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Sample records for lymnaea cubensis molusco

  1. Characterisation of Lymnaea cubensis, L. viatrix and L. neotropica n. sp., the main vectors of Fasciola hepatica in Latin America, by analysis of their ribosomal and mitochondrial DNA.

    PubMed

    Bargues, M D; Artigas, P; Mera Y Sierra, R L; Pointier, J P; Mas-Coma, S

    2007-10-01

    Although, in the endemic areas throughout the world, human fascioliasis presents varying patterns in its epidemiology, the species of lymnaeid snail that act as intermediate hosts and vectors are always crucial in the transmission of the causative parasites. Species in the Galba/Fossaria group of snails, such as Lymnaea cubensis, L. viatrix var. A ventricosa, L. viatrix var. B elongata and Galba truncatula, appear to be frequently involved in the transmission of Fasciola hepatica in Central and South America, although specific classification within this morphologically and anatomically confusing group is often very difficult. To explore the potential use of molecular analyses in the identification of vector snails, regions of the ribosomal DNA - the small subunit (18S) gene and internal transcribed spacers (ITS-2 and ITS-1) - and of the mitochondrial DNA - the cytochrome c oxidase subunit I (COI) - of wild-caught lymnaeid snails of L. cubensis, L. viatrix var. A ventricosa, L. viatrix var. B elongata and G. truncatula have been sequenced. The samples of the Latin American species included specimens from the respective type localities. The genetic distances observed and the results of phylogenetic analyses demonstrate that two different species exist within L. viatrix. Lymnaea neotropica n. sp. (=L. viatrix var. B elongata) is here proposed for specimens from Lima, Peru, and is differentiated from L. viatrix (=L. viatrix var. A ventricosa), L. cubensis and G. truncatula. The data collected on the 18S ribosomal-RNA gene indicate that the snails investigated may cover more than one supraspecific taxon. The ITS-2, ITS-1 and COI nucleotide sequences are clearly useful markers for the differentiation of these morpho-anatomically similar lymnaeid species. The numerous microsatellite repeats found within ITS-2 are potential tools for differentiation at population level.

  2. Seed Transmission of Pseudoperonospora cubensis

    PubMed Central

    Cohen, Yigal; Rubin, Avia E.; Galperin, Mariana; Ploch, Sebastian; Runge, Fabian; Thines, Marco

    2014-01-01

    Pseudoperonospora cubensis, an obligate biotrophic oomycete causing devastating foliar disease in species of the Cucurbitaceae family, was never reported in seeds or transmitted by seeds. We now show that P. cubensis occurs in fruits and seeds of downy mildew-infected plants but not in fruits or seeds of healthy plants. About 6.7% of the fruits collected during 2012–2014 have developed downy mildew when homogenized and inoculated onto detached leaves and 0.9% of the seeds collected developed downy mildew when grown to the seedling stage. This is the first report showing that P. cubensis has become seed-transmitted in cucurbits. Species-specific PCR assays showed that P. cubensis occurs in ovaries, fruit seed cavity and seed embryos of cucurbits. We propose that international trade of fruits or seeds of cucurbits might be associated with the recent global change in the population structure of P. cubensis. PMID:25329308

  3. Genetic and pathogenic relatedness of Pseudoperonospora cubensis and P. humuli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The most economically important plant pathogens in the genus Pseudoperonospora (Peronosporaceae) are P. cubensis and P. humuli, causal agents of downy mildew on cucurbits and hop, respectively. Recently, P. humuli was reduced to a taxonomic synonym of P. cubensis based on internal transcribed spacer...

  4. Field response of cucurbit hosts to Pseudoperonospora cubensis in Michigan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Downy mildew, caused by Pseudoperonospora cubensis, is a severe foliar disease of many cucurbit crops worldwide. Forty-one cucurbit cultigens (commercial cultivars and plant introductions) from five genera (Cucumis, Citrullus, Cucurbita, Lagenaria, and Luffa) were assessed for susceptibility to Ps....

  5. Ecological observations on Lymnaea (Bullastra) cumingiana.

    PubMed

    Monzon, R B; Kitikoon, V; Thammapalerd, N; Temcharoen, P; Sornmani, S; Viyanant, V

    1993-09-01

    Field surveys conducted at Echague, Isabela and San Pablo, Laguna revealed that Lymnaea (Bullastra) cumingiana, the natural second snail intermediate host of Echinostoma malayanum in the Philippines, exhibits a moderate degree of diversity in its choice of habitats. Rice fields of all stages of development, stagnant shallow streams and springs are the main areas where the snail can be collected from at Echague, Isabela. However, they were absent in rice fields that had been extensively sprayed with molluscicides to control the "golden apple snail" (Ampullarius canaliculatus). In contrast, they were also very abundant in the highly eutrophic waters of Sampaloc lake, San Pablo, Laguna. L. cumingiana co-exists with various species of insects, snails, fish and plants in these habitats. Information on ecological characteristics affecting its distribution will be useful for those who wish to collect and study this species in the future.

  6. Resurgence of Pseudoperonospora cubensis: The Causal Agent of Cucurbit Downy Mildew.

    PubMed

    Cohen, Yigal; Van den Langenberg, Kyle M; Wehner, Todd C; Ojiambo, Peter S; Hausbeck, Mary; Quesada-Ocampo, Lina M; Lebeda, Aleš; Sierotzki, Helge; Gisi, Ulrich

    2015-07-01

    The downy mildew pathogen, Pseudoperonospora cubensis, which infects plant species in the family Cucurbitaceae, has undergone major changes during the last decade. Disease severity and epidemics are far more destructive than previously reported, and new genotypes, races, pathotypes, and mating types of the pathogen have been discovered in populations from around the globe as a result of the resurgence of the disease. Consequently, disease control through host plant resistance and fungicide applications has become more complex. This resurgence of P. cubensis offers challenges to scientists in many research areas including pathogen biology, epidemiology and dispersal, population structure and population genetics, host preference, host-pathogen interactions and gene expression, genetic host plant resistance, inheritance of host and fungicide resistance, and chemical disease control. This review serves to summarize the current status of this major pathogen and to guide future management and research efforts within this pathosystem.

  7. Identification of genetic variation between obligate plant pathogens Psuedoperonospora cubensis and P. humuli using RNA sequencing and genotyping-by-sequencing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    RNA sequencing (RNA-seq) and genotyping-by-sequencing (GBS) were used for single nucleotide polymorphism (SNP) identification from two economically important obligate plant pathogens, Pseudoperonospora cubensis and P. humuli. Twenty isolates of P. cubensis and 19 isolates of P. humuli were genotyped...

  8. Octopamine: a new feeding modulator in Lymnaea

    PubMed Central

    Vehovszky, Á.

    1998-01-01

    The role of octopamine (OA) in the feeding system of the pond snail, Lymnaea stagnalis, was studied by applying behavioural tests on intact animals, and a combination of electrophysiological analysis and morphological labelling in the isolated central nervous system. OA antagonists phentolamine, demethylchlordimeform (DCDM) and 2-chloro-4-methyl-2-(phenylimino)-imidazolidine (NC-7) were injected into intact snails and the sucrose-induced feeding response of animals was monitored. Snails that received 25 to 50 mg kg-1 phentolamine did not start feeding in sucrose, and the same dose of NC-7 reduced the number of feeding animals by 80 to 90% 1 to 3 hours after injection. DCDM treatment reduced feeding by 20 to 60%. In addition, both phentolamine and NC-7 significantly decreased the feeding rate of those animals that still accepted food after 1 to 6 hours of injection. In the central nervous system a pair of buccal neurons was identified by electrophysiological and morphological criteria. After double labelling (intracellular staining with Lucifer yellow followed by OA-immunocytochemistry) these neurons were shown to be OA immunoreactive, and electrophysiological experiments confirmed that they are members of the buccal feeding system. Therefore the newly identified buccal neurons were called OC neurons (putative octopamine containing neurons or octopaminergic cells). Synchronous intracellular recordings demonstrated that the OC neurons share a common rhythm with feeding neurons either appearing spontaneously or evoked by intracellularly stimulated feeding interneurons. OC neurons also have synaptic connections with identified members of the feeding network: electrical coupling was demonstrated between OC neurons and members of the B4 cluster motoneurons, furthermore, chemically transmitted synaptic responses were recorded both on feeding motoneurons (B1, B2 cells) and the SO modulatory interneuron after the stimulation of OC neurons. However, elementary synaptic

  9. [Anatomy and morphometry of Physa cubensis Pfeiffer, 1839 (Pulmonata: Physidae) in Cuba].

    PubMed

    Cong, M Y; Ferrer López, J R; Perera de Puga, G

    1994-01-01

    The anatomic description of Physa cubensis, based on specimens collected in 5 different sites in Cuba, is presented. The anatomic characteristics of the reproductive system as well as mantle digitations give ground for affirming that this is a unique species with external morphological variations and thus it should be kept under the Physa genre. Besides, dispersion diagrams and regression lines of the length in the width were analyzed in the five sites; the L/A (length/width) variation quotient was compared using a variance analysis. Significant differences were observed in the different sites probably due to the feeding sources of each biotope.

  10. Central Localization of Plasticity Involved in Appetitive Conditioning in "Lymnaea"

    ERIC Educational Resources Information Center

    Straub, Volko A.; Styles, Benjamin J.; Ireland, Julie S.; O'Shea, Michael; Benjamin, Paul R.

    2004-01-01

    Learning to associate a conditioned (CS) and unconditioned stimulus (US) results in changes in the processing of CS information. Here, we address directly the question whether chemical appetitive conditioning of "Lymnaea" feeding behavior involves changes in the peripheral and/or central processing of the CS by using extracellular recording…

  11. Pseudoperonospora cubensis and P. humuli detection using species-specific probes and high definition melt curve analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three assays were developed for molecular differentiation of Pseudoperonospora cubensis and P. humuli, causal agents of cucurbit and hop downy mildew, respectively, for detection of airborne sporangia and diagnosis of symptomatic leaf tissue. The assays were based on previously identified single nuc...

  12. A multiplex PCR for the detection of Fasciola hepatica in the intermediate snail host Galba cubensis.

    PubMed

    Alba, Annia; Vázquez, Antonio A; Hernández, Hilda; Sánchez, Jorge; Marcet, Ricardo; Figueredo, Mabel; Sarracent, Jorge; Fraga, Jorge

    2015-07-30

    Fasciolosis is a snail-borne trematode infection that has re-emerged as a human disease, and is considered a significant problem for veterinary medicine worldwide. The evaluation of the transmission risk of fasciolosis as well as the efficacy of the strategies for its control could be carried out through epidemiological surveillance of the snails that act as intermediate hosts of the parasites. The present study aimed to develop the first multiplex PCR to detect Fasciola hepatica in Galba cubensis, an important intermediate host of the parasite in the Americas and especially in the Caribbean basin. The multiplex PCR was optimized for the amplification of a 340 bp fragment of the second internal transcribed spacer (ITS-2) of F. hepatica rDNA, while another set of primers was designed and used to amplify a conserved segment of the nuclear 18S rDNA of the snail (451 bp), as an internal control of the reaction. The assay was able to detect up to 100 pg of the parasite even at high concentrations of snail DNA, an analytical sensitivity that allows the detection of less than a single miracidium, which is the minimal biological infestation unit. A controlled laboratory-reared G. cubensis - F. hepatica system was used for the evaluation of the developed multiplex PCR, and 100% sensitivity and specificity was achieved. This assay constitutes a novel, useful and suitable technique for the survey of fasciolosis transmission through one of the main intermediate hosts in the Western hemisphere.

  13. The effect of particle size on the leaching of Scirpus cubensis Poepp & Kunth.

    PubMed

    Bianchini Júnior, I; Antonio, R M

    2003-05-01

    An investigation was made on the effects of detritus particle size on leaching rates in organic matter, and the associated environmental changes caused by detritus re-cycling in an oxbow lake (Lagoa do Infernão). Experiments were conducted during the decay of an aquatic macrophyte specie, S. cubensis, which in turn led to the formation of colored compounds. The S. cubensis were collected from the Lagoa do Infernão and taken to the laboratory where they were washed, dried, and fractionated using a sieve pedological set. The detritus was classified into six groups according to size, viz. 100, 10, 1.13, 0.78, 0.61, and 0.25 mm. Overall, the fragmentation process tended to increase the detritus fraction to be dissolved and to decrease the leaching rates owing to the possible dissolution of refracting matter. Fragmentation also caused the amount of colored compounds to increase and appeared to favor dissolved electrolyte release. Finally, in Lagoa do Infernão fragmentation is probably mediated by the metabolic action of benthic communities.

  14. Occurrence and Distribution of Mating Types of Pseudoperonospora cubensis in the United States.

    PubMed

    Thomas, Anna; Carbone, Ignazio; Cohen, Yigal; Ojiambo, Peter S

    2017-03-01

    During the past two decades, a resurgence of cucurbit downy mildew has occurred around the world, resulting in severe disease epidemics. In the United States, resurgence of the disease occurred in 2004 and several hypotheses, including introduction of a new genetic recombinant or pathotype of the pathogen, have been suggested as potential causes for this resurgence. Occurrence and distribution of mating types of Pseudoperonospora cubensis in the United States were investigated using 40 isolates collected from cucurbits across 11 states from 2005 to 2013. Pairing of unknown isolates with known mating-type tester strains on detached leaves of cantaloupe or cucumber resulted in oospore formation 8 to 10 days after inoculation. Isolates differed in their ability to form oospores across all coinoculation pairings, with oospore numbers ranging from 280 to 1,000 oospores/cm(2) of leaf tissue. Oospores were hyaline to golden-yellow, spherical, and approximately 36 μm in diameter. Of the 40 isolates tested, 24 were found to be of the A1 mating type, while 16 were of the A2 mating type. Mating type was significantly (P < 0.0001) associated with host type, whereby all isolates collected from cucumber were of the A1 mating type, while isolates from squash and watermelon were of the A2 mating type. Similarly, mating type was significantly (P = 0.0287) associated with geographical region, where isolates from northern-tier states of Michigan, New Jersey, New York, and Ohio were all A1, while isolates belonging to either A1 or A2 mating type were present in equal proportions in southern-tier states of Alabama, Florida, Georgia, North Carolina, South Carolina, and Texas. Viability assays showed that oospores were viable and, on average, approximately 40% of the oospores produced were viable as determined by the plasmolysis method. This study showed that A1 and A2 mating types of P. cubensis are present and the pathogen could potentially reproduce sexually in cucurbits within the

  15. Distributed network organization underlying feeding behavior in the mollusk Lymnaea.

    PubMed

    Benjamin, Paul R

    2012-04-17

    The aim of the work reviewed here is to relate the properties of individual neurons to network organization and behavior using the feeding system of the gastropod mollusk, Lymnaea. Food ingestion in this animal involves sequences of rhythmic biting movements that are initiated by the application of a chemical food stimulus to the lips and esophagus. We investigated how individual neurons contribute to various network functions that are required for the generation of feeding behavior such as rhythm generation, initiation ('decision making'), modulation and hunger and satiety. The data support the view that feeding behavior is generated by a distributed type of network organization with individual neurons often contributing to more than one network function, sharing roles with other neurons. Multitasking in a distributed type of network would be 'economically' sensible in the Lymnaea feeding system where only about 100 neurons are available to carry out a variety of complex tasks performed by millions of neurons in the vertebrate nervous system. Having complementary and potentially alternative mechanisms for network functions would also add robustness to what is a 'noisy' network where variable firing rates and synaptic strengths are commonly encountered in electrophysiological recording experiments.

  16. Expression profiling of Cucumis sativus in response to infection by Pseudoperonospora cubensis.

    PubMed

    Adhikari, Bishwo N; Savory, Elizabeth A; Vaillancourt, Brieanne; Childs, Kevin L; Hamilton, John P; Day, Brad; Buell, C Robin

    2012-01-01

    The oomycete pathogen, Pseudoperonospora cubensis, is the causal agent of downy mildew on cucurbits, and at present, no effective resistance to this pathogen is available in cultivated cucumber (Cucumis sativus). To better understand the host response to a virulent pathogen, we performed expression profiling throughout a time course of a compatible interaction using whole transcriptome sequencing. As described herein, we were able to detect the expression of 15,286 cucumber genes, of which 14,476 were expressed throughout the infection process from 1 day post-inoculation (dpi) to 8 dpi. A large number of genes, 1,612 to 3,286, were differentially expressed in pair-wise comparisons between time points. We observed the rapid induction of key defense related genes, including catalases, chitinases, lipoxygenases, peroxidases, and protease inhibitors within 1 dpi, suggesting detection of the pathogen by the host. Co-expression network analyses revealed transcriptional networks with distinct patterns of expression including down-regulation at 2 dpi of known defense response genes suggesting coordinated suppression of host responses by the pathogen. Comparative analyses of cucumber gene expression patterns with that of orthologous Arabidopsis thaliana genes following challenge with Hyaloperonospora arabidopsidis revealed correlated expression patterns of single copy orthologs suggesting that these two dicot hosts have similar transcriptional responses to related pathogens. In total, the work described herein presents an in-depth analysis of the interplay between host susceptibility and pathogen virulence in an agriculturally important pathosystem.

  17. Alternative Splicing of a Multi-Drug Transporter from Pseudoperonospora cubensis Generates an RXLR Effector Protein That Elicits a Rapid Cell Death

    PubMed Central

    Savory, Elizabeth A.; Zou, Cheng; Adhikari, Bishwo N.; Hamilton, John P.; Buell, C. Robin; Shiu, Shin-Han; Day, Brad

    2012-01-01

    Pseudoperonospora cubensis, an obligate oomycete pathogen, is the causal agent of cucurbit downy mildew, a foliar disease of global economic importance. Similar to other oomycete plant pathogens, Ps. cubensis has a suite of RXLR and RXLR-like effector proteins, which likely function as virulence or avirulence determinants during the course of host infection. Using in silico analyses, we identified 271 candidate effector proteins within the Ps. cubensis genome with variable RXLR motifs. In extending this analysis, we present the functional characterization of one Ps. cubensis effector protein, RXLR protein 1 (PscRXLR1), and its closest Phytophthora infestans ortholog, PITG_17484, a member of the Drug/Metabolite Transporter (DMT) superfamily. To assess if such effector-non-effector pairs are common among oomycete plant pathogens, we examined the relationship(s) among putative ortholog pairs in Ps. cubensis and P. infestans. Of 271 predicted Ps. cubensis effector proteins, only 109 (41%) had a putative ortholog in P. infestans and evolutionary rate analysis of these orthologs shows that they are evolving significantly faster than most other genes. We found that PscRXLR1 was up-regulated during the early stages of infection of plants, and, moreover, that heterologous expression of PscRXLR1 in Nicotiana benthamiana elicits a rapid necrosis. More interestingly, we also demonstrate that PscRXLR1 arises as a product of alternative splicing, making this the first example of an alternative splicing event in plant pathogenic oomycetes transforming a non-effector gene to a functional effector protein. Taken together, these data suggest a role for PscRXLR1 in pathogenicity, and, in total, our data provide a basis for comparative analysis of candidate effector proteins and their non-effector orthologs as a means of understanding function and evolutionary history of pathogen effectors. PMID:22496844

  18. Which Morphological Characteristics Are Most Influenced by the Host Matrix in Downy Mildews? A Case Study in Pseudoperonospora cubensis

    PubMed Central

    Runge, Fabian; Ndambi, Beninweck; Thines, Marco

    2012-01-01

    Before the advent of molecular phylogenetics, species concepts in the downy mildews, an economically important group of obligate biotrophic oomycete pathogens, have mostly been based upon host range and morphology. While molecular phylogenetic studies have confirmed a narrow host range for many downy mildew species, others, like Pseudoperonospora cubensis affect even different genera. Although often morphological differences were found for new, phylogenetically distinct species, uncertainty prevails regarding their host ranges, especially regarding related plants that have been reported as downy mildew hosts, but were not included in the phylogenetic studies. In these cases, the basis for deciding if the divergence in some morphological characters can be deemed sufficient for designation as separate species is uncertain, as observed morphological divergence could be due to different host matrices colonised. The broad host range of P. cubensis (ca. 60 host species) renders this pathogen an ideal model organism for the investigation of morphological variations in relation to the host matrix and to evaluate which characteristics are best indicators for conspecificity or distinctiveness. On the basis of twelve morphological characterisitcs and a set of twelve cucurbits from five different Cucurbitaceae tribes, including the two species, Cyclanthera pedata and Thladiantha dubia, hitherto not reported as hosts of P. cubensis, a significant influence of the host matrix on pathogen morphology was found. Given the high intraspecific variation of some characteristics, also their plasticity has to be taken into account. The implications for morphological species determination and the confidence limits of morphological characteristics are discussed. For species delimitations in Pseudoperonospora it is shown that the ratio of the height of the first ramification to the sporangiophore length, ratio of the longer to the shorter ultimate branchlet, and especially the length and

  19. Copper toxicity to the fresh water snail, Lymnaea luteola

    SciTech Connect

    Reddy, N.M.; Rao, P.V.

    1987-07-01

    Haemocyanins are found in arthropoda and mollusca and show a copper content characteristic for each phylum. Heavy metal accumulation by mollusks is widely reported. Approximately one third of the enzymes either required addition of a metal ion as a cofactor in order to exhibit maximum activity or contained a slightly bound metal ion which appeared to be involved in the catalytic process. Copper is the only metal which has been detected in significant amounts in amino oxidase. The present study is designed to evaluate the influence of such copper, which is of such common occurrence in biological material, on some of the lipolytic enzymes of fresh water pulmonate snail, Lymnaea luteola when added to ambient medium. The present study also highlights the possible detoxification mechanism prevailing in this fresh water mollusk.

