), total acidity (TA), color, total phenolic content (TPC), total monomeric anthocyanin content (TMC), antioxidant activity, organic acid profile, and microbial analysis were evaluated at regular intervals during the storage. Results indicated that irradiation did not have any significant effect on TSS, while level of TA increased significantly at the dose of 6 kGy (p<0.05). Furthermore, irradiation treatment and storage time led to a significant increase in L* and b* values and a decrease in a* values. Total monomeric anthocyanin content of the irradiated SCJ was lower than that of the non-irradiated one (24% at 3.0 kGy) and also changed toward a more negative direction during the storage (63% at 3.0 kGy for 60 days). There was a significant decrease in the antioxidant activity (DPPH radical scavenging and FRAP assay) in both irradiated and stored SCJs. After irradiation (0-6 kGy), the results showed that the concentration of malic and oxalic acid significantly increased; but, the concentration of ascorbic, citric, fumaric, and succinic acids significantly decreased. Gamma irradiation with doses of ≥3 kGy resulted in overall reduction in microbial loads. Based on the results obtained from the changes of physicochemical properties, antioxidant activity, and microbial analysis, irradiation of SCJ at doses of higher than 3.0 kGy is not recommended.

  8. Thermal and high pressure inactivation kinetics of blueberry peroxidase.

    PubMed

    Terefe, Netsanet Shiferaw; Delon, Antoine; Versteeg, Cornelis

    2017-10-01

    This study for the first time investigated the stability and inactivation kinetics of blueberry peroxidase in model systems (McIlvaine buffer, pH=3.6, the typical pH of blueberry juice) during thermal (40-80°C) and combined high pressure-thermal processing (0.1-690MPa, 30-90°C). At 70-80°C, the thermal inactivation kinetics was best described by a biphasic model with ∼61% labile and ∼39% stable fractions at temperature between 70 and 75°C. High pressure inhibited the inactivation of the enzyme with no inactivation at pressures as high as 690MPa and temperatures less than 50°C. The inactivation kinetics of the enzyme at 60-70°C, and pressures higher than 500MPa was best described by a first order biphasic model with ∼25% labile fraction and 75% stable fraction. The activation energy values at atmospheric pressure were 548.6kJ/mol and 324.5kJ/mol respectively for the stable and the labile fractions. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  9. Ribosome-inactivating proteins: potent poisons and molecular tools.

    PubMed

    Walsh, Matthew J; Dodd, Jennifer E; Hautbergue, Guillaume M

    2013-11-15

    Ribosome-inactivating proteins (RIPs) were first isolated over a century ago and have been shown to be catalytic toxins that irreversibly inactivate protein synthesis. Elucidation of atomic structures and molecular mechanism has revealed these proteins to be a diverse group subdivided into two classes. RIPs have been shown to exhibit RNA N-glycosidase activity and depurinate the 28S rRNA of the eukaryotic 60S ribosomal subunit. In this review, we compare archetypal RIP family members with other potent toxins that abolish protein synthesis: the fungal ribotoxins which directly cleave the 28S rRNA and the newly discovered Burkholderia lethal factor 1 (BLF1). BLF1 presents additional challenges to the current classification system since, like the ribotoxins, it does not possess RNA N-glycosidase activity but does irreversibly inactivate ribosomes. We further discuss whether the RIP classification should be broadened to include toxins achieving irreversible ribosome inactivation with similar turnovers to RIPs, but through different enzymatic mechanisms.

  10. Thermal inactivation kinetics of β-galactosidase during bread baking.

    PubMed

    Zhang, Lu; Chen, Xiao Dong; Boom, Remko M; Schutyser, Maarten A I

    2017-06-15

    In this study, β-galactosidase was utilized as a model enzyme to investigate the mechanism of enzyme inactivation during bread baking. Thermal inactivation of β-galactosidase was investigated in a wheat flour/water system at varying temperature-moisture content combinations, and in bread during baking at 175 or 205°C. In the wheat flour/water system, the thermostability of β-galactosidase increased with decreased moisture content, and a kinetic model was accurately fitted to the corresponding inactivation data (R 2 =0.99). Interestingly, the residual enzyme activity in the bread crust (about 30%) was hundredfold higher than that in the crumb (about 0.3%) after baking, despite the higher temperature in the crust throughout baking. This result suggested that the reduced moisture content in the crust increased the thermostability of the enzyme. Subsequently, the kinetic model reasonably predicted the enzyme inactivation in the crumb using the same parameters derived from the wheat flour/water system. However, the model predicted a lower residual enzyme activity in the crust compared with the experimental result, which indicated that the structure of the crust may influence the enzyme inactivation mechanism during baking. The results reported can provide a quantitative understanding of the thermal inactivation kinetics of enzyme during baking, which is essential to better retain enzymatic activity in bakery products supplemented with heat-sensitive enzymes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Selective inactivation of glutaredoxin by sporidesmin and other epidithiopiperazinediones.

    PubMed

    Srinivasan, Usha; Bala, Aveenash; Jao, Shu-chuan; Starke, David W; Jordan, T William; Mieyal, John J

    2006-07-25

    Glutaredoxin (thioltransferase) is a thiol-disulfide oxidoreductase that displays efficient and specific catalysis of protein-SSG deglutathionylation and is thereby implicated in homeostatic regulation of the thiol-disulfide status of cellular proteins. Sporidesmin is an epidithiopiperazine-2,5-dione (ETP) fungal toxin that disrupts cellular functions likely via oxidative alteration of cysteine residues on key proteins. In the current study sporidesmin inactivated human glutaredoxin in a time- and concentration-dependent manner. Under comparable conditions other thiol-disulfide oxidoreductase enzymes, glutathione reductase, thioredoxin, and thioredoxin reductase, were unaffected by sporidesmin. Inactivation of glutaredoxin required the reduced (dithiol) form of the enzyme, the oxidized (intramolecular disulfide) form of sporidesmin, and molecular oxygen. The inactivated glutaredoxin could be reactivated by dithiothreitol only in the presence of urea, followed by removal of the denaturant, indicating that inactivation of the enzyme involves a conformationally inaccessible disulfide bond(s). Various cysteine-to-serine mutants of glutaredoxin were resistant to inactivation by sporidesmin, suggesting that the inactivation reaction specifically involves at least two of the five cysteine residues in human glutaredoxin. The relative ability of various epidithiopiperazine-2,5-diones to inactivate glutaredoxin indicated that at least one phenyl substituent was required in addition to the epidithiodioxopiperazine moiety for inhibitory activity. Mass spectrometry of the modified protein is consistent with formation of intermolecular disulfides, containing one adducted toxin per glutaredoxin but with elimination of two sulfur atoms from the detected product. We suggest that the initial reaction is between the toxin sulfurs and cysteine 22 in the glutaredoxin active site. This study implicates selective modification of sulfhydryls of target proteins in some of the cytotoxic

  12. Inactivation of particle-associated coliforms by chlorine and monochloramine.

    PubMed Central

    Berman, D; Rice, E W; Hoff, J C

    1988-01-01

    Sieves and nylon screens were used to separate primary sewage effluent solids into particle fractions of less than 7- or greater than 7-micron size. The efficiency of separation was determined by using a particle counter. Indigenous coliforms associated with the particle fractions were tested for their resistance to chlorine and monochloramine. Coliforms associated with the less than 7-microns fraction were inactivated more rapidly by 0.5 mg of chlorine per liter at 5 degrees C and pH 7 than coliforms associated with the greater than 7-microns fraction. Homogenization of the greater than 7-microns fraction not only resulted in an increase in the number of less than 7-microns particles, but also increased the rate of inactivation to a rate similar to that of the less than 7-microns fraction. With 1 mg of monochloramine per liter at 5 degrees C and pH 7, particle size had no appreciable effect on the rate of inactivation. At pH 8, however, the less than 7-micron fraction was inactivated more rapidly than the greater than 7-micron fraction. The time required for 99% inactivation of the particle fractions with monochloramine at pH 7 or 8 was 20- to 50-fold greater than the time required for the same amount of inactivation with chlorine at pH 7. The results indicate that coliforms associated with sewage effluent particles are inactivated more rapidly with 0.5 mg of chlorine per liter than with 1.0 mg of monochloramine per liter. However, greater than 7-micron particles can have a protective effect against the disinfecting action of chlorine. PMID:3355136

  13. The Risk of Microbial Contamination in Multiple-Dose Preservative-Free Ophthalmic Preparations.

    PubMed

    Saisyo, Atsuyuki; Shimono, Rima; Oie, Shigeharu; Kimura, Kazuhiro; Furukawa, Hiroyuki

    2017-01-01

    Multiple-dose ophthalmic preparations that do not contain preservatives carry high risks of microbial contamination. However, there are various types of hospital preparations, with different physicochemical properties. In the present study, we evaluated the association between physicochemical properties and microbial contamination in ophthalmic preparations. The investigated hospital preparations included ophthalmic preparations of physiological saline, 0.2% fluconazole, 0.5% vancomycin hydrochloride, and 2% cyclosporine. We investigated the microbial dynamics of each ophthalmic preparation and microbial contamination in ophthalmic preparations used by patients. Remarkable growth of Pseudomonas aeruginosa, Burkholderia cepacia, and Serratia marcescens was observed in ophthalmic preparations of physiological saline and 0.2% fluconazole. All tested microorganisms displayed decreased counts after inoculation in 0.5% vancomycin hydrochloride. In 2% cyclosporine, all investigated microorganisms were below the limit of detection after inoculation for 6 h. The microbial contamination rates of ophthalmic preparations used by patients were 16.7% (3/18 samples) for 0.5% vancomycin hydrochloride and 0% (0/30 samples) for 2% cyclosporine. All detected contaminants in 0.5% vancomycin hydrochloride were Candida spp., one of which was present at a level of 1×10 4 colony-forming units/mL. The storage method for in-use ophthalmic preparations should be considered on the basis of their physicochemical properties.

  14. Physiology of the Inactivation of Vegetative Bacteria by Thermal Treatments: Mode of Action, Influence of Environmental Factors and Inactivation Kinetics

    PubMed Central

    Condón, Santiago; Mañas, Pilar

    2017-01-01

    Heat has been used extensively in the food industry as a preservation method, especially due to its ability to inactivate microorganisms present in foods. However, many aspects regarding the mechanisms of bacterial inactivation by heat and the factors affecting this process are still not fully understood. The purpose of this review is to offer a general overview of the most important aspects of the physiology of the inactivation or survival of microorganisms, particularly vegetative bacteria, submitted to heat treatments. This could help improve the design of current heat processes methods in order to apply milder and/or more effective treatments that could fulfill consumer requirements for fresh-like foods while maintaining the advantages of traditional heat treatments. PMID:29189748

  15. Thermoradiation inactivation of naturally occurring organisms in soil

    NASA Technical Reports Server (NTRS)

    Reynolds, M. C.; Lindell, K. F.; David, T. J.

    1973-01-01

    Samples of soil collected from Kennedy Space Center near spacecraft assembly facilities were found to contain microorganisms very resistant to conventional sterilization techniques. The inactivation behavior of the naturally occurring spores in soil was investigated using dry heat and ionizing radiation, first separately, then in combination. Dry heat inactivation rates of spores were determined for 105 and 125 C. Radiation inactivation rates were determined for dose rates of 660 and 76 krad/hr at 25 C. Simultaneous combinations of heat and radiation were then investigated at 105, 110, 115, 120, and 125 C. Combined treatment was found to be highly synergistic requiring greatly reduced radiation doses to accomplish sterilization.

  16. Kinetics of inactivation of indicator pathogens during thermophilic anaerobic digestion.

    PubMed

    Popat, Sudeep C; Yates, Marylynn V; Deshusses, Marc A

    2010-12-01

    Thermophilic anaerobic sludge digestion is a promising process to divert waste to beneficial use, but an important question is the required temperature and holding time to achieve a given degree of pathogen inactivation. In this study, the kinetics of inactivation of Ascaris suum and vaccine strain poliovirus type 1 (PVS-1), selected as indicators for helminth ova and enteric viruses respectively, were determined during anaerobic digestion at temperatures ranging from 51 to 56 °C. Inactivation of both indicator organisms was fast with greater than two log reductions achieved within 2 h for A. suum and three log reductions for PVS-1, suggesting that the current U.S. regulations are largely conservative. The first-order inactivation rate constants k followed Arrhenius relationship with activation energies of 105 and 39 KJ mol(-1) for A. suum and PVS-1, respectively indicating that A. suum was more sensitive to temperature. Although inactivation was fast, the presence of compounds in the sludge that are known to be protective of pathogen inactivation was observed, suggesting that composition-dependent time-temperature relationships are necessary. Copyright © 2010 Elsevier Ltd. All rights reserved.

  17. The microbial community structure in petroleum-contaminated sediments corresponds to geophysical signatures

    USGS Publications Warehouse

    Allen, J.P.; Atekwana, E.A.; Duris, J.W.; Werkema, D.D.; Rossbach, S.

    2007-01-01

    The interdependence between geoelectrical signatures at underground petroleum plumes and the structures of subsurface microbial communities was investigated. For sediments contaminated with light non-aqueousphase liquids, anomalous high conductivity values have been observed. Vertical changes in the geoelectrical properties of the sediments were concomitant with significant changes in the microbial community structures as determined by the construction and evaluation of 16S rRNA gene libraries. DNA sequencing of clones from four 16S rRNA gene libraries from different depths of a contaminated field site and two libraries from an uncontaminated background site revealed spatial heterogeneity in the microbial community structures. Correspondence analysis showed that the presence of distinct microbial populations, including the various hydrocarbon-degrading, syntrophic, sulfate-reducing, and dissimilatory-iron-reducing populations, was a contributing factor to the elevated geoelectrical measurements. Thus, through their growth and metabolic activities, microbial populations that have adapted to the use of petroleum as a carbon source can strongly influence their geophysical surroundings. Since changes in the geophysical properties of contaminated sediments parallel changes in the microbial community compositions, it is suggested that geoelectrical measurements can be a cost-efficient tool to guide microbiological sampling for microbial ecology studies during the monitoring of natural or engineered bioremediation processes. Copyright ?? 2007, American Society for Microbiology. All Rights Reserved.

  18. Growth and inactivation of Salmonella at low refrigerated storage temperatures and thermal inactivation on raw chicken meat and laboratory media: mixed effect meta-analysis.

    PubMed

    Smadi, Hanan; Sargeant, Jan M; Shannon, Harry S; Raina, Parminder

    2012-12-01

    Growth and inactivation regression equations were developed to describe the effects of temperature on Salmonella concentration on chicken meat for refrigerated temperatures (⩽10°C) and for thermal treatment temperatures (55-70°C). The main objectives were: (i) to compare Salmonella growth/inactivation in chicken meat versus laboratory media; (ii) to create regression equations to estimate Salmonella growth in chicken meat that can be used in quantitative risk assessment (QRA) modeling; and (iii) to create regression equations to estimate D-values needed to inactivate Salmonella in chicken meat. A systematic approach was used to identify the articles, critically appraise them, and pool outcomes across studies. Growth represented in density (Log10CFU/g) and D-values (min) as a function of temperature were modeled using hierarchical mixed effects regression models. The current meta-analysis analysis found a significant difference (P⩽0.05) between the two matrices - chicken meat and laboratory media - for both growth at refrigerated temperatures and inactivation by thermal treatment. Growth and inactivation were significantly influenced by temperature after controlling for other variables; however, no consistent pattern in growth was found. Validation of growth and inactivation equations against data not used in their development is needed. Copyright © 2012 Ministry of Health, Saudi Arabia. Published by Elsevier Ltd. All rights reserved.

  19. Biogeochemical Processes in Microbial Ecosystems

    NASA Technical Reports Server (NTRS)

    DesMarais, David J.; DeVincenzi, Donald L. (Technical Monitor)

    2001-01-01

    The hierarchical organization of microbial ecosystems determines process rates that shape Earth's environment, create the biomarker sedimentary and atmospheric signatures of life and define the stage upon which major evolutionary events occurred. In order to understand how microorganisms have shaped the global environment of Earth and potentially, other worlds, we must develop an experimental paradigm that links biogeochemical processes with ever-changing temporal and spatial distributions of microbial population, and their metabolic properties. Photosynthetic microbial mats offer an opportunity to define holistic functionality at the millimeter scale. At the same time, their Biogeochemistry contributes to environmental processes on a planetary scale. These mats are possibly direct descendents of the most ancient biological communities; communities in which oxygenic photosynthesis might have been invented. Mats provide one of the best natural systems to study how microbial populations associate to control dynamic biogeochemical gradients. These are self-sustaining, complete ecosystems in which light energy absorbed over a diel (24 hour) cycle drives the synthesis of spatially-organized, diverse biomass. Tightly-coupled microorganisms in the mat have specialized metabolisms that catalyze transformations of carbon, nitrogen. sulfur, and a host of other elements.

  20. Thermal inactivation of enzymes and pathogens in biosamples for MS analysis.

    PubMed

    Ahnoff, Martin; Cazares, Lisa H; Sköld, Karl

    2015-01-01

    Protein denaturation is the common basis for enzyme inactivation and inactivation of pathogens, necessary for preservation and safe handling of biosamples for downstream analysis. While heat-stabilization technology has been used in proteomic and peptidomic research since its introduction in 2009, the advantages of using the technique for simultaneous pathogen inactivation have only recently been addressed. The time required for enzyme inactivation by heat (≈1 min) is short compared with chemical treatments, and inactivation is irreversible in contrast to freezing. Heat stabilization thus facilitates mass spectrometric studies of biomolecules with a fast conversion rate, and expands the chemical space of potential biomarkers to include more short-lived entities, such as phosphorylated proteins, in tissue samples as well as whole-blood (dried blood sample) samples.

  1. Kinetic analysis of Legionella inactivation using ozone in wastewater.

    PubMed

    Li, Jun; Li, Kunquan; Zhou, Yan; Li, Xuebin; Tao, Tao

    2017-02-01

    Legionella inactivation using ozone was studied in wastewater using kinetic analysis and modeling. The experimental results indicate that the relationship between the ozone concentration, germ concentration, and chemical oxygen demand (COD) can be used to predict variations in germ and COD concentrations. The ozone reaction with COD and inactivation of Legionella occurred simultaneously, but the reaction with COD likely occurred at a higher rate than the inactivation, as COD is more easily oxidized by ozone than Legionella. Higher initial COD concentrations resulted in a lower inactivation rate and higher lnN/N 0 . Higher temperature led to a higher inactivation efficiency. The relationship of the initial O 3 concentration and Legionella inactivation rate was not linear, and thus, the Ct value required for a 99.99% reduction was not constant. The initial O 3 concentration was more important than the contact time, and a reduction of the initial O 3 concentration could not be compensated by increasing the contact time. The Ct values were compared over a narrow range of initial concentrations; the Ct values could only be contrasted when the initial O 3 concentrations were very similar. A higher initial O 3 concentration led to a higher inflection point value for the lnN/N 0 vs C 0 t curve. Energy consumption using a plasma corona was lower than when using boron-doped diamond electrodes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Antimicrobial property and microstructure of micro-emulsion edible composite films against Listeria.