  20. Characterization of pre- and postsynaptic dopamine receptors in Lymnaea.

    PubMed

    Audesirk, T E

    1989-01-01

    1. The effects of dopamine and several synthetic agonists and antagonists were studied using two identified neurons of the snail Lymnaea stagnalis. 2. In both the buccal-2 (B-2) neurons and the pedal giant (RPeD1) neuron dopamine elicited a hyperpolarizing response at least partly due to potassium efflux. RPeD1 is itself dopaminergic, implicating autoreceptors in its response to dopamine. 3. The following agents were tested: agonists--LY171555, pergolide, SKF38393, (-)-3-PPP, R(-)NPA and dopamine; antagonists--SCH23390, sulpiride, and metaclopramide. Dibutyryl cAMP was applied to determine whether the response is cAMP-mediated. 4. Results indicate that the pharmacological profiles of dopamine receptors on these neurons are inconsistent with those of either D-1, D-2 or autoreceptors in mammals.

  1. Effects of millimeter waves on ionic currents of Lymnaea neurons.

    PubMed

    Alekseev, S I; Ziskin, M C

    1999-01-01

    The effects of mm-waves 60.22-62.22 GHz and 75 GHz on A-type K+ currents and the effects of 61.22 GHz on Ca2+ currents of Lymnaea neurons were investigated using a whole-cell voltage-clamp technique. The open end of a rectangular waveguide covered with a thin Teflon film served as a radiator. Specific absorption rates at the waveguide outlet, inserted into physiological solution, were in the range of 0-2400 W/kg. Millimeter wave irradiation increased the peak amplitudes, activation rates, and inactivation rates of both ion currents. The changes in A-type K+ current were not dependent on the irradiation frequency. It was shown that the changes in the amplitudes and kinetics of both currents resulted from the temperature rise produced by irradiation. No additional effects of irradiation on A-type K+ current other than thermal were found when tested at the phase transition temperature or in the presence of ethanol. Ethanol reduced the thermal effect of irradiation. Millimeter waves had no effect on the steady-state activation and inactivation curves, suggesting that the membrane surface charge and binding of calcium ions to the membrane in the area of channel locations did not change.

  2. Susceptibility of two-week old Lymnaea natalensis to some plant extracts.

    PubMed

    Kela, S L; Ogunsusi, R A; Ogbogu, V C; Nwude, N

    1989-01-01

    The molluscacidal potency of 17 Nigerian plants extracted by the unevaporated crude water (UECW) method was evaluated on two-week old Lymnaea natalensis Krauss. Five extracts were not active but extracts of Balanites aegytiaca, Blighia sapida, Boswellia dalzielii, Cissampelos mucronata, Detarium microcarpum, Kigelia africana, Opilia celtidifolia, Parkia clappertoniana, Polygonum limbatum, Pseudocedrela kotschyi, Nauclea latifolia and Securidaca longipedunculata were molluscacidal. There is potential for their future use in the integrated control of Lymnaea natalensis, as well as other snails. Mortality data for lethal concentration values for all extracts were analysed by use of probit transformation. The upper and lower fiducial limits of the LC50 (P = 0.05) were also determined.

  3. Identification of Genetic Variation between Obligate Plant Pathogens Pseudoperonospora cubensis and P. humuli Using RNA Sequencing and Genotyping-By-Sequencing.

    PubMed

    Summers, Carly F; Gulliford, Colwyn M; Carlson, Craig H; Lillis, Jacquelyn A; Carlson, Maryn O; Cadle-Davidson, Lance; Gent, David H; Smart, Christine D

    2015-01-01

    RNA sequencing (RNA-seq) and genotyping-by-sequencing (GBS) were used for single nucleotide polymorphism (SNP) identification from two economically important obligate plant pathogens, Pseudoperonospora cubensis and P. humuli. Twenty isolates of P. cubensis and 19 isolates of P. humuli were genotyped using RNA-seq and GBS. Principle components analysis (PCA) of each data set showed genetic separation between the two species. Additionally, results supported previous findings that P. cubensis isolates from squash are genetically distinct from cucumber and cantaloupe isolates. A PCA-based procedure was used to identify SNPs correlated with the separation of the two species, with 994 and 4,231 PCA-correlated SNPs found within the RNA-seq and GBS data, respectively. The corresponding unigenes (n = 800) containing these potential species-specific SNPs were then annotated and 135 putative pathogenicity genes, including 3 effectors, were identified. The characterization of genes containing SNPs differentiating these two closely related downy mildew species may contribute to the development of improved detection and diagnosis strategies and improve our understanding of host specificity pathways.

  4. The Novel Oomycide Oxathiapiprolin Inhibits All Stages in the Asexual Life Cycle of Pseudoperonospora cubensis - Causal Agent of Cucurbit Downy Mildew

    PubMed Central

    Cohen, Yigal

    2015-01-01

    Oxathiapiprolin is a new oomycide (piperidinyl thiazole isoxazoline class) discovered by DuPont which controls diseases caused by oomycete plant pathogens. It binds in the oxysterol-binding protein domain of Oomycetes. Growth chambers studies with detached leaves and potted plants showed remarkable activity of oxathiapiprolin against Pseudoperonospora cubensis in cucurbits. The compound affected all stages in the asexual life cycle of the pathogen. It inhibited zoospore release, cystospore germination, lesion formation, lesion expansion, sporangiophore development and sporangial production. When applied to the foliage as a preventive spray no lesions developed due to inhibition of zoospore release and cystospore germination, and when applied curatively, at one or two days after inoculation, small restricted lesions developed but no sporulation occurred. When applied later to mature lesions, sporulation was strongly inhibited. Oxathiapiprolin suppressed sporulation of P. cubensis in naturally-infected leaves. It exhibited trans-laminar activity, translocated acropetaly from older to younger leaves, and moved from the root system to the foliage. Seed coating was highly effective in protecting the developed cucumber plants against downy mildew. UV microscopy observations made with cucumber leaves infected with P. cubensis revealed that inhibition of mycelium growth and sporulation induced by oxathiapiprolin was associated with callose encasement of the haustoria. PMID:26452052

  5. Identification of Genetic Variation between Obligate Plant Pathogens Pseudoperonospora cubensis and P. humuli Using RNA Sequencing and Genotyping-By-Sequencing

    PubMed Central

    Summers, Carly F.; Gulliford, Colwyn M.; Carlson, Craig H.; Lillis, Jacquelyn A.; Carlson, Maryn O.; Cadle-Davidson, Lance; Gent, David H.; Smart, Christine D.

    2015-01-01

    RNA sequencing (RNA-seq) and genotyping-by-sequencing (GBS) were used for single nucleotide polymorphism (SNP) identification from two economically important obligate plant pathogens, Pseudoperonospora cubensis and P. humuli. Twenty isolates of P. cubensis and 19 isolates of P. humuli were genotyped using RNA-seq and GBS. Principle components analysis (PCA) of each data set showed genetic separation between the two species. Additionally, results supported previous findings that P. cubensis isolates from squash are genetically distinct from cucumber and cantaloupe isolates. A PCA-based procedure was used to identify SNPs correlated with the separation of the two species, with 994 and 4,231 PCA-correlated SNPs found within the RNA-seq and GBS data, respectively. The corresponding unigenes (n = 800) containing these potential species-specific SNPs were then annotated and 135 putative pathogenicity genes, including 3 effectors, were identified. The characterization of genes containing SNPs differentiating these two closely related downy mildew species may contribute to the development of improved detection and diagnosis strategies and improve our understanding of host specificity pathways. PMID:26599440

  6. Predator detection enables juvenile Lymnaea to form long-term memory.

    PubMed

    Orr, M V; Hittel, K; Lukowiak, K

    2010-01-15

    Learning and memory provide the flexibility an organism requires to respond to changing social and ecological conditions. Juvenile Lymnaea have previously been shown to have a diminished capacity to form long-term memory (LTM) following operant conditioning of aerial respiratory behavior. Juvenile Lymnaea, however, can form LTM following classical conditioning of appetitive behaviors. Here, we demonstrate that laboratory-reared juvenile Lymnaea have the ability to detect the presence of a sympatric predator (i.e. crayfish) and respond to the predator by altering their aerial respiratory behavior. In addition to increasing their total breathing time, predator detection confers on juvenile Lymnaea an enhanced capability to form LTM following operant conditioning of aerial respiratory behavior. That is, these juveniles now have the ability to form long-lasting memory. These data support the hypothesis that biologically relevant levels of stress associated with predator detection induce behavioral phenotypic alterations (i.e. enhanced LTM formation) in juveniles, which may increase their fitness. These data also support the notion that learning and memory formation in conjunction with predator detection is a form of inducible defense.

  7. An octopaminergic system in the CNS of the snails, Lymnaea stagnalis and Helix pomatia

    PubMed Central

    Hiripi, L.

    1998-01-01

    Octopamine (OA) levels in each ganglion of the terrestrial snail, Helix pomatia, and the pond snail, Lymnaea stagnalis, were measured by using the HPLC technique. In both species an inhomogeneous distribution of OA was found in the central nervous system. The buccal ganglia contained a concentration of OA (12.6 pmol mg-1 and 18.8 pmol mg-1) that was two to three times higher than the pedal (4.93 pmol mg-1 and 9.2 pmol mg-1) or cerebral (4.46 pmol mg-1 and 4.9 pmol mg-1) ganglia of Helix and Lymnaea, respectively, whereas no detectable amount of OA could be assayed in the visceroparietal complex. In Lymnaea ganglia, the OA uptake into the synaptosomal fraction had a high (Km1 = 4.07 ± 0.51 μM, Vmax1 = 0.56 ± 0.11 pmol mg-1 per 20 min), and a low (Km2 = 47.6 ± 5.2 μM, Vmax2 = 4.2 ± 0.27 pmol mg-1 per 20 min), affinity component. A specific and dissociable 3H-OA binding to the membrane pellet prepared from the CNS of both Helix and Lymnaea was demonstrated. The Scatchard analysis of the ligand binding data showed a one-binding site, representing a single receptor site. The Kd and Bmax values were found to be 33.7 ± 5.95 nM and 1678 ± 179 fmol g-1 tissue in Helix and 84.9 ± 17.4 nM and 3803 ± 515 fmol g-1 tissue in Lymnaea preparation. The pharmacological properties of the putative molluscan OA receptor were characterized in both species and it was demonstrated that the receptor resembled the insect OA2 rather than to the cloned Lymnaea OA receptor. Immunocytochemical labelling demonstrated the presence of OA-immunoreactive neurons and fibres in the buccal, cerebral and pedal ganglia in the central nervous system of both species investigated. Electrophysiological experiments also suggested that the Lymnaea brain possessed specific receptors for OA. Local application of OA onto the identified buccal B2 neuron evoked a hyperpolarization which could selectively be inhibited by the OAergic agents phentolamine, demethylchlordimeform and 2-chloro-4-methyl-2

  8. Metabolic acceleration in the pond snail Lymnaea stagnalis?

    NASA Astrophysics Data System (ADS)

    Zimmer, Elke I.; Ducrot, V.; Jager, T.; Koene, J.; Lagadic, L.; Kooijman, S. A. L. M.

    2014-11-01

    Under constant environmental conditions, most animals tend to grow following the von Bertalanffy growth curve. Deviations from this curve can point to changes in the environment that the animals experience, such as food limitation when the available food is not sufficient or suitable. However, such deviations can also point to a phenomenon called metabolic acceleration, which is receiving increasing attention in the field of Dynamic Energy Budget (DEB) modeling. Reasons for such an acceleration are usually changes in shape during ontogeny, which cause changes in the surface area to volume ratio of the organism. Those changes, in turn, lead to changes in some of the model parameters that have length in their dimension. The life-history consequences of metabolic acceleration as implemented in the DEB theory are an s-shaped growth curve (when body size is expressed as a length measure) and a prolongation of the hatching time. The great pond snail Lymnaea stagnalis was earlier found to be food limited during the juvenile phase in laboratory experiments conducted under classical ecotoxicity test protocols. The pond snail has isomorphic shell growth but yet does not exhibit the expected von Bertalanffy growth curve under food limitation. When applying the standard DEB model to data from such life-cycle experiments, we also found that the hatching time is consistently underestimated, which could be a sign of metabolic acceleration. We here present an application of the DEB model including metabolic acceleration to the great pond snail. We account for the simultaneous hermaphroditism of the snail by including a model extension that describes the relative investment into the male and female function. This model allowed us to adequately predict the life history of the snail over the entire life cycle. However, the pond snail does not change in shape substantially after birth, so the original explanation for the metabolic acceleration does not hold. Since the change in shape

  9. Resurrection of Scolopendra longipes Wood and Scolopendra cubensis Saussure from synonymy with Scolopendra alternans Leach (Chilopoda, Scolopendromorpha, Scolopendridae): an enigmatic species-group needing phylogeographic analysis, with an overview on the origin and distribution of centipedes in the Caribbean region.

    PubMed

    Mercurio, Randy J

    2016-05-12

    Resurrection of Scolopendra longipes Wood, 1862, and Scolopendra cubensis Saussure, 1860, from junior synonymy with Scolopendra alternans Leach, 1815, is proposed. A neotype specimen of Scolopendra longipes is designated. Scolopendra longipes has a restricted range from the Dry Tortugas up through the Florida Keys of Monroe County into the mainland Florida counties of Collier and Dade southeast to the Bahamas, while Scolopendra cubensis is endemic to Cuba. Characters distinguishing S. longipes, and S. cubensis from S. alternans are illustrated and compared using digital photography, micrography and morphometric data. It is suggested that what has been considered Scolopendra alternans from southern Florida through the Caribbean and into northern South America is probably an evolving species-group that has undergone major diversification sometime during the Paleocene and early Eocene ~65.5-50 million years ago (Ma), mainly due to geographic isolation caused by a combination of plate tectonics and 100,000 year cycles of glaciation/deglaciation.

  10. Behavioural responses of the snail Lymnaea acuminata to carbohydrates in snail-attractant pellets

    NASA Astrophysics Data System (ADS)

    Tiwari, Farindra; Singh, D. K.

    Snail control is one of the most important tools in the campaign to reduce the incidence of fascioliasis. In order to attain this objective, the method of bait formulation in order to contain an attractant and a molluscicide is an expedient approach to lure the target snail population to the molluscicide. This study identifies certain carbohydrates, namely sucrose, maltose, glucose, fructose and starch, for preparing such baits. These were tested on Lymnaea acuminata, an intermediate host of the digenean trematodes Fasciola hepatica and Fasciola gigantica. The behavioural responses of snails to these carbohydrates were examined. Significant variations in behavioural responses were observed in the snail even when the five carbohydrates were used in low concentrations in snail-attractant pellets. Starch emerged as the strongest attractant for Lymnaea acuminata, followed by maltose.

  11. Polyethylene mulch modifies greenhouse microclimate and reduces infection of phytophthora infestans in tomato and Pseudoperonospora cubensis in cucumber.

    PubMed

    Shtienberg, D; Elad, Y; Bornstein, M; Ziv, G; Grava, A; Cohen, S

    2010-01-01

    The individual and joint effects of covering the soil with polyethylene mulch before planting and fungicides commonly used by organic growers on tomato late blight (caused by Phytophthora infestans) were studied in three experiments conducted from 2002 to 2005. Application of fungicides resulted in inconsistent and insufficient late blight suppression (control efficacy +/- standard error of 34.5 +/- 14.3%) but the polyethylene mulch resulted in consistent, effective, and highly significant suppression (control efficacy of 83.6 +/- 5.5%) of the disease. The combined effect of the two measures was additive. In a second set of three experiments carried out between 2004 and 2006, it was found that the type of polyethylene mulch used (bicolor aluminized, clear, or black) did not affect the efficacy of late blight suppression (control efficacy of 60.1 to 95.8%) and the differences in the effects among the different polyethylene mulches used were insignificant. Next, the ability of the mulch to suppress cucumber downy mildew (caused by Pseudoperonospora cubensis) was studied in four experiments carried out between 2006 and 2008. The mulch effectively suppressed cucumber downy mildew but the effect was less substantial (control efficacy of 34.9 +/- 4.8%) than that achieved for tomato late blight. The disease-suppressing effect of mulch appeared to come from a reduction in leaf wetness duration, because mulching led to reductions in both the frequency of nights when dew formed and the number of dew hours per night when it formed. Mulching also reduced relative humidity in the canopy, which may have reduced sporulation.

  12. Enzyme Inhibition by Molluscicidal Components of Myristica fragrans Houtt. in the Nervous Tissue of Snail Lymnaea acuminata

    PubMed Central

    Jaiswal, Preetee; Kumar, Pradeep; Singh, V. K.; Singh, D. K.

    2010-01-01

    This study was designed to investigate the effects of molluscicidal components of Myristica fragrans Houtt. (Myristicaceae) on certain enzymes in the nervous tissue of freshwater snail Lymnaea acuminata Lamarck (Lymnaeidae). In vivo and in vitro treatments of trimyristin and myristicin (active molluscicidal components of Myristica fragrans Houtt.) significantly inhibited the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the nervous tissue of Lymnaea acuminata. The inhibition kinetics of these enzymes indicates that both the trimyristin and myristicin caused competitive noncompetitive inhibition of AChE. Trimyristin caused uncompetitive and competitive/noncompetitive inhibitions of ACP and ALP, respectively whereas the myristicin caused competitive and uncompetitive inhibition of ACP and ALP, respectively. Thus results from the present study suggest that inhibition of AChE, ACP, and ALP by trimyristin and myristicin in the snail Lymnaea acuminata may be the cause of the molluscicidal activity of Myristica fragrans. PMID:21048864

  13. Enzyme Inhibition by Molluscicidal Components of Myristica fragrans Houtt. in the Nervous Tissue of Snail Lymnaea acuminata.

    PubMed

    Jaiswal, Preetee; Kumar, Pradeep; Singh, V K; Singh, D K

    2010-01-01

    This study was designed to investigate the effects of molluscicidal components of Myristica fragrans Houtt. (Myristicaceae) on certain enzymes in the nervous tissue of freshwater snail Lymnaea acuminata Lamarck (Lymnaeidae). In vivo and in vitro treatments of trimyristin and myristicin (active molluscicidal components of Myristica fragrans Houtt.) significantly inhibited the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the nervous tissue of Lymnaea acuminata. The inhibition kinetics of these enzymes indicates that both the trimyristin and myristicin caused competitive noncompetitive inhibition of AChE. Trimyristin caused uncompetitive and competitive/noncompetitive inhibitions of ACP and ALP, respectively whereas the myristicin caused competitive and uncompetitive inhibition of ACP and ALP, respectively. Thus results from the present study suggest that inhibition of AChE, ACP, and ALP by trimyristin and myristicin in the snail Lymnaea acuminata may be the cause of the molluscicidal activity of Myristica fragrans.

  14. Origin of automaticity and neural regulation of peristalsis in the gastrointestinal tract of Aplysia and Lymnaea. Short communication.

    PubMed

    Kurokawa, M; Kasuya, Y; Okamoto, T

    2012-01-01

    We examined whether the enteric nervous system (ENS) is capable of controlling autonomous peristalsis, which occurs in the crop of Aplysia as well as in the esophagus of Lymnaea. Interestingly, "pacemaker neurons", which lead peristaltic rhythm, were found in the gizzard in Aplysia and in the crop in Lymnaea; both of these structures are located distal to the regions exhibiting peristalsis. Thus, the bursting activity of the ENS first occurred in lower regions and then progressed in an ascending direction (i.e. in the opposite direction of peristalsis). The two species are thought to differ in terms of the mechanisms involved in producing peristalsis.

  15. Effects of Developmental Temperature on Gametocysts and Oocysts of Two Species of Gregarines Blabericola migrator and Blabericola cubensis (Apicomplexa: Eugregarinida: Blabericolidae) Parasitizing Blaberid Cockroaches (Dictyoptera: Blaberidae).