    PubMed

    Guo, Mingming; Jin, Tony Z; Yadav, Madhav P; Yang, Ruijin

    2015-09-02

    Edible antimicrobial composite films from micro-emulsions containing all natural compounds were developed and their antimicrobial properties and microstructures were investigated. Chitosan, allyl isothiocyanate (AIT), barley straw arabinoxylan (BSAX), and organic acids (acetic, lactic and levulinic acids) were used as film-forming agent, antimicrobial agent, emulsifier, and solvent, respectively. Micro-emulsions were obtained using high pressure homogenization (HPH) processing at 138MPa for 3cycles. The composite films made from the micro-emulsions significantly (p<0.05) inactivated Listeria innocua in tryptic soy broth (TSB) and on the surface of ready-to-eat (RTE) meat samples, achieving microbial reductions of over 4logCFU/ml in TSB after 2days at 22°C and on meat samples after 35days at 10°C. AIT was a major contributor to the antimicrobial property of the films and HPH processing further enhanced its antimicrobial efficacy, while the increase of chitosan from 1.5% to 3%, or addition of acetic acid to the formulations didn't result in additional antimicrobial effects. This study demonstrated an effective approach to developing new edible antimicrobial films and coatings used for food applications. Published by Elsevier B.V.

  3. Efficacy of ultraviolet radiation as an alternative technology to inactivate microorganisms in grape juices and wines.

    PubMed

    Fredericks, Ilse N; du Toit, Maret; Krügel, Maricel

    2011-05-01

    Since sulphur dioxide (SO(2)) is associated with health risks, the wine industry endeavours to reduce SO(2) levels in wines with new innovative techniques. The aim of this study was, therefore, to investigate the efficacy of ultraviolet radiation (UV)-C (254 nm) as an alternative technology to inactivate microorganisms in grape juices and wines. A pilot-scale UV-C technology (SurePure, South Africa) consisting of an UV-C germicidal lamp (100 W output; 30 W UV-C output) was used to apply UV-C dosages ranging from 0 to 3672 J l(-1), at a constant flow rate of 4000 l h(-1) (Re > 7500). Yeasts, lactic and acetic acid bacteria were singly and co-inoculated into 20 l batches of Chenin blanc juice, Shiraz juice, Chardonnay wine and Pinotage wine, respectively. A dosage of 3672 J l(-1), resulted in an average log(10) microbial reduction of 4.97 and 4.89 in Chardonnay and Pinotage, respectively. In Chenin blanc and Shiraz juice, an average log(10) reduction of 4.48 and 4.25 was obtained, respectively. UV-C efficacy may be influenced by liquid properties such as colour and turbidity. These results had clearly indicated significant (p < 0.05) germicidal effect against wine-specific microorganisms; hence, UV-C radiation may stabilize grape juice and wine microbiologically in conjunction with reduced SO(2) levels. Copyright © 2010 Elsevier Ltd. All rights reserved.

  4. Inactivation of a subpopulation of human neutrophils by exposure to ultrahigh-molecular-weight polyethylene wear debris.

    PubMed

    Bernard, Louis; Vaudaux, Pierre; Huggler, Elzbieta; Stern, Richard; Fréhel, Claude; Francois, Patrice; Lew, Daniel; Hoffmeyer, Pierre

    2007-04-01

    Polymorphonuclear neutrophils, a first line of defence against invading microbial pathogens, may be attracted by inflammatory mediators triggered by ultrahigh-molecular-weight polyethylene (UHMWPE) wear particles released from orthopaedic prostheses. Phagocytosis of UHMWPE particles by neutrophils may indirectly compromise their phagocytic-bactericidal mechanisms, thus enhancing host susceptibility to microbial infections. In an in vitro assay, pre-exposure of purified human neutrophils to UHMWPE micrometre- and submicrometre-sized wear particles interfered with subsequent Staphylococcos aureus uptake in a heterogeneous way, as assessed by a dual label fluorescence microscopic assay that discriminated intracellular rhodamine-labelled UHMWPE particles from fluorescein isothiocyanate-labelled S. aureus. Indeed, a higher percentage (44%) of neutrophils having engulfed UHMWPE particles lost the ability to phagocytize S. aureus, compared with UHMWPE-free neutrophils (<3%). Pre-exposure of neutrophils to UHMWPE wear particles did not impair but rather stimulated their oxidative burst response in a chemoluminescence assay. The presence of UHMWPE wear particles did not lead to significant overall consumption of complement-mediated opsonic factors nor decreased surface membrane display of neutrophil complement receptors. In conclusion, engulfment of UHMWPE wear particles led to inactivation of S. aureus uptake in nearly half of the neutrophil population, which may potentially impair host clearance mechanisms against pyogenic infections.

  5. Use of PRD1 bacteriophage in groundwater viral transport, inactivation, and attachment studies

    USGS Publications Warehouse

    Harvey, R.W.; Ryan, J.N.

    2004-01-01

    PRD1, an icosahedra-shaped, 62 nm (diameter), double-stranded DNA bacteriophage with an internal membrane, has emerged as an important model virus for studying the manner in which microorganisms are transported through a variety of groundwater environments. The popularity of this phage for use in transport studies involving geologic media is due, in part, to its relative stability over a range of temperatures and low degree of attachment in aquifer sediments. Laboratory and field investigations employing PRD1 are leading to a better understanding of viral attachment and transport behaviors in saturated geologic media and to improved methods for describing mathematically subsurface microbial transport at environmentally significant field scales. Radioisotopic labeling of PRD1 is facilitating additional information about the nature of viral interactions with solid surfaces in geologic media, the importance of iron oxide surfaces, and allowing differentiation between inactivation and attachment in field-scale tracer tests. ?? 2004 Published by Elsevier B.V. on behalf of the Federation of European Microbiological Societies.

  6. Impact of Substratum Surface on Microbial Community Structure and Treatment Performance in Biological Aerated Filters

    PubMed Central

    Kim, Lavane; Pagaling, Eulyn; Zuo, Yi Y.

    2014-01-01

    The impact of substratum surface property change on biofilm community structure was investigated using laboratory biological aerated filter (BAF) reactors and molecular microbial community analysis. Two substratum surfaces that differed in surface properties were created via surface coating and used to develop biofilms in test (modified surface) and control (original surface) BAF reactors. Microbial community analysis by 16S rRNA gene-based PCR-denaturing gradient gel electrophoresis (DGGE) showed that the surface property change consistently resulted in distinct profiles of microbial populations during replicate reactor start-ups. Pyrosequencing of the bar-coded 16S rRNA gene amplicons surveyed more than 90% of the microbial diversity in the microbial communities and identified 72 unique bacterial species within 19 bacterial orders. Among the 19 orders of bacteria detected, Burkholderiales and Rhodocyclales of the Betaproteobacteria class were numerically dominant and accounted for 90.5 to 97.4% of the sequence reads, and their relative abundances in the test and control BAF reactors were different in consistent patterns during the two reactor start-ups. Three of the five dominant bacterial species also showed consistent relative abundance changes between the test and control BAF reactors. The different biofilm microbial communities led to different treatment efficiencies, with consistently higher total organic carbon (TOC) removal in the test reactor than in the control reactor. Further understanding of how surface properties affect biofilm microbial communities and functional performance would enable the rational design of new generations of substrata for the improvement of biofilm-based biological treatment processes. PMID:24141134

  7. Microbial biofilms in intertidal systems: an overview

    NASA Astrophysics Data System (ADS)

    Decho, Alan W.

    2000-07-01

    Intertidal marine systems are highly dynamic systems which are characterized by periodic fluctuations in environmental parameters. Microbial processes play critical roles in the remineralization of nutrients and primary production in intertidal systems. Many of the geochemical and biological processes which are mediated by microorganisms occur within microenvironments which can be measured over micrometer spatial scales. These processes are localized by cells within a matrix of extracellular polymeric secretions (EPS), collectively called a "microbial biofilm". Recent examinations of intertidal systems by a range of investigators using new approaches show an abundance of biofilm communities. The purpose of this overview is to examine recent information concerning the roles of microbial biofilms in intertidal systems. The microbial biofilm is a common adaptation of natural bacteria and other microorganisms. In the fluctuating environments of intertidal systems, biofilms form protective microenvironments and may structure a range of microbial processes. The EPS matrix of biofilm forms sticky coatings on individual sediment particles and detrital surfaces, which act as a stabilizing anchor to buffer cells and their extracellular processes during the frequent physical stresses (e.g., changes in salinity and temperature, UV irradiation, dessication). EPS is an operational definition designed to encompass a range of large microbially-secreted molecules having widely varying physical and chemical properties, and a range of biological roles. Examinations of EPS using Raman and Fourier-transform infared spectroscopy, and atomic-force microscopy suggest that some EPS gels possess physical and chemical properties which may hasten the development of sharp geochemical gradients, and contribute a protective effect to cells. Biofilm polymers act as a sorptive sponge which binds and concentrates organic molecules and ions close to cells. Concurrently, the EPS appear to localize

  8. Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests.

    PubMed

    Rosenstierne, Maiken Worsøe; Karlberg, Helen; Bragstad, Karoline; Lindegren, Gunnel; Stoltz, Malin Lundahl; Salata, Cristiano; Kran, Anne-Marte Bakken; Dudman, Susanne Gjeruldsen; Mirazimi, Ali; Fomsgaard, Anders

    2016-10-01

    Rapid bedside inactivation of Ebola virus would be a solution for the safety of medical and technical staff, risk containment, sample transport, and high-throughput or rapid diagnostic testing during an outbreak. We show that the commercially available Magna Pure lysis/binding buffer used for nucleic acid extraction inactivates Ebola virus. A rapid bedside inactivation method for nucleic acid tests is obtained by simply adding Magna Pure lysis/binding buffer directly into vacuum blood collection EDTA tubes using a thin needle and syringe prior to sampling. The ready-to-use inactivation vacuum tubes are stable for more than 4 months, and Ebola virus RNA is preserved in the Magna Pure lysis/binding buffer for at least 5 weeks independent of the storage temperature. We also show that Ebola virus RNA can be manually extracted from Magna Pure lysis/binding buffer-inactivated samples using the QIAamp viral RNA minikit. We present an easy and convenient method for bedside inactivation using available blood collection vacuum tubes and reagents. We propose to use this simple method for fast, safe, and easy bedside inactivation of Ebola virus for safe transport and routine nucleic acid detection. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  9. Stratosphere Conditions Inactivate Bacterial Endospores from a Mars Spacecraft Assembly Facility

    NASA Astrophysics Data System (ADS)

    Khodadad, Christina L.; Wong, Gregory M.; James, Leandro M.; Thakrar, Prital J.; Lane, Michael A.; Catechis, John A.; Smith, David J.

    2017-04-01

    Every spacecraft sent to Mars is allowed to land viable microbial bioburden, including hardy endospore-forming bacteria resistant to environmental extremes. Earth's stratosphere is severely cold, dry, irradiated, and oligotrophic; it can be used as a stand-in location for predicting how stowaway microbes might respond to the martian surface. We launched E-MIST, a high-altitude NASA balloon payload on 10 October 2015 carrying known quantities of viable Bacillus pumilus SAFR-032 (4.07 × 107 spores per sample), a radiation-tolerant strain collected from a spacecraft assembly facility. The payload spent 8 h at ˜31 km above sea level, exposing bacterial spores to the stratosphere. We found that within 120 and 240 min, spore viability was significantly reduced by 2 and 4 orders of magnitude, respectively. By 480 min, <0.001% of spores carried to the stratosphere remained viable. Our balloon flight results predict that most terrestrial bacteria would be inactivated within the first sol on Mars if contaminated spacecraft surfaces receive direct sunlight. Unfortunately, an instrument malfunction prevented the acquisition of UV light measurements during our balloon mission. To make up for the absence of radiometer data, we calculated a stratosphere UV model and conducted ground tests with a 271.1 nm UVC light source (0.5 W/m2), observing a similarly rapid inactivation rate when using a lower number of contaminants (640 spores per sample). The starting concentration of spores and microconfiguration on hardware surfaces appeared to influence survivability outcomes in both experiments. With the relatively few spores that survived the stratosphere, we performed a resequencing analysis and identified three single nucleotide polymorphisms compared to unexposed controls. It is therefore plausible that bacteria enduring radiation-rich environments (e.g., Earth's upper atmosphere, interplanetary space, or the surface of Mars) may be pushed in evolutionarily consequential directions.

  10. Phthalocyanine-assisted photodynamic inactivation of pathogenic microorganisms

    NASA Astrophysics Data System (ADS)

    Mantareva, Vanya; Angelov, Ivan; Borissova, Ekaterina; Avramov, Latchezar; Kussovski, Vesselin

    2007-03-01

    The phthalocyanine zinc(II) and aluminum (III) complexes were studied to photoinactivate the bacterial strains, Staphylococcus aureus, methacillin-sensitive and methacillin-resistant, Pseudomonas aeruginosa and one yeast Candida albicans. The binding of phthalocyanines to bacteria and fungi cells was evaluated by the means of laserinduced fluorescence technique. The fluorescent spectra of dyes (650 - 800 nm) after direct excitation (635 nm) were measured as follows: 1. for the aqua supernatants obtained after 10 min cell incubation with the respected phthalocyanines (1.6 μmol.l -1), 2. for the washed from the unbound dye cells, and 3. for the organic extracts from the three times washed cells. Fluorescent intensities at the emission maximum (~690 nm) were compared to the spectra of the phthalocyanines in organic solutions. The phthalocyanines uptake data for bacteria and fungi were determined at different cell densities. Nevertheless the better fluorescence properties of AlPc (fluorescent quantum yield of 0.4 towards 0.3 for ZnPcs) the lower drug accumulation in microorganisms was obtained. PDI results indicated an intensive lowering of the bacterial survival of both strains of S. aureus treated with cationic ZnPcMe followed by the anionic ZnPcS, at irradiance of 100 mW cm -2 and fluence rate of 60 J cm -2. More resistant to phototreatment P. aeruginosa and morphologically complicated yeast C. albicans were successfully inactivated only with cationic ZnPcMe. These data indicate the promising future application of cationic phthalocyanine in photodynamic inactivation of pathogenic microorganisms.

  11. DNA Polymerase λ Inactivation by Oxidized Abasic Sites&

    PubMed Central

    Stevens, Adam J.; Guan, Lirui; Bebenek, Katarzyna; Kunkel, Thomas A.; Greenberg, Marc M.

    2013-01-01

    Base excision repair plays a vital role in maintaining genomic integrity in mammalian cells. DNA polymerase λ is believed to play a backup role to DNA polymerase β in base excision repair. Two oxidized abasic lesions that are produced by a variety of DNA damaging agents, including several antitumor antibiotics, the C4′-oxidized abasic site following Ape1 incision (pC4-AP) and 5′-(2-phosphoryl-1,4-dioxobutane) (DOB), irreversibly inactivate Pol β and Pol λ. The interactions of DOB and pC4-AP with Pol λ are examined in detail using DNA substrates containing these lesions at defined sites. Single turnover kinetic experiments show that Pol λ excises DOB almost 13-times more slowly than a 5′-phosphorylated 2-deoxyribose (dRP). pC4-AP is excised approximately twice as fast as DOB. The absolute rate constants are considerably slower than those reported for Pol β at the respective reactions, suggesting that Pol λ may be an inefficient backup in BER. DOB inactivates Pol λ approximately 3-fold less efficiently than it does Pol β and the difference is attributable to a higher KI (33 ± 7 nM). Inactivation of Pol λ’s lyase activity by DOB also prevents the enzyme from carrying out polymerization following preincubation of the protein and DNA. Mass spectral analysis of GluC digested Pol λ inactivated by DOB shows that Lys324 is modified. There is inferential support that Lys312 may also be modified. Both residues are within the Pol λ lyase active site. Protein modification involves reaction with released but-2-ene-1,4-dial. When acting on pC4-AP, Pol λ achieves approximately 4 turnovers on average before being inactivated. Lyase inactivation by pC4-AP is also accompanied by loss of polymerase activity and mass spectrometry indicates that Lys312 and Lys324 are modified by the lesion. The ability of DOB and pC4-AP to inactivate Pol λ provides additional evidence that these lesions are significant sources of the cytotoxicity of DNA damaging agents that

  12. Inactivation of Gating Currents of L-Type Calcium Channels

    PubMed Central

    Shirokov, Roman; Ferreira, Gonzalo; Yi, Jianxun; Ríos, Eduardo

    1998-01-01

    In studies of gating currents of rabbit cardiac Ca channels expressed as α1C/β2a or α1C/β2a/α2δ subunit combinations in tsA201 cells, we found that long-lasting depolarization shifted the distribution of mobile charge to very negative potentials. The phenomenon has been termed charge interconversion in native skeletal muscle (Brum, G., and E. Ríos. 1987. J. Physiol. (Camb.). 387:489–517) and cardiac Ca channels (Shirokov, R., R. Levis, N. Shirokova, and E. Ríos. 1992. J. Gen. Physiol. 99:863–895). Charge 1 (voltage of half-maximal transfer, V1/2 ≃ 0 mV) gates noninactivated channels, while charge 2 (V1/2 ≃ −90 mV) is generated in inactivated channels. In α1C/β2a cells, the available charge 1 decreased upon inactivating depolarization with a time constant τ ≃ 8, while the available charge 2 decreased upon recovery from inactivation (at −200 mV) with τ ≃ 0.3 s. These processes therefore are much slower than charge movement, which takes <50 ms. This separation between the time scale of measurable charge movement and that of changes in their availability, which was even wider in the presence of α2δ, implies that charges 1 and 2 originate from separate channel modes. Because clear modal separation characterizes slow (C-type) inactivation of Na and K channels, this observation establishes the nature of voltage-dependent inactivation of L-type Ca channels as slow or C-type. The presence of the α2δ subunit did not change the V1/2 of charge 2, but sped up the reduction of charge 1 upon inactivation at 40 mV (to τ ≃ 2 s), while slowing the reduction of charge 2 upon recovery (τ ≃ 2 s). The observations were well simulated with a model that describes activation as continuous electrodiffusion (Levitt, D. 1989. Biophys. J. 55:489–498) and inactivation as discrete modal change. The effects of α2δ are reproduced assuming that the subunit lowers the free energy of the inactivated mode. PMID:9607938