    PubMed

    Kolman, Jon A; Clopton, Richard E; Clopton, Debra T

    2015-12-01

    Abiotic environmental conditions, especially temperature and humidity, have profound effects on the growth and development of gregarines, but these effects remain largely undocumented. Quantifying the effects of environmental conditions on the growth and development of exogenous gregarine ontogenetic stages is an important first step in understanding the transmission, population dynamics, and environmental persistence of gregarine infection. In this study, we examined the effect of 6 environmental temperatures (10, 18, 22, 27, 35, and 40 C) at constant humidity (0 mmHg vapor pressure deficit) on gametocyst development and oocyst viability in 2 gregarine species: Blabericola migrator and Blabericola cubensis parasitizing the Tiger-striped Hissing Cockroach, Princisia vanwaerebecki, and the Discoid Cockroach, Blaberus discoidalis, respectively. Temperature has a significant effect on gametocyst development and oocyst viability for both gregarine species. Gametocyst development for both gregarine species displays a similar threshold response to environmental temperature: 10 and 40 C represent extremes outside their developmental range, but within these extremes, the relationship between gametocyst development and temperature is weakly direct. Dehiscence increased with temperature from 68% at 18 C to 93% at 22 C and remained at that level through 35 C. Developmental temperature also has a meaningful but inverse effect on oocyst viability of both B. migrator and B. cubensis. For both species, oocyst viability is highest at 18 and 22 C and is significantly reduced at 27 and 35 C. Thus oocyst production and sporozoite viability are linked but environmentally independent phenomena. Overall, there is an acceptable developmental temperature zone for B. migrator and B. cubensis that ranges from 18 to 27 C, but production of viable sporozoites is greatest in a relatively narrow zone around 22 C. Prior studies have postulated that mechanisms that concentrate oocysts and hosts

  16. Activities and functions of peripheral neurons in the enteric nervous system of Aplysia and Lymnaea.

    PubMed

    Kurokawa, M; Ito, S; Okamoto, T

    2008-01-01

    In order to explore the functions of the peripheral neurons in the enteric nervous system (ENS) of the gastropods, Aplysia and Lymnaea, we investigated the correlation between peripheral neuronal activities and movements of the digestive tract. In Aplysia, movements of the gizzard were distinguished into two types of contraction: a large constriction of the whole gizzard following bursting activities of the neurons on the gizzard and EJP-like potentials in the musculature; and a small contraction of a restricted part of the gizzard following a slow muscle potential. When TTX was applied to isolated gizzard preparation, the bursting activities were blocked and the EJP-like potentials and the subsequent constriction disappeared, whereas the slow potentials in the musculature and partial contractions appeared to be unaffected. Therefore, it was suggested that the peripheral neurons on the gizzard were motor neurons for constriction, while the partial contraction was thought to be myogenic. In Lymnaea, we recorded periodic bursting activities in the enteric nervous system that were followed by EJP-like potentials and gastrointestinal movements. The results show that, in both species, there may exist motor neurons in the ENS that are responsible for neurogenic movements of the digestive tract.

  17. Methamphetamine enhances memory of operantly conditioned respiratory behavior in the snail Lymnaea stagnalis.

    PubMed

    Kennedy, Colin D; Houmes, Stephen W; Wyrick, Katherine L; Kammerzell, Samuel M; Lukowiak, Ken; Sorg, Barbara A

    2010-06-15

    Amphetamines have been used as cognitive enhancers to promote learning and memory. Amphetamines are also drugs of abuse that may promote the initiation of strong memories that ultimately lead to addiction. To understand how methamphetamine (Meth) may be augmenting learning and memory, we chose a relatively simple system, the pond snail, Lymnaea stagnalis. We studied the effects of Meth exposure on the long-term memory (LTM), extinction and reinstatement of operantly conditioned aerial respiratory behavior in Lymnaea. We first determined doses of Meth that would acutely alter respiratory behavior. Next, we measured the impact of training snails in Meth solution or water (control group) using a training procedure that produces LTM (>6 h) in control conditions. Meth exposure impaired the expression of LTM 21 h after two training sessions, but this appeared to be a context-dependent effect only. However, snails exposed to 3.3 mumol l(-1) Meth during training had a decreased rate of extinction of the operantly conditioned memory. We then tested whether this decreased ability of snails to extinguish memory was due to enhanced LTM or impaired extinction of that memory. Snails were operantly conditioned in water and exposed to Meth 16 h after their last trial but 4-5 h prior to extinction. Meth produced an increase rather than a decrease in extinction rate. Thus, Meth impaired extinction only when snails were exposed to Meth during training. Last, we tested the effect of Meth on the ability to form LTM using a single training procedure that is suboptimal for LTM formation. Control snails did not demonstrate LTM, as expected, but pre-exposure of snails to 3.3 micromol l(-1) Meth 24 h prior to the single training session produced LTM 24 h later, indicating that Meth pre-exposure primed snails for LTM formation. Taken together, our studies suggest that LTM is strengthened by Meth such that extinction training is less effective. Lymnaea provides a simple and useful model

  18. [The growth and energy metabolism of Lymnaea stagnalis (Lymnaeidae, Gastropoda): I. Early postlarval period].

    PubMed

    Zotin, A A

    2009-01-01

    The growth and the oxygen uptake rate of Lymnaea stagnalis were studied during the first ten days after hatching. It is shown that these processes are atypical during early ontogenesis in comparison with adult mollusks. The obtained data on linear (height of shell) and weight growth can be equally well approximated with the von Bertalanffy equation or exponential and polynomial equations. Both linear and weight growth are characterized by an approximately constant specific rate associated with synchronous oscillations of a two-week period. The oscillations were observed also for the oxygen uptake rate, but of another period (about 2.6 weeks). On average the metabolic rate after the initial triple increase during the first three days remains stable. The polynomial coefficient of the allometric dependence of the total weight on the shell height is less reliable than that of the adult.

  19. Behavioural responses of the snail Lymnaea acuminata to carbohydrates and amino acids in bait pellets.

    PubMed

    Agrahari, P; Singh, D K

    2010-12-01

    Snail control could play an important role in programmes against fascioliasis, especially if the methods used for molluscicide delivery could be improved, such as by the development of bait formulations containing both an effective attractant and a molluscicide, to ensure good levels of contact between the molluscicide and the target snail populations. In a recent study, the attractiveness to Lymnaea acuminata (an intermediate host of the digenean trematode Fasciola gigantica) of potential components of snail-attractant pellets was investigated. Carbohydrates (glucose, maltose, sucrose or starch, each at 10 mM) and amino acids (citrulline, tryptophan, proline or serine, each at 20 mM), were tested in aquaria, with the snails initially placed 22.5, 30 or 45 cm from an agar pellet containing the component under test. Under these conditions, starch and proline emerged as the strongest attractants for L. acuminata, followed by maltose and serine.

  20. The participation of NMDA receptors, PKC, and MAPK in Lymnaea memory extinction.

    PubMed

    Rosenegger, David; Lukowiak, Ken

    2013-02-01

    The aerial respiratory behavior of Lymnaea can be operantly conditioned to form a long-term memory (LTM) that will persist for >24h. LTM formation is dependent on altered gene activity and new protein synthesis, with the N-methyl-D-aspartate (NMDA) receptors, mitogen activated protein kinase (MAPK), and protein kinase C (PKC) pathways playing a critical role. LTM can also undergo extinction, whereby the original memory is temporarily masked by a new memory. Here we investigate if the formation of an extinction memory uses similar molecular pathways to those required for LTM formation. We find that the formation of the extinction memory can be blocked by inhibitors of NMDA receptors, PKC, and MAPK suggesting that extinction memory formation uses similar mechanisms to that of 'normal' memory formation.

  1. Growth, fecundity and glycogen utilization in Lymnaea palustris exposed to atrazine and hexachlorobenzene in freshwater mesocosms

    SciTech Connect

    Baturo, W.; Lagadic, L.; Caquet, T.

    1995-03-01

    Freshwater mesocosms were used to study the long-term sublethal effects of atrazine and hexachlorobenzene (HCB) on a basommatophoran gastropod, Lymnaea palustris (Mueller). Growth, fecundity, and biochemical parameters related to polysaccharide metabolism of pesticide-exposed snails were compared with those of control animals maintained in untreated mesocosms. HCB inhibited body growth and stimulated egg production, whereas atrazine had no relevant effect on these physiological parameters. Also, HCB stimulated the activity of polysaccharide-hydrolyzing enzymes, suggesting that changes in the metabolism of reserve polysaccharides (glycogen) may be involved in the inhibition of growth and increase of fecundity. In contrast, atrazine had no effect on the metabolism of polysaccharides. It is concluded that the effects of HCB are related to its neurotoxicity that would have affected the neurohormonal control of growth and reproduction of exposed snails. It is suggested that polysaccharide-hydrolyzing enzymes may be used as biomarkers to predict the effects of neurotoxic pesticides on freshwater snail populations.

  2. Evidence for genetic influences on neurotransmitter content of identified neurones of Lymnaea stagnalis.

    PubMed

    Audesirk, G; Audesirk, T; McCaman, R E; Ono, J K

    1985-01-01

    Neurotransmitter content was measured in two identified giant neurones in isogenic and wild-type populations of the freshwater pond snail Lymnaea stagnalis. The paired serotonergic cerebral giant neurones (LC1 and RC1) have higher transmitter levels and less variability in inbred animals than in wild-type animals. The transmitter content of the unpaired dopaminergic right pedal giant neurone (RPeD1) does not differ between inbred and wild-type animals in either level or variability. It is proposed that serotonin content of the cerebral giant neurones is under partial genetic control, and that animals of the wild-type population may possess a number of different alleles for the genes influencing serotonin levels. Inbreeding resulted in fixation of an allele promoting high serotonin levels. This particular wild-type population is probably already isogenic for genes influencing dopamine content in the right pedal giant neurone.

  3. Amine-containing neurons in the brain of Lymnaea stagnalis: distribution and effects of precursors.

    PubMed

    Audesirk, G

    1985-01-01

    Glyoxylic acid-induced fluorescence in whole-brain preparations of the central nervous system of the freshwater pond snail, Lymnaea stagnalis, was used to map the distribution of serotonin-and dopamine-containing neurons. Serotonin and dopamine were easily distinguishable by differences in color of fluorescence. Serotonin-containing neurons were consistently found in the cerebral, pedal, right parietal and visceral ganglia. Dopamine-containing neurons were found in the pedal, and buccal ganglia. Prior incubation of brains in 5-hydroxytryptophan (5-HTP), the immediate precursor to serotonin, produced serotonin-like fluoresence in neurons which do not normally fluoresce. These neurons thus probably possess specific uptake mechanisms for 5-HTP. Since 5-HTP itself fluoresces yellow, the glyoxylic acid technique cannot determine if these neurons contain the enzyme aromatic amino acid decarboxylase, which converts 5-HTP to serotonin, or merely fluoresce because of the 5-HTP taken into the cells.

  4. Juveniles of Lymnaea 'smart' snails do not perseverate and have the capacity to form LTM.

    PubMed

    Shymansky, Tamila; Protheroe, Amy; Hughes, Emily; Swinton, Cayley; Swinton, Erin; Lukowiak, Kai S; Phillips, Iain; Lukowiak, Ken

    2017-02-01

    Previously, it was concluded that the nervous systems of juvenile snails were not capable of mediating long-term memory (LTM). However, exposure and training of those juvenile snails in the presence of a predator cue significantly altered their ability to learn and form LTM. In addition, there are some strains of Lymnaea which have been identified as 'smart'. These snails form LTM significantly better than the lab-bred strain. Here, we show that juveniles of two smart snail strains not only are capable of associative learning but also have the capacity to form LTM following a single 0.5 h training session. We also show that freshly collected 'wild' 'average' juveniles are also not able to form LTM. Thus, the smart snail phenotype in these strains is expressed in juveniles.

  5. Embryonic exposure to model naphthenic acids delays growth and hatching in the pond snail Lymnaea stagnalis.

    PubMed

    Johnston, Christina U; Clothier, Lindsay N; Quesnel, Dean M; Gieg, Lisa M; Chua, Gordon; Hermann, Petra M; Wildering, Willem C

    2017-02-01

    Naphthenic acids (NAs), a class of structurally diverse carboxylic acids with often complex ring structures and large aliphatic tail groups, are important by-products of many petrochemical processes including the oil sands mining activity of Northern Alberta. While it is evident that NAs have both acute and chronic harmful effects on many organisms, many aspects of their toxicity remain to be clarified. Particularly, while substantive data sets have been collected on NA toxicity in aquatic prokaryote and vertebrate model systems, to date, nothing is known about the toxic effects of these compounds on the embryonic development of aquatic invertebrate taxa, including freshwater mollusks. This study examines under laboratory conditions the toxicity of NAs extracted from oil sands process water (OSPW) and the low-molecular weight model NAs cyclohexylsuccinic acid (CHSA), cyclohexanebutyric acid (CHBA), and 4-tert-butylcyclohexane carboxylic acid (4-TBCA) on embryonic development of the snail Lymnaea stagnalis, a common freshwater gastropod with a broad Palearctic distribution. Evidence is provided for concentration-dependent teratogenic effects of both OSPW-derived and model NAs with remarkably similar nominal threshold concentrations between 15 and 20 mg/L and 28d EC50 of 31 mg/L. In addition, the data provide evidence for substantial toxicokinetic differences between CHSA, CHBA and 4-TBCA. Together, our study introduces Lymnaea stagnalis embryonic development as an effective model to assay NA-toxicity and identifies molecular architecture as a potentially important toxicokinetic parameter in the toxicity of low-molecular weight NA in embryonic development of aquatic gastropods.

  6. A cholinergic modulatory interneuron in the feeding system of the snail, Lymnaea.

    PubMed

    Yeoman, M S; Parish, D C; Benjamin, P R

    1993-07-01

    1. Pharmacological and physiological methods were used to examine the role of acetylcholine (ACh) in modulation of the Lymnaea feeding central pattern generator (CPG) by the slow oscillator (SO) interneuron. 2. Extracts of dissected SO cell bodies inhibited spontaneous ventricular contractions of the clam Mya arenaria, indicating the presence of ACh. These effects were blocked by the specific antagonist benzoquinonium chloride (10(-7) M). 3. Isolated SO cells grown in culture synthesized ACh from tritiated choline. 4. High [K+] saline induced release of synthesized ACh from cultured SO cells into the medium. 5. The specific ACh antagonist phenyltrimethylammonium (10(-4) M) blocked both excitatory, biphasic (inhibitory-excitatory) and inhibitory monosynaptic connections from the SO to feeding CPG interneurons and motor neurons. Less specific cholinergic antagonists blocked either excitatory (hexamethonium, 10(-4) M) or both excitatory and inhibitory connections (d-tubo-curarine, 10(-4) M). 6. The synaptic responses of the SO could be mimicked by brief (20 ms) pressure-pulsed application of ACh onto the cell bodies of the postsynaptic cells in high-Mg2+ saline. In normal saline, ACh elicited bursts of spikes in the N1 cells, indicating that a fictive feeding pattern had been induced in the CPG. This mimics the main mechanism by which the SO activates the CPG, which is by exciting the N1s. 7. The frequency of SO-induced fictive feeding rhythm was reduced by bath application of hexamethonium chloride to the buccal ganglia. This reduced the amplitude of the SO-->N1 excitatory synaptic response (30% of controls) and is probably the main mechanism for the reduction in the frequency of the rhythm. 8. The evidence suggests that ACh is the main neurochemical involved in allowing the SO to initiate and control the frequency of the Lymnaea feeding CPG.

  7. Electrophysiological characteristics of feeding-related neurons after taste avoidance Pavlovian conditioning in Lymnaea stagnalis.

    PubMed

    Sunada, Hiroshi; Takigami, Satoshi; Lukowiak, Ken; Sakakibara, Manabu

    2014-01-01

    Taste avoidance conditioning (TAC) was carried out on the pond snail, Lymnaea stagnalis. The conditional stimulus (CS) was sucrose which elicits feeding behavior; while the unconditional stimulus (US) was a tactile stimulus to the head which causes feeding to be suppressed. The neuronal circuit that drives feeding behavior in Lymnaea is well worked out. We therefore compared the physiological characteristics on 3 classes of neurons involved with feeding behavior especially in response to the CS in conditioned vs. control snails. The cerebral giant cell (CGC) modulates feeding behavior, N1 medial neuron (N1M) is one of the central pattern generator neurons that organizes feeding behavior, while B3 is a motor neuron active during the rasp phase of feeding. We found the resting membrane potential in CGC was hyperpolarized significantly in conditioned snails but impulse activity remained the same between conditioned vs. control snails. There was, however, a significant increase in spontaneous activity and a significant depolarization of N1M's resting membrane potential in conditioned snails. These changes in N1M activity as a result of training are thought to be due to withdrawal interneuron RPeD11 altering the activity of the CGCs. Finally, in B3 there was: 1) a significant decrease in the amplitude and the frequency of the post-synaptic potentials; 2) a significant hyperpolarization of resting membrane potential in conditioned snails; and 3) a disappearance of bursting activity typically initiated by the CS. These neuronal modifications are consistent with the behavioral phenotype elicited by the CS following conditioning.

  8. /sup 45/Ca uptake from water by snails (Lymnaea vulgaris) in control and detergent-polluted samples

    SciTech Connect

    Misra, V.; Lal, H.; Viswanathan, P.N.; Murti, C.R.

    1984-02-01

    A biostatic assay method involving /sup 45/Ca uptake into shells and tissues of snails (Lymnaea vulgaris) in 72 hr was developed to follow the effect of detergent-polluted water on ecosystems. There was a marked decrease in the /sup 45/Ca uptake by shells and tissues of linear alkyl benzene sulfonate-exposed animals as compared to controls. No change in /sup 45/Ca uptake was observed in dead shells, thereby excluding the possibility of passive exchange.

  9. Characterisation of fascioliasis lymnaeid intermediate hosts from Chile by DNA sequencing, with emphasis on Lymnaea viator and Galba truncatula.

    PubMed

    Artigas, Patricio; Bargues, M Dolores; Mera y Sierra, Roberto L; Agramunt, Veronica H; Mas-Coma, Santiago

    2011-12-01

    In South America, Fasciola hepatica infection poses serious health problems in both humans and livestock. In Chile, the medical impact appears yearly stable and mainly concentrated in central regions, where the veterinary problem is highlighted by higher animal prevalences. Studies were undertaken by rDNA ITS-2 and ITS-1 and mtDNA cox1 sequencing to clarify the specific status of the lymnaeids, their geographical distribution and fascioliasis transmission capacity in Chile, by comparison with other American countries and continents. Results change the lymnaeid scenario known so far. The lymnaeid fauna of mainland Chile shows to be poor, including only two authochthonous species, Lymnaea viator and Pectinidens diaphana, and a third introduced species of Palaearctic origin Galba truncatula. Both Lymnaea lebruni and Lymnaea patagonica proved to be synonyms of P. diaphana. G. truncatula appears to have always been confused with L. viator and seems distributed from Región VI to Región IX, overlapping with human endemic areas. DNA sequencing results suggest that the absence of correlation between remote sensing data and disease prevalences could be due to transmission capacity differences between L. viator and G. truncatula. Results furnish a new baseline on which to undertake future appropriate studies on transmission, epidemiology and control.

  10. Environmental Persistence and Infectivity of Oocysts of Two Species of Gregarines, Blabericola migrator and Blabericola cubensis (Apicomplexa: Eugregarinida: Blabericolidae), Parasitizing Blaberid Cockroaches (Dictyoptera: Blaberidae).

    PubMed

    Clopton, Richard E; Steele, Shelby M; Clopton, Debra T

    2016-04-01

    For apicomplexan parasites using an oral-fecal transmission route with significant environmental exposure, the environmental persistence and infectivity of the oocyst has a direct impact on local infection dynamics, including the ability to withstand extended periods without readily available hosts. Herein we quantify the environmental persistence and infectivity of the oocysts of 2 septate gregarine species at controlled temperature and humidity and demonstrate that they can persist over multiple generational time spans. Species of Blabericola generally complete their endogenous life cycles from oocyst to oocyst within 10 days. The median residual environmental oocyst lifetime for Blabericola oocysts in this study is 21-28 days, but a significant number of oocysts of Blabericola migrator persisted and remained infective in the environment for up to 39 days while those of Blabericola cubensis persisted and remained infective for up to 92 days. Although long-lived relative to their own generational time, the oocysts of Blabericola species infecting cockroaches are short-lived relative to gregarines infecting tenebrionid beetles. For these gregarines, oocysts can persist in the environment and remain infective for up to 787 days. Mechanistically, environmental persistence and infectivity are probably energy-limited phenomena related to the amount of stored amylopectin and the basal metabolic rate of quiescent oocysts.

  11. Natural prevalence in Cuban populations of the lymnaeid snail Galba cubensis infected with the liver fluke Fasciola hepatica: small values do matter.

    PubMed

    Vázquez, Antonio A; Sánchez, Jorge; Alba, Annia; Pointier, Jean-Pierre; Hurtrez-Boussès, Sylvie

    2015-11-01

    Natural infections of lymnaeid snails by Fasciola hepatica are of primary importance to study transmission. Also, infected snails in the field can be used to explore the existing compatibility in host-parasite interactions. This paper aimed to describe the infection rate of Galba cubensis populations in fasciolosis transmission areas. Eight sites were sampled in western Cuba and 24 infected snails at six sites were found. The mean prevalence was 2.94% and the maximum value was 11.4%. The intensity of parasite infection was assessed as the number of rediae inside a single snail. High variation within the sites examined was observed, but a maximum of 76 rediae was recovered from one individual. Although the presence of two other trematode families (Schistosomatidae and Paramphistomatidae) was discovered in dissected individuals, no co-infection with F. hepatica was observed. This is the first time a study of natural prevalence of F. hepatica infection is carried out in Cuba, considered a hyper endemic country for bovine fasciolosis. Our results suggest that fasciolosis transmission may occur even when the number of infected snails remains relatively low.