  13. Thermal Inactivation of Bacillus anthracis Spores Using Rapid Resistive Heating

    DTIC Science & Technology

    2016-03-24

    thermal inactivation research. However, the research conducted to support this thesis utilizes the B.a. Sterne strain which is used in livestock vaccines...methodology conducted for this research including hard surface recovery, thermal inactivation of Bacillus anthracis spores, and the rapid resistive heating...to 500°C range but again, many of the thermal inactivation studies were conducted in the 350 to 2000°C range. Sample plots will be discussed in

  14. Mechanism of Cd2+-coordination during Slow Inactivation in Potassium Channels

    PubMed Central

    Raghuraman, H.; Cordero-Morales, Julio F.; Jogini, Vishwanath; Pan, Albert C.; Kollewe, Astrid; Roux, Benoît; Perozo, Eduardo

    2013-01-01

    Summary In K+ channels, rearrangements of the pore outer-vestibule have been associated with C-type inactivation gating. Paradoxically, the crystal structure of Open/C-type inactivated KcsA suggest these movements to be modest in magnitude. Here, we show that under physiological conditions, the KcsA outer-vestibule undergoes relatively large dynamic rearrangements upon inactivation. External Cd2+ enhances the rate of C-type inactivation in an outer-vestibule cysteine mutant (Y82C) via metal-bridge formation. This effect is not present in a non-inactivating mutant (E71A/Y82C). Tandem dimer and tandem tetramer constructs of equivalent cysteine mutants in KcsA and Shaker K+ channels demonstrate that these Cd2+ metal bridges are formed only between adjacent subunits. This is well supported by molecular dynamics simulations. Based on the crystal structure of Cd2+-bound Y82C-KcsA in the closed state, together with EPR distance measurements in the KcsA outer-vestibule, we suggest that subunits must dynamically come in close proximity as the channels undergo inactivation. PMID:22771214

  15. Inactivation of biofilm bacteria.

    PubMed Central

    LeChevallier, M W; Cawthon, C D; Lee, R G

    1988-01-01

    The current project was developed to examine inactivation of biofilm bacteria and to characterize the interaction of biocides with pipe surfaces. Unattached bacteria were quite susceptible to the variety of disinfectants tested. Viable bacterial counts were reduced 99% by exposure to 0.08 mg of hypochlorous acid (pH 7.0) per liter (1 to 2 degrees C) for 1 min. For monochloramine, 94 mg/liter was required to kill 99% of the bacteria within 1 min. These results were consistent with those found by other investigators. Biofilm bacteria grown on the surfaces of granular activated carbon particles, metal coupons, or glass microscope slides were 150 to more than 3,000 times more resistant to hypochlorous acid (free chlorine, pH 7.0) than were unattached cells. In contrast, resistance of biofilm bacteria to monochloramine disinfection ranged from 2- to 100-fold more than that of unattached cells. The results suggested that, relative to inactivation of unattached bacteria, monochloramine was better able to penetrate and kill biofilm bacteria than free chlorine. For free chlorine, the data indicated that transport of the disinfectant into the biofilm was a major rate-limiting factor. Because of this phenomenon, increasing the level of free chlorine did not increase disinfection efficiency. Experiments where equal weights of disinfectants were used suggested that the greater penetrating power of monochloramine compensated for its limited disinfection activity. These studies showed that monochloramine was as effective as free chlorine for inactivation of biofilm bacteria. The research provides important insights into strategies for control of biofilm bacteria. Images PMID:2849380

  16. Effect of thyme/cumin essential oils and butylated hydroxyl anisole/butylated hydroxyl toluene on physicochemical properties and oxidative/microbial stability of chicken patties.

    PubMed

    Sariçoban, Cemalettin; Yilmaz, Mustafa Tahsin

    2014-02-01

    In this study, effects of thyme/cumin essential oils (EO) and butylated hydroxyl anisole (BHA)/butylated hydroxyl toluene (BHT) on physicochemical properties and storage stability of chicken patties were compared in different storage periods (0, 3, 7, 14, 21, and 28 d). It was found that there were significant (P < 0.05) differences between physicochemical properties of patty samples treated with EO and the synthetic antioxidants. The EO showed similar performance to those of BHA and BHT in limiting TBARS values of chicken patty samples. Similarity in performance was also the case for microbial stability (total aerobic mesophilic, psychrotrophic, lactic acid, and coliform bacteria as well as molds and yeasts); namely, their effects were significant (P < 0.05). Effect of thyme EO was significant (P < 0.05) and remarkable, not allowing any coliform bacteria to grow in the samples. Given that EO were obtained from natural sources, the data suggested that the EO might be more useful than their synthetic counterparts, BHA and BHT, as additives for chicken patties to maintain oxidative/microbial stability and increase shelf life.

  17. THE ANTIGENIC POTENCY OF EPIDEMIC INFLUENZA VIRUS FOLLOWING INACTIVATION BY ULTRAVIOLET RADIATION

    PubMed Central

    Salk, Jonas E.; Lavin, G. I.; Francis, Thomas

    1940-01-01

    A study of the antigenic potency of influenza virus inactivated by ultraviolet radiation has been made. Virus so inactivated is still capable of functioning as an immunizing agent when given to mice by the intraperitoneal route. In high concentrations inactivated virus appears to be nearly as effective as active virus but when quantitative comparisons of the immunity induced by different dilutions are made, it is seen that a hundredfold loss in immunizing capacity occurs during inactivation. Virus in suspensions prepared from the lungs of infected mice is inactivated more rapidly than virus in tissue culture medium. A standard for the comparison of vaccines of epidemic influenza virus is proposed. PMID:19871057

  18. Microbial safety of air-dried and rewetted biosolids.

    PubMed

    Rouch, Duncan A; Mondal, Tania; Pai, Sneha; Glauche, Florian; Fleming, Vennessa A; Thurbon, Nerida; Blackbeard, Judy; Smith, Stephen R; Deighton, Margaret

    2011-06-01

    To assess microbial safety of treated sewage sludge (biosolids), we examined the inactivation of microbial indicators for potential bacterial, viral and protozoan pathogens. The levels of indicators were determined throughout the air-drying and storage phases of anaerobically digested sewage sludge. Samples were collected from two wastewater treatment plants (WWTPS) in Victoria, Australia. Established methods were applied for analysis of bacteria and coliphages, based on membrane filtration and layered plates, respectively. In the pan drying phase, the prevalence of Escherichia coli was reduced by >5 log10 compared with sludge entering the pan. Thus, after pan drying of 8-11 months at WWTP A and 15 months at WWTP B, the numbers of E. coli were reduced to below 10(2) cfu/g dry solids (DS). This level is acceptable for unrestricted use in agriculture in Australia (P1 treatment grade), the UK (enhanced treatment status) and the USA (Class A pathogen reduction). Coliphage numbers also decreased substantially during the air-drying phase, indicating that enteric viruses are also likely to be destroyed during this phase. Clostridium perfringens appeared to be an overly conservative indicator. Survival, but not regrowth, of E. coli or Salmonella was observed in rewetted biosolids (15-20% moisture content), after being seeded with these species, indicating a degree of safety of stored biosolids upon rewetting by rain.

  19. Short-time effect of salvage harvesting on microbial soil properties in a Mediterranean area affected by a wildfire: preliminary results

    NASA Astrophysics Data System (ADS)

    Moltó, Jorge; Mataix-Solera, Jorge; Arcenegui, Victoria; Morugan, Alicia; Girona, Antonio; Garcia-orenes, Fuensanta

    2014-05-01

    In the Mediterranean region, wildfires are considered one of the main ecological factors, which, in addition to and in relation to changes in soil use, may cause soil loss and degradation, one of the most important environmental problems that humanity must face up to. As is well known, the soil-plant system is one of the key factors determining ecological recovery after the occurrence of a wildfire. Traditionally, a variety of forestry practices have been implemented on spanish sites after the incidence of a wildfire. Among them stands out the complete extraction of the burned wood, which consist in getting rid of the branches and other wooden debris using small controlled bonfires, splintering or mechanical extraction. This set of post-fire management practices is known as salvage logging or salvage harvesting. Despite the remarkable relevance and influence that this conjunction of techniques has on land management after a wildfire, very little experimental research focused on assessing the impact of salvage logging on the vegetal community has been done. Furthermore, even less research inquiring into the mode and grade of incidence that the salvage logging produces on soil properties has taken place. The aim of this research is to assess the effects that the salvage harvesting has on different soil microbial properties and other related properties. The study area is located in the Natural Park of the "Sierra de Mariola" in the province of Alicante, southeastern Spain. This location was affected by a wildfire whose extension reached more than 500 Ha in July 2012. Different post-fire treatments were proposed by the authorities, including salvage harvesting in some areas. Two different treatments were distinguished for the study, "control" (without any kind of burned wood removal) and "harvest" (where salvage logging was carried out), in each area three 4 m2 sampling plots were set up. These two treatments were established on the same slope with the same orography

  20. Effects of irradiation and fumaric acid treatment on the inactivation of Listeria monocytogenes and Salmonella typhimurium inoculated on sliced ham

    NASA Astrophysics Data System (ADS)

    Song, Hyeon-Jeong; Lee, Ji-Hye; Song, Kyung Bin

    2011-11-01

    To examine the effects of fumaric acid and electron beam irradiation on the inactivation of foodborne pathogens in ready-to-eat meat products, sliced ham was inoculated with Listeria monocytogenes and Salmonella typhimurium. The inoculated ham slices were treated with 0.5% fumaric acid or electron beam irradiation at 2 kGy. Fumaric acid treatment reduced the populations of L. monocytogenes and S. typhimurium by approximately 1 log CFU/g compared to control populations. In contrast, electron beam irradiation decreased the populations of S. typhimurium and L. monocytogenes by 3.78 and 2.42 log CFU/g, respectively. These results suggest that electron beam irradiation is a better and appropriate technique for improving the microbial safety of sliced ham.

  1. Mechanism of poliovirus inactivation by ammonia

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

    Ward, R.L.

    1978-05-01

    Poliovirus inactivation by ammonia causes a slight reduction in the sedimentation coefficients of viral particles, but has no detectable effect on either the electrophoretic pattern of viral capsid proteins or the isoelectric points of inactivated particles. These virions still attach to cells, but are unable to repress host translation or stimulate the synthesis of detectable amounts of viral RNA. Although ammonia has no detectable effect on naked poliovirus RNA, it causes cleavage of this RNA when still within viral particles. Therefore, the RNA genome appears to be the only component of poliovirus significantly affected by ammonia.

  2. Method for determining virus inactivation during sludge treatment processes.

    PubMed Central

    Traub, F; Spillmann, S K; Wyler, R

    1986-01-01

    A simple and reliable method is described which allows determination of virus inactivation rates during sludge treatment processes in situ. Bacteriophage f2 was adsorbed onto an electropositive membrane filter which was then sandwiched between two polycarbonate membranes with pores smaller than the virus diameter. The resulting sandwich was fixed in an open filter holder, and several such devices were connected before being exposed in sludge-digesting tanks. The device described prevented uncontrolled virus escape, but allowed direct contact of the various inactivating or stabilizing substances present in the environment tested with the virus adsorbed to the carrier membrane. After exposure to an environment, the surviving fraction of virus was eluted from the inner filter and determined by plaque counting. By using polycarbonate membranes without pores for sandwiching, the influence of temperature alone on virus inactivation could be measured. Thermophilic fermentation at 60 degrees C and at 65 kPa pressure led to a bacteriophage f2 titer reduction of 3.5 log10 units per h, whereas during thermophilic digestion at 54.5 degrees C titers decreased 1.2 log10 units per h. During mesophilic digestion an inactivation rate of only 0.04 log10 units per h was observed. Under these latter conditions, temperature had only a minor effect (19%) on virus inactivation, whereas at 54.5 degrees C during thermophilic digestion heat accounted for 32% of the total inactivation, and during thermophilic fermentation at 60 degrees C temperature and pressure were 100% responsible for virus denaturation. PMID:3532955

  3. Infectious pancreatic necrosis virus in fish by-products is inactivated with inorganic acid (pH 1) and base (pH 12).

    PubMed

    Myrmel, M; Modahl, I; Nygaard, H; Lie, K M

    2014-04-01

    The aquaculture industry needs a simple, inexpensive and safe method for the treatment of fish waste without heat. Microbial inactivation by inorganic acid (HCl) or base (KOH) was determined using infectious pancreatic necrosis virus (IPNV) as a model organism for fish pathogens. Salmonella and spores of Clostridium perfringens were general hygiene indicators in supplementary examinations. IPNV, which is considered to be among the most chemical- and heat-resistant fish pathogens, was reduced by more than 3 log in 4 h at pH 1.0 and pH 12.0. Salmonella was rapidly inactivated by the same treatment, whereas spores of C. perfringens were hardly affected. The results indicate that low and high pH treatment could be particularly suitable for fish waste destined for biogas production. pH treatment at aquaculture production sites could reduce the spread of fish pathogens during storage and transportation without disturbing the anaerobic digestion process. The treatment could also be an alternative to the current energy-intensive steam pressure sterilization of fish waste to be used by the bioenergy, fertilizer and soil improver industries. © 2013 John Wiley & Sons Ltd.

  4. Synthesis and characterization of PLGA nanoparticles containing mixture of curcuminoids for optimization of photodynamic inactivation

    NASA Astrophysics Data System (ADS)

    Suzuki, Isabella L.; Inada, Natália M.; Marangoni, Valéria S.; Corrêa, Thaila Q.; Zucolotto, Valtencir; Kurachi, Cristina; Bagnato, Vanderlei S.

    2016-03-01

    Because of excessive use of antibiotics there is a growth in the number of resistant strains. Due to this growth of multiresistant bacteria, the number of searches looking for alternatives antibacterial therapeutic has increased, and among them is the antimicrobial photodynamic therapy (aPDT) or photodynamic inactivation (PDI). The photodynamic inactivation involves the action of a photosensitizer (PS), activated by a specific wavelength, in the present of oxygen, resulting in cytotoxic effect. Natural curcumin, consists of a mixture of three curcuminoids: curcumin, demethoxycurcumin and bis-demethoxycurcumin. Curcumin has various pharmacological properties, however, has extremely low solubility in aqueous solutions, which difficult the use as therapeutic agent. The present study aims to develop polymeric PLGA nanoparticles containing curcuminoids to improve water solubility, increase bioavailability providing protection from degradation (chemistry and physics), and to verify the efficacy in photodynamic inactivation of microorganisms. The PLGA-CURC were synthesized by nanoprecipitation, resulting in two different systems, with an average size of 172 nm and 70% encapsulation efficiency for PLGA-CURC1, and 215 nm and 80% for PLGA-CURC2. Stability tests showed the polymer protected the curcuminoids against premature degradation. Microbiological tests in vitro with curcuminoids water solution and both suspension of PLGA-CURC were efficient in Gram-positive bacterium and fungus. However, the solution presented dark toxicity at high concentrations, unlike the nanoparticles. Thus, it was concluded that it was possible to let curcuminoids water soluble by encapsulation in PLGA nanoparticles, to ensure improved stability in aqueous medium (storage), and to inactivate bacteria and fungus.

  5. Mathematical Modeling of Microbial Community Dynamics: A Methodological Review

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

    Song, Hyun-Seob; Cannon, William R.; Beliaev, Alex S.

    Microorganisms in nature form diverse communities that dynamically change in structure and function in response to environmental variations. As a complex adaptive system, microbial communities show higher-order properties that are not present in individual microbes, but arise from their interactions. Predictive mathematical models not only help to understand the underlying principles of the dynamics and emergent properties of natural and synthetic microbial communities, but also provide key knowledge required for engineering them. In this article, we provide an overview of mathematical tools that include not only current mainstream approaches, but also less traditional approaches that, in our opinion, can bemore » potentially useful. We discuss a broad range of methods ranging from low-resolution supra-organismal to high-resolution individual-based modeling. Particularly, we highlight the integrative approaches that synergistically combine disparate methods. In conclusion, we provide our outlook for the key aspects that should be further developed to move microbial community modeling towards greater predictive power.« less

  6. Fullerene C60 and graphene photosensibiles for photodynamic virus inactivation

    NASA Astrophysics Data System (ADS)

    Belousova, I.; Hvorostovsky, A.; Kiselev, V.; Zarubaev, V.; Kiselev, O.; Piotrovsky, L.; Anfimov, P.; Krisko, T.; Muraviova, T.; Rylkov, V.; Starodubzev, A.; Sirotkin, A.; Grishkanich, A.; Kudashev, I.; Kancer, A.; Kustikova, M.; Bykovskaya, E.; Mayurova, A.; Stupnikov, A.; Ruzankina, J.; Afanasyev, M.; Lukyanov, N.; Redka, D.; Paklinov, N.

    2018-02-01

    A solid-phase photosensitizer based on aggregated C60 fullerene and graphene oxide for photodynamic inactivation of pathogens in biological fluids was studied. The most promising technologies of inactivation include the photodynamic effect, which consists in the inactivation of infectious agents by active oxygen forms (including singlet oxygen), formed when light is activated by the photosensitizer introduced into the plasma. Research shows features of solid-phase systems based on graphene and fullerene C60 oxide, which is a combination of an effective inactivating pathogens (for example, influenza viruses) reactive oxygen species formed upon irradiation of the photosensitizer in aqueous and biological fluids, a high photostability fullerene coatings and the possibility of full recovery photosensitizer from the biological environment after the photodynamic action.

  7. Inactivation of the Prelimbic Cortex Attenuates Context-Dependent Operant Responding

    PubMed Central

    Shipman, Megan L.; Bouton, Mark E.