  12. DIFFERENTIAL TRACER COUPLING BETWEEN PAIRS OF IDENTIFIED NEURONES OF THE MOLLUSC LYMNAEA STAGNALIS

    PubMed

    Ewadinger; Syed; Lukowiak; Bulloch

    1994-07-01

    Electrical coupling is a common means of cell-to-cell communication in both neuronal and non-neuronal tissues (Lowenstein, 1985). Within the nervous system, many electrically coupled neurones exhibit dye coupling (Bennett, 1973; Stewart, 1978; Glantz and Kirk, 1980; Spencer and Satterlie, 1980; Fraser and Heitler, 1993); however, some electrically coupled cells do not dye-couple (Audesirk et al. 1982; Murphy et al. 1983; Berdan, 1987; Robinson et al. 1993; Veenstra et al. 1993). Electrical coupling and dye coupling, often considered in parallel, are in fact two different parameters that can vary independently (e.g. Audesirk et al. 1982; Perez-Armendariz et al. 1991). The giant identified neurones of pulmonate and opisthobranch molluscs have frequently been used for studies of neuronal communication and its plasticity (Winlow and McCrohan, 1987; Bulloch, 1989). In the present study, we explored the relationship between electrical and tracer coupling in both strongly and weakly coupled pairs of molluscan neurones. Specifically, we examined electrically coupled, identified neurones in a freshwater pond snail, Lymnaea stagnalis L., and tested for tracer coupling with Lucifer Yellow CH and biocytin. The cells examined were the strongly electrically coupled neurones, visceral dorsal 1 (VD1) and right parietal dorsal 2 (RPD2) (Boer et al. 1979; Benjamin and Pilkington, 1986), and the weakly coupled neurones, left buccal 1 (LB1) and right buccal 1 (RB1) (Benjamin and Rose, 1979). The use of these particular neurones made it possible to compare electrical coupling with tracer coupling in the molluscan central nervous system (CNS). All experiments were performed on laboratory-bred Lymnaea stagnalis (Mollusca, Pulmonata), maintained as previously described (Ridgway et al. 1991). The CNS was dissected from mature animals (16­18 mm shell length) and pinned to the silicone rubber (RTV 616 GE) base of a recording dish in normal saline (51.3 mmol l-1 NaCl, 1.7 mmol l-1 KCl, 4

  13. Protein kinase C mediates memory consolidation of taste avoidance conditioning in Lymnaea stagnalis.

    PubMed

    Takigami, Satoshi; Sunada, Hiroshi; Lukowiak, Ken; Kuzirian, Alan M; Alkon, Daniel L; Sakakibara, Manabu

    2014-05-01

    In Lymnaea stagnalis, in order to obtain a 10 min short-term memory (STM) of taste avoidance conditioning (TAC) at least 10 paired presentations of a conditioned stimulus (CS), sucrose, and an unconditioned stimulus (US), tactile stimulation to the animal's head, are required. Pre-exposure of snails to the protein kinase C (PKC) α and ε activator bryostatin (Bryo) facilitated STM formation in that only 5 paired CS-US trials were required. Typically 20 paired presentations of the CS-US are required for formation of STM and LTM. However, 20 paired presentations do not result in STM or LTM if snails are pre-incubated with a PKC inhibitor, Ro-32-0432. We also found that LTM lasting longer than 48 h was acquired with Bryo incubation for 45 min even after termination of the conditioning paradigm. These data suggest that activation of the α and ε isozymes of PKC is crucially involved in the formation of LTM and provide further support for a mechanism that has been conserved across the evolution of species ranging from invertebrate molluscs to higher mammals.

  14. A two-neuron system for adaptive goal-directed decision-making in Lymnaea

    PubMed Central

    Crossley, Michael; Staras, Kevin; Kemenes, György

    2016-01-01

    During goal-directed decision-making, animals must integrate information from the external environment and their internal state to maximize resource localization while minimizing energy expenditure. How this complex problem is solved by the nervous system remains poorly understood. Here, using a combined behavioural and neurophysiological approach, we demonstrate that the mollusc Lymnaea performs a sophisticated form of decision-making during food-searching behaviour, using a core system consisting of just two neuron types. The first reports the presence of food and the second encodes motivational state acting as a gain controller for adaptive behaviour in the absence of food. Using an in vitro analogue of the decision-making process, we show that the system employs an energy management strategy, switching between a low- and high-use mode depending on the outcome of the decision. Our study reveals a parsimonious mechanism that drives a complex decision-making process via regulation of levels of tonic inhibition and phasic excitation. PMID:27257106

  15. Environmental versus anthropogenic effects on population adaptive divergence in the freshwater snail Lymnaea stagnalis.

    PubMed

    Bouétard, Anthony; Côte, Jessica; Besnard, Anne-Laure; Collinet, Marc; Coutellec, Marie-Agnès

    2014-01-01

    Repeated pesticide contaminations of lentic freshwater systems located within agricultural landscapes may affect population evolution in non-target organisms, especially in species with a fully aquatic life cycle and low dispersal ability. The issue of evolutionary impact of pollutants is therefore conceptually important for ecotoxicologists. The impact of historical exposure to pesticides on genetic divergence was investigated in the freshwater gastropod Lymnaea stagnalis, using a set of 14 populations from contrasted environments in terms of pesticide and other anthropogenic pressures. The hypothesis of population adaptive divergence was tested on 11 life-history traits, using Q(ST)-F(ST) comparisons. Despite strong neutral differentiation (mean F(ST) = 0.291), five adult traits or parameters were found to be under divergent selection. Conversely, two early expressed traits showed a pattern consistent with uniform selection or trait canalization, and four adult traits appeared to evolve neutrally. Divergent selection patterns were mostly consistent with a habitat effect, opposing pond to ditch and channel populations. Comparatively, pesticide and other human pressures had little correspondence with evolutionary patterns, despite hatching rate impairment associated with global anthropogenic pressure. Globally, analyses revealed high genetic variation both at neutral markers and fitness-related traits in a species used as model in ecotoxicology, providing empirical support for the need to account for genetic and evolutionary components of population response in ecological risk assessment.

  16. Critical Period of Memory Enhancement during Taste Avoidance Conditioning in Lymnaea stagnalis

    PubMed Central

    Sunada, Hiroshi; Lukowiak, Ken; Sakakibara, Manabu

    2013-01-01

    The present study investigated the optimal training procedure leading to long-lasting taste avoidance behavior in Lymnaea. A training procedure comprising 5 repeated pairings of a conditional stimulus (CS, sucrose), with an unconditional stimulus (US, a tactile stimulation to the animal’s head), over a 4-day period resulted in an enhanced memory formation than 10 CS-US repeated pairings over a 2-day period or 20 CS-US repeated pairings on a single day. Backward conditioning (US-CS) pairings did not result in conditioning. Thus, this taste avoidance conditioning was CS-US pairing specific. Food avoidance behavior was not observed following training, however, if snails were immediately subjected to a cold-block (4°C for 10 min). It was critical that the cold-block be applied within 10 min to block long-term memory (LTM) formation. Further, exposure to the cold-block 180 min after training also blocked both STM and LTM formation. The effects of the cold-block on subsequent learning and memory formation were also examined. We found no long lasting effects of the cold-block on subsequent memory formation. If protein kinase C was activated before the conditioning paradigm, snails could still acquire STM despite exposure to the cold-block. PMID:24098373

  17. Seasonal and Geographic Distribution of Cercarial Infection in Lymnaea gedrosiana (Pulmunata: Lymnaeidae) In North West Iran

    PubMed Central

    IMANI-BARAN, Abbas; YAKHCHALI, Mohammad; MALEKZADEH-VIAYEH, Reza; FARAHNAK, Ali

    2013-01-01

    Background Trematodes are a diverse group of endoparasites which require molluscan and vertebrate animals as intermediate and definitive hosts in their life cycle. The present study was carried out to determine the diversity and geographic distribution of infection with trematodes'cercariae in the snail Lymnaea gedrosiana from north-west Iran. Methods A total number of 6759 Lymnaeidae snails were collected from 28 snail habitats; of these L. gedrosiana was the prevalent snail (74.37%) which examined for cercarial infection by shedding method. Results The overall infection rate was 8.03%. The most frequent trematodes cercariae in the snail were xiphidiocercariae (81.98%), furcocercariae (32.26%), echinostome cercariae (5.19%), and monostome cercariae (1.24%). The highest infection rate in L. gedrosiana (100%) was with echinostome cercariae from Golestaneh in autumn. Conclusion Due to the important role of pond snails in transmission of cercariae to fish as a source of zoonotic diseases, it is essential to estimate the distribution and abundance of the snails and the rate of their infection with different trematodes’ cercariae, and establish control programs in each region. PMID:24454436

  18. Molluscicidal activity of Saraca asoca and Thuja orientalis against the fresh water snail Lymnaea acuminata.

    PubMed

    Singh, Arundhati; Singh, V K

    2009-10-14

    The molluscicidal activity of bark powder of Saraca asoca, leaf powder of Thuja orientalis against the snail Lymnaea acuminata was studied. The molluscicidal activity of all the plant products was found to be both time and concentration dependent. The 96 h LC(50) of T. orientalis leaf powder against L. acuminata was 250.5mg/l. Ethanol extracts were more toxic than other organic extracts. The ethanol extract of T. orientalis leaf (24h LC(50): 32.74 mg/l) was more effective than that of S. asoca bark (24h LC(50): 82.38 mg/l). The 24h LC(50) of column purified fraction of T. orientalis leaf and S. asoca bark powder was 29.25 and 64.89 mg/l, respectively. Saponin and thujone were identified as active molluscicide components in the bark of S. asoca and leaf of T. orientalis, respectively. The product of S. asoca and T. orientalis may be used as potent molluscicides.

  19. High sensitivity of spontaneous spike frequency to sodium leak current in a Lymnaea pacemaker neuron.

    PubMed

    Lu, T Z; Kostelecki, W; Sun, C L F; Dong, N; Pérez Velázquez, J L; Feng, Z-P

    2016-12-01

    The spontaneous rhythmic firing of action potentials in pacemaker neurons depends on the biophysical properties of voltage-gated ion channels and background leak currents. The background leak current includes a large K(+) and a small Na(+) component. We previously reported that a Na(+) -leak current via U-type channels is required to generate spontaneous action potential firing in the identified respiratory pacemaker neuron, RPeD1, in the freshwater pond snail Lymnaea stagnalis. We further investigated the functional significance of the background Na(+) current in rhythmic spiking of RPeD1 neurons. Whole-cell patch-clamp recording and computational modeling approaches were carried out in isolated RPeD1 neurons. The whole-cell current of the major ion channel components in RPeD1 neurons were characterized, and a conductance-based computational model of the rhythmic pacemaker activity was simulated with the experimental measurements. We found that the spiking rate is more sensitive to changes in the Na(+) leak current as compared to the K(+) leak current, suggesting a robust function of Na(+) leak current in regulating spontaneous neuronal firing activity. Our study provides new insight into our current understanding of the role of Na(+) leak current in intrinsic properties of pacemaker neurons.

  20. Reproductive impacts of tributyltin (TBT) and triphenyltin (TPT) in the hermaphroditic freshwater gastropod Lymnaea stagnalis.

    PubMed

    Giusti, Arnaud; Barsi, Alpar; Dugué, Maël; Collinet, Marc; Thomé, Jean-Pierre; Joaquim-Justo, Célia; Roig, Benoit; Lagadic, Laurent; Ducrot, Virginie

    2013-07-01

    Tributyltin (TBT) and triphenyltin (TPT) are emblematic endocrine disruptors, which have been mostly studied in gonochoric prosobranchs. Although both compounds can simultaneously occur in the environment, they have mainly been tested separately for their effects on snail reproduction. Because large discrepancies in experimental conditions occurred in these tests, the present study aimed to compare the relative toxicity of TBT and TPT under similar laboratory conditions in the range of 0 ng Sn/L to 600 ng Sn/L. Tests were performed on the simultaneous hermaphrodite Lymnaea stagnalis, a freshwater snail in which effects of TPT were unknown. Survival, shell length, and reproduction were monitored in a 21-d semistatic test. Frequency of abnormal eggs was assessed as an additional endpoint. Triphenyltin hampered survival while TBT did not. Major effects on shell solidity and reproduction were observed for both compounds, reproductive outputs being more severely hampered by TBT than by TPT. Considering the frequency of abnormal eggs allowed increasing test sensitivity, because snail responses to TBT could be detected at concentrations as low as 19 ng Sn/L. However, the putative mode of action of the 2 compounds could not be deduced from the structure of the molecules or from the response of apical endpoints. Sensitivity of L. stagnalis to TBT and TPT was compared with the sensitivity of prosobranch mollusks with different habitats and different reproductive strategies.

  1. Lymnaea glabra: progressive increase in susceptibility to Fasciola hepatica through successive generations of experimentally infected snails.

    PubMed

    Rondelaud, D; Teukeng, F F Djuikwo; Vignoles, P; Dreyfuss, G

    2015-07-01

    Experimental infections of Lymnaea glabra (two populations) with Fasciola hepatica were carried out during seven successive snail generations, to determine if prevalence and intensity of snail infection increased over time through descendants of snails already infected with F. hepatica. Controls were descendants coming from uninfected parents and infected according to the same protocol. No larval forms were found in the bodies of control snails coming from uninfected parents. In contrast, prevalence and intensity of F. hepatica infection in snails originating from infected parents progressively increased from the F2 or F3 to the F6 generation of L. glabra. In another experiment carried out with the F7 generations of L. glabra and a single generation of Galba truncatula (as controls), the prevalence of F. hepatica infection and the total number of cercariae were lower in L. glabra (without significant differences between both populations). If the number of cercariae shed by infected snails was compared to overall cercarial production noted in snails containing cercariae but dying without emission, the percentage was greater in G. truncatula (69% instead of 52-54% in L. glabra). Even if most characteristics of F. hepatica infection were lower in L. glabra, prevalence and intensity of parasite infection increased with snail generation when tested snails came from infected parents. This mode of snail infection with F. hepatica suggests an explanation for cases of fasciolosis occurring in cattle-breeding farms where paramphistomosis is lacking and G. truncatula is absent.

  2. Optimizing the design of a reproduction toxicity test with the pond snail Lymnaea stagnalis.

    PubMed

    Charles, Sandrine; Ducrot, Virginie; Azam, Didier; Benstead, Rachel; Brettschneider, Denise; De Schamphelaere, Karel; Filipe Goncalves, Sandra; Green, John W; Holbech, Henrik; Hutchinson, Thomas H; Faber, Daniel; Laranjeiro, Filipe; Matthiessen, Peter; Norrgren, Leif; Oehlmann, Jörg; Reategui-Zirena, Evelyn; Seeland-Fremer, Anne; Teigeler, Matthias; Thome, Jean-Pierre; Tobor Kaplon, Marysia; Weltje, Lennart; Lagadic, Laurent

    2016-11-01

    This paper presents the results from two ring-tests addressing the feasibility, robustness and reproducibility of a reproduction toxicity test with the freshwater gastropod Lymnaea stagnalis (RENILYS strain). Sixteen laboratories (from inexperienced to expert laboratories in mollusc testing) from nine countries participated in these ring-tests. Survival and reproduction were evaluated in L. stagnalis exposed to cadmium, tributyltin, prochloraz and trenbolone according to an OECD draft Test Guideline. In total, 49 datasets were analysed to assess the practicability of the proposed experimental protocol, and to estimate the between-laboratory reproducibility of toxicity endpoint values. The statistical analysis of count data (number of clutches or eggs per individual-day) leading to ECx estimation was specifically developed and automated through a free web-interface. Based on a complementary statistical analysis, the optimal test duration was established and the most sensitive and cost-effective reproduction toxicity endpoint was identified, to be used as the core endpoint. This validation process and the resulting optimized protocol were used to consolidate the OECD Test Guideline for the evaluation of reproductive effects of chemicals in L. stagnalis.

  3. Crystal structures of Lymnaea stagnalis AChBP in complex with neonicotinoid insecticides imidacloprid and clothianidin

    PubMed Central

    Ihara, Makoto; Okajima, Toshihide; Yamashita, Atsuko; Oda, Takuma; Hirata, Koichi; Nishiwaki, Hisashi; Morimoto, Takako; Akamatsu, Miki; Ashikawa, Yuji; Kuroda, Shun’ichi; Mega, Ryosuke; Kuramitsu, Seiki; Sattelle, David B.

    2008-01-01

    Neonicotinoid insecticides, which act on nicotinic acetylcholine receptors (nAChRs) in a variety of ways, have extremely low mammalian toxicity, yet the molecular basis of such actions is poorly understood. To elucidate the molecular basis for nAChR–neonicotinoid interactions, a surrogate protein, acetylcholine binding protein from Lymnaea stagnalis (Ls-AChBP) was crystallized in complex with neonicotinoid insecticides imidacloprid (IMI) or clothianidin (CTD). The crystal structures suggested that the guanidine moiety of IMI and CTD stacks with Tyr185, while the nitro group of IMI but not of CTD makes a hydrogen bond with Gln55. IMI showed higher binding affinity for Ls-AChBP than that of CTD, consistent with weaker CH–π interactions in the Ls-AChBP–CTD complex than in the Ls-AChBP–IMI complex and the lack of the nitro group-Gln55 hydrogen bond in CTD. Yet, the NH at position 1 of CTD makes a hydrogen bond with the backbone carbonyl of Trp143, offering an explanation for the diverse actions of neonicotinoids on nAChRs. PMID:18338186

  4. NCS-1 differentially regulates growth cone and somata calcium channels in Lymnaea neurons.

    PubMed

    Hui, Kwokyin; Feng, Zhong-Ping

    2008-02-01

    Local voltage-gated calcium channels, which regulate intracellular Ca2+ levels by allowing Ca2+ influx, play an important role in guiding and shaping growth cones, and in regulating the outgrowth and branching of neurites. Therefore, elucidating the mechanisms that regulate the biophysical properties of whole-cell calcium currents in the growth cones and somata of growing neurons is important to improving our understanding of neuronal development and regeneration. In this study, taking advantage of the large size of the pedal A (PeA) neurons in Lymnaea stagnalis, we compared the biophysical properties of somata and growth cone whole-cell calcium channel currents using Ba2+ and Ca2+ as current carriers. We found that somata and growth cone currents exhibit similar high-voltage activation properties. However, Ba2+ and Ca2+ currents in growth cones and somata are differentially affected by a dominant-negative peptide containing the C-terminal amino acid sequence of neuronal calcium sensor-1 (NCS-1). The peptide selectively reduces the peak and sustained components of current densities and the slope conductance in growth cones, and shifts the reversal potential of the growth cone currents to more hyperpolarized voltages. In contrast, the peptide had no significant effect on the somata calcium channels. Thus, we conclude that NCS-1 differentially modulates Ca2+ currents in the somata and growth cones of regenerating neurons, and may serve as a key regulator to facilitate the growth cone calcium channel activity.

  5. Mucus secretion by the freshwater snail Lymnaea stagnalis limits aluminum concentrations of the aqueous environment

    SciTech Connect

    Jugdaohsingh, R.; Thompson, R.P.H.; Powell, J.J.; Campbell, M.M.; Mccrohan, C.R.; White, K.N.

    1998-09-01

    Extracellular mucopolysaccharide (EPS) is a significant component in many waters. Its role in the cycling and mobilization of metals is unclear. In vitro studies were conducted to examine the influence of EPS, secreted by the freshwater pond snail, Lymnaea stagnalis, on soluble water Al concentrations at near-neutral pH. Snails maintained in aerated water of known ion content and added aluminum reduced Al in solution as compared to controls. Although snails accumulated Al into soft tissue, this only accounted for a small percentage of the total reduction. The remaining Al was recovered following acidification of the water. This observation was attributed to pedal EPS secreted by L. stagnalis which is chiefly insoluble and substrate bound. The Al that remained in solution was more labile, possibly due to the influence of soluble EPS. Further experiments with isolated EPS, confirmed that this poorly soluble film binds and reduces Al in solution. The influence of EPS on the solution chemistry and bioavailability of Al and possibly other metals may be important in natural waters.

  6. Comparative toxicity of Paraquat herbicide and some plant extracts in Lymnaea natalensis snails.

    PubMed

    Bakry, Fayez A; Eleiwa, Mona E; Taha, Samir A; Ismil, Somya M

    2016-01-01

    Paraquat has been shown to be a highly toxic compound for humans and animals, and many cases of acute poisoning and death have been reported over the past few decades. The present study was undertaken to evaluate comprehensively herbicides (Paraquat) and some plant extracts to biochemical aspects of Lymnaea natalensis snails. It was found that the exposure of L. natalensis to Paraquat and plant extracts led to a significant reduction in the infectivity of Fasciola gigantica miracidia to the snail. The glucose level in hemolymph of exposed snails was elevated, while the glycogen showed a decrease in soft tissues when compared with the control group. In addition, the activity level of some enzymes representing glycolytic enzymes as hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK), lactate dehydrogenase (LDH), and glucose phosphate isomerase (GPI) in snail's tissues were reduced in response to the treatment. It was concluded that the pollution of the aquatic environment by herbicide would adversely affect the metabolism of the L. natalensis snails. Snails treated with Agave attenuate, Ammi visnaga, and Canna iridiflora plant had less toxic effect compared to snails treated with Paraquat.

  7. Echinostomatid larval stages in Lymnaea viatrix (Gastropoda: Pulmonata) from southwest Patagonia, Argentina.

    PubMed

    Prepelitchi, Lucila; de Núñez, Margarita Ostrowski

    2007-04-01

    The partial life cycle of an echinostomatid found in Lymnaea viatrix from Patagonia, Argentina, was experimentally clarified. Emerging cercariae were exposed to laboratory-reared specimens of Biomphalaria sp. Metacercariae obtained from both naturally and experimentally infected snails were force-fed to chicks. Specimens recovered from the chicks belong to Echinoparyphium sp. on the basis of morphological features. The studied species possesses 43 collar spines arranged in 4-4-27-4-4 at all stages, a cercariae with over 100 small corpuscles in the excretory system, a cercariae tail without finfolds, and a metacercariae with a thin cyst wall. The present species cannot be assigned to Echinoparyphium megacirrus despite their morphological similarity because of differences in the habitats of L. viatrix and the intermediate hosts of E. megacirrus, namely Chilina dombeiana, Diplodon chilensis, and Temnocephala chilensis. More information on some life cycle stages and on the ecology of the intermediate hosts is needed to clarify the taxonomic status of the parasite. This study represents the first detailed description of parasites other than Fasciola hepatica in L. viatrix from Argentina.