    2017-01-01

    Operant responding in rats provides an analog to voluntary behavior in humans and is used to study maladaptive behaviors, such as overeating, drug taking, or relapse. In renewal paradigms, extinguished behavior recovers when tested outside the context where extinction was learned. Inactivation of the prelimbic (PL) region of the medial prefrontal cortex by baclofen/muscimol (B/M) during testing attenuates renewal when tested in the original acquisition context after extinction in another context (ABA renewal). Two experiments tested the hypothesis that the PL is important in context-dependent responding learned during conditioning. In the first, rats learned to lever-press for a sucrose-pellet reward. Following acquisition, animals were infused with either B/M or vehicle in the PL and tested in the acquisition context (A) and in a different context (B). All rats showed a decrement in responding when switched from Context A to Context B, but PL inactivation decreased responding only in Context A. Experiment 2a examined the effects of PL inactivation on ABC renewal in the same rats. Here, following reacquisition of the response, responding was extinguished in a new context (C). Following infusions of B/M or vehicle in the PL, responding was tested in Context C and another new context (D). The rats exhibited ACD renewal regardless of PL inactivation. Experiment 2b demonstrated that PL inactivation attenuated the ABA renewal effect in the same animals, replicating earlier results and demonstrating that cannulae were still functional. The results suggest that, rather than attenuating renewal generally, PL inactivation specifically affects ABA renewal by reducing responding in the conditioning context. SIGNIFICANCE STATEMENT Extinguished operant behavior can recover (“renew”) when tested outside the extinction context. This suggests that behaviors, such as overeating or drug taking, might be especially prone to relapse following treatment. In rats, inactivation of

  8. Resistance of Bovine Spongiform Encephalopathy (BSE) Prions to Inactivation

    PubMed Central

    Giles, Kurt; Glidden, David V.; Beckwith, Robyn; Seoanes, Rose; Peretz, David; DeArmond, Stephen J.; Prusiner, Stanley B.

    2008-01-01

    Distinct prion strains often exhibit different incubation periods and patterns of neuropathological lesions. Strain characteristics are generally retained upon intraspecies transmission, but may change on transmission to another species. We investigated the inactivation of two related prions strains: BSE prions from cattle and mouse-passaged BSE prions, termed 301V. Inactivation was manipulated by exposure to sodium dodecyl sulfate (SDS), variations in pH, and different temperatures. Infectivity was measured using transgenic mouse lines that are highly susceptible to either BSE or 301V prions. Bioassays demonstrated that BSE prions are up to 1,000-fold more resistant to inactivation than 301V prions while Western immunoblotting showed that short acidic SDS treatments reduced protease-resistant PrPSc from BSE prions and 301V prions at similar rates. Our findings argue that despite being derived from BSE prions, mouse 301V prions are not necessarily a reliable model for cattle BSE prions. Extending these comparisons to human sporadic Creutzfeldt-Jakob disease and hamster Sc237 prions, we found that BSE prions were 10- and 106-fold more resistant to inactivation, respectively. Our studies contend that any prion inactivation procedures must be validated by bioassay against the prion strain for which they are intended to be used. PMID:19008948

  9. Thermal inactivation of the wine spoilage yeasts Dekkera/Brettanomyces.

    PubMed

    Couto, José António; Neves, Filipe; Campos, Francisco; Hogg, Tim

    2005-10-25

    The heat resistance of three strains of Dekkera/Brettanomyces (Dekkera anomala PYCC 5,153, Dekkera bruxellensis PYCC 4,801 and Dekkera/Brettanomyces 093) was evaluated at different temperatures between 32.5 and 55 degrees C. Thermal inactivation tests were performed in tartrate buffer solution (pH 4.0) and in wines. In the studies employing buffer as the heating menstruum, measurable thermal inactivation began only at temperatures of 50 degrees C. When heating was performed in wine, significant inactivation begins at 35 degrees C. Subsequent thermal inactivation tests were performed in buffer at various levels of pH, ethanol concentration, and various phenolic acids. Results from experiments in buffer with added ethanol suggest that the greater heat sensitivity shown in wines can be largely attributed to ethanol, although potentiation of this effect might be due to the phenolic content, particularly from ferulic acid. In the range of pH values tested (2.5-4.5), this factor had no influence in the heat inactivation kinetics. Relevant data, in the form of D and Z values calculated in the various environments, potentially useful for the establishment of regimes of thermal control of Dekkera/Brettanomyces yeasts in wine and contaminated equipment is presented.

  10. Effects of processing parameters on the inactivation of Bacillus cereus spores on red pepper (Capsicum annum L.) flakes by microwave-combined cold plasma treatment.

    PubMed

    Kim, Jung Eun; Choi, Hyeon-Son; Lee, Dong-Un; Min, Sea C

    2017-12-18

    The efficacy of microwave-combined cold plasma treatment (MCPT) for inactivating Bacillus cereus spores contaminating red pepper (Capsicum annum L.) flakes was investigated. The effects of red pepper drying method, particle size, and water activity (a w ) were also evaluated at two levels of microwave power (1700 and 2500W/cm 2 ). The inactivation effect of MCPT was higher at higher microwave power. Spore reduction was more effective with vacuum-dried red pepper than far-infrared-dried flakes. A significantly higher level of spore reduction was observed with the red pepper sample with a smaller surface to volume ratio when one surface (exterior surface) was inoculated (p<0.05). Spore reduction by MCPT at high microwave power increased from 1.7 to 2.6logspores/cm 2 when the a w of flake increased from 0.4 to 0.9 (p<0.05). MCPT did not change the color of red pepper flakes. MCPT demonstrated potential as a microbial decontaminating technology for red pepper flakes. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Virus inactivation by silver doped titanium dioxide nanoparticles for drinking water treatment.

    PubMed

    Liga, Michael V; Bryant, Erika L; Colvin, Vicki L; Li, Qilin

    2011-01-01

    Photocatalytic inactivation of viruses and other microorganisms is a promising technology that has been increasingly utilized in recent years. In this study, photocatalytic silver doped titanium dioxide nanoparticles (nAg/TiO(2)) were investigated for their capability of inactivating Bacteriophage MS2 in aqueous media. Nano-sized Ag deposits were formed on two commercial TiO(2) nanopowders using a photochemical reduction method. The MS2 inactivation kinetics of nAg/TiO(2) was compared to the base TiO(2) material and silver ions leached from the catalyst. The inactivation rate of MS2 was enhanced by more than 5 fold depending on the base TiO(2) material, and the inactivation efficiency increased with increasing silver content. The increased production of hydroxyl free radicals was found to be responsible for the enhanced viral inactivation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  12. Pretreatment to avoid positive RT-PCR results with inactivated viruses.

    PubMed

    Nuanualsuwan, Suphachai; Cliver, Dean O

    2002-07-01

    Enteric viruses that are important causes of human disease must often be detected by reverse transcription-polymerase chain reaction (RT-PCR), a method that commonly yields positive results with samples that contain only inactivated virus. This study was intended to develop a pretreatment for samples, so that inactivated viruses would not be detected by the RT-PCR procedure. Model viruses were human hepatitis A virus, vaccine poliovirus 1 and feline calicivirus as a surrogate for the Norwalk-like viruses. Each virus was inactivated (from an initial titer of approximately 10(3) PFU/ml) by ultraviolet light, hypochlorite or heating at 72 degrees C. Inactivated viruses, that were treated with proteinase K and ribonuclease for 30 min at 37 degrees C before RT-PCR, gave a negative result, which is to say that no amplicon was detected after the reaction was completed. This antecedent to the RT-PCR method may be applicable to other types of viruses, to viruses inactivated in other ways and to other molecular methods of virus detection.

  13. Response surface methodology as a tool for modeling and optimization of Bacillus subtilis spores inactivation by UV/ nano-Fe0 process for safe water production.

    PubMed

    Yousefzadeh, Samira; Matin, Atiyeh Rajabi; Ahmadi, Ehsan; Sabeti, Zahra; Alimohammadi, Mahmood; Aslani, Hassan; Nabizadeh, Ramin

    2018-04-01

    One of the most important aspects of environmental issues is the demand for clean and safe water. Meanwhile, disinfection process is one of the most important steps in safe water production. The present study aims at estimating the performance of UV, nano Zero-Valent Iron particles (nZVI, nano-Fe 0 ), and UV treatment with the addition of nZVI (combined process) for Bacillus subtilis spores inactivation. Effects of different factors on inactivation including contact time, initial nZVI concentration, UV irradiance and various aerations conditions were investigated. Response surface methodology, based on a five-level, two variable central composite design, was used to optimize target microorganism reduction and the experimental parameters. The results indicated that the disinfection time had the greatest positive impact on disinfection ability among the different selected independent variables. According to the results, it can be concluded that microbial reduction by UV alone was more effective than nZVI while the combined UV/nZVI process demonstrated the maximum log reduction. The optimum reduction of about 4 logs was observed at 491 mg/L of nZVI and 60 min of contact time when spores were exposed to UV radiation under deaerated condition. Therefore, UV/nZVI process can be suggested as a reliable method for Bacillus subtilis spores inactivation. Copyright © 2018. Published by Elsevier Ltd.

  14. Effect of atmospheric pressure plasma on inactivation of pathogens inoculated onto bacon using two different gas compositions.

    PubMed

    Kim, Binna; Yun, Hyejeong; Jung, Samooel; Jung, Yeonkook; Jung, Heesoo; Choe, Wonho; Jo, Cheorun

    2011-02-01

    Atmospheric pressure plasma (APP) is an emerging non-thermal pasteurization method for the enhancement of food safety. In this study, the effect of APP on the inactivation of pathogens inoculated onto bacon was observed. Sliced bacon was inoculated with Listeria monocytogenes (KCTC 3596), Escherichia coli (KCTC 1682), and Salmonella Typhimurium (KCTC 1925). The samples were treated with APP at 75, 100, and 125 W of input power for 60 and 90 s. Two gases, helium (10 lpm) or a mixture of helium and oxygen, (10 lpm and 10 sccm, respectively) were used for the plasma generation. Plasma with helium could only reduce the number of inoculated pathogens by about 1-2 Log cycles. On the other hand, the helium/oxygen gas mixture was able to achieve microbial reduction of about 2-3 Log cycles. The number of total aerobic bacteria showed 1.89 and 4.58 decimal reductions after plasma treatment with helium and the helium/oxygen mixture, respectively. Microscopic observation of the bacon after plasma treatment did not find any significant changes, except that the L∗-value of the bacon surface was increased. These results clearly indicate that APP treatment is effective for the inactivation of the three pathogens used in this study, although further investigation is needed for elucidating quality changes after treatment. Copyright © 2010 Elsevier Ltd. All rights reserved.

  15. Standardization of Spore Inactivation Method for PMA-PhyloChip Analysis

    NASA Technical Reports Server (NTRS)

    Schrader, Michael

    2011-01-01

    In compliance with the Committee on Space Research (COSPAR) planetary protection policy, National Aeronautics and Space Administration (NASA) monitors the total microbial burden of spacecraft as a means for minimizing the inadvertent transfer of viable contaminant microorganisms to extraterrestrial environments (forward contamination). NASA standard assay-based counts are used both as a proxy for relative surface cleanliness and to estimate overall microbial burden as well as to assess whether forward planetary protection risk criteria are met for a given mission, which vary by the planetary body to be explored and whether or not life detection missions are present. Despite efforts to reduce presence of microorganisms from spacecraft prior to launch, microbes have been isolated from spacecraft and associated surfaces within the extreme conditions of clean room facilities using state of the art molecular technologies. Development of a more sensitive method that will better enumerate all viable microorganisms from spacecraft and associated surfaces could support future life detection missions. Current culture-based (NASA standard spore assay) and nucleic-acid-based polymerase chain reaction (PCR) methods have significant shortcomings in this type of analysis. The overall goal of this project is to evaluate and validate a new molecular method based on the use of a deoxyribonucleic acid (DNA) intercalating agent propidium monoazide (PMA). This is used in combination with DNA microarray (PhyloChip) which has been shown to identify very low levels of organisms on spacecraft associated surfaces. PMA can only penetrate the membrane of dead cells. Once penetrated, it intercalates the DNA and, upon photolysis using visible light it produces stable DNA monoadducts. This allows DNA to be unavailable for further PCR analysis. The specific aim of this study is to standardize the spore inactivation method for PMA-PhyloChip analysis. We have used the bacterial spores Bacillus

  16. Archean Microbial Mat Communities

    NASA Astrophysics Data System (ADS)

    Tice, Michael M.; Thornton, Daniel C. O.; Pope, Michael C.; Olszewski, Thomas D.; Gong, Jian

    2011-05-01

    Much of the Archean record of microbial communities consists of fossil mats and stromatolites. Critical physical emergent properties governing the evolution of large-scale (centimeters to meters) topographic relief on the mat landscape are (a) mat surface roughness relative to the laminar sublayer and (b) cohesion. These properties can be estimated for fossil samples under many circumstances. A preliminary analysis of Archean mat cohesion suggests that mats growing in shallow marine environments from throughout this time had cohesions similar to those of modern shallow marine mats. There may have been a significant increase in mat strength at the end of the Archean.

  17. Removal of inactivation causes time-invariant sodium current decays

    PubMed Central

    1988-01-01

    The kinetic properties of the closing of Na channels were studied in frog skeletal muscle to obtain information about the dependence of channel closing on the past history of the channel. Channel closing was studied in normal and modified channels. Chloramine-T was used to modify the channels so that inactivation was virtually removed. A series of depolarizing prepulse potentials was used to activate Na channels, and a -140-mV postpulse was used to monitor the closing of the channels. Unmodified channels decay via a biexponential process with time constants of 72 and 534 microseconds at 12 degrees C. The observed time constants do not depend upon the potential used to activate the channels. The contribution of the slow component to the total decay increases as the activating prepulse is lengthened. After inactivation is removed, the biexponential character of the decay is retained, with no change in the magnitude of the time constants. However, increases in the duration of the activating prepulse over the range where the current is maximal 1-75 ms) produce identical biexponential decays. The presence of biexponential decays suggests that either two subtypes of Na channels are found in muscle, or Na channels can exist in one of two equally conductive states. The time- invariant decays observed suggest that channel closure does not depend upon their past history. PMID:2852208

  18. Inactivation Strategies for Clostridium perfringens Spores and Vegetative Cells.

    PubMed

    Talukdar, Prabhat K; Udompijitkul, Pathima; Hossain, Ashfaque; Sarker, Mahfuzur R

    2017-01-01

    Clostridium perfringens is an important pathogen to human and animals and causes a wide array of diseases, including histotoxic and gastrointestinal illnesses. C. perfringens spores are crucial in terms of the pathogenicity of this bacterium because they can survive in a dormant state in the environment and return to being live bacteria when they come in contact with nutrients in food or the human body. Although the strategies to inactivate C. perfringens vegetative cells are effective, the inactivation of C. perfringens spores is still a great challenge. A number of studies have been conducted in the past decade or so toward developing efficient inactivation strategies for C. perfringens spores and vegetative cells, which include physical approaches and the use of chemical preservatives and naturally derived antimicrobial agents. In this review, different inactivation strategies applied to control C. perfringens cells and spores are summarized, and the potential limitations and challenges of these strategies are discussed. Copyright © 2016 American Society for Microbiology.

  19. Inactivation Strategies for Clostridium perfringens Spores and Vegetative Cells

    PubMed Central

    Talukdar, Prabhat K.; Udompijitkul, Pathima; Hossain, Ashfaque

    2016-01-01

    ABSTRACT Clostridium perfringens is an important pathogen to human and animals and causes a wide array of diseases, including histotoxic and gastrointestinal illnesses. C. perfringens spores are crucial in terms of the pathogenicity of this bacterium because they can survive in a dormant state in the environment and return to being live bacteria when they come in contact with nutrients in food or the human body. Although the strategies to inactivate C. perfringens vegetative cells are effective, the inactivation of C. perfringens spores is still a great challenge. A number of studies have been conducted in the past decade or so toward developing efficient inactivation strategies for C. perfringens spores and vegetative cells, which include physical approaches and the use of chemical preservatives and naturally derived antimicrobial agents. In this review, different inactivation strategies applied to control C. perfringens cells and spores are summarized, and the potential limitations and challenges of these strategies are discussed. PMID:27795314

  20. Casein Kinase 2 Reverses Tail-Independent Inactivation of Kinesin-1

    NASA Astrophysics Data System (ADS)

    Xu, Jing

    2013-03-01

    Kinesin-1 is a plus-end microtubule-based motor, and defects in kinesin-based transport are linked to diseases including neurodegeneration. Kinesin can auto-inhibit via a head-tail interaction, but is believed to be active otherwise. Here we report a tail-independent inactivation of kinesin, reversible by the disease-relevant signalling protein, casein kinase 2 (CK2). The majority of initially active kinesin (native or tail-less) loses its ability to interact with microtubules in vitro, and CK2 reverses this inactivation (approximately fourfold) without altering kinesin's single motor properties. This activation pathway does not require motor phosphorylation, and is independent of head-tail auto-inhibition. In cultured mammalian cells, reducing CK2 expression, but not its kinase activity, decreases the force required to stall lipid droplet transport, consistent with a decreased number of active kinesin motors. Our results (Nat. Commun., 3:754, 2012) provide the first direct evidence of a protein kinase upregulating kinesin-based transport, and suggest a novel pathway for regulating the activity of cargo-bound kinesin. Work supported by NIGMS grants GM64624 to SPG, GM74830-06A1 to LH, GM76516 to LB, NS048501 to SJK, and AHA grant 825278F to JX.