  8. Molluscicidal activity of Sapindus mukorossi and Terminalia chebula against the freshwater snail Lymnaea acuminata.

    PubMed

    Upadhyay, Aparna; Singh, D K

    2011-04-01

    The molluscicidal activity of Sapindus mukorossi and Terminalia chebula fruit powder against the vector snail Lymnaea acuminata was time and concentration dependent. The molluscicidal activity of T. chebula fruit powder (96 h LC(50):93.59 mg L(-1)) was more pronounced than that of S. mukorossi fruit powder (96 h LC(50):119.57 mg L(-1)). Ethanolic extracts of S. mukorossi and T. chebula fruit powder were more toxic than their other organic solvent extracts. The molluscicidal activity of ethanolic extract of S. mukorossi fruit powder (24h LC(50):2.75 mg L(-1)) was more effective than the ethanolic extract of T. chebula fruit powder (24h LC(50):124.06 mg L(-1)). The 96 h LC(50) of column-purified fraction of S. mukorossi fruit powder was 5.43 mg L(-1) whereas those of T. chebula fruit powder was 7.49 mg L(-1). Column, thin layer and high performance liquid chromatography analysis demonstrates that the active molluscicidal component in S. mukorossi and T. chebula is saponin (96 h LC(50):1.31 mg L(-1)) and tannic acid (96 h LC(50):1.64 mg L(-1)), respectively. These plants may be used as potent source of molluscicides against the snail L. acuminata.

  9. Millimeter waves thermally alter the firing rate of the Lymnaea pacemaker neuron

    SciTech Connect

    Alekseev, S.I.; Kochetkova, N.V.; Ziskin, M.C.; Bolshakov, M.A.

    1997-05-01

    The effects of millimeter waves (mm-waves, 75 GHz) and temperature elevation on the firing rate of the BP-4 pacemaker neuron of the pond snail Lymnaea stagnalis were studied by using microelectrode techniques. The open end of a rectangular waveguide covered with a thin Teflon film served as a radiator. Specific absorption rates (SARs), measured in physiological solution at the radiator outlet, ranged from 600 to 4,200 W/kg, causing temperature rises from 0.3 to 2.2 C, respectively. Irradiation at an SAR of 4,200 W/kg caused a biphasic change in the firing rate, i.e., a transient decrease in the firing rate followed by a gradual increase to a new level that was 68 {+-} 21% above control. The biphasic changes in the firing rate were reproduced by heating under the condition that the magnitude (2 C) and the rate of temperature rise were equal to those produced by the irradiation. The addition of 0.05 mM of ouabain caused the disappearance of transient responses of the neuron to the irradiation. It was shown that the rate of temperature rise played an important role in the development of a transient neuronal response. The threshold stimulus for a transient response of the BP-4 neutron found in warming experiments was a temperature rise of 0.0025 C/s.

  10. A two-neuron system for adaptive goal-directed decision-making in Lymnaea.

    PubMed

    Crossley, Michael; Staras, Kevin; Kemenes, György

    2016-06-03

    During goal-directed decision-making, animals must integrate information from the external environment and their internal state to maximize resource localization while minimizing energy expenditure. How this complex problem is solved by the nervous system remains poorly understood. Here, using a combined behavioural and neurophysiological approach, we demonstrate that the mollusc Lymnaea performs a sophisticated form of decision-making during food-searching behaviour, using a core system consisting of just two neuron types. The first reports the presence of food and the second encodes motivational state acting as a gain controller for adaptive behaviour in the absence of food. Using an in vitro analogue of the decision-making process, we show that the system employs an energy management strategy, switching between a low- and high-use mode depending on the outcome of the decision. Our study reveals a parsimonious mechanism that drives a complex decision-making process via regulation of levels of tonic inhibition and phasic excitation.

  11. Variability and frequent failure of lucifer yellow to pass between two electrically coupled neurons in Lymnaea stagnalis.

    PubMed

    Audesirk, G; Audesirk, T; Bowsher, P

    1982-07-01

    The electrically coupled giant neurosecretory neurons VD1 and RPD2 of Lymnaea stagnalis were found to have coupling coefficients ranging from ca. 0.1-0.6. When the fluoroescent dye Lucifer Yellow was injected intracellularly into one of the neurons, in most preparations no dye was observed to pass through into the coupled cell body or the process leading to it. There was no apparent correlation between the amount of dye coupling and the length of time allowed for diffusion of the dye in the cells. In eight preparations, the electrical coupling coefficient was measured before dye was injected. There was no correlation between dye coupling and the electrical coupling coefficient.

  12. Toxicity of Chlorophyllin against Lymnaea acuminata at Different Wavelengths of Visible Light

    PubMed Central

    Chaturvedi, Divya; Singh, Vinay Kumar

    2016-01-01

    Fasciolosis is a water and food-borne disease caused by the liver fluke Fasciola hepatica and Fasciola gigantica. This disease is widespread in different parts of the world. Lymnaeidae and Planorbidae snails are the intermediate hosts of these flukes. Snail population management is a good tool to control fasciolosis because gastropods represent the weakest link in the life-cycle of trematodes. Chlorophyll can be extracted from any green plant. Chlorophyllin was prepared from spinach in 100% ethanol by using different types of chemicals. The chlorophyll obtained from spinach was transformed into water-soluble chlorophyllin. In the present paper, toxicity of chlorophyllin against the snail Lymnaea acuminata was time and concentration dependent. The toxicity of extracted and pure chlorophyllin at continuous 4 h exposure of sunlight was highest with lethal concentration (LC50) of 331.01 mg/L and 2.60 mg/L, respectively, than discontinuous exposure of sunlight up to 8 h with LC50 of 357.04 mg/L and 4.94 mg/L, respectively. Toxicity of extracted chlorophyllin was noted in the presence of different monochromatic visible lights. The highest toxicity was noted in yellow light (96 h, LC50 392.77 mg/L) and the lowest in green light (96 h, LC50 833.02 mg/L). Chlorophyllin in combination with solar radiation or different wavelength of monochromatic visible lights may become a latent remedy against the snail L. acuminata. It was demonstrated that chlorophyllin was more toxic in sunlight. Chlorophyllin is ecologically safe and more economical than synthetic molluscicides which have the potential to control the incidence of fasciolosis in developing countries. PMID:27688849

  13. Enhanced memory persistence is blocked by a DNA methyltransferase inhibitor in the snail Lymnaea stagnalis.

    PubMed

    Lukowiak, Ken; Heckler, Benjamin; Bennett, Thomas E; Schriner, Ellen K; Wyrick, Kathryn; Jewett, Cynthia; Todd, Ryan P; Sorg, Barbara A

    2014-08-15

    Lymnaea stagnalis provides an excellent model system for studying memory because these snails have a well-described set of neurons, a single one of which controls expression of long-term memory of operantly conditioned respiratory behavior. We have shown that several different manipulations, including pre-training exposure to serotonin (5-HT) or methamphetamine, submersion of snails after training to prevent memory interference, and exposure to effluent from predatory crayfish (CE), enhance memory persistence. Changes in DNA methylation underlie formation of strong memories in mammals and 5-HT-enhanced long-term facilitation in Aplysia. Here we determined the impact of the DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5-AZA; 87 μmol l(-1)), on enhanced memory persistence by all four manipulations. We found that 5-HT (100 μmol l(-1)) enhanced memory persistence, which was blocked by 5-AZA pretreatment. Snails pre-exposed to 3.3 μmol l(-1) Meth 4 h prior to training demonstrated memory 72 h later, which was not present in controls. This memory-enhancing effect was blocked by pre-treatment with 87 μmol l(-1) 5-AZA. Similarly, submersion to prevent interference learning as well as training in CE produced memory that was not present in controls, and these effects were blocked by pre-treatment with 87 μmol l(-1) 5-AZA. In contrast, 5-AZA injection did not alter expression of normal (non-enhanced) memory, suggesting that these four stimuli enhance memory persistence by increasing DNA methyltransferase activity, which, in turn, increases expression of memory-enhancing genes and/or inhibits memory suppressor genes. These studies lay important groundwork for delineating gene methylation changes that are common to persistent memory produced by different stimuli.

  14. An ancient process in a modern mollusc: early development of the shell in Lymnaea stagnalis

    PubMed Central

    2013-01-01

    Background The morphological variety displayed by the molluscan shell underlies much of the evolutionary success of this phylum. However, the broad diversity of shell forms, sizes, ornamentations and functions contrasts with a deep conservation of early cell movements associated with the initiation of shell construction. This process begins during early embryogenesis with a thickening of an ectodermal, ‘dorsal’ (opposite the blastopore) population of cells, which then invaginates into the blastocoel to form the shell gland. The shell gland evaginates to form the shell field, which then expands and further differentiates to eventually become the adult shell-secreting organ commonly known as the mantle. Despite the deep conservation of the early shell forming developmental program across molluscan classes, little is known about the fine-scale cellular or molecular processes that underlie molluscan shell development. Results Using modern imaging techniques we provide here a description of the morphogenesis of a gastropod shell gland and shell field using the pulmonate gastropod Lymnaea stagnalis as a model. We find supporting evidence for a hypothesis of molluscan shell gland specification proposed over 60 years ago, and present histochemical assays that can be used to identify a variety of larval shell stages and distinct cell populations in whole mounts. Conclusions By providing a detailed spatial and temporal map of cell movements and differentiation events during early shell development in L. stagnalis we have established a platform for future work aimed at elucidation of the molecular mechanisms and regulatory networks that underlie the evo-devo of the molluscan shell. PMID:23845038

  15. Toxicity of Chlorophyllin against Lymnaea acuminata at Different Wavelengths of Visible Light.

    PubMed

    Chaturvedi, Divya; Singh, Vinay Kumar

    2016-08-01

    Fasciolosis is a water and food-borne disease caused by the liver fluke Fasciola hepatica and Fasciola gigantica. This disease is widespread in different parts of the world. Lymnaeidae and Planorbidae snails are the intermediate hosts of these flukes. Snail population management is a good tool to control fasciolosis because gastropods represent the weakest link in the life-cycle of trematodes. Chlorophyll can be extracted from any green plant. Chlorophyllin was prepared from spinach in 100% ethanol by using different types of chemicals. The chlorophyll obtained from spinach was transformed into water-soluble chlorophyllin. In the present paper, toxicity of chlorophyllin against the snail Lymnaea acuminata was time and concentration dependent. The toxicity of extracted and pure chlorophyllin at continuous 4 h exposure of sunlight was highest with lethal concentration (LC50) of 331.01 mg/L and 2.60 mg/L, respectively, than discontinuous exposure of sunlight up to 8 h with LC50 of 357.04 mg/L and 4.94 mg/L, respectively. Toxicity of extracted chlorophyllin was noted in the presence of different monochromatic visible lights. The highest toxicity was noted in yellow light (96 h, LC50 392.77 mg/L) and the lowest in green light (96 h, LC50 833.02 mg/L). Chlorophyllin in combination with solar radiation or different wavelength of monochromatic visible lights may become a latent remedy against the snail L. acuminata. It was demonstrated that chlorophyllin was more toxic in sunlight. Chlorophyllin is ecologically safe and more economical than synthetic molluscicides which have the potential to control the incidence of fasciolosis in developing countries.

  16. Pronase acutely modifies high voltage-activated calcium currents and cell properties of Lymnaea neurons.

    PubMed

    Hermann, P M; Lukowiak, K; Wildering, W C; Bulloch, A G

    1997-12-01

    Pronase E ('pronase') is one of the proteolytic enzymes that are used in preparative procedures such as cell isolation and to soften the sheath of invertebrate ganglia. Although several effects of proteolytic enzymes on the physiology of non-neuronal tissues have been described, the effects of these enzymes on central neurons have received little attention. We examined the effects of bath-applied pronase on neurons in the Lymnaea central nervous system and in vitro. Pronase caused action potential broadening in neurons that exhibit a shoulder on the repolarization phase of their action potentials. This effect of pronase was accompanied by, although unrelated to, a depolarization and decrease in action potential interval. Some, but not all, effects of pronase in the central nervous system were reversible. For example, the decreases in membrane potential and action potential interval were both reversed after approximately 1 h of washing with saline. However, the effect of pronase on the action potential duration was not reversed after a period of 90 min. The modulation of action potential width prompted us to examine Ca2+ currents. Exposure to pronase resulted in an increase in both peak and late high voltage-activated Ca2+ currents in isolated neurons. Pronase neither changed the inactivation rate nor caused a shift in the current-voltage relationship of the current. The changes in action potential duration could be prevented by application of 0.1 mM Cd2+, indicating that the action potential broadening caused by pronase depends on Ca2+ influx. This is the first systematic study of the acute and direct actions of pronase on Ca2+ currents and cell properties both in the CNS and in vitro.

  17. Evidence of trophic transfer of microcystins from the gastropod Lymnaea stagnalis to the fish Gasterosteus aculeatus.

    PubMed

    Lance, Emilie; Petit, Anais; Sanchez, Wilfried; Paty, Christelle; Gérard, Claudia; Bormans, Myriam

    2014-01-01

    According to our previous results the gastropod Lymnaea stagnalis exposed to MC-producing cyanobacteria accumulates microcystins (MCs) both as free and covalently bound forms in its tissues, therefore representing a potential risk of MC transfer through the food web. This study demonstrates in a laboratory experiment the transfer of free and bound MCs from L. stagnalis intoxicated by MC-producing Planktothrix agardhii ingestion to the fish Gasterosteus aculeatus. Fish were fed during five days with digestive glands of L. stagnalis containing various concentrations of free and bound MCs, then with toxin-free digestive glands during a 5-day depuration period. MC accumulation was measured in gastropod digestive gland and in various fish organs (liver, muscle, kidney, and gills). The impact on fish was evaluated through detoxification enzyme (glutathion-S-transferase, glutathion peroxydase and superoxyde dismutase) activities, hepatic histopathology, and modifications in gill ventilation, feeding and locomotion. G. aculeatus ingestion rate was similar with intoxicated and toxin-free diet. Fish accumulated MCs (up to 3.96±0.14μgg(-1)DW) in all organs and in decreasing order in liver, muscle, kidney and gills. Hepatic histopathology was moderate. Glutathion peroxydase was activated in gills during intoxication suggesting a slight reactive oxygen species production, but without any impact on gill ventilation. Intoxication via ingestion of MC-intoxicated snails impacted fish locomotion. Intoxicated fish remained significantly less mobile than controls during the intoxication period possibly due to a lower health condition, whereas they showed a greater mobility during the depuration period that might be related to an acute foraging for food. During depuration, MC elimination was total in gills and kidney, but partial in liver and muscle. Our results assess the MC transfer from gastropods to fish and the potential risk induced by bound MCs in the food web.

  18. Pituitary adenylate cyclase activating polypeptide (PACAP) and its receptors are present and biochemically active in the central nervous system of the pond snail Lymnaea stagnalis.

    PubMed

    Pirger, Zsolt; Laszlo, Zita; Hiripi, Laszlo; Hernadi, Laszlo; Toth, Gabor; Lubics, Andrea; Reglodi, Dora; Kemenes, Gyorgy; Mark, Laszlo

    2010-11-01

    PACAP is a highly conserved adenylate cyclase (AC) activating polypeptide, which, along with its receptors (PAC1-R, VPAC1, and VPAC2), is expressed in both vertebrate and invertebrate nervous systems. In vertebrates, PACAP has been shown to be involved in associative learning, but it is not known if it plays a similar role in invertebrates. To prepare the way for a detailed investigation into the possible role of PACAP and its receptors in a suitable invertebrate model of learning and memory, here, we undertook a study of their expression and biochemical role in the central nervous system of the pond snail Lymnaea stagnalis. Lymnaea is one of the best established invertebrate model systems to study the molecular mechanisms of learning and memory, including the role of cyclic AMP-activated signaling mechanisms, which crucially depend on the learning-induced activation of AC. However, there was no information available on the expression of PACAP and its receptors in sensory structures and central ganglia of the Lymnaea nervous system known to be involved in associative learning or whether or not PACAP can actually activate AC in these ganglia. Here, using matrix-assisted laser desorption ionization time of flight (MALDI-TOF) and immunohistochemistry, we established the presence of PACAP-like peptides in the cerebral ganglia and the lip region of Lymnaea. The MALDI-TOF data indicated an identity with mammalian PACAP-27 and the presence of a squid-like PACAP-38 highly homologous to vertebrate PACAP-38. We also showed that PACAP, VIP, and maxadilan stimulated the synthesis of cAMP in Lymnaea cerebral ganglion homogenates and that this effect was blocked by the appropriate general and selective PACAP receptor antagonists.

  19. Investigations into the mechanism of lead toxicity to the freshwater pulmonate snail, Lymnaea stagnalis.

    PubMed

    Brix, Kevin V; Esbaugh, Andrew J; Munley, Kathleen M; Grosell, Martin

    2012-01-15

    The freshwater pulmonate snail, Lymnaea stagnalis, is the most sensitive aquatic organism tested to date for Pb with an estimated EC20 for juvenile snail growth of 3 μg l⁻¹. A previous study supported the hypothesis that this hypersensitivity to Pb was due to an extremely high Ca²⁺ uptake rate needed to support shell formation. The current study sought to build upon this working hypothesis and develop a mechanistic predictive model for inhibition of snail growth as a function of Pb exposure. Initial experiments confirmed previous predictions that juvenile snails have net Ca²⁺ uptake rates of 7000-8000 nmol g⁻¹ h⁻¹, approximately 100-fold higher than observed in a typical freshwater fish. However, an initial time course study revealed that the onset of growth inhibition occurs at least 4d prior to inhibition of net Ca²⁺ flux in Pb-exposed snails indicating the latter is not the primary mechanism of action. Qualitative observations during this experiment indicated snail feeding was inhibited in a dose-dependent manner. A subsequent experiment demonstrated that when food is withheld from snails for even 24 h, net Ca²⁺ uptake is significantly (∼50%) reduced. A second time course study demonstrated quantitatively that snail feeding is inhibited by Pb exposure by up to 98% at relatively high Pb concentrations (57 μg l⁻¹) but no inhibition was observed at ≤ 10 μg l⁻¹ Pb indicating feeding inhibition is not causing observed growth effects at concentrations approximating the EC20 of 3 μg l⁻¹ Pb. A final experiment testing whether Pb-induced growth effects are related to inhibition of carbonic anhydrase activity in the snail mantle also failed to demonstrate an effect. We conclude that while both feeding and net Ca²⁺ uptake in snails are affected by Pb exposure, they appear to be secondary effects. The primary mechanism of action explaining L. stagnalis hypersensitivity to Pb remains to be identified.

  20. Lymnaea schirazensis, an Overlooked Snail Distorting Fascioliasis Data: Genotype, Phenotype, Ecology, Worldwide Spread, Susceptibility, Applicability

    PubMed Central

    Bargues, María Dolores; Artigas, Patricio; Khoubbane, Messaoud; Flores, Rosmary; Glöer, Peter; Rojas-García, Raúl; Ashrafi, Keyhan; Falkner, Gerhard; Mas-Coma, Santiago

    2011-01-01

    Background Lymnaeid snails transmit medical and veterinary important trematodiases, mainly fascioliasis. Vector specificity of fasciolid parasites defines disease distribution and characteristics. Different lymnaeid species appear linked to different transmission and epidemiological patterns. Pronounced susceptibility differences to absolute resistance have been described among lymnaeid populations. When assessing disease characteristics in different endemic areas, unexpected results were obtained in studies on lymnaeid susceptibility to Fasciola. We undertook studies to understand this disease transmission heterogeneity. Methodology/Principal Findings A ten-year study in Iran, Egypt, Spain, the Dominican Republic, Mexico, Venezuela, Ecuador and Peru, demonstrated that such heterogeneity is not due to susceptibility differences, but to a hitherto overlooked cryptic species, Lymnaea schirazensis, confused with the main vector Galba truncatula and/or other Galba/Fossaria vectors. Nuclear rDNA and mtDNA sequences and phylogenetic reconstruction highlighted an old evolutionary divergence from other Galba/Fossaria species, and a low intraspecific variability suggesting a recent spread from one geographical source. Morphometry, anatomy and egg cluster analyses allowed for phenotypic differentiation. Selfing, egg laying, and habitat characteristics indicated a migration capacity by passive transport. Studies showed that it is not a vector species (n = 8572 field collected, 20 populations): snail finding and penetration by F. hepatica miracidium occur but never lead to cercarial production (n = 338 experimentally infected). Conclusions/Significance This species has been distorting fasciolid specificity/susceptibility and fascioliasis geographical distribution data. Hence, a large body of literature on G. truncatula should be revised. Its existence has henceforth to be considered in research. Genetic data on livestock, archeology and history along the 10,000-year

  1. Phyto-adaptogens protect against environmental stress-induced death of embryos from the freshwater snail Lymnaea stagnalis.

    PubMed

    Boon-Niermeijer, E K; van den Berg, A; Wikman, G; Wiegant, F A

    2000-10-01

    The main purpose of the studies presented in this paper is twofold: 1) to evaluate whether phyto-adaptogens (Acanthopanax senticosus and Rhodiola rosea) are able to exert a protective action against stress-induced death of embryos of the pond snail Lymnaea stagnalis; and 2) whether a possible protective action by phyto-adaptogens can be explained by the induction of heat shock proteins. Enhancement in resistance by phyto-adaptogens was studied by applying plant extracts for a period of 20 hours to 3-day old larvae of the pond snail Lymnaea stagnalis. Subsequently they were exposed to a high and toxic dose of different environmental stressors. The following stress conditions were selected: a physical stress condition (heat shock: 43 degrees C for 4 minutes), an oxidative stress condition (superoxide radicals induced by menadione (600 microM for 2 hours)) and heavy metal-induced stress (copper (150 microM for 1 hour) or cadmium (20 microM during 1 hour)). Both Acanthopanax and Rhodiola exert a strong protective action against a lethal heat shock. These adaptogens also significantly protect against the negative effect of superoxide radicals as induced by menadione. With respect to the protective action against exposure to heavy metals a small but significant protection was observed against intoxication with copper or cadmium by the phyto-adaptogens. In summary, there appears to be a difference in efficiency in enhancing resistance to the various stress conditions used (heat shock>menadione>copper>cadmium). Based on the results presented in this paper, we can conclude that phyto-adaptogens are able to enhance the resistance against the different stress conditions tested in developing individuals of Lymnaea. Although the degree to which resistance is enhanced appears to depend on the type of stressor applied, our results confirm the definition of phyto-adaptogens as being universal enhancers of non-specific resistance against different kinds of stress conditions. With

  2. Binary Combination of Carica papaya, Areca catechu and Myristica fragrans with Piperonyl Butoxide / MGK-264 against Freshwater Snail Lymnaea acuminata

    PubMed Central

    Hanif, Farhat; Singh, Dinesh Kumar

    2013-01-01

    Piperonyl butoxide (PB) and MGK-264 were used to enhance the toxicity of the active components papain, arecoline and myristicin from the plants Carica papaya, Areca catechu and Myristica fragrans, respectively, against the vector snail Lymnaea acuminata. A time- and dose-dependent relationship was observed for the toxicity of these combinations. The toxic effects of these plant-derived molluscicides in combination with the synergists PB and MGK-264 were several times higher than the effect of the individual treatments. The highest degree of synergism was observed when MGK-264 was used in combination with C. papaya latex (10.47-fold increase) and PB was used with papain (8.35-fold increase). PMID:24575245

  3. Binary Combination of Carica papaya, Areca catechu and Myristica fragrans with Piperonyl Butoxide / MGK-264 against Freshwater Snail Lymnaea acuminata.

    PubMed

    Hanif, Farhat; Singh, Dinesh Kumar

    2013-12-01

    Piperonyl butoxide (PB) and MGK-264 were used to enhance the toxicity of the active components papain, arecoline and myristicin from the plants Carica papaya, Areca catechu and Myristica fragrans, respectively, against the vector snail Lymnaea acuminata. A time- and dose-dependent relationship was observed for the toxicity of these combinations. The toxic effects of these plant-derived molluscicides in combination with the synergists PB and MGK-264 were several times higher than the effect of the individual treatments. The highest degree of synergism was observed when MGK-264 was used in combination with C. papaya latex (10.47-fold increase) and PB was used with papain (8.35-fold increase).

  4. Effects of DDT and permethrin on neurite growth in cultured neurons of chick embryo brain and Lymnaea stagnalis.

    PubMed

    Ferguson, C A; Audesirk, G

    1990-01-01

    The pesticides permethrin and 1,1-bis(4-chlorophenyl)-2,2,2-trichloroethane (DDT), dissolved in either ethanol (EtOH) or dimethylsulphoxide (DMSO), were studied to determine their effect on neurite growth from cultured neurons of Lymnaea stagnalis and embryonic chicks. Both of these toxins decreased the percentage of neurons growing neurites, mean neurite length, and number of neurites/cell in a dose-dependent manner. DMSO increased the toxicity of permethrin and DDT in L. stagnalis neurons. EtOH was not used as a solvent with the embryonic chick cultures. Pre-existing neurites of L. stagnalis neurons exposed to permethrin regressed in a dose- and time-dependent manner. These two toxins may affect neurite outgrowth through interference with intracellular calcium regulation.

  5. Effect of DDT and MCPA (4-chloro-2-methylphenoxyacetic acid) on reproduction of the pond snail, Lymnaea stagnalis L

    SciTech Connect

    Woin, P.; Broenmark, C. )

    1992-01-01

    Reproduction is the single most important function in the life cycle of an organism. Successful reproduction determines fitness of organisms. The inability of an organism to complete any one stage of the reproductive process severely reduces its lifetime reproductive success. Disruptions in the reproduction will ultimately affect the abundance and distribution of the species. Therefore, laboratory tests of long-term impact of sublethal pollutant concentrations on organisms preferably is done on the reproductive success. Pollutants of diverse structure may affect the reproductive system which is sensitive to toxic agents. Certain pollutants, notably the organochlorine compounds, have been shown to affect the male and female reproductive systems. The authors have studied the effect of sublethal concentrations of DDT and the herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) on the reproductive output of the pulmonate snail Lymnaea stagnalis under a 2-mon exposure period.

  6. Kinetics of enzyme inhibition by active molluscicidal agents ferulic acid, umbelliferone, eugenol and limonene in the nervous tissue of snail Lymnaea acuminata.

    PubMed

    Kumar, Pradeep; Singh, V K; Singh, D K

    2009-02-01

    Ferulic acid, umbelliferone (Ferula asafoetida), eugenol (Syzygium aromaticum) and limonene (Carum carvi) are active molluscicidal components that inhibited the activity of alkaline phosphatase and acetylcholinesterase in in vivo and in vitro exposure of Lymnaea acuminata. It was observed that ferulic acid, umbelliferone and eugenol are competitive and limonene is a competitive-non-competitive inhibitor of alkaline phosphatase. Ferulic acid and umbelliferone are competitive, whereas eugenol and limonene are competitive-non-competitive and uncompetitive inhibitors of acetylcholinesterase, respectively.

  7. Effects of in vitro lead exposure on voltage-sensitive calcium channels differ among cell types in central neurons of Lymnaea stagnalis.

    PubMed

    Audesirk, G; Audesirk, T

    1989-01-01

    The effects of acute in vitro lead exposure on slowly inactivating voltage-sensitive calcium channels in central neurons of the freshwater pond snail Lymnaea stagnalis were studied under voltage clamp. Three physiologically distinct cell types were used: two subsets of the B cell cluster (Bpos and Bneg) and the pedal giant neuron (RPeD1). In Bpos neurons, 5 nM free Pb2+ irreversibly inhibited current flow through calcium channels by 38 +/- 10%. In Bneg neurons, 5 nM free Pb2+ slightly inhibited inward currents (12 +/- 6%) and may have shifted their voltage dependence to more depolarized voltages. The inhibition and voltage shift were irreversible. In RPeD1 neurons, Pb2+ caused a small, statistically insignificant inhibition of inward current (5 nM free Pb2+; 18 +/- 19%; 30 nM free Pb2+: 31 +/- 23%). The effects of Pb2+ were fully reversible. These data indicate that (1) voltage-sensitive calcium channels in Lymnaea neurons are inhibited by nanomolar concentrations of free Pb2+; (2) there are multiple types of calcium channels in Lymnaea neurons; and (3) the effects of in vitro lead exposure differ qualitatively among channel types.

  8. The Early Worm Catches the Bird? Productivity and Patterns of Trichobilharzia szidati Cercarial Emission from Lymnaea stagnalis

    PubMed Central

    Sures, Bernd

    2016-01-01

    Digenean trematodes are common and abundant in aquatic habitats and their free-living larvae, the cercariae, have recently been recognized as important components of ecosystems in terms of comprising a significant proportion of biomass and in having a potentially strong influence on food web dynamics. One strategy to enhance their transmission success is to produce high numbers of cercariae which are available during the activity peak of the next host. In laboratory experiments with 13 Lymnaea stagnalis snails infected with Trichobilharzia szidati the average daily emergence rate per snail was determined as 2,621 cercariae, with a maximum of 29,560. During a snail’s lifetime this summed up to a mass equivalent of or even exceeding the snail’s own body mass. Extrapolated for the eutrophic pond where the snails were collected, annual T. szidati biomass may reach 4.65 tons, a value equivalent to a large Asian elephant. Emission peaks were observed after the onset of illumination, indicating emission synchronizing with the high morning activities of the definitive hosts, ducks. However, high cercarial emission is possible throughout the day under favorable lightning conditions. Therefore, although bird schistosomes, such as T. szidati constitute only a fraction of the diverse trematode communities in the studied aquatic ecosystem, their cercariae can still pose a considerable risk for humans of getting cercarial dermatitis (swimmer's itch) due to the high number of cercariae emitted from infected snails. PMID:26895541

  9. Octopamine-containing (OC) interneurons enhance central pattern generator activity in sucrose-induced feeding in the snail Lymnaea.

    PubMed

    Vehovszky, Agnes; Szabó, Henriette; Elliott, Christopher J H

    2004-10-01

    In the pond snail Lymnaea stagnalis octopamine-containing (OC) interneurons trigger and reconfigure the feeding pattern in isolated CNS by excitation of the central pattern generator. In semi-intact (lip-mouth-CNS) preparations, this central pattern generator is activated by chemosensory inputs. We now test if sucrose application to the lips activates the OC neurons independently of the rest of the feeding central pattern generator, or if the OC interneuron is activated by inputs from the feeding network. In 66% of experiments, sucrose stimulated feeding rhythms and OC interneurons received regular synaptic inputs. Only rarely (14%) did the OC interneuron fire action potentials, proving that firing of OC interneurons is not necessary for the sucrose-induced feeding. Prestimulation of OC neurons increased the intensity and duration of the feeding rhythm evoked by subsequent sucrose presentations. One micromolar octopamine in the CNS bath mimicked the effect of OC interneuron stimulation, enhancing the feeding response when sucrose is applied to the lips. We conclude that the modulatory OC neurons are not independently excited by chemosensory inputs to the lips, but rather from the buccal central pattern generator network. However, when OC neurons fire, they release modulatory octopamine, which provides a positive feedback to the network to enhance the sucrose-activated central pattern generator rhythm.

  10. Inverse Relationship between Basal Pacemaker Neuron Activity and Aversive Long-Term Memory Formation in Lymnaea stagnalis

    PubMed Central

    Dong, Nancy; Feng, Zhong-Ping

    2017-01-01

    Learning and memory formation are essential physiological functions. While quiescent neurons have long been the focus of investigations into the mechanisms of memory formation, there is increasing evidence that spontaneously active neurons also play key roles in this process and possess distinct rules of activity-dependent plasticity. In this study, we used a well-defined aversive learning model of aerial respiration in the mollusk Lymnaea stagnalis (L. stagnalis) to study the role of basal firing activity of the respiratory pacemaker neuron Right Pedal Dorsal 1 (RPeD1) as a determinant of aversive long-term memory (LTM) formation. We investigated the relationship between basal aerial respiration behavior and RPeD1 firing activity, and examined aversive LTM formation and neuronal plasticity in animals exhibiting different basal aerial respiration behavior. We report that animals with higher basal aerial respiration behavior exhibited early responses to operant conditioning and better aversive LTM formation. Early behavioral response to the conditioning procedure was associated with biphasic enhancements in the membrane potential, spontaneous firing activity and gain of firing response, with an early phase spanning the first 2 h after conditioning and a late phase that is observed at 24 h. Taken together, we provide the first evidence suggesting that lower neuronal activity at the time of learning may be correlated with better memory formation in spontaneously active neurons. Our findings provide new insights into the diversity of cellular rules of plasticity underlying memory formation. PMID:28101006

  11. Testosterone levels and fecundity in the hermaphroditic aquatic snail Lymnaea stagnalis exposed to testosterone and endocrine disruptors.

    PubMed

    Giusti, Arnaud; Ducrot, Virginie; Joaquim-Justo, Célia; Lagadic, Laurent

    2013-08-01

    Endocrine disruptors are known to alter endogenous free and esterified levels of androgenic and estrogenic steroid hormones in aquatic mollusks. The origin of steroids in these animals, however, remains controversial. In the present study, free and esterified testosterone concentrations were measured in the hermaphroditic aquatic gastropod Lymnaea stagnalis exposed to molecules known for their androgenic (testosterone and tributyltin), anti-androgenic (cyproterone-acetate), and estrogenic (chlordecone) properties, by reference to their mode of action in vertebrates. In parallel, snail oviposition and fecundity were followed over a 21-d exposure period. Testosterone exposure resulted in increased esterified testosterone levels, whereas free testosterone concentrations remained stable. In contrast, cyproterone-acetate significantly increased the free form of testosterone with no changes in the esterified form, whereas chlordecone showed a tendency to reduce (though not significantly) esterified testosterone concentrations without changing free testosterone levels. Finally, tributyltin did not alter testosterone homeostasis. The production of egg clutches and eggs was significantly reduced only in the snails exposed to the highest concentrations of chlordecone (19.6 µg/L) and tributyltin (94.2 ng Sn/L). Overall, the present study demonstrates that uptake of testosterone from the exposure medium occurs in L. stagnalis. Moreover, it shows that cyproterone-acetate and, to a lesser extent, chlordecone can alter endogenous testosterone levels in this freshwater snail. However, the relationship between hormonal changes and snail reproduction has not been established. Environ Toxicol Chem 2013;32:1740-1745. © 2013 SETAC.

  12. Relationship between the grades of a learned aversive-feeding response and the dopamine contents in Lymnaea

    PubMed Central

    Aonuma, Hitoshi; Kaneda, Mugiho; Hatakeyama, Dai; Watanabe, Takayuki; Lukowiak, Ken

    2016-01-01

    ABSTRACT The pond snail Lymnaea learns conditioned taste aversion (CTA) and remembers not to respond to food substances that initially cause a feeding response. The possible relationship between how well snails learn to follow taste-aversion training and brain dopamine contents is not known. We examined this relationship and found the following: first, snails in the act of eating just before the commencement of CTA training were poor learners and had the highest dopamine contents in the brain; second, snails which had an ad libitum access to food, but were not eating just before training, were average learners and had lower dopamine contents; third, snails food-deprived for one day before training were the best learners and had significantly lower contents of dopamine compared to the previous two cohorts. There was a negative correlation between the CTA grades and the brain dopamine contents in these three cohorts. Fourth, snails food-deprived for five days before training were poor learners and had higher dopamine contents. Thus, severe hunger increased the dopamine content in the brain. Because dopamine functions as a reward transmitter, CTA in the severely deprived snails (i.e. the fourth cohort) was thought to be mitigated by a high dopamine content. PMID:27815244

  13. Molluscicidal activity of Ferula asafoetida, Syzygium aromaticum and Carum carvi and their active components against the snail Lymnaea acuminata.

    PubMed

    Kumar, Pradeep; Singh, D K

    2006-06-01

    The molluscicidal activity of dried root latex powder of Ferula asafoetida, flower-bud powder of Syzygium aromaticum and seed powder of Carum carvi against the snail Lymnaea acuminata was studied. The molluscicidal activity of all the three plant products was found to be both time and concentration dependent. The toxicity of S. aromaticum flower-bud powder (96 h LC(50):51.98 mg/l) was more pronounced than that of root latex powder of F. asafoetida (96 h LC(50):82.71 mg/l) and seed powder of C. carvi (96 h LC(50):140.58 mg/l). Ethanol extract was more toxic than other organic extracts. The ethanol extract of S. aromaticum (24h LC(50):83.53 mg/l) was more effective than that of F. asafoetida (24h LC(50):132.31 mg/l) and C. carvi (24h LC(50):130.61 mg/l) in killing the test animals. The 96 h LC(50) of column purified fraction of seed powder of C. carvi was 5.40 mg/l whereas those of flower-bud powder of S. aromaticum and dried root latex powder of F. asafoetida were 7.87 and 9.67 mg/l, respectively. The product of F. asafoetida, S. aromaticum and C. carvi may be used as potent molluscicides.

  14. Feeding of Bait to Snail Lymnaea acuminata and Their Effect on Certain Enzyme in the Nervous Tissue

    PubMed Central

    Kumar, Pradeep; Singh, V. K.; Singh, D. K.

    2012-01-01

    Fascioliasis, a snail-borne parasitic zoonosis, has been recognized for a long time because of its major veterinary and human impact. Different Bait formulations were fed to the snail Lymnaea acuminata in clear glass aquaria having diameter of 30 cm. Snail attractant containing bait formulations was prepared from different binary combination (1 : 1 ratio) of carbohydrates (glucose, starch 10 mM) and amino acid (methionine, histidine 10 mM) in 100 ml of 2% agar solution + sublethal (20% and 60% of 24 h and 96 h LC50) doses of different molluscicides (eugenol, ferulic acid, umbelliferone, and limonene). Snails fed on bait containing sub-lethal concentration of different molluscicides and the snail attractant, causing a significant inhibition in alkaline phosphatase (ALP) and acetylcholinesterase (AChE) activity in the nervous tissue of the vector snail L. acuminata. Maximum inhibition in ALP (20% of control) and AChE (49.49% of control) activity was observed in the nervous tissue of the L. acuminata exposed to 60% of 96 h LC50 of eugenol in the bait pellets containing starch + histidine, starch + methionine, respectively. PMID:25969756

  15. Modelling effects of diquat under realistic exposure patterns in genetically differentiated populations of the gastropod Lymnaea stagnalis

    PubMed Central

    Ducrot, Virginie; Péry, Alexandre R. R.; Lagadic, Laurent

    2010-01-01

    Pesticide use leads to complex exposure and response patterns in non-target aquatic species, so that the analysis of data from standard toxicity tests may result in unrealistic risk forecasts. Developing models that are able to capture such complexity from toxicity test data is thus a crucial issue for pesticide risk assessment. In this study, freshwater snails from two genetically differentiated populations of Lymnaea stagnalis were exposed to repeated acute applications of environmentally realistic concentrations of the herbicide diquat, from the embryo to the adult stage. Hatching rate, embryonic development duration, juvenile mortality, feeding rate and age at first spawning were investigated during both exposure and recovery periods. Effects of diquat on mortality were analysed using a threshold hazard model accounting for time-varying herbicide concentrations. All endpoints were significantly impaired at diquat environmental concentrations in both populations. Snail evolutionary history had no significant impact on their sensitivity and responsiveness to diquat, whereas food acted as a modulating factor of toxicant-induced mortality. The time course of effects was adequately described by the model, which thus appears suitable to analyse long-term effects of complex exposure patterns based upon full life cycle experiment data. Obtained model outputs (e.g. no-effect concentrations) could be directly used for chemical risk assessment. PMID:20921047

  16. Two proteolytic fragments of menin coordinate the nuclear transcription and postsynaptic clustering of neurotransmitter receptors during synaptogenesis between Lymnaea neurons

    PubMed Central

    Getz, Angela M.; Visser, Frank; Bell, Erin M.; Xu, Fenglian; Flynn, Nichole M.; Zaidi, Wali; Syed, Naweed I.

    2016-01-01

    Synapse formation and plasticity depend on nuclear transcription and site-specific protein targeting, but the molecular mechanisms that coordinate these steps have not been well defined. The MEN1 tumor suppressor gene, which encodes the protein menin, is known to induce synapse formation and plasticity in the CNS. This synaptogenic function has been conserved across evolution, however the underlying molecular mechanisms remain unidentified. Here, using central neurons from the invertebrate Lymnaea stagnalis, we demonstrate that menin coordinates subunit-specific transcriptional regulation and synaptic clustering of nicotinic acetylcholine receptors (nAChR) during neurotrophic factor (NTF)-dependent excitatory synaptogenesis, via two proteolytic fragments generated by calpain cleavage. Whereas menin is largely regarded as a nuclear protein, our data demonstrate a novel cytoplasmic function at central synapses. Furthermore, this study identifies a novel synaptogenic mechanism in which a single gene product coordinates the nuclear transcription and postsynaptic targeting of neurotransmitter receptors through distinct molecular functions of differentially localized proteolytic fragments. PMID:27538741

  17. Small-scale to large-scale and back: larval trematodes in Lymnaea stagnalis and Planorbarius corneus in Central Europe.

    PubMed

    Brown, Rebecca; Soldánová, Miroslava; Barrett, John; Kostadinova, Aneta

    2011-01-01

    We examined the small-scale temporal and spatial variability in composition and structure of larval trematode communities in Lymnaea stagnalis and Planorbarius corneus in two fish ponds in the Czech Republic and compared the patterns of richness and similarity to continental and regional trematode faunas of these hosts. The levels of parasitism in the populations of both hosts were high, the former parasitized predominantly by allogenic species maturing in a wide range of birds and the latter infected by relatively more species completing their life cycles in micromammals. Communities in both hosts exhibited a congruent pattern of seasonal change in overall infection rates and community composition with lower levels of infection in spring. Both temporal and spatial variation was closely related to the structure of snail populations, and no significant differentiation of community composition with respect to pond was observed. Comparisons with large-scale inventories revealed overall congruent patterns of decreased richness and similarity and increased variability at the smaller scales in both host-parasite systems. The relative compositional homogeneity of larval communities in both snail hosts irrespective of scale suggests that historical data at small to medium regional scales may provide useful estimates of past richness and composition of larval trematode communities in these snail hosts.