  1. Strategy to inactivate Clostridium perfringens spores in meat products.

    PubMed

    Akhtar, Saeed; Paredes-Sabja, Daniel; Torres, J Antonio; Sarker, Mahfuzur R

    2009-05-01

    The current study aimed to develop an inactivation strategy for Clostridium perfringens spores in meat through a combination of spore activation at low pressure (100-200 MPa, 7 min) and elevated temperature (80 degrees C, 10 min); spore germination at high temperatures (55, 60 or 65 degrees C); and inactivation of germinated spores with elevated temperatures (80 and 90 degrees C, 10 and 20 min) and high pressure (586 MPa, at 23 and 73 degrees C, 10 min). Low pressures (100-200 MPa) were insufficient to efficiently activate C. perfringens spores for germination. However, C. perfringens spores were efficiently activated with elevated temperature (80 degrees C, 10 min), and germinated at temperatures lethal for vegetative cells (>or= 55 degrees C) when incubated for 60 min with a mixture of L-asparagine and KCl (AK) in phosphate buffer (pH 7) and in poultry meat. Inactivation of spores (approximately 4 decimal reduction) in meat by elevated temperatures (80-90 degrees C for 20 min) required a long germination period (55 degrees C for 60 min). However, similar inactivation level was reached with shorter germination period (55 degrees C for 15 min) when spore contaminated-meat was treated with pressure-assisted thermal processing (568 MPa, 73 degrees C, 10 min). Therefore, the most efficient strategy to inactivate C. perfringens spores in poultry meat containing 50 mM AK consisted: (i) a primary heat treatment (80 degrees C, 10 min) to pasteurize and denature the meat proteins and to activate C. perfringens spores for germination; (ii) cooling of the product to 55 degrees C in about 20 min and further incubation at 55 degrees C for about 15 min for spore germination; and (iii) inactivation of germinated spores by pressure-assisted thermal processing (586 MPa at 73 degrees C for 10 min). Collectively, this study demonstrates the feasibility of an alternative and novel strategy to inactivate C. perfringens spores in meat products formulated with germinants specific for C

  2. Cryptococcus neoformans capsule protects cell from oxygen reactive species generated by antimicrobial photodynamic inactivation

    NASA Astrophysics Data System (ADS)

    Prates, Renato Araujo; Hamblin, Michael R.; Kato, Ilka T.; Fuchs, Beth; Mylonakis, Eleytherios; Simões Ribeiro, Martha; Tegos, George

    2011-03-01

    Antimicrobial photodynamic inactivation (APDI) is based on the utilization of substances that can photosensitize biological tissues and are capable of being activated in the presence of light. Cryptococcus neoformans is an yeast surrounded by a capsule composed primarily of glucoronoxylomannan that plays an important role in its virulence. This yeast causes infection on skin, lungs and brain that can be associated with neurological sequelae and neurosurgical interventions, and its conventional treatment requires prolonged antifungal therapy, which presents important adverse effects. The aim of this study was to evaluate the protective effect of Cryptococcus neoformans capsule against reactive oxygen species generated by APDI. Cryptococcus neoformans KN99α, which is a strain able to produce capsule, and CAP59 that does not present capsule production were submitted to APDI using methylene blue (MB), rose bengal (RB), and pL-ce6 as photosensitizers (PS). Then microbial inactivation was evaluated by counting colony form units following APDI and confocal laser scanning microscopy (CLSM) illustrated localization as well as the preferential accumulation of PS into the fungal cells. C. neoformans KN99α was more resistant to APDI than CAP59 for all PSs tested. CLSM showed incorporation of MB and RB into the cytoplasm and a preferential uptake in mitochondria. A nuclear accumulation of MB was also observed. Contrarily, pL-ce6 appears accumulated in cell wall and cell membrane and minimal florescence was observed inside the fungal cells. In conclusion, the ability of C. neoformans to form capsule enhances survival following APDI.

  3. Heat-Denatured Lysozyme Inactivates Murine Norovirus as a Surrogate Human Norovirus.

    PubMed

    Takahashi, Hajime; Nakazawa, Moemi; Ohshima, Chihiro; Sato, Miki; Tsuchiya, Tomoki; Takeuchi, Akira; Kunou, Masaaki; Kuda, Takashi; Kimura, Bon

    2015-07-02

    Human norovirus infects humans through the consumption of contaminated food, contact with the excrement or vomit of an infected person, and through airborne droplets that scatter the virus through the air. Being highly infectious and highly viable in the environment, inactivation of the norovirus requires a highly effective inactivating agent. In this study, we have discovered the thermal denaturing capacity of a lysozyme with known antimicrobial activity against gram-positive bacteria, as well as its inactivating effect on murine norovirus. This study is the first report on the norovirus-inactivating effects of a thermally denatured lysozyme. We observed that lysozymes heat-treated for 40 min at 100 °C caused a 4.5 log reduction in infectivity of norovirus. Transmission electron microscope analysis showed that virus particles exposed to thermally denatured lysozymes were expanded, compared to the virus before exposure. The amino acid sequence of the lysozyme was divided into three sections and the peptides of each artificially synthesised, in order to determine the region responsible for the inactivating effect. These results suggest that thermal denaturation of the lysozyme changes the protein structure, activating the region responsible for imparting an inactivating effect against the virus.

  4. Design and mechanism of tetrahydrothiophene-based γ-aminobutyric acid aminotransferase inactivators.

    PubMed

    Le, Hoang V; Hawker, Dustin D; Wu, Rui; Doud, Emma; Widom, Julia; Sanishvili, Ruslan; Liu, Dali; Kelleher, Neil L; Silverman, Richard B

    2015-04-08

    Low levels of γ-aminobutyric acid (GABA), one of two major neurotransmitters that regulate brain neuronal activity, are associated with many neurological disorders, such as epilepsy, Parkinson's disease, Alzheimer's disease, Huntington's disease, and cocaine addiction. One of the main methods to raise the GABA level in human brain is to use small molecules that cross the blood-brain barrier and inhibit the activity of γ-aminobutyric acid aminotransferase (GABA-AT), the enzyme that degrades GABA. We have designed a series of conformationally restricted tetrahydrothiophene-based GABA analogues with a properly positioned leaving group that could facilitate a ring-opening mechanism, leading to inactivation of GABA-AT. One compound in the series is 8 times more efficient an inactivator of GABA-AT than vigabatrin, the only FDA-approved inactivator of GABA-AT. Our mechanistic studies show that the compound inactivates GABA-AT by a new mechanism. The metabolite resulting from inactivation does not covalently bind to amino acid residues of GABA-AT but stays in the active site via H-bonding interactions with Arg-192, a π-π interaction with Phe-189, and a weak nonbonded S···O═C interaction with Glu-270, thereby inactivating the enzyme.

  5. Removal of microbial pathogens in a sandy gravel aquifer under forced-gradient subsurface flow conditions

    NASA Astrophysics Data System (ADS)

    Oudega, Thomas James; Derx, Julia; van Driezum, Inge; Cisneros, Anibal; Sommer, Regina; Kirschner, Alexander; Farnleitner, Andreas; Blaschke, Alfred Paul

    2017-04-01

    Subsurface media are being used around the world as a means to mitigate microbial contamination, but vary widely in their ability to remove pathogens. To help to provide accurate risk assessments of microbial contamination of groundwaters, and establish safe setback distances between receiving waters and disposal fields, this study aims to use aquifer tracer tests to evaluate the ability of subsurface media to attenuate these pathogens. The novelty of this work is the use of a variety of different tracer substances (e.g. phages, spores, microspheres, conservative tracers) together in field experiments. This will be done by means of injecting these substances under a forced gradient in a sandy gravel aquifer in Lobau, Austria. The extraction of the tracers will be monitored in a pumping well at a distrance of 50m downgradient. This will be able to provide us with insight to the characteristics of microbial transport and how the microorganisms react to the subsurface in the study site. Subsequent numerical modelling of the experiments can tell us more about quantification of subsurface processes such as attachment/detachment, inactivation and die-off of these substances. The first field experiment with conservative tracers (NaCl) has been carried out in December 2016, and subsequent tests are being planned for the next months.

  6. Dog response to inactivated canine parvovirus and feline panleukopenia virus vaccines.

    PubMed

    Pollock, R V; Carmichael, L E

    1982-01-01

    Inactivated canine parvovirus (CPV) and inactivated feline panleukopenia virus (FPV) vaccines were evaluated in dogs. Maximal serologic response occurred within 1-2 weeks after vaccination. Antibody titers then declined rapidly to low levels that persisted at least 20 weeks. Immunity to CPV, defined as complete resistance to infection, was correlated with serum antibody titer and did not persist longer than 6 weeks after vaccination with inactivated virus. However, protection against generalized infection was demonstrated 20 weeks after vaccination. In unvaccinated dogs, viremia and generalized infection occurred after oronasal challenge with virulent CPV. In contrast, viral replication was restricted to the intestinal tract and gut-associated lymphoid tissue of vaccinated dogs. Canine parvovirus was inactivated by formalin, beta-propiolactone (BPL), and binary ethylenimine (BEI) in serum-free media; inactivation kinetics were determined. Formalin resulted in a greater loss of viral HA than either BEI of BPL, and antigenicity was correspondingly reduced.

  7. Mouse cones require an arrestin for normal inactivation of phototransduction.

    PubMed

    Nikonov, Sergei S; Brown, Bruce M; Davis, Jason A; Zuniga, Freddi I; Bragin, Alvina; Pugh, Edward N; Craft, Cheryl M

    2008-08-14

    Arrestins are proteins that arrest the activity of G protein-coupled receptors (GPCRs). While it is well established that normal inactivation of photoexcited rhodopsin, the GPCR of rod phototransduction, requires arrestin (Arr1), it has been controversial whether the same requirement holds for cone opsin inactivation. Mouse cone photoreceptors express two distinct visual arrestins: Arr1 and Arr4. By means of recordings from cones of mice with one or both arrestins knocked out, this investigation establishes that a visual arrestin is required for normal cone inactivation. Arrestin-independent inactivation is 70-fold more rapid in cones than in rods, however. Dual arrestin expression in cones could be a holdover from ancient genome duplication events that led to multiple isoforms of arrestin, allowing evolutionary specialization of one form while the other maintains the basic function.

  8. Effects of biochar blends on microbial community composition in two coastal plain soils

    EPA Science Inventory

    The amendment of soil with biochar has been demonstrated to have an effect not only on the soil physicochemical properties, but also on soil microbial community composition and activity. Previous reports have demonstrated significant impacts on soil microbial community structure....

  9. Heat inactivation of poliovirus in wastewater sludge

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

    Ward, R.L.; Ashley, C.S.; Moseley, R.H.

    1976-09-01

    The effect of raw and anaerobically digested sludge on heat inactivation of poliovirus was investigated. Raw sludge was found to be very protective of poliovirus plaque-forming ability at all temperatures studied, but digested sludge had variable effects that were highly dependent upon the experimental conditions. In low concentrations and at relatively low inactivation temperatures, digested sludge is nearly as protective of poliovirus as raw sludge. However, at higher temperatures and concentrations, digested sludge caused a significant acceleration of poliovirus inactivation. The difference between the protective capability of raw and digested sludge is not due to loss of protective material, becausemore » this component is present in the solids of digested sludge as well as in those of raw sludge. Instead, the difference is due to a virucidal agent acquired during digestion. Addition of this agent to the solids of either raw or digested sludge reverses the protective potential of these solids during heat treatment of poliovirus.« less

  10. Calmodulin-dependent activation and inactivation of anoctamin calcium-gated chloride channels

    PubMed Central

    Vocke, Kerstin; Dauner, Kristin; Hahn, Anne; Ulbrich, Anne; Broecker, Jana; Keller, Sandro; Frings, Stephan

    2013-01-01

    Calcium-dependent chloride channels serve critical functions in diverse biological systems. Driven by cellular calcium signals, the channels codetermine excitatory processes and promote solute transport. The anoctamin (ANO) family of membrane proteins encodes three calcium-activated chloride channels, named ANO 1 (also TMEM16A), ANO 2 (also TMEM16B), and ANO 6 (also TMEM16F). Here we examined how ANO 1 and ANO 2 interact with Ca2+/calmodulin using nonstationary current analysis during channel activation. We identified a putative calmodulin-binding domain in the N-terminal region of the channel proteins that is involved in channel activation. Binding studies with peptides indicated that this domain, a regulatory calmodulin-binding motif (RCBM), provides two distinct modes of interaction with Ca2+/calmodulin, one at submicromolar Ca2+ concentrations and one in the micromolar Ca2+ range. Functional, structural, and pharmacological data support the concept that calmodulin serves as a calcium sensor that is stably associated with the RCBM domain and regulates the activation of ANO 1 and ANO 2 channels. Moreover, the predominant splice variant of ANO 2 in the brain exhibits Ca2+/calmodulin-dependent inactivation, a loss of channel activity within 30 s. This property may curtail ANO 2 activity during persistent Ca2+ signals in neurons. Mutagenesis data indicated that the RCBM domain is also involved in ANO 2 inactivation, and that inactivation is suppressed in the retinal ANO 2 splice variant. These results advance the understanding of Ca2+ regulation in anoctamin Cl− channels and its significance for the physiological function that anoctamin channels subserve in neurons and other cell types. PMID:24081981

  11. Microbial lipid extraction from Lipomyces starkeyi using irreversible electroporation.

    PubMed

    Karim, Ahasanul; Yousuf, Abu; Islam, M Amirul; Naif, Yasir H; Faizal, Che Ku Mohammad; Alam, Md Zahangir; Pirozzi, Domenico

    2018-02-21

    The aim of the study was to investigate the feasibility of using irreversible electroporation (EP) as a microbial cell disruption technique to extract intracellular lipid within short time and in an eco-friendly manner. An EP circuit was designed and fabricated to obtain 4 kV with frequency of 100 Hz of square waves. The yeast cells of Lipomyces starkeyi (L. starkeyi) were treated by EP for 2-10 min where the distance between electrodes was maintained at 2, 4, and 6 cm. Colony forming units (CFU) were counted to observe the cell viability under the high voltage electric field. The forces of the pulsing electric field caused significant damage to the cell wall of L. starkeyi and the disruption of microbial cells was visualized by field emission scanning electron microscopic (FESEM) image. After breaking the cell wall, lipid was extracted and measured to assess the efficiency of EP over other techniques. The extent of cell inactivation was up to 95% when the electrodes were placed at the distance of 2 cm, which provided high treatment intensity (36.7 kWh m -3 ). At this condition, maximum lipid (63 mg g -1 ) was extracted when the biomass was treated for 10 min. During the comparison, EP could extract 31.88% lipid while the amount was 11.89% for ultrasonic and 16.8% for Fenton's reagent. The results recommend that the EP is a promising technique for lowering the time and solvent usage for lipid extraction from microbial biomass. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

  12. Proteins regulating the biosynthesis and inactivation of neuromodulatory fatty acid amides.

    PubMed

    Patricelli, M P; Cravatt, B F

    2001-01-01

    Fatty acid amides (FAAs) represent a growing family of biologically active lipids implicated in a diverse range of cellular and physiological processes. At present, two general types of fatty acid amides, the N-acylethanolamines (NAEs) and the fatty acid primary amides (FAPAs), have been identified as potential physiological neuromodulators/neurotransmitters in mammals. Representative members of these two subfamilies include the endocannabinoid NAE anandamide and the sleep-inducing FAPA oleamide. In this Chapter, molecular mechanisms proposed for the biosynthesis and inactivation of FAAs are critically evaluated, with an emphasis placed on the biochemical and cell biological properties of proteins thought to mediate these processes.

  13. Comparative study on the mechanisms of rotavirus inactivation by sodium dodecyl sulfate and ethylenediaminetetraacetate

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

    Ward, R.L.; Ashley, C.S.

    1980-06-01

    This report describes a comparative study on the effects of the anionic detergent sodium dodecyl sulfate and the chelating agent ethylenediaminetetraacetate on purified rotavirus SA-11 particles. Both chemicals readily inactivated rotavirus at quite low concentrations and under very mild conditions. In addition, both agents modified the viral capsid and prevented the adsorption of inactivated virions to cells. Capsid damage by ethylenediaminetetraacetate caused a shift in the densities of rotavirions from about l.35 to about 1.37 g/ml and a reduction in their sedimentation coefficients. Sodium dodcyl sulfate, on the other hand, did not detectably alter either of these physical properties ofmore » rotavirions. Both agents caused some alteration of the isoelectric points of the virions. Finally, analysis of rotavirus proteins showed that ethylenediaminetetraacetate caused the loss of two protein peaks from the electrophoretic pattern of virions but sodium dodecyl sulfate caused the loss of only one of these same protein peaks.« less

  14. Inactivation of bacterial quorum sensing signals N-acyl homoserine lactones is widespread in yeasts.

    PubMed

    Leguina, Ana Carolina Del V; Nieto, Carolina; Pajot, Hipólito M; Bertini, Elisa V; Mac Cormack, Walter; Castellanos de Figueroa, Lucía I; Nieto-Peñalver, Carlos G

    2018-01-01

    The inactivation of quorum sensing signals, a phenomenon known as quorum quenching, has been described in diverse microorganisms, though it remains almost unexplored in yeasts. Beyond the well-known properties of these microorganisms for the industry or as eukaryotic models, the role of yeasts in soil or in the inner tissues of a plant is largely unknown. In this report, the wider survey of quorum quenching activities in yeasts isolated from Antarctic soil and the inner tissues of sugarcane, a tropical crop, is presented. Results show that, independently of their niche, quorum quenching activities are broadly present in unicellular fungi. Although yeasts showing a broad range of quorum quenching activity are present in the two niches, at the same time specific AHL inactivation profiles can also be found. Furthermore, yeasts from both sampling sites show quorum quenching activities compatible with lactonase-like and acylase-like inactivations of AHLs. Interestingly, the characterization of Rhodotorula mucilaginosa 7Apo1 showed that the presence of a particular AHL does not interfere with the quenching of a second molecule. Evidence suggests that yeasts could play a role in the modulation of the quorum sensing activity of bacteria. The relationship among phylogeny, sampling sites and yeast quorum quenching activities of the isolates is analyzed. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  15. Increased skewing of X chromosome inactivation in Rett syndrome patients and their mothers.

    PubMed

    Knudsen, Gun Peggy S; Neilson, Tracey C S; Pedersen, June; Kerr, Alison; Schwartz, Marianne; Hulten, Maj; Bailey, Mark E S; Orstavik, Karen Helene

    2006-11-01

    Rett syndrome is a largely sporadic, X-linked neurological disorder with a characteristic phenotype, but which exhibits substantial phenotypic variability. This variability has been partly attributed to an effect of X chromosome inactivation (XCI). There have been conflicting reports regarding incidence of skewed X inactivation in Rett syndrome. In rare familial cases of Rett syndrome, favourably skewed X inactivation has been found in phenotypically normal carrier mothers. We have investigated the X inactivation pattern in DNA from blood and buccal cells of sporadic Rett patients (n=96) and their mothers (n=84). The mean degree of skewing in blood was higher in patients (70.7%) than controls (64.9%). Unexpectedly, the mothers of these patients also had a higher mean degree of skewing in blood (70.8%) than controls. In accordance with these findings, the frequency of skewed (XCI > or =80%) X inactivation in blood was also higher in both patients (25%) and mothers (30%) than in controls (11%). To test whether the Rett patients with skewed X inactivation were daughters of skewed mothers, 49 mother-daughter pairs were analysed. Of 14 patients with skewed X inactivation, only three had a mother with skewed X inactivation. Among patients, mildly affected cases were shown to be more skewed than more severely affected cases, and there was a trend towards preferential inactivation of the paternally inherited X chromosome in skewed cases. These findings, particularly the greater degree of X inactivation skewing in Rett syndrome patients, are of potential significance in the analysis of genotype-phenotype correlations in Rett syndrome.

  16. Microbial exopolysaccharides as determinants of geomorphological, hydrological and optical properties of soil crusts from the Precambrian till today

    NASA Astrophysics Data System (ADS)

    Garcia-Pichel, F.