  18. Does water chemistry affect the dietary uptake and toxicity of silver nanoparticles by the freshwater snail Lymnaea stagnalis?

    USGS Publications Warehouse

    López-Serrano Oliver, Ana; Croteau, Marie-Noële; Stoiber, Tasha L.; Tejamaya, Mila; Römer, Isabella; Lead, Jamie R.; Luoma, Samuel N.

    2014-01-01

    Silver nanoparticles (AgNPs) are widely used in many applications and likely released into the aquatic environment. There is increasing evidence that Ag is efficiently delivered to aquatic organisms from AgNPs after aqueous and dietary exposures. Accumulation of AgNPs through the diet can damage digestion and adversely affect growth. It is well recognized that aspects of water quality, such as hardness, affect the bioavailability and toxicity of waterborne Ag. However, the influence of water chemistry on the bioavailability and toxicity of dietborne AgNPs to aquatic invertebrates is largely unknown. Here we characterize for the first time the effects of water hardness and humic acids on the bioaccumulation and toxicity of AgNPs coated with polyvinyl pyrrolidone (PVP) to the freshwater snail Lymnaea stagnalis after dietary exposures. Our results indicate that bioaccumulation and toxicity of Ag from PVP-AgNPs ingested with food are not affected by water hardness and by humic acids, although both could affect interactions with the biological membrane and trigger nanoparticle transformations. Snails efficiently assimilated Ag from the PVP-AgNPs mixed with diatoms (Ag assimilation efficiencies ranged from 82 to 93%). Rate constants of Ag uptake from food were similar across the entire range of water hardness and humic acid concentrations. These results suggest that correcting regulations for water quality could be irrelevant and ineffective where dietary exposure is important.

  19. Characterization of the molluscicidal activity of Bauhinia variegata and Mimusops elengi plant extracts against the fasciola vector Lymnaea acuminata.

    PubMed

    Singh, Kanchan Lata; Singh, D K; Singh, Vinay Kumar

    2012-01-01

    The molluscicidal activity of Bauhinia variegata leaf and Mimusops elengi bark was studied against vector snail Lymnaea acuminata. The toxicity of both plants was time and concentration-dependent. Among organic extracts, ethanol extracts of both plants were more toxic. Toxicity of B. variegata leaf ethanolic extract (96h LC50- 14.4 mg/L) was more pronounced than M. elengi bark ethanolic extract (96h LC50-15.0 mg/L). The 24h LC50 of column purified fraction of B. variegata and M. elengi bark were 20.3 mg/L and 18.3 mg/L, respectively. Saponin and quercetin were characterized and identified as active molluscicidal component. Co-migration of saponin (Rf 0.48) and quercetin (Rf 0.52) with column purified bark of M. elengi and leaf of B. variegata on thin layer chromatography demonstrate same Rf value i.e. 0.48 and 0.52, respectively. The present study clearly indicates the possibility of using M. elengi and/or B. variegata as potent molluscicide.

  20. Metabolomics to Explore Imidacloprid-Induced Toxicity in the Central Nervous System of the Freshwater Snail Lymnaea stagnalis.

    PubMed

    Tufi, Sara; Stel, Jente M; de Boer, Jacob; Lamoree, Marja H; Leonards, Pim E G

    2015-12-15

    Modern toxicology is seeking new testing methods to better understand toxicological effects. One of the most concerning chemicals is the neonicotinoid pesticide imidacloprid. Although imidacloprid is designed to target insects, recent studies have shown adverse effects on nontarget species. Metabolomics was applied to investigate imidacloprid-induced sublethal toxicity in the central nervous system of the freshwater snail Lymnaea stagnalis. The snails (n = 10 snails) were exposed for 10 days to increasing imidacloprid concentrations (0.1, 1, 10, and 100 μg/L). The comparison between control and exposure groups highlighted the involvement and perturbation of many biological pathways. The levels of several metabolites belonging to different metabolite classes were significantly changed by imidacloprid exposure. A change in the amino acids and nucleotide metabolites like tryptophan, proline, phenylalanine, uridine, and guanosine was found. Many fatty acids were down-regulated, and the levels of the polyamines, spermidine and putrescine, were found to be increased which is an indication of neuron cell injury. A turnover increase between choline and acetylcholine led us to hypothesize an increase in cholinergic gene expression to overcome imidacloprid binding to the nicotinic acetylcholine receptors. Metabolomics revealed imidacloprid induced metabolic changes at low and environmentally relevant concentration in a nontarget species and generated a novel mechanistic hypothesis.

  1. Molecular Evidence of Trichobilharzia Species (Digenea: Schistosomatidae) in the Snails of Lymnaea auricularia from Urmia Suburb, North West Iran

    PubMed Central

    YAKHCHALI, Mohammad; HOSSEINPANAHI, Asaad; MALEKZADEH-VIAYEH, Reza

    2016-01-01

    Background: The present study was carried out to detect the infection of larval stages of Trichobilharzia species in the snail Lymnaea auricularia in northwestern Iran based on DNA analysis. Methods: A total number of 320 snails of L. auricularia were sampled from four water-bodies located in the suburb of Urmia City, North West Iran, during May to November 2011. The snails were first microscopically inspected for the infection with larval stages of trematodes. Genomic DNA was extracted from the snails and PCR was performed to amplify a fragment of the ribosomal DNA of Trichobilharzia species in the infected snails. Results: Microscopic examinations indicated that 11.25% (36 out of 320) of the snails were infected with larval stages of trematodes, while the PCR patterns showed a much higher infection rate (31.25%, 100/320). According to the PCR, the infections were caused by the larval stages of T. szidati (21.56%, 69/320) and T. franki (9.69%, 31/320) or both of them (8.44%, 27/320). The infected snails were observed in three out of the four studied sites. The highest infection rate in a single site was 50% (25/50). Only 7.81% (25 out of 320) of the infected snails were from the plain areas, while the remaining was from high altitudes. Conclusion: Results of this study contribute the utility of the employed technique for quick and accurate detection of the infection with trichobilharzian species in their intermediate host snails, which may have potential zoonotic role in the region. PMID:28127334

  2. Proteomic Analysis of the Reproductive Organs of the Hermaphroditic Gastropod Lymnaea stagnalis Exposed to Different Endocrine Disrupting Chemicals

    PubMed Central

    Giusti, Arnaud; Leprince, Pierre; Mazzucchelli, Gabriel; Thomé, Jean-Pierre; Lagadic, Laurent; Ducrot, Virginie; Joaquim-Justo, Célia

    2013-01-01

    Many studies have reported perturbations of mollusc reproduction following exposure to low concentrations (ng/L range) of endocrine disrupting chemicals (EDCs). However, the mechanisms of action of these molecules on molluscs are still poorly understood. Investigation of the modifications of protein expression in organisms exposed to chemicals using proteomic methods can provide a broader and more comprehensive understanding of adverse impacts of pollution on organisms than conventional biochemical biomarkers (e.g., heat-shock proteins, metallothioneins, GST, EROD). In this study we have investigated the impacts of four chemicals, which exhibit different endocrine disrupting properties in vertebrates, on the proteome of the hermaphroditic freshwater pulmonate gastropod Lymnaea stagnalis after 21 days of exposure. Testosterone, tributyltin, chlordecone and cyproterone acetate were chosen as tested compounds as they can induce adverse effects on the reproduction of this snail. The 2D-DIGE method was used to identify proteins whose expression was affected by these compounds. In addition to modifying the expression of proteins involved in the structure and function of the cytoskeleton, chemicals had impacts on the expression of proteins involved in the reproduction of L. stagnalis. Exposure to 19.2 µg/L of chlordecone increased the abundance of ovipostatin, a peptide transmitted during mating through seminal fluid, which reduces oviposition in this species. The expression of yolk ferritin, the vitellogenin equivalent in L. stagnalis, was reduced after exposure to 94.2 ng Sn/L of tributyltin. The identification of yolk ferritin and the modification of its expression in snails exposed to chemicals were refined using western blot analysis. Our results showed that the tested compounds influenced the abundance of yolk ferritin in the reproductive organs. Alteration in proteins involved in reproductive pathways (e.g., ovipostatin and yolk ferritin) could constitute relevant

  3. The electrotonic location of low-resistance intercellular junctions between a pair of giant neurones in the snail Lymnaea.

    PubMed Central

    Benjamin, P R; Pilkington, J B

    1986-01-01

    The passive electrotonic properties of neurones VD1 and RPD2 in the brain of the snail Lymnaea can be represented by a soma-finite cable model with closed-circuit axon termination. There is a considerable individual variation in input resistance, membrane time constant, electrotonic length and axon-soma conductance ratio, but the average values for these parameters are similar in the two neurones. The cells are tightly coupled by an electrotonic synapse giving an average steady-state coupling coefficient of 0.68 and an average resistance measured between recording sites in the cell bodies of 20 M omega. Calculations using a model consisting of a symmetrical pair of cells with standard values for the electrotonic parameters show that in this system, for a soma-soma resistance of 20 M omega, the junction cannot be more than 0.16 length constants from the cell bodies. Reduction in coupling due to membrane current losses in such short proximal axon segments is insignificant. Intra-axonal recordings indicate that most of the coupling resistance is located at the junction between VD1 and RPD2, which must therefore be closer to the cell bodies than the limiting value of 0.16 length constants assuming an electrical equivalent model which includes the standard electrotonic parameters. If all the soma-soma resistance is located at the junction, then it could be physically a single array of gap-junction particles. Despite its low conductance (1/20 M omega = 50 nS) and possibly small physical dimensions, the electrotonic synapse is more than sufficient to ensure spike synchrony in the two cells. PMID:3958976

  4. Direct mass spectrometric peptide profiling and sequencing of nervous tissues to identify peptides involved in male copulatory behavior in Lymnaea stagnalis

    NASA Astrophysics Data System (ADS)

    Dreisewerd, Klaus; Kingston, Robert; Geraerts, Wijnand P. M.; Li, Ka Wan

    1997-12-01

    Matrix-assisted laser desorption mass spectrometry (MALDI-MS) was performed directly on a small piece of single penis nerve of the pond snail, Lymnaea stagnalis, and reveals the presence of complex peptide profiles, including many hitherto undescribed peptides. Two of the peptides have molecular weights corresponding exactly to the previously described Lymnaea small cardioactive peptides (SCP) A and B. We confirmed their identities by structural characterization of the two peptides directly from a single penis nerve by matrix-assisted laser desorption ionization high-energy collision tandem MS analysis. MALDI-MS of nervous tissues also demonstrates that a cluster of central neurons, which send their axons to the penis nerve, contain the two peptides. As the penis nerve is the nerve that innervates the penis complex, we propose that the peptides are involved in the modulation of male copulatory processes. A bioassay indeed showed that the peptides increase the contraction frequency of the vas deference in a dose-dependent manner. The results demonstrate the potential of direct MALDI-MS analysis of nervous tissue to complement or substitute conventional biochemical techniques for the identification and localization of neuropeptides.

  5. The effect of abiotic factors on the toxicity of cypermethrin against the snail Lymnaea acuminata in the control of fascioliasis.

    PubMed

    Singh, V; Singh, D K

    2009-03-01

    Every month during the year 2006-2007, the 24, 48, 72 and 96 h LC50 values of a molluscicide, cypermethrin, were determined for a snail Lymnaea acuminata, with concomitant estimation of levels of temperature, pH, dissolved oxygen and carbon dioxide and electrical conductivity, both in control and test water. On the basis of a 24 h toxicity assay, it was noted that LC50 values of 10.39, 10.90 and 11.19 mg l- 1 during the months of May, June and July, respectively, were most effective in killing the snails, while the molluscicide was least effective in the month of January, when its 24 h LC50 was 65.84 mg l- 1.There was a significant positive correlation between LC50 of cypermethrin and levels of dissolved O2/pH of water in corresponding months. On the contrary, a negative correlation was observed between LC50 and dissolved CO2/temperature of test water in the same months. In order to ascertain that such a relationship between toxicity and abiotic factors is not coincidental, the nervous tissue of the snail was assayed for the activity of acetylcholinesterase (AChE), acid phosphatase (ACP) and alkaline phosphatase (ALP) to sublethal concentrations (40% and 80%) of 24 h LC50 during each of the 12 months of the same year. The findings confirmed that abiotic factors indeed influence toxicity of cypermethrin in the snail. A significant positive rank correlation between AChE, ACP and ALP activity did exist following exposure to the corresponding sublethal concentrations. Moreover, there was a maximum inhibition of 61.29 and 76.16% of AChE and ACP, respectively, in snails exposed to 80% of the 24 h LC50 in the month of May. A similar treatment caused a maximum inhibition of 70.53% of ALP activity in the month of June. This work shows conclusively that the best time to control the snail population with cypermethrin is during the months of May and June.

  6. EFFECTS OF SINGLE, BINARY AND TERTIARY COMBINATIONS WITH Jatropha gossypifolia AND OTHER PLANT-DERIVED MOLLUSCICIDES ON REPRODUCTION AND SURVIVAL OF THE SNAIL Lymnaea acuminata

    PubMed Central

    Yadav, Ram P.; Singh, Ajay

    2014-01-01

    The effect of sub-lethal doses (40% and 80% of LC50/24h) of plant derived molluscicides of singly, binary (1:1) and tertiary (1:1:1) combinations of the Rutin, Ellagic acid, Betulin and taraxerol with J. gossypifolia latex, leaf and stem bark powder extracts and their active component on the reproduction of freshwater snail Lymnaea acuminata have been studied. It was observed that the J. gossypifolia latex, stem bark, individual leaf and their combinations with other plant derived active molluscicidal components caused a significant reduction in fecundity, hatchability and survival of young snails. It is believed that sub-lethal exposure of these molluscicides on snail reproduction is a complex process involving more than one factor in reducing the reproductive capacity. PMID:25229223

  7. Effect of binary combination of some plant-derived molluscicides with MGK-264 or piperonyl butoxide on the reproduction of the snail Lymnaea acuminata.

    PubMed

    Singh, Priyamvada; Singh, Vinay K; Singh, Dinesh K

    2005-02-01

    The effects of sub-lethal treatments (20 and 60% of 24-h LC(50)) with plant-derived molluscicides Annona squamosa, acetogenins, Argemone mexicana seed and protopine, in combination (1 + 5) with MGK-264 (ENT 8184) or piperonyl butoxide on the reproduction of Lymnaea acuminata has been studied. The plant-derived molluscicides and their active molluscicidal components, protopine and acetogenins, in combination with ENT 8184 or piperonyl butoxide caused a significant reduction in the fecundity, hatchability and survival of young snails. Combination of A squamosa seed powder with piperonyl butoxide was very effective as it caused a complete arrest of snail fecundity within 24 h of treatment. Removal of the snails to fresh water after the 96-h treatments caused a significant recovery in the fecundity of L acuminata.

  8. Effects of single, binary and tertiary combinations with Jatropha gossypifolia and other plant-derived molluscicides on reproduction and survival of the snail Lymnaea acuminata.

    PubMed

    Yadav, Ram P; Singh, Ajay

    2014-01-01

    The effect of sub-lethal doses (40% and 80% of LC(50)/24h) of plant derived molluscicides of singly, binary (1:1) and tertiary (1:1:1) combinations of the Rutin, Ellagic acid, Betulin and taraxerol with J. gossypifolia latex, leaf and stem bark powder extracts and their active component on the reproduction of freshwater snail Lymnaea acuminata have been studied. It was observed that the J. gossypifolia latex, stem bark, individual leaf and their combinations with other plant derived active molluscicidal components caused a significant reduction in fecundity, hatchability and survival of young snails. It is believed that sub-lethal exposure of these molluscicides on snail reproduction is a complex process involving more than one factor in reducing the reproductive capacity.

  9. Effect of active molluscicidal component of spices on different enzyme activities and biogenic amine levels in the nervous tissue of Lymnaea acuminata.

    PubMed

    Singh, V K; Singh, S; Singh, D K

    1999-12-01

    In vivo exposure of Lymnaea acuminata to thymol and [6]-gingerol (active molluscicidal components of Trachyspermum ammi and Zingiber officinale, respectively) indicates that they significantly alter acetylcholinesterase, lactic dehydrogenase, succinic dehydrogenase and cyto-oxidase activity in the nervous -tissue of snails. In vitro exposure showed that, except for acetylcholinesterase and lactic dehydrogenase, no significant changes were observed in cyto-oxidase and succinic dehydrogenase activity in the nervous tissue of L. acuminata. Sublethal exposure to thymol and [6]-gingerol reduced the levels of 5-hydroxytryptamine (5-HT) and dopamine (DA) in the nervous tissue of L. acuminata. There was, however, no significant change in the level of 5-hydroxy indol acetic acid (5-HIAA). Thymol and [6]-gingerol thus affects all the known neurotransmission mechanisms in the snail either separately or through a complex interaction between the different neurotransmitters. This may account for their toxicity to snails.

  10. GABA(A)- and AMPA-like receptors modulate the activity of an identified neuron within the central pattern generator of the pond snail Lymnaea stagnalis.

    PubMed

    Moccia, Francesco; Di Cristo, Carlo; Winlow, William; Di Cosmo, Anna

    2009-03-01

    To examine the neurochemistry underlying the firing of the RPeD1 neuron in the respiratory central pattern generator of the pond snail, Lymnaea stagnalis, we examined electrophysiologically and pharmacologically either "active" or "silent" preparations by intracellular recording and pharmacology. GABA inhibited electrical firing by hyperpolarizing RPeD1, while picrotoxin, an antagonist of GABA(A) receptors, excited silent cells and reversed GABA-induced inhibition. Action potential activity was terminated by 1 mM glutamate (Glu) while silent cells were depolarized by the GluR agonists, AMPA, and NMDA. Kainate exerted a complex triphasic effect on membrane potential. However, only bath application of AMPA desensitized the firing. These data indicate that GABA inhibits RPeD1 via activation of GABA(A) receptors, while Glu stimulates the neuron by activating AMPA-sensitive GluRs.

  11. Inhibition kinetics of certain enzymes in the nervous tissue of vector snail Lymnaea acuminata by active molluscicidal components of Sapindus mukorossi and Terminalia chebula.

    PubMed

    Upadhyay, Aparna; Singh, Dinesh K

    2011-10-01

    Effect of active molluscicidal components of Sapindus mukorossi and Terminalia chebula on the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activity in the nervous tissue of freshwater snail Lymnaea acuminata were studied. In vivo and in vitro exposure of saponin (active component of S. mukorossi pericarp) and tannic acid (active component of T. chebula) significantly inhibited the AChE, ACP and ALP activity in the nervous tissue of L. acuminata. The inhibition kinetics of these enzymes indicate that saponin and tannic acid caused competitive and competitive-non-competitive inhibition of AChE, respectively. Saponin also caused competitive and competitive-non-competitive inhibition of ACP and ALP, respectively, whereas tannic acid caused competitive-non-competitive inhibition of ACP and ALP. Thus the inhibition of AChE, ACP and ALP by saponin and tannic acid in the nervous tissue of L. acuminata may be the cause of molluscicidal activity of S. mukorossi and T. chebula.

  12. DNA multigene characterization of Fasciola hepatica and Lymnaea neotropica and its fascioliasis transmission capacity in Uruguay, with historical correlation, human report review and infection risk analysis

    PubMed Central

    Gayo, Valeria; Sanchis, Jaime; Artigas, Patricio; Khoubbane, Messaoud; Birriel, Soledad; Mas-Coma, Santiago

    2017-01-01

    Background Fascioliasis is a pathogenic disease transmitted by lymnaeid snails and recently emerging in humans, in part due to effects of climate changes, anthropogenic environment modifications, import/export and movements of livestock. South America is the continent presenting more human fascioliasis hyperendemic areas and the highest prevalences and intensities known. These scenarios appear mainly linked to altitude areas in Andean countries, whereas lowland areas of non-Andean countries, such as Uruguay, only show sporadic human cases or outbreaks. A study including DNA marker sequencing of fasciolids and lymnaeids, an experimental study of the life cycle in Uruguay, and a review of human fascioliasis in Uruguay, are performed. Methodology/Principal findings The characterization of Fasciola hepatica from cattle and horses of Uruguay included the complete sequences of the ribosomal DNA ITS-2 and ITS-1 and mitochondrial DNA cox1 and nad1. ITS-2, ITS-1, partial cox1 and rDNA 16S gene of mtDNA were used for lymnaeids. Results indicated that vectors belong to Lymnaea neotropica instead of to Lymnaea viator, as always reported from Uruguay. The life cycle and transmission features of F. hepatica by L. neotropica of Uruguay were studied under standardized experimental conditions to enable a comparison with the transmission capacity of F. hepatica by Galba truncatula at very high altitude in Bolivia. On this baseline, we reviewed the 95 human fascioliasis cases reported in Uruguay and analyzed the risk of human infection in front of future climate change estimations. Conclusions/Significance The correlation of fasciolid and lymnaeid haplotypes with historical data on the introduction and spread of livestock into Uruguay allowed to understand the molecular diversity detected. Although Uruguayan L. neotropica is a highly efficient vector, its transmission capacity is markedly lower than that of Bolivian G. truncatula. This allows to understand the transmission and

  13. DNA sequence characterisation and phylogeography of Lymnaea cousini and related species, vectors of fascioliasis in northern Andean countries, with description of L. meridensis n. sp. (Gastropoda: Lymnaeidae)

    PubMed Central

    2011-01-01

    Background Livestock fascioliasis is a problem throughout Ecuador, Colombia and Venezuela, mainly in Andean areas where the disease also appears to affect humans. Transmission patterns and epidemiological scenarios of liver fluke infection have shown to differ according to the lymnaeid vector snail species involved. These Andean countries present the vectors Lymnaea cousini, L. bogotensis and L. ubaquensis, unknown in the rest of Latin America. An exhaustive combined haplotype study of these species is performed by means of DNA sequencing of the nuclear ribosomal 18S RNA gene, ITS-2 and ITS-1, and mitochondrial DNA cox1 gene. Results The conserved 5.8S rDNA sequence corroborated that no pseudogenes are involved in the numerous non-microsatellite/minisatellite-related indels appearing between the ITS-2 and ITS-1 sequences when comparing different L. cousini - L. bogotensis populations. Sequence analyses and phylogenetic reconstruction methods including other lymnaeid vector species show that (i) L. bogotensis is a synonym of L. cousini, (ii) L. ubaquensis is a synonym of Pseudosuccinea columella, and (iii) populations of L. cousini hitherto known from Venezuelan highlands indeed belong to a new species for which the name L. meridensis n. sp. is proposed. This new species is described and a complete phenotypic differentiation provided. Conclusions ITS-2, ITS-1 and cox1 prove to be good markers for specimen classification and haplotype characterisation of these morphologically similar lymnaeids in endemic areas. Analysis of the 18S gene and phylogenetic reconstructions indicate that L. cousini and L. meridensis n. sp. cluster in an evolutionary line different from the one of P. columella, despite their external resemblance. This suggests an evolutionary phenotypic convergence related to similar environments and which has given rise to frequent specimen misclassification. Body size and phylogenetic relationships of L. meridensis n. sp. with well-known vectors as

  14. Chlorophyllin Bait Formulation and Exposure to Different Spectrum of Visible Light on the Reproduction of Infected/Uninfected Snail Lymnaea acuminata

    PubMed Central

    Kumar, Navneet; Singh, D. K.; Singh, Vinay Kumar

    2016-01-01

    Fasciolosis is a waterborne disease, caused by Fasciola species. Snail Lymnaea acuminata is an intermediate host of these flukes. Control of snail population is major tool in reducing the incidences. Variation in light intensity and wavelength caused significant changes in reproduction pattern of snails. Maximum fecundity was noted with bait containing carbohydrate (starch, 468 ± 0.10/20 snails) or amino acid (serine, 319 ± 0.29/20 snails) as attractant. Sublethal feeding of chlorophyllin bait with starch or serine attractant to infected and uninfected snails caused significant reduction in fecundity, hatchability, and survivability. These significant changes are observed in snails exposed to different spectral band of visible light and sunlight. Maximum fecundity of 536 ± 2.0 and minimum of 89.3 ± 0.4 were noted in snails not fed with bait and exposed to sunlight and red spectral band, respectively. There was complete arrest in the fecundity of infected and uninfected snails and no survivability of uninfected snails after 48 h feeding with bait containing chlorophyllin + attractant. Minimum hatchability (9.25 ± 0.5) was noted in red light exposed, chlorophyllin + starch fed infected snails and hatching period of bait fed snails was prolonged. Conclusively, chlorophyllin bait and red light reduce reproduction capacity in snails. PMID:26925296

  15. Effect of the photoperiod on the time schedule of egg mass production in Lymnaea stagnalis, as induced by ovulation hormone injections.

    PubMed

    Dogterom, G E; Bohlken, S; Joosse, J

    1983-02-01

    In the pulmonate snail Lymnaea stagnalis the neurosecretory caudo-dorsal cells (CDC) produce an ovulation hormone (CDCH) which is released at the periphery of the cerebral commissure (COM). Time schedules of the successive stages of the egg mass production following CDCH injection (COM extract) were determined at 20 degrees. Ovulation is performed rapidly, within 10 to 20 min. The latencies of the other stages are: egg formation, 20-30 min; egg mass formation, 60-90 min; and oviposition, about 120 min. The duration of oviposition is dependent on the size of the egg mass and varies from 5 to 20 min. At a 16-hr photoperiod all stages start 10 min earlier than at a 12-hr photoperiod. It is suggested that the effect of the photoperiod is achieved by a change of release activities of dorsal bodies and/or of CDC. The possibility that a nervous mechanism is involved in the control of the packaging of the egg cells is discussed.

  16. The population density of Lymnaea columella (Say, 1817) (Mollusca, Lymnaeidae) an intermediate host of Fasciola hepatica (Linnaeus, 1758), in the Caparaó microregion, ES, Brazil.

    PubMed

    D'Almeida, S C G; Freitas, D F; Carneiro, M B; Camargo, P F; Azevedo, J C; Martins, I V F

    2016-06-01

    The aim of this study was to monitor the population density of Lymnaea columella, an intermediate host of Fasciola hepatica, in various aquatic habitats and in drinking water in the area of the Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, on Caparaó Microregion, municipality of Alegre, state of Espírito Santo, Brazil. Monthly samplings were performed at certain points between drainage areas and drinking water in cattle and goat production systems during the years 2010 to 2013. The mean temperature, precipitation and the frequency of samples of L. columella were analysed graphically according the monthly average during the study period. A total of 2,038 molluscs were collected, 1558 of which were L. columella, that predominated in all sampled points. The highest average of specimens observed for L. columella was in the years 2010 and 2013 (51.0), and occurred decreased in 2011 (19.8). The temperature and precipitation averaged is 23.7 °C and 141 mm/year, respectively. Rainfall peak occurred in March (2011, 2013) and November (2012), during these periods the population of L. columella growth. There was no significant difference in the relationship between the specimens observed with seasons (dry-wet), thus the population of L. columella remained stable and can be found throughout the year.

  17. Chlorophyllin Bait Formulation and Exposure to Different Spectrum of Visible Light on the Reproduction of Infected/Uninfected Snail Lymnaea acuminata.

    PubMed

    Kumar, Navneet; Singh, D K; Singh, Vinay Kumar

    2016-01-01

    Fasciolosis is a waterborne disease, caused by Fasciola species. Snail Lymnaea acuminata is an intermediate host of these flukes. Control of snail population is major tool in reducing the incidences. Variation in light intensity and wavelength caused significant changes in reproduction pattern of snails. Maximum fecundity was noted with bait containing carbohydrate (starch, 468 ± 0.10/20 snails) or amino acid (serine, 319 ± 0.29/20 snails) as attractant. Sublethal feeding of chlorophyllin bait with starch or serine attractant to infected and uninfected snails caused significant reduction in fecundity, hatchability, and survivability. These significant changes are observed in snails exposed to different spectral band of visible light and sunlight. Maximum fecundity of 536 ± 2.0 and minimum of 89.3 ± 0.4 were noted in snails not fed with bait and exposed to sunlight and red spectral band, respectively. There was complete arrest in the fecundity of infected and uninfected snails and no survivability of uninfected snails after 48 h feeding with bait containing chlorophyllin + attractant. Minimum hatchability (9.25 ± 0.5) was noted in red light exposed, chlorophyllin + starch fed infected snails and hatching period of bait fed snails was prolonged. Conclusively, chlorophyllin bait and red light reduce reproduction capacity in snails.

  18. Effects of in vitro and in vivo lead exposure on voltage-dependent calcium channels in central neurons of Lymnaea stagnalis.

    PubMed

    Audesirk, G

    1987-01-01

    Currents through calcium channels of members of an identified cluster of neurons (B cells) in the pond snail Lymnaea stagnalis were studied under voltage clamp. The normal physiological saline was modified to maximize the visibility of voltage-dependent calcium currents and minimize contamination by other currents. Barium was used as the charge carrier for the calcium channels. Depolarizing voltage steps induce an inward current, the magnitude of which varies with the barium concentration. In brains taken from animals not exposed in vivo to lead, in vitro addition of lead acetate to the recording medium (0.25 to 14 microM) inhibits the barium current by 59 +/- 14% (mean +/- s.d.), in a manner that is independent of the lead concentration. The magnitude of the residual current still varies with the barium concentration. The voltage dependence of the current appears to be unaffected by lead. In contrast to some other calcium-channel blockers, such as cobalt, the inhibition of barium currents by in vitro lead exposure is irreversible, at least in short-term experiments. Contrary to expectations based on these in vitro results, barium currents in B cells of animals exposed to 5 microM lead for 6 to 12 weeks in vivo were approximately twice as large as barium currents in B cells from unexposed controls, when both were recorded in lead-free saline. It is possible that chronic in vivo lead exposure causes an increase in the number of calcium channels in these neurons.

  19. The role of spatial and temporal heterogeneity and competition in structuring trematode communities in the great pond snail, Lymnaea stagnalis (L.)

    USGS Publications Warehouse

    Soldánová, Miroslava; Kuris, Armand M.; Scholz, Tomáš; Lafferty, Kevin D.

    2012-01-01

    We assessed how spatial and temporal heterogeneity and competition structure larval trematode communities in the pulmonate snail Lymnaea stagnalis. To postulate a dominance hierarchy, mark-release-recapture was used to monitor replacements of trematode species within snails over time. In addition, we sampled the trematode community in snails in different ponds in 3 consecutive years. A total of 7,623 snails (10,382 capture events) was sampled in 7 fishponds in the Jindřichův Hradec and Třeboň areas in South Bohemia (Czech Republic) from August 2006 to October 2008. Overall, 39% of snails were infected by a community of 14 trematode species; 7% of snails were infected with more than 1 trematode species (constituting 16 double- and 4 triple-species combinations). Results of the null-model analyses suggested that spatial heterogeneity in recruitment among ponds isolated trematode species from each other, whereas seasonal pulses in recruitment increased species interactions in some ponds. Competitive exclusion among trematodes led to a rarity of multiple infections compared to null-model expectations. Competitive relationships among trematode species were hypothesized as a dominance hierarchy based on direct evidence of replacement and invasion and on indirect evidence. Seven top dominant species with putatively similar competitive abilities (6 rediae and 1 sporocyst species) reduced the prevalence of the other trematode species developing in sporocysts only.

  20. A comparison of metal levels and antioxidant enzymes in freshwater snails, Lymnaea natalensis, exposed to sediment and water collected from Wright Dam and Lower Mguza Dam, Bulawayo, Zimbabwe.

    PubMed

    Siwela, A H; Nyathi, C B; Naik, Y S

    2010-10-01

    We compared the bioaccumulation of lead (Pb), cadmium (Cd), zinc (Zn), copper (Cu), nickel (Ni) and iron (Fe) with antioxidant enzyme activity in tissues of the snails, Lymnaea natalensis, exposed to elements of two differently polluted dams. 45 snails were exposed to sediment and water collected from Wight Dam (reference) whilst another 45 snails were also exposed to sediment and water collected from Lower Mguza Dam (polluted dam). Except for Fe in sediment and Pb in water, metal concentrations were statistically higher in sediment and water collected from Lower Mguza Dam. Lead, Cd and Zn were two times higher in tissues of snails exposed to Lower Mguza Dam elements. On one hand, superoxide dismutase (SOD), diphosphotriphosphodiaphorase (DTD) and catalase (CAT) activities were significantly lower whilst malondialdehyde (MDA) levels were significantly higher in tissues of snails exposed to Lower Mguza Dam sediment and water. On the other hand, selenium-dependent glutathione peroxidase (Se-GPX) activity was significantly elevated in tissues of snails exposed to Lower Mguza Dam sediment and water. Snails exposed to Lower Mguza Dam elements seem to have responded to pollution by increasing CAT and Se-GPX specific activity in an effort to detoxify peroxides produced as a result of metal induced oxidative stress.

  1. Effects of parasitism and pesticide exposure on characteristics and functions of hemocyte populations in the freshwater snail Lymnaea palustris (Gastropoda, Pulmonata).

    PubMed

    Russo, J; Lagadic, L

    2000-01-01

    Morphological characteristics and functions of hemocytes were used to compare the immunological effects of biological and chemical stress in the freshwater snail Lymnaea palustris. Animals were either infected by a trematode parasite (Metaleptocephalus sp.), or exposed to environmental contaminants, namely atrazine and hexachlorobenzene (HCB). Three populations of circulating hemocytes, morphologically and cytochemically distinct (round cells, hyalinocytes, granulocytes), were identified in both control and parasitized or pesticide-exposed snails. After 6 h of exposure, HCB and atrazine resulted in 8-fold increases in the mean total number of hemocytes, whereas only a 2.2-fold increase was observed 6 h after cercaria emission in parasitized snails. The impact of HCB was limited to the first 24 h of exposure, whereas long-lasting effects of atrazine were observed. Hyalinocytes and, to a lesser extent, round cells contributed most to the increases in hemocyte density in pesticide-exposed snails. Parasitism and atrazine treatment resulted in significant increases of lectin-stained hemocytes, whereas exposure to HCB did not affect the percentages of stained and unstained cells. Hemocyte phagocytic activity increased in HCB-exposed snails but with no concomitant change of the oxidative burst. Opposite results were obtained in atrazine-treated snail hemocytes, with unchanged phagocytosis and decreased phorbol 12-myristate 13-acetate-stimulated production of reactive oxygen intermediates. No increase in phagocytosis, or in the production of reactive oxygen intermediates, was observed in hemocytes from parasitized snails. Infection with the immunologically compatible trematode parasite Metaleptocephalus sp. and exposure to atrazine generated similar reactions from circulating hemocytes, whereas a different response pattern was observed in HCB-exposed snails.

  2. Cross-phylum extrapolation of the Daphnia magna chronic biotic ligand model for zinc to the snail Lymnaea stagnalis and the rotifer Brachionus calyciflorus.

    PubMed

    De Schamphelaere, Karel A C; Janssen, Colin R

    2010-10-15

    We investigated if the chronic zinc biotic ligand model (BLM) developed earlier for the arthropod Daphnia magna could be extrapolated to predict chronic ecotoxicity of zinc as a function of water chemistry to two species from other phyla, i.e. the mollusc Lymnaea stagnalis and the rotifer Brachionus calyciflorus. We chronically exposed these two species to zinc in six natural surface waters. These water covered a wide range of pH (6.8-8.3), dissolved organic carbon (1.2-12.7mg/L) and Ca (8.8-118mg/L). Across all waters tested, the 28d-EC10s (200-1629μg Zn/L) and EC50s (382-2026μg Zn/L) for L. stagnalis spanned a 8.1-fold and 5.3-fold range, respectively. The 2d-EC10s (142-550μg Zn/L) and 2d-EC50s (195-1104μg Zn/L) for B. calyciflorus spanned a 3.9-fold and 5.7-fold range, respectively. The data indicated that higher pH and higher concentrations of Ca and DOC were generally associated with lower toxicity (higher ECx values). Furthermore, the chronic Zn BLM for D. magna, when calibrated only to reflect the intrinsic sensitivity of L. stagnalis and B. calyciflorus, was able to predict all ECx values with a less than 1.6-fold error, which demonstrates that the chronic D. magna Zn BLM can be extrapolated to other invertebrate phyla. This lends further support to the use of the chronic Zn BLM to account for bioavailability of zinc in aquatic risk assessment and the derivation of environmental quality standards.

  3. Seasonal variation in abiotic factors and ferulic acid toxicity in snail-attractant pellets against the intermediate host snail Lymnaea acuminata.

    PubMed

    Agrahari, P; Singh, D K

    2013-11-01

    Laboratory evaluation was made to access the seasonal variations in abiotic environmental factors temperature, pH, dissolved oxygen, carbon dioxide, electrical conductivity and ferulic acid toxicity in snail-attractant pellets (SAP) against the intermediate host snail Lymnaea acuminata in each month of the years 2010 and 2011. On the basis of a 24-h toxicity assay, it was noted that lethal concentration values of 4.03, 3.73% and 4.45% in SAP containing starch and 4.16, 4.23% and 4.29% in SAP containing proline during the months of May, June and September, respectively, were most effective in killing the snails, while SAP containing starch/proline + ferulic acid was least effective in the month of January/February (24-h lethal concentration value was 7.67%/7.63% in SAP). There was a significant positive correlation between lethal concentration value of ferulic acid containing SAP and levels of dissolved O2 /pH of water in corresponding months. On the contrary, a negative correlation was observed between lethal concentration value and dissolved CO2 /temperature of test water in the same months. To ascertain that such a relationship between toxicity and abiotic factors is not co-incidental, the nervous tissue of treated (40% and 80% of 24-h lethal concentration value) and control group of snails was assayed for the activity of acetylcholinesterase (AChE) in each of the 12 months of the same year. There was a maximum inhibition of 58.43% of AChE, in snails exposed to 80% of the 24-h lethal concentration value of ferulic acid + starch in the month of May. This work shows conclusively that the best time to control snail population with SAP containing ferulic acid is during the months of May, June and September.

  4. Accumulation and detoxication responses of the gastropod Lymnaea stagnalis to single and combined exposures to natural (cyanobacteria) and anthropogenic (the herbicide RoundUp(®) Flash) stressors.

    PubMed

    Lance, Emilie; Desprat, Julia; Holbech, Bente Frost; Gérard, Claudia; Bormans, Myriam; Lawton, Linda A; Edwards, Christine; Wiegand, Claudia

    2016-08-01

    Freshwater gastropods are increasingly exposed to multiple stressors in the field such as the herbicide glyphosate in Roundup formulations and cyanobacterial blooms either producing or not producing microcystins (MCs), potentially leading to interacting effects. Here, the responses of Lymnaea stagnalis to a 21-day exposure to non-MC or MC-producing (33μgL(-1)) Planktothrix agardhii alone or in combination with the commercial formulation RoundUp(®) Flash at a concentration of 1μgL(-1) glyphosate, followed by 14days of depuration, were studied via i) accumulation of free and bound MCs in tissues, and ii) activities of anti-oxidant (catalase CAT) and biotransformation (glutathione-S-transferase GST) enzymes. During the intoxication, the cyanobacterial exposure induced an early increase of CAT activity, independently of the MC content, probably related to the production of secondary cyanobacterial metabolites. The GST activity was induced by RoundUp(®) Flash alone or in combination with non MC-producing cyanobacteria, but was inhibited by MC-producing cyanobacteria with or without RoundUp(®) Flash. Moreover, MC accumulation in L. stagnalis was 3.2 times increased when snails were concomitantly exposed to MC-producing cyanobacteria with RoundUp(®), suggesting interacting effects of MCs on biotransformation processes. The potent inhibition of detoxication systems by MCs and RoundUp(®) Flash was reversible during the depuration, during which CAT and GST activities were significantly higher in snails previously exposed to MC-producing cyanobacteria with or without RoundUp(®) Flash than in other conditions, probably related to the oxidative stress caused by accumulated MCs remaining in tissues.

  5. The effects of host size and temperature on the emergence of Echinoparyphium recurvatum cercariae from Lymnaea peregra under natural light conditions.

    PubMed

    Morley, N J; Adam, M E; Lewis, J W

    2010-09-01

    The production of cercariae from their snail host is a fundamental component of transmission success in trematodes. The emergence of Echinoparyphium recurvatum (Trematoda: Echinostomatidae) cercariae from Lymnaea peregra was studied under natural sunlight conditions, using naturally infected snails of different sizes (10-17 mm) within a temperature range of 10-29 degrees C. There was a single photoperiodic circadian cycle of emergence with one peak, which correlated with the maximum diffuse sunlight irradiation. At 21 degrees C the daily number of emerging cercariae increased with increasing host snail size, but variations in cercarial emergence did occur between both individual snails and different days. There was only limited evidence of cyclic emergence patterns over a 3-week period, probably due to extensive snail mortality, particularly those in the larger size classes. Very few cercariae emerged in all snail size classes at the lowest temperature studied (10 degrees C), but at increasingly higher temperatures elevated numbers of cercariae emerged, reaching an optimum between 17 and 25 degrees C. Above this range emergence was reduced. At all temperatures more cercariae emerged from larger snails. Analysis of emergence using the Q10 value, a measure of physiological processes over temperature ranges, showed that between 10 and 21 degrees C (approximately 15 degrees C) Q10 values exceeded 100 for all snail size classes, indicating a substantially greater emergence than would be expected for normal physiological rates. From 14 to 25 degrees C (approximately 20 degrees C) cercarial emergence in most snail size classes showed little change in Q10, although in the smallest size class emergence was still substantially greater than the typical Q10 increase expected over this temperature range. At the highest range of 21-29 degrees C (approximately 25 degrees C), Q10 was much reduced. The importance of these results for cercarial emergence under global climate change

  6. Calicophoron daubneyi and Fasciola hepatica: characteristics of natural and experimental co-infections of these digeneans in the snail Lymnaea glabra.

    PubMed

    Vignoles, P; Titi, A; Mekroud, A; Rondelaud, D; Dreyfuss, G

    2017-01-01

    A retrospective study on different Lymnaea glabra samples collected from central France between 1993 and 2010 was carried out to determine the prevalence of natural co-infections with Calicophoron daubneyi and Fasciola hepatica, and to specify the composition of redial burdens. Experimental infections of L. glabra performed during the same period of time were also analysed to study metacercarial production of each digenean in co-infected snails. Controls were naturally or experimentally co-infected Galba truncatula. In natural co-infections, prevalence was 0.7% in L. glabra (186/25,128) and 0.4% in G. truncatula (137/31,345). Low redial burdens were found in these snails, with F. hepatica rediae significantly more numerous in L. glabra than in G. truncatula (7.5 per snail instead of 5.2). In contrast, the total numbers of C. daubneyi rediae in both lymnaeids were close to each other (4.3 and 3.0 rediae, respectively