    2012-04-01

    The presence of microbial extracellular polysaccharides (EPS) in the soil solution and/or in association with particular microbial types can impart novel properties to biological soil crust (BSC), and hence to soil surfaces. For the most part these properties are of a geobiological relevance that exceeds what one could surmise from its relatively low specific mass content. I will review some examples that range from the mundane to the unexpected. EPS associated with filamentous cyanobacteria can effectively and in the long term stabilize the soil surface against erosive forces, even after the microbes are long gone. Electrostatic interactions between EPS and blowing dust may help retain dust particles, enriching the soil with new nutrient sources. In a telltale sign of BSC presence, EPS is the agent that allows sandy soils to fold and curl-up, to form pee-tee's and elephant-skin surfaces, and to crack into polygons like clays would. EPS in large quantities in flat crusts can retain fluids (both liquid and gaseous) resulting in the alteration of hydrological flow and in the formation of internal vesicular horizons, gas bubbles, pock-marked surfaces and other characteristic structures. Yet, in some settings, EPS plays an architectural role in creating a "spongy" texture that increases hydraulic conductivity. This architectural role can indirectly result in significant increases of a crust's albedo. While the diversity of consequences of EPS presence is far from understood, evidence for its sustained role through Earth's history can be found in the form of sedimentary bio-signatures as far back as the Proterozoic.

  17. Emulsion properties of pork myofibrillar protein in combination with microbial transglutaminase and calcium alginate under various pH conditions.

    PubMed

    Hong, Geun Pyo; Min, Sang-Gi; Chin, Koo Bok

    2012-01-01

    In this study, the effects of microbial transglutaminase (MTG) and calcium alginate (CA) systems in combination with soybean oil on the emulsion properties of porcine myofibrillar protein (MP) were evaluated under various pH conditions. MTG was shown to improve emulsifying capacity and creaming stability, which increased with increasing pH values up to 6.5. The CA did not influence emulsifying capacity, but it improved the creaming stability of the MP-stabilized emulsions. Both MTG and CA enhanced the rheological properties, but their effects on the physical characteristics of the protein evidenced an opposite trend in relation to pH, i.e., the MTG system improved both the emulsion and gelling properties with increasing pH, whereas the CA system was effective when the pH was lowered. By combining the two MP gelling systems, a stable and pH-insensible emulsion could be produced. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Effects of application of corn straw on soil microbial community structure during the maize growing season.

    PubMed

    Lu, Ping; Lin, Yin-Hua; Yang, Zhong-Qi; Xu, Yan-Peng; Tan, Fei; Jia, Xu-Dong; Wang, Miao; Xu, De-Rong; Wang, Xi-Zhuo

    2015-01-01

    This study investigated the influence of corn straw application on soil microbial communities and the relationship between such communities and soil properties in black soil. The crop used in this study was maize (Zea mays L.). The five treatments consisted of applying a gradient (50, 100, 150, and 200%) of shattered corn straw residue to the soil. Soil samples were taken from May through September during the 2012 maize growing season. The microbial community structure was determined using phospholipid fatty acid (PLFA) analysis. Our results revealed that the application of corn straw influenced the soil properties and increased the soil organic carbon and total nitrogen. Applying corn straw to fields also influenced the variation in soil microbial biomass and community composition, which is consistent with the variations found in soil total nitrogen (TN) and soil respiration (SR). However, the soil carbon-to-nitrogen ratio had no effect on soil microbial communities. The abundance of PLFAs, TN, and SR was higher in C1.5 than those in other treatments, suggesting that the soil properties and soil microbial community composition were affected positively by the application of corn straw to black soil. A Principal Component Analysis indicated that soil microbial communities were different in the straw decomposition processes. Moreover, the soil microbial communities from C1.5 were significantly different from those of CK (p < 0.05). We also found a high ratio of fungal-to-bacterial PLFAs in black soil and significant variations in the ratio of monounsaturated-to-branched fatty acids with different straw treatments that correlated with SR (p < 0.05). These results indicated that the application of corn straw positively influences soil properties and soil microbial communities and that these properties affect these communities. The individual PLFA signatures were sensitive indicators that reflected the changes in the soil environment condition. © 2015 WILEY

  19. Biotic and Abiotic Properties Mediating Plant Diversity Effects on Soil Microbial Communities in an Experimental Grassland

    PubMed Central

    Lange, Markus; Habekost, Maike; Eisenhauer, Nico; Roscher, Christiane; Bessler, Holger; Engels, Christof; Oelmann, Yvonne; Scheu, Stefan; Wilcke, Wolfgang; Schulze, Ernst-Detlef; Gleixner, Gerd

    2014-01-01

    Plant diversity drives changes in the soil microbial community which may result in alterations in ecosystem functions. However, the governing factors between the composition of soil microbial communities and plant diversity are not well understood. We investigated the impact of plant diversity (plant species richness and functional group richness) and plant functional group identity on soil microbial biomass and soil microbial community structure in experimental grassland ecosystems. Total microbial biomass and community structure were determined by phospholipid fatty acid (PLFA) analysis. The diversity gradient covered 1, 2, 4, 8, 16 and 60 plant species and 1, 2, 3 and 4 plant functional groups (grasses, legumes, small herbs and tall herbs). In May 2007, soil samples were taken from experimental plots and from nearby fields and meadows. Beside soil texture, plant species richness was the main driver of soil microbial biomass. Structural equation modeling revealed that the positive plant diversity effect was mainly mediated by higher leaf area index resulting in higher soil moisture in the top soil layer. The fungal-to-bacterial biomass ratio was positively affected by plant functional group richness and negatively by the presence of legumes. Bacteria were more closely related to abiotic differences caused by plant diversity, while fungi were more affected by plant-derived organic matter inputs. We found diverse plant communities promoted faster transition of soil microbial communities typical for arable land towards grassland communities. Although some mechanisms underlying the plant diversity effect on soil microorganisms could be identified, future studies have to determine plant traits shaping soil microbial community structure. We suspect differences in root traits among different plant communities, such as root turnover rates and chemical composition of root exudates, to structure soil microbial communities. PMID:24816860

  20. Whole-Genome Sequencing in Microbial Forensic Analysis of Gamma-Irradiated Microbial Materials.

    PubMed

    Broomall, Stacey M; Ait Ichou, Mohamed; Krepps, Michael D; Johnsky, Lauren A; Karavis, Mark A; Hubbard, Kyle S; Insalaco, Joseph M; Betters, Janet L; Redmond, Brady W; Rivers, Bryan A; Liem, Alvin T; Hill, Jessica M; Fochler, Edward T; Roth, Pierce A; Rosenzweig, C Nicole; Skowronski, Evan W; Gibbons, Henry S

    2016-01-15

    Effective microbial forensic analysis of materials used in a potential biological attack requires robust methods of morphological and genetic characterization of the attack materials in order to enable the attribution of the materials to potential sources and to exclude other potential sources. The genetic homogeneity and potential intersample variability of many of the category A to C bioterrorism agents offer a particular challenge to the generation of attributive signatures, potentially requiring whole-genome or proteomic approaches to be utilized. Currently, irradiation of mail is standard practice at several government facilities judged to be at particularly high risk. Thus, initial forensic signatures would need to be recovered from inactivated (nonviable) material. In the study described in this report, we determined the effects of high-dose gamma irradiation on forensic markers of bacterial biothreat agent surrogate organisms with a particular emphasis on the suitability of genomic DNA (gDNA) recovered from such sources as a template for whole-genome analysis. While irradiation of spores and vegetative cells affected the retention of Gram and spore stains and sheared gDNA into small fragments, we found that irradiated material could be utilized to generate accurate whole-genome sequence data on the Illumina and Roche 454 sequencing platforms. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  1. Microbial Relevant Fouling in Membrane Bioreactors: Influencing Factors, Characterization, and Fouling Control

    PubMed Central

    Wu, Bing; Fane, Anthony G.

    2012-01-01

    Microorganisms in membrane bioreactors (MBRs) play important roles on degradation of organic/inorganic substances in wastewaters, while microbial deposition/growth and microbial product accumulation on membranes potentially induce membrane fouling. Generally, there is a need to characterize membrane foulants and to determine their relations to the evolution of membrane fouling in order to identify a suitable fouling control approach in MBRs. This review summarized the factors in MBRs that influence microbial behaviors (community compositions, physical properties, and microbial products). The state-of-the-art techniques to characterize biofoulants in MBRs were reported. The strategies for controlling microbial relevant fouling were discussed and the future studies on membrane fouling mechanisms in MBRs were proposed. PMID:24958297

  2. Nutrient Limitation of Microbial Mediated Decomposition and Arctic Soil Chronology

    NASA Astrophysics Data System (ADS)

    Melle, C. J.; Darrouzet-Nardi, A.; Wallenstein, M. D.

    2012-12-01

    Soils of northern permafrost regions currently contain twice as much carbon as the entire Earth's atmosphere. Traditionally, environmental constraints have limited microbial activity resulting in restricted decomposition of soil organic matter in these systems and accumulation of massive amounts of soil organic carbon (SOC), however climate change is reducing the constraints of decomposition in arctic permafrost regions. Carbon cycling in nutrient poor, arctic ecosystems is tightly coupled to other biogeochemical cycles. Several studies have suggested strong nitrogen limitations of primary productivity and potentially warm-season microbial activity in these nutrient deficient soils. Nitrogen is required for microbial extracellular enzyme production which drives the decomposition of soil organic matter (SOM). Nitrogen limited arctic soils may also experience limitation via labile carbon availability despite the SOM rich environment due to low extracellular enzyme production. Few studies have directly addressed nutrient induced microbial limitation in SOC rich arctic tundra soils, and even less is known about the potential for nutrient co-limitation. Additionally, through the process of becoming deglaciated, sites within close proximity to one another may have experienced drastic differences in their effective soil ages due to the varied length of their active histories. Many soil properties and nutrient deficiencies are directly related to soil age, however this chronology has not previously been a focus of research on nutrient limitation of arctic soil microbial activity. Understanding of nutrient limitations, as well as potential co-limitation, on arctic soil microbial activity has important implications for carbon cycling and the ultimate fate of the current arctic SOC reservoir. Analyses of nutrient limitation on soils of a single site are not adequate for fully understanding the controls on soil microbial activity across a vast land mass with large variation in

  3. Effect of PLA films containing propolis ethanolic extract, cellulose nanoparticle and Ziziphora clinopodioides essential oil on chemical, microbial and sensory properties of minced beef.

    PubMed

    Shavisi, Nassim; Khanjari, Ali; Basti, Afshin Akhondzadeh; Misaghi, Ali; Shahbazi, Yasser

    2017-02-01

    This study was conducted to examine the effects of polylactic acid (PLA) film containing propolis ethanolic extract (PE), cellulose nanoparticle (CN) and Ziziphora clinopodioides essential oil (ZEO) on chemical, microbial and sensory properties of minced beef during storage at refrigerated temperature for 11days. The initial total volatile base nitrogen (TVB-N) was 8.2mg/100g and after 7days reached to 29.1mg/100g in control, while it was lower than 25mg/100g for treated samples. At the end of storage time in control samples peroxide value (PV) reached to 2.01meqperoxide/1000g lipid, while the values for the treated samples remained lower than 2meqperoxide/1000g lipid. Final microbial population decreased approximately 1-3logCFU/g in treated samples compared to control (P<0.05). Films containing 2% ZEO alone and in combination with different concentrations of PE and CN extended the shelf life of minced beef during storage in refrigerated condition for at least 11days without any unfavorable organoleptic properties. Copyright © 2016. Published by Elsevier Ltd.

  4. Application of atomic force microscopy to microbial surfaces: from reconstituted cell surface layers to living cells.

    PubMed

    Dufrêne, Y F

    2001-02-01

    The application of atomic force microscopy (AFM) to probe the ultrastructure and physical properties of microbial cell surfaces is reviewed. The unique capabilities of AFM can be summarized as follows: imaging surface topography with (sub)nanometer lateral resolution; examining biological specimens under physiological conditions; measuring local properties and interaction forces. AFM is being used increasingly for: (i) visualizing the surface ultrastructure of microbial cell surface layers, including bacterial S-layers, purple membranes, porin OmpF crystals and fungal rodlet layers; (ii) monitoring conformational changes of individual membrane proteins; (iii) examining the morphology of bacterial biofilms, (iv) revealing the nanoscale structure of living microbial cells, including fungi, yeasts and bacteria, (v) mapping interaction forces at microbial surfaces, such as van der Waals and electrostatic forces, solvation forces, and steric/bridging forces; and (vi) probing the local mechanical properties of cell surface layers and of single cells.

  5. Inactivation of Heterosigma akashiwo in ballast water by circular orifice plate-generated hydrodynamic cavitation.

    PubMed

    Feng, Daolun; Zhao, Jie; Liu, Tian

    2016-01-01

    The discharge of alien ballast water is a well-known, major reason for marine species invasion. Here, circular orifice plate-generated hydrodynamic cavitation was used to inactivate Heterosigma akashiwo in ballast water. In comparison with single- and multihole orifice plates, the conical-hole orifice plate yielded the highest inactivation percentage, 51.12%, and consumed only 6.84% energy (based on a 50% inactivation percentage). Repeating treatment, either using double series-connection or circling inactivation, elevated the inactivation percentage, yet consumed much more energy. The results indicate that conical-hole-generated hydrodynamic cavitation shows great potential as a pre-inactivation method for ballast water treatment.

  6. Inactivation of the Prelimbic Cortex Attenuates Context-Dependent Operant Responding.

    PubMed

    Trask, Sydney; Shipman, Megan L; Green, John T; Bouton, Mark E

    2017-03-01

    Operant responding in rats provides an analog to voluntary behavior in humans and is used to study maladaptive behaviors, such as overeating, drug taking, or relapse. In renewal paradigms, extinguished behavior recovers when tested outside the context where extinction was learned. Inactivation of the prelimbic (PL) region of the medial prefrontal cortex by baclofen/muscimol (B/M) during testing attenuates renewal when tested in the original acquisition context after extinction in another context (ABA renewal). Two experiments tested the hypothesis that the PL is important in context-dependent responding learned during conditioning. In the first, rats learned to lever-press for a sucrose-pellet reward. Following acquisition, animals were infused with either B/M or vehicle in the PL and tested in the acquisition context (A) and in a different context (B). All rats showed a decrement in responding when switched from Context A to Context B, but PL inactivation decreased responding only in Context A. Experiment 2a examined the effects of PL inactivation on ABC renewal in the same rats. Here, following reacquisition of the response, responding was extinguished in a new context (C). Following infusions of B/M or vehicle in the PL, responding was tested in Context C and another new context (D). The rats exhibited ACD renewal regardless of PL inactivation. Experiment 2b demonstrated that PL inactivation attenuated the ABA renewal effect in the same animals, replicating earlier results and demonstrating that cannulae were still functional. The results suggest that, rather than attenuating renewal generally, PL inactivation specifically affects ABA renewal by reducing responding in the conditioning context. SIGNIFICANCE STATEMENT Extinguished operant behavior can recover ("renew") when tested outside the extinction context. This suggests that behaviors, such as overeating or drug taking, might be especially prone to relapse following treatment. In rats, inactivation of the

  7. Inactivation of foodborne microorganisms using engineered water nanostructures (EWNS).

    PubMed

    Pyrgiotakis, Georgios; Vasanthakumar, Archana; Gao, Ya; Eleftheriadou, Mary; Toledo, Eduardo; DeAraujo, Alice; McDevitt, James; Han, Taewon; Mainelis, Gediminas; Mitchell, Ralph; Demokritou, Philip

    2015-03-17

    Foodborne diseases caused by the consumption of food contaminated with pathogenic microorganisms or their toxins have very serious economic and public health consequences. Here, we explored the effectiveness of a recently developed intervention method for inactivation of microorganisms on fresh produce, and food production surfaces. This method utilizes Engineered Water Nanostructures (EWNS) produced by electrospraying of water vapor. EWNS possess unique properties; they are 25 nm in diameter, remain airborne in indoor conditions for hours, contain Reactive Oxygen Species (ROS) and have very strong surface charge (on average 10 e/structure). Here, their efficacy in inactivating representative foodborne bacteria such as Escherichia coli, Salmonella enterica, and Listeria innocua, on stainless steel surfaces and on organic tomatoes, was assessed. The inactivation was facilitated using two different exposure approaches in order to optimize the delivery of EWNS to bacteria: (1) EWNS were delivered on the surfaces by diffusion and (2) a "draw through" Electrostatic Precipitator Exposure System (EPES) was developed and characterized for EWNS delivery to surfaces. Using the diffusion approach and an EWNS concentration of 24,000 #/cm3, the bacterial concentrations on the surfaces were reduced, depending on the bacterium and the surface type, by values ranging between 0.7 to 1.8 logs. Using the EPES approach and for an aerosol concentration of 50,000 #/cm3 at 90 min of exposure, results show a 1.4 log reduction for E. coli on organic tomato surfaces, as compared to the control (same conditions in regards to temperature and Relative Humidity). Furthermore, for L. innocua, the dose-response relationship was demonstrated and found to be a 0.7 and 1.2 logs removal at 12,000 and 23,000 #/cm3, respectively. The results presented here indicate that this novel, chemical-free, and environmentally friendly intervention method holds potential for development and application in the

  8. Ion channels to inactivate neurons in Drosophila.

    PubMed

    Hodge, James J L

    2009-01-01

    Ion channels are the determinants of excitability; therefore, manipulation of their levels and properties provides an opportunity for the investigator to modulate neuronal and circuit function. There are a number of ways to suppress electrical activity in Drosophila neurons, for instance, over-expression of potassium channels (i.e. Shaker Kv1, Shaw Kv3, Kir2.1 and DORK) that are open at resting membrane potential. This will result in increased potassium efflux and membrane hyperpolarisation setting resting membrane potential below the threshold required to fire action potentials. Alternatively over-expression of other channels, pumps or co-transporters that result in a hyperpolarised membrane potential will also prevent firing. Lastly, neurons can be inactivated by, disrupting or reducing the level of functional voltage-gated sodium (Nav1 paralytic) or calcium (Cav2 cacophony) channels that mediate the depolarisation phase of action potentials. Similarly, strategies involving the opposite channel manipulation should allow net depolarisation and hyperexcitation in a given neuron. These changes in ion channel expression can be brought about by the versatile transgenic (i.e. Gal4/UAS based) systems available in Drosophila allowing fine temporal and spatial control of (channel) transgene expression. These systems are making it possible to electrically inactivate (or hyperexcite) any neuron or neural circuit in the fly brain, and much like an exquisite lesion experiment, potentially elucidate whatever interesting behaviour or phenotype each network mediates. These techniques are now being used in Drosophila to reprogram electrical activity of well-defined circuits and bring about robust and easily quantifiable changes in behaviour, allowing different models and hypotheses to be rapidly tested.

  9. Natural inactivation of Escherichia coli in anaerobic and reduced groundwater.

    PubMed

    Lisle, J T

    2016-06-01

    Inactivation rates of Escherichia coli in groundwater have most often been determined in aerobic and oxidized systems. This study examined E. coli inactivation rates in anaerobic and extremely reduced groundwater systems that have been identified as recharge zones. Groundwater from six artesian wells was diverted to above-ground, flow-through mesocosms that contained laboratory grown E. coli in diffusion chambers. All groundwater was anaerobic and extremely reduced (ORP < -300 mV). Cells were plated onto mTEC agar during 21-day incubation periods. All data fit a bi-phasic inactivation model, with >95% of the E. coli population being inactivated <11·0 h (mean k = 0·488 ±0·188 h(-1) ). The groundwater geochemical conditions enhanced the inactivation of E. coli to rates approx. 21-fold greater than previously published inactivation rate in groundwater (mean k = 0·023 ± 0·030 h(-1) ). Also, mTEC agar inhibits E. coli growth following exposure to anaerobic and reduced groundwater. Aquifer recharge zones with geochemical characteristics observed in this study complement above-ground engineered processes (e.g. filtration, disinfection), while increasing the overall indicator micro-organism log-reduction rate of a facility. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  10. Long-lasting potassium channel inactivation in myoepithelial fibres is related to characteristics of swimming in diphyid siphonophores.

    PubMed

    Inoue, Isao; Tsutsui, Izuo; Bone, Quentin

    2005-12-01

    Diphyid siphonophores swim using bursts of propulsive jets, which are produced by contractions of a monolayer of subumbrellar myoepithelial fibres lining the nectophore. This swimming behaviour is characterised by successive increases in the force generating the jets during the initial jets of the burst. Action potentials that generate the contractions propagate throughout the myoepithelial layer: both their amplitude and duration successively increase during the first part of the burst. To investigate the ionic mechanism of this action potential augmentation, single myoepithelial cells were enzymatically dissociated and whole-cell voltage clamped. Na+, Ca2+ and K+ currents were recorded under different internal and external salt compositions. The Na+ current was blocked by a relatively high concentration (4 micromol l-1 or higher) of tetrodotoxin (TTX), indicating that the Na+ channel belongs to a group of TTX-resistant Na+ channels. The Ca2+ current was blocked by nifedipine (10 micromol l-1) and Co2+ (5 mmol l-1), indicating that the Ca2+ channel is L-type. The K+ current possessed a unique property of long-lasting inactivation. The K+ current fully inactivated during a depolarisation to +30 mV with a time-constant of approximately 9 ms, and the time constant of recovery from inactivation at -70 mV was 13.2 s. This long-lasting inactivation of the K+ channel was the major factor in the augmentation of both action potentials and contractions of the myoepithelial sheet during the initial part of the burst.

  11. X inactivation in a mammal species with three sex chromosomes.

    PubMed

    Veyrunes, Frédéric; Perez, Julie

    2018-06-01

    X inactivation is a fundamental mechanism in eutherian mammals to restore a balance of X-linked gene products between XY males and XX females. However, it has never been extensively studied in a eutherian species with a sex determination system that deviates from the ubiquitous XX/XY. In this study, we explore the X inactivation process in the African pygmy mouse Mus minutoides, that harbours a polygenic sex determination with three sex chromosomes: Y, X, and a feminizing mutant X, named X*; females can thus be XX, XX*, or X*Y, and all males are XY. Using immunofluorescence, we investigated histone modification patterns between the two X chromosome types. We found that the X and X* chromosomes are randomly inactivated in XX* females, while no histone modifications were detected in X*Y females. Furthermore, in M. minutoides, X and X* chromosomes are fused to different autosomes, and we were able to show that the X inactivation never spreads into the autosomal segments. Evaluation of X inactivation by immunofluorescence is an excellent quantitative procedure, but it is only applicable when there is a structural difference between the two chromosomes that allows them to be distinguished.

  12. A new dual-collimation batch reactor for determination of ultraviolet inactivation rate constants for microorganisms in aqueous suspensions

    PubMed Central

    Martin, Stephen B.; Schauer, Elizabeth S.; Blum, David H.; Kremer, Paul A.; Bahnfleth, William P.; Freihaut, James D.

    2017-01-01

    We developed, characterized, and tested a new dual-collimation aqueous UV reactor to improve the accuracy and consistency of aqueous k-value determinations. This new system is unique because it collimates UV energy from a single lamp in two opposite directions. The design provides two distinct advantages over traditional single-collimation systems: 1) real-time UV dose (fluence) determination; and 2) simple actinometric determination of a reactor factor that relates measured irradiance levels to actual irradiance levels experienced by the microbial suspension. This reactor factor replaces three of the four typical correction factors required for single-collimation reactors. Using this dual-collimation reactor, Bacillus subtilis spores demonstrated inactivation following the classic multi-hit model with k = 0.1471 cm2/mJ (with 95% confidence bounds of 0.1426 to 0.1516). PMID:27498232

  13. Survey of microbial degradation of asphalts with notes on relationship to nuclear waste management

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

    ZoBell, C.E.; Molecke, M.A.

    1978-12-01

    A survey has been made of the microbial degradation of asphalts. Topics covered include chemical and physical properties of asphalts, their chemical stability, methods of demonstrating their microbial degradation, and environmental extremes for microbial activity based on existing literature. Specific concerns for the use of asphalt in nuclear waste management, plus potential effects and consequences thereof are discussed. 82 references.

  14. Feedback-Based, System-Level Properties of Vertebrate-Microbial Interactions

    PubMed Central

    Rivas, Ariel L.; Jankowski, Mark D.; Piccinini, Renata; Leitner, Gabriel; Schwarz, Daniel; Anderson, Kevin L.; Fair, Jeanne M.; Hoogesteijn, Almira L.; Wolter, Wilfried; Chaffer, Marcelo; Blum, Shlomo; Were, Tom; Konah, Stephen N.; Kempaiah, Prakash; Ong’echa, John M.; Diesterbeck, Ulrike S.; Pilla, Rachel; Czerny, Claus-Peter; Hittner, James B.; Hyman, James M.; Perkins, Douglas J.

    2013-01-01

    Background Improved characterization of infectious disease dynamics is required. To that end, three-dimensional (3D) data analysis of feedback-like processes may be considered. Methods To detect infectious disease data patterns, a systems biology (SB) and evolutionary biology (EB) approach was evaluated, which utilizes leukocyte data structures designed to diminish data variability and enhance discrimination. Using data collected from one avian and two mammalian (human and bovine) species infected with viral, parasite, or bacterial agents (both sensitive and resistant to antimicrobials), four data structures were explored: (i) counts or percentages of a single leukocyte type, such as lymphocytes, neutrophils, or macrophages (the classic approach), and three levels of the SB/EB approach, which assessed (ii) 2D, (iii) 3D, and (iv) multi-dimensional (rotating 3D) host-microbial interactions. Results In all studies, no classic data structure discriminated disease-positive (D+, or observations in which a microbe was isolated) from disease-negative (D–, or microbial-negative) groups: D+ and D– data distributions overlapped. In contrast, multi-dimensional analysis of indicators designed to possess desirable features, such as a single line of observations, displayed a continuous, circular data structure, whose abrupt inflections facilitated partitioning into subsets statistically significantly different from one another. In all studies, the 3D, SB/EB approach distinguished three (steady, positive, and negative) feedback phases, in which D– data characterized the steady state phase, and D+ data were found in the positive and negative phases. In humans, spatial patterns revealed false-negative observations and three malaria-positive data classes. In both humans and bovines, methicillin-resistant Staphylococcus aureus (MRSA) infections were discriminated from non-MRSA infections. Conclusions More information can be extracted, from the same data, provided that data are

  15. Feedback-based, system-level properties of vertebrate-microbial interactions.

    PubMed

    Rivas, Ariel L; Jankowski, Mark D; Piccinini, Renata; Leitner, Gabriel; Schwarz, Daniel; Anderson, Kevin L; Fair, Jeanne M; Hoogesteijn, Almira L; Wolter, Wilfried; Chaffer, Marcelo; Blum, Shlomo; Were, Tom; Konah, Stephen N; Kempaiah, Prakash; Ong'echa, John M; Diesterbeck, Ulrike S; Pilla, Rachel; Czerny, Claus-Peter; Hittner, James B; Hyman, James M; Perkins, Douglas J

    2013-01-01

    Improved characterization of infectious disease dynamics is required. To that end, three-dimensional (3D) data analysis of feedback-like processes may be considered. To detect infectious disease data patterns, a systems biology (SB) and evolutionary biology (EB) approach was evaluated, which utilizes leukocyte data structures designed to diminish data variability and enhance discrimination. Using data collected from one avian and two mammalian (human and bovine) species infected with viral, parasite, or bacterial agents (both sensitive and resistant to antimicrobials), four data structures were explored: (i) counts or percentages of a single leukocyte type, such as lymphocytes, neutrophils, or macrophages (the classic approach), and three levels of the SB/EB approach, which assessed (ii) 2D, (iii) 3D, and (iv) multi-dimensional (rotating 3D) host-microbial interactions. In all studies, no classic data structure discriminated disease-positive (D+, or observations in which a microbe was isolated) from disease-negative (D-, or microbial-negative) groups: D+ and D- data distributions overlapped. In contrast, multi-dimensional analysis of indicators designed to possess desirable features, such as a single line of observations, displayed a continuous, circular data structure, whose abrupt inflections facilitated partitioning into subsets statistically significantly different from one another. In all studies, the 3D, SB/EB approach distinguished three (steady, positive, and negative) feedback phases, in which D- data characterized the steady state phase, and D+ data were found in the positive and negative phases. In humans, spatial patterns revealed false-negative observations and three malaria-positive data classes. In both humans and bovines, methicillin-resistant Staphylococcus aureus (MRSA) infections were discriminated from non-MRSA infections. More information can be extracted, from the same data, provided that data are structured, their 3D relationships are

  16. Inactivation of enterococci and fecal coliforms from sewage and meatworks effluents in seawater chambers.

    PubMed Central

    Sinton, L W; Davies-Colley, R J; Bell, R G

    1994-01-01

    Inactivation in sunlight of fecal coliforms (FC) and enterococci (Ent) from sewage and meatworks effluents was measured in 300-liter effluent-seawater mixtures (2% vol/vol) held in open-topped chambers. Dark inactivation rates (kDs) were measured (from log-linear survival curves) in enclosed chambers and 6-liter pots. The kD for FC was 2 to 4 times that for Ent, and inactivation was generally slower at lower temperatures. Sunlight inactivation was described in terms of shoulder size (n) and the slope (k) of the log-linear portion of the survival curve as a function of global solar insolation and UV-B fluence. The n values tended to be larger for Ent than for FC, and the k values for FC were around twice those for Ent in both effluent-seawater mixtures. The combined sunlight data showed a general inactivation rate (k) ranking in effluent-seawater mixtures of meatworks FC > sewage FC > meatworks Ent > sewage Ent. Describing 90% inactivation in terms of insolation (S90) gave far less seasonal variation than T90 (time-dependent) values. However, there were significant differences in inactivation rates between experiments, indicating the contribution to inactivation of factors other than insolation. Inactivation rates under different long-pass optical filters decreased with the increase in the spectral cutoff wavelength (lambda 50) of the filters and indicated little contribution by UV-B to total inactivation. Most inactivation appeared to be caused by two main regions of the solar spectrum--between 318 and 340 nm in the UV region and > 400 nm in the visible region. PMID:8031097

  17. Chromosomal inactivation of Bacillus subtilis exfusants: a prokaryotic model of epigenetic regulation.

    PubMed

    Grandjean, V; Hauck, Y; Beloin, C; Le Hégarat, F; Hirschbein, L

    1998-01-01

    Epigenetic mechanisms are not exclusively reserved to eukaryotic organisms. They are also observed in prokaryotes. As described first by Hotchkiss and Gabor, protoplast fusion between strains of Bacillus subtilis produces heterodiploid cells. Heterodiploidy is associated with the inactivation of one of the chromosomes. To study the physical structure of the fusion product and the molecular mechanisms of inactivation, we constructed heterodiploid clones containing two chromosomes labeled by a NotI restriction fragment length polymorphism. In the progeny, we identified haploid recombinant clones that contain a chromosome carrying large regions of inactivated DNA. Studies of both recombinants of the latter kind and heterodiploid cells indicated that chromosomal inactivation was not determined by alteration of the inactivated nucleotide sequence, but was probably due to a modification in the structure of the bacterial chromatin.

  18. [Mechanism of action of neurotoxins acting on the inactivation of voltage-gated sodium channels].

    PubMed

    Benoit, E

    1998-01-01

    This review focuses on the mechanism(s) of action of neurotoxins acting on the inactivation of voltage-gated Na channels. Na channels are transmembrane proteins which are fundamental for cellular communication. These proteins form pores in the plasma membrane allowing passive ionic movements to occur. Their opening and closing are controlled by gating systems which depend on both membrane potential and time. Na channels have three functional properties, mainly studied using electrophysiological and biochemical techniques, to ensure their role in the generation and propagation of action potentials: 1) a highly selectivity for Na ions, 2) a rapid opening ("activation"), responsible for the depolarizing phase of the action potential, and 3) a late closing ("inactivation") involved in the repolarizing phase of the action potential. As an essential protein for membrane excitability, the Na channel is the specific target of a number of vegetal and animal toxins which, by binding to the channel, alter its activity by affecting one or more of its properties. At least six toxin receptor sites have been identified on the neuronal Na channel on the basis of binding studies. However, only toxins interacting with four of these sites (sites 2, 3, 5 et 6) produce alterations of channel inactivation. The maximal percentage of Na channels modified by the binding of neurotoxins to sites 2 (batrachotoxin and some alkaloids), 3 (alpha-scorpion and sea anemone toxins), 5 (brevetoxins and ciguatoxins) et 6 (delta-conotoxins) is different according to the site considered. However, in all cases, these channels do not inactivate. Moreover, Na channels modified by toxins which bind to sites 2, 5 and 6 activate at membrane potentials more negative than do unmodified channels. The physiological consequences of Na channel modifications, induced by the binding of neurotoxins to sites 2, 3, 5 and 6, are (i) an inhibition of cellular excitability due to an important membrane depolarization (site

  19. Inactivation of infectious hematopoietic necrosis virus by low levels of iodine

    USGS Publications Warehouse

    Batts, William N.; Landolt, Marsha L.; Winton, James R.

    1991-01-01

    The fish rhabdovirus infectious hematopoietic necrosis virus (IHNV) was rapidly inactivated by extremely low concentrations of iodine in water. A 99.9% virus reduction was obtained in 7.5 s when virus (105PFU/ml) and iodine (0.1 mg/liter, final concentration) were combined in distilled-deionized or hatchery water. Iodine efficacy decreased at pHs greater than 7.5 or when proteinaceous material was added to the water. Bovine serum albumin blocked iodine inactivation of the virus more effectively than did equal concentrations of fetal bovine serum or river sediment. Sodium thiosulfate effectively neutralized free iodine. Powder, iodophor, and crystalline iodine solutions inactivated IHNV equally. Iodine rapidly inactivated IHNV isolates representing each of the five electropherotypes. Under the conditions used in this study, inactivation was not affected by temperature, salinity, or water hardness. When Dworshak National Fish Hatchery water was continuously treated to provide a free iodine concentration of 0.14 mg/liter, a 7.5-s exposure to iodine was sufficient to inactivate 99.9% of the IHNV. Iodine added to water that contained IHNV prevented infection of rainbow trout (Oncorhynchus mykiss) fry. These results suggest that the waterborne route of IHNV transmission can be blocked by adding low iodine concentrations to the water supplies of hatcheries.

  20. Design and Mechanism of Tetrahydrothiophene-Based γ-Aminobutyric Acid Aminotransferase Inactivators

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

    Le, Hoang V.; Hawker, Dustin D.; Wu, Rui

    Low levels of gamma-aminobutyric acid (GABA), one of two major neurotransmitters that regulate brain neuronal activity, are associated with many neurological disorders, such as epilepsy, Parkinsons disease, Alzheimers disease, Huntingtons disease, and cocaine addiction. One of the main methods to raise the GABA level in human brain is to use small molecules that cross the bloodbrain barrier and inhibit the activity of gamma-aminobutyric acid aminotransferase (GABA-AT), the enzyme that degrades GABA. We have designed a series of conformationally restricted tetrahydrothiophene-based GABA analogues with a properly positioned leaving group that could facilitate a ring-opening mechanism, leading to inactivation of GABA-AT. Onemore » compound in the series is 8 times more efficient an inactivator of GABA-AT than vigabatrin, the only FDA-approved inactivator of GABA-AT. Our mechanistic studies show that the compound inactivates GABA-AT by a new mechanism. The metabolite resulting from inactivation does not covalently bind to amino acid residues of GABA-AT but stays in the active site via H-bonding interactions with Arg-192, a pi-pi interaction with Phe-189, and a weak nonbonded (SO)-O-...=C interaction with Glu-270, thereby inactivating the enzyme.« less

  1. Degradation of microbial polyesters.

    PubMed

    Tokiwa, Yutaka; Calabia, Buenaventurada P

    2004-08-01

    Microbial polyhydroxyalkanoates (PHAs), one of the largest groups of thermoplastic polyesters are receiving much attention as biodegradable substitutes for non-degradable plastics. Poly(D-3-hydroxybutyrate) (PHB) is the most ubiquitous and most intensively studied PHA. Microorganisms degrading these polyesters are widely distributed in various environments. Although various PHB-degrading microorganisms and PHB depolymerases have been studied and characterized, there are still many groups of microorganisms and enzymes with varying properties awaiting various applications. Distributions of PHB-degrading microorganisms, factors affecting the biodegradability of PHB, and microbial and enzymatic degradation of PHB are discussed in this review. We also propose an application of a new isolated, thermophilic PHB-degrading microorganism, Streptomyces strain MG, for producing pure monomers of PHA and useful chemicals, including D-3-hydroxycarboxylic acids such as D-3-hydroxybutyric acid, by enzymatic degradation of PHB.

  2. A specific inactivator of mammalian C'4 isolated from nurse shark (Ginglymostoma cirratum) serum.

    PubMed

    Jensen, J A

    1969-08-01

    A material which specifically inactivates mammalian C'4 was isolated from low ionic strength precipitates of nurse shark serum. The C'4 inactivator was not detected in whole serum. The conditions of its generation and its immunoelectrophoretic behavior seem to indicate that it is an enzymatically formed cleavage product of a precursor contained in whole shark serum. The inactivator was partially purified and characterized. It had an S-value of 3.3 (sucrose gradient) which was in agreement with its retardation on gel filtration, was stable between pH 5.0 and 10.0, had a half-life of 5 min at 56 degrees C, pH 7.5, was inactivated by trypsin and was nontoxic. Its powerful anticomplementary activity in vitro and in vivo was solely due to the rapid inactivation of C'4; no other complement components were affected. No cofactor requirement was observed for the equally rapid inactivation of highly purified human and guinea pig C'4. The kinetics of C'4 inactivation and TAME hydrolysis, the greater anodic mobility of inactivated human C'4, and the influence of temperature on the rate of inactivation suggest that the inactivator is an enzyme and C'4 its substrate. This conclusion was supported by the more recent detection of a split product of C'4. Intravenous administration of the C'4 inactivator could prevent lethal Forssman shock and suppress the Arthus reaction in guinea pigs; it prolonged significantly the rejection time of renal xenografts but had no detectable effect on passive cutaneous anaphylaxis. Anaphylatoxin could be generated in C'4 depleted guinea pig serum with the cobra venom factor, but not with immune precipitates. The possible relationship between C'1 esterase and the C'4 inactivator is discussed on the basis of similarities and dissimilarities.

  3. The prognostic relevance of p16 inactivation in head and neck cancer.

    PubMed

    Koscielny, Sven; Dahse, Regine; Ernst, Gunther; von Eggeling, Ferdinand

    2007-01-01

    The inactivation of the tumor suppressor gene p16 plays an important role in the development of malignant tumors. p16 loss can result from point mutations, loss of heterozygosity (LOH) or methylation of the promoter region. A total of 67 samples of tumor tissue from squamous cell carcinoma of the oral cavity, the pharynx and the larynx were analyzed for an inactivation of p16. The results of the molecular-biological investigations were correlated with the known clinical prognostic parameters after a follow-up period of approximately 3 years. Methylation of the promoter region and LOH were the main mechanisms of p16 inactivation. Point mutations presented as rare events. An inactivation of p16 did not have any statistical influence on tumor prognosis. Patients with a p16 gene inactivated by promoter methylation appeared to have a slightly lower tendency for local and regional recurrences. The inactivation of the tumor suppressor gene p16 plays a role in the carcinogenesis of head and neck cancer. Copyright (c) 2007 S. Karger AG, Basel.

  4. Fast- or Slow-inactivated State Preference of Na+ Channel Inhibitors: A Simulation and Experimental Study

    PubMed Central

    Karoly, Robert; Lenkey, Nora; Juhasz, Andras O.; Vizi, E. Sylvester; Mike, Arpad

    2010-01-01

    Sodium channels are one of the most intensively studied drug targets. Sodium channel inhibitors (e.g., local anesthetics, anticonvulsants, antiarrhythmics and analgesics) exert their effect by stabilizing an inactivated conformation of the channels. Besides the fast-inactivated conformation, sodium channels have several distinct slow-inactivated conformational states. Stabilization of a slow-inactivated state has been proposed to be advantageous for certain therapeutic applications. Special voltage protocols are used to evoke slow inactivation of sodium channels. It is assumed that efficacy of a drug in these protocols indicates slow-inactivated state preference. We tested this assumption in simulations using four prototypical drug inhibitory mechanisms (fast or slow-inactivated state preference, with either fast or slow binding kinetics) and a kinetic model for sodium channels. Unexpectedly, we found that efficacy in these protocols (e.g., a shift of the “steady-state slow inactivation curve”), was not a reliable indicator of slow-inactivated state preference. Slowly associating fast-inactivated state-preferring drugs were indistinguishable from slow-inactivated state-preferring drugs. On the other hand, fast- and slow-inactivated state-preferring drugs tended to preferentially affect onset and recovery, respectively. The robustness of these observations was verified: i) by performing a Monte Carlo study on the effects of randomly modifying model parameters, ii) by testing the same drugs in a fundamentally different model and iii) by an analysis of the effect of systematically changing drug-specific parameters. In patch clamp electrophysiology experiments we tested five sodium channel inhibitor drugs on native sodium channels of cultured hippocampal neurons. For lidocaine, phenytoin and carbamazepine our data indicate a preference for the fast-inactivated state, while the results for fluoxetine and desipramine are inconclusive. We suggest that conclusions

  5. Inactivation of neurotensin and neuromedin N by Zn metallopeptidases.

    PubMed

    Kitabgi, Patrick

    2006-10-01

    The two related peptides neurotensin (NT) and neuromedin N (NN) are efficiently inactivated by peptidases in vitro. Whereas NT is primarily degraded by a combination of three Zn metallo-endopeptidases, namely endopeptidases 24.11, 24.15 and 24.16, in all systems examined, NN is essentially inactivated by the Zn metallo-exopeptidase aminopeptidase M. In this paper we review the work that has led to the identification of the NT- and NN-degrading enzymes and to the purification and cloning of EP 24.16, a previously unidentified peptidase. We provide a brief description of the three NT-inactivating endopeptidases and of their specific and mixed inhibitors, some of them developed in the course of studying NT degradation. Finally, we review in vivo data obtained with these inhibitors that strongly support a physiological role for EP 24.11, 24.15 and 24.16 in the termination of NT-generated signals and for aminopeptidase in terminating NN action. Knowledge of the NT and NN inactivation mechanisms offers the perspective to develop metabolically stable analogs of these peptides with potential therapeutic value.

  6. Inactivation of human and simian rotaviruses by chlorine dioxide

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

    Chen, Yu-Shiaw; Vaughn, J.M.

    1990-05-01

    The inactivation of single-particle stocks of human (type 2, Wa) and simian (SA-11) rotaviruses by chlorine dioxide was investigated. Experiments were conducted at 4{degree}C in a standard phosphate-carbonate buffer. Both virus types were rapidly inactivated, within 20 s under alkaline conditions, when chlorine dioxide concentrations ranging from 0.05 to 0.2 mg/liter were used. Similar reductions of 10{sup 5}-fold in infectivity required additional exposure time of 120 s at 0.2 mg/liter for Wa and at 0.5 mg/liter for SA-11, respectively, at pH 6.0. The inactivation of both virus types was moderate a neutral pH, and the sensitivities to chlorine dioxide weremore » similar. The observed enhancement of virucidal efficiency with increasing pH was contrary to earlier findings with chlorine- and ozone-treated rotavirus particles, where efficiencies decreased with increasing alkalinity. Comparison of 99.9% virus inactivation times revealed ozone to be the most effective virucidal agent among these three disinfectants.« less

  7. Microbial astronauts: assembling microbial communities for advanced life support systems

    NASA Technical Reports Server (NTRS)

    Roberts, M. S.; Garland, J. L.; Mills, A. L.

    2004-01-01

    Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem

  8. Microbial astronauts: assembling microbial communities for advanced life support systems.

    PubMed

    Roberts, M S; Garland, J L; Mills, A L

    2004-02-01

    Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem

  9. [Carbon sources metabolic characteristics of airborne microbial communities in constructed wetlands].

    PubMed

    Song, Zhi-Wen; Wang, Lin; Xu, Ai-Ling; Wu, Deng-Deng; Xia, Yan

    2015-02-01

    Using BIOLOG-GN plates, this article describes the carbon sources metabolic characteristics of airborne microbial communities in a free surface-flow constructed wetland in different seasons and clarify the correlation between airborne microbial metabolic functions and environmental factors. The average well color development (AWCD), carbon metabolic profiles and McIntosh values of airborne microbial communities in different seasons were quite different. Analysis of the variations showed that AWCD in spring and summer differed significantly from that in autumn and winter (P < 0.01). In the same season, the degree of utilization of different types of carbon by airborne microbes was different. Summer had a significant difference from other seasons (P < 0.05). Dominant communities of airborne microbes in four seasons were carboxylic acids metabolic community, carbohydrates metabolic community, polymers metabolic community and carboxylic acids metabolic community respectively. Principal component analysis showed that the carbon metabolic characteristics of airborne microbial community in autumn were similar to those in winter but different from those in spring and summer. The characteristics of carbon metabolism revealed differences between summer and spring, autumn, or winter. These differences were mainly caused by amines or amides while the differences between spring and autumn or winter were mainly caused by carboxylic acids. Environmental factors, including changes in wind speed, temperature, and humidity acted to influence the carbon sources metabolic properties of airborne microbial community. The dominant environmental factors that acted to influence the carbon sources metabolic properties of airborne microbial community varied between different seasons.

  10. Anti-oxidant and anti-microbial properties of some ethno-therapeutically important medicinal plants of Indian Himalayan Region.

    PubMed

    Rawat, Sandeep; Jugran, Arun K; Bahukhandi, Amit; Bahuguna, Asutosh; Bhatt, Indra D; Rawal, Ranbeer S; Dhar, Uppeandra

    2016-12-01

    Therapeutic potential of medicinal plants as a source of noble natural anti-oxidants and anti-microbial agents has been well recognised all across the globe. In this study, phenolic compounds, in vitro anti-oxidant activity and anti-microbial properties have been investigated in five Himalayan medicinal plants, (e.g., Acorus calamus, Habenaria intermedia, Hedychium spicatum, Roscoea procera and Valeriana jatamansi) in different solvent systems. R. procera exhibited significantly (p < 0.05) higher phenolics; while H. spicatum was rich in flavonoids and V. jatamansi in anti-oxidant activity. Also, R. procera and H. spicatum were found rich in gallic acid; V. jatamansi in catechin, hydroxylbenzoic acid and caffeic acid and H. intermedia in hydroxyl benzoic acid. Solvent systems showed species specific response for extraction of total flavonoids and anti-oxidant activity. All the extracts were found effective against different bacterial and fungal strains in a dose dependent manner and maximum antimicrobial activity was found in R. procera as compared to other species. All the plant extracts showed greater activity against bacterial strains as compared to fungal strains. The results of this study suggest that extract of these species can be used as natural anti-oxidant to reduce free radical mediated disorders and as natural alternative for food preservation.

  11. Microbial Quality and Shelf Life of Blueberry Purée Developed Using Cavitation Technology.

    PubMed

    Fan, Lihua; Martynenko, Alex; Doucette, Craig; Hughes, Timothy; Fillmore, Sherry

    2018-03-01

    Blueberry purée was developed using hydrodynamic cavitation technology. The product was made from entire blueberries without adding any food additives. In this study, microbial reduction following each processing stage (at the industry setting) and after product pasteurization at 86, 88, 90, 92, 94, and 96 °C was investigated. Microbial quality including total plate counts, yeast and molds, and heat-resistant molds counts was determined. Shelf life of pasteurized products stored for up to 24 weeks at room temperature were assessed for microbial quality, soluble solids (°Brix), titratable acidity (citric acid %), pH, viscosity (cP) and flow rate (cm/30 s). Our results indicated that heat-resistant molds, initially present in frozen blueberries with counts at 2.03 log CFU/200g, were totally inactivated at 94 to 96 °C with 1 to 2 min holding time. Shelf life study showed that no product spoilage was caused by bacteria, yeasts and heat-resistant molds along with non-significant changes of textural characteristics. This study provided useful information for the food industry to develop variety of fruit purée products with no wastes of fruit materials. This study provides useful information for the food industry to develop safe liquid food products using cavitation technology without wasting any raw materials. © 2018 Institute of Food Technologists®.

  12. A novel method for bacterial inactivation using electrosprayed water nanostructures

    NASA Astrophysics Data System (ADS)

    Pyrgiotakis, Georgios; McDevitt, James; Yamauchi, Toshiyuki; Demokritou, Philip

    2012-08-01

    This is a study focusing on the potential to deactivate biological agents (bacteria and endospores) using engineered water nanostructures (EWNS). The EWNS were generated using an electrospray device that collects water by condensing atmospheric water vapor on a Peltier-cooled electrode. A high voltage is applied between the collection electrode and a grounded electrode resulting in aerosolization of the condensed water and a constant generation of EWNS. Gram-negative Serratia marcescens, gram-positive Staphylococcus aureus, and Bacillus atrophaeus endospores were placed on stainless steel coupons and exposed to generated EWNS at multiple time intervals. Upon exposures, the bacteria were recovered and placed on nutrient agar to grow, and the colony forming units were counted. Ozone levels as well as air temperature and relative humidity were monitored during the experiments. Qualitative confirmation of bacterial destruction was also obtained by transmission electron microscopy. In addition, important EWNS aerosol properties such as particle number concentration as a function of size as well as the average surface charge of the generated EWNS were measured using real-time instrumentation. It was shown that the novel electrospray method can generate over time a constant flux of EWNS. EWNS have a peak number concentration of 8,000 particles per cubic centimeter with a modal peak size around 20 nm. The average surface charge of the generated EWNS was found to be 10 ± 2 electrons per particle. In addition, it was shown that the EWNS have the potential to deactivate both bacteria types from surfaces. At the same administrate dose, however, the endospores were not inactivated. This novel method and the unique properties of the generated EWNS could potentially be used to develop an effective, environmentally friendly, and inexpensive method for bacteria inactivation.

  13. Influenza (flu) vaccine (Inactivated or Recombinant): What you need to know

    MedlinePlus

    ... taken in its entirety from the CDC Inactivated Influenza Vaccine Information Statement (VIS) www.cdc.gov/vaccines/hcp/vis/vis-statements/flu.html CDC review information for Inactivated Influenza VIS: ...

  14. Histone underacetylation is an ancient component of mammalian X chromosome inactivation

    PubMed Central

    Wakefield, Matthew J.; Keohane, Ann M.; Turner, Bryan M.; Graves, Jennifer A. Marshall

    1997-01-01

    Underacetylation of histone H4 is thought to be involved in the molecular mechanism of mammalian X chromosome inactivation, which is an important model system for large-scale genetic control in eukaryotes. However, it has not been established whether histone underacetylation plays a critical role in the multistep inactivation pathway. Here we demonstrate differential histone H4 acetylation between the X chromosomes of a female marsupial, Macropus eugenii. Histone underacetylation is the only molecular aspect of X inactivation known to be shared by marsupial and eutherian mammals. Its strong evolutionary conservation implies that, unlike DNA methylation, histone underacetylation was a feature of dosage compensation in a common mammalian ancestor, and is therefore likely to play a central role in X chromosome inactivation in all mammals. PMID:9275180

  15. Acoustic and Electrical Property Changes Due to Microbial Growth and Biofilm Formation in Porous Media

    EPA Science Inventory

    A laboratory study was conducted to investigate the effect of microbial growth and biofilm formation on compressional waves, and complex conductivity during stimulated microbial growth. Over the 29 day duration of the experiment, compressional wave amplitudes and arrival times f...

  16. Biophysical processes supporting the diversity of microbial life in soil

    PubMed Central

    Tecon, Robin

    2017-01-01

    Abstract Soil, the living terrestrial skin of the Earth, plays a central role in supporting life and is home to an unimaginable diversity of microorganisms. This review explores key drivers for microbial life in soils under different climates and land-use practices at scales ranging from soil pores to landscapes. We delineate special features of soil as a microbial habitat (focusing on bacteria) and the consequences for microbial communities. This review covers recent modeling advances that link soil physical processes with microbial life (termed biophysical processes). Readers are introduced to concepts governing water organization in soil pores and associated transport properties and microbial dispersion ranges often determined by the spatial organization of a highly dynamic soil aqueous phase. The narrow hydrological windows of wetting and aqueous phase connectedness are crucial for resource distribution and longer range transport of microorganisms. Feedbacks between microbial activity and their immediate environment are responsible for emergence and stabilization of soil structure—the scaffolding for soil ecological functioning. We synthesize insights from historical and contemporary studies to provide an outlook for the challenges and opportunities for developing a quantitative ecological framework to delineate and predict the microbial component of soil functioning. PMID:28961933

  17. CRYPTOSPORIDIUM INACTIVATION AND REMOVAL RESEARCH

    EPA Science Inventory

    Bench- and pilot-scale tests were performed to assess the ability of conventional treatment, ozonation and chlorine dioxide to remove and inactivate Cryptosporidium oocysts. The impacts of coagulant type, coagulant dose, raw water quality, filter loading rates and filter media w...

  18. CHLORINE INACTIVATION OF BACILLUS ENDOSPORES

    EPA Science Inventory

    The possibility of a bioterrorism event resulting in the release of Bacillus anthracis endospores into a drinking water distribution system necessitates research into means by which these endospores can be inactivated. This study was designed to determine the chlorine resistance...

  19. Chemical-free inactivated whole influenza virus vaccine prepared by ultrashort pulsed laser treatment

    NASA Astrophysics Data System (ADS)

    Tsen, Shaw-Wei David; Donthi, Nisha; La, Victor; Hsieh, Wen-Han; Li, Yen-Der; Knoff, Jayne; Chen, Alexander; Wu, Tzyy-Choou; Hung, Chien-Fu; Achilefu, Samuel; Tsen, Kong-Thon

    2015-05-01

    There is an urgent need for rapid methods to develop vaccines in response to emerging viral pathogens. Whole inactivated virus (WIV) vaccines represent an ideal strategy for this purpose; however, a universal method for producing safe and immunogenic inactivated vaccines is lacking. Conventional pathogen inactivation methods such as formalin, heat, ultraviolet light, and gamma rays cause structural alterations in vaccines that lead to reduced neutralizing antibody specificity, and in some cases, disastrous T helper type 2-mediated immune pathology. We have evaluated the potential of a visible ultrashort pulsed (USP) laser method to generate safe and immunogenic WIV vaccines without adjuvants. Specifically, we demonstrate that vaccination of mice with laser-inactivated H1N1 influenza virus at about a 10-fold lower dose than that required using conventional formalin-inactivated influenza vaccines results in protection against lethal H1N1 challenge in mice. The virus, inactivated by the USP laser irradiation, has been shown to retain its surface protein structure through hemagglutination assay. Unlike conventional inactivation methods, laser treatment did not generate carbonyl groups in protein, thereby reducing the risk of adverse vaccine-elicited T helper type 2 responses. Therefore, USP laser treatment is an attractive potential strategy to generate WIV vaccines with greater potency and safety than vaccines produced by current inactivation techniques.

  20. Ultraviolet-C light inactivation of Penicillium expansum on fruit surfaces

    USDA-ARS?s Scientific Manuscript database

    Understanding the influence of fruit surface morphology on ultraviolet-C (UV-C 254 nm) inactivation of microorganisms is required for designing effective treatment systems. In this study, we analyzed UV-C inactivation of Penicillium expansum that was inoculated onto the surface of organic fruits. Re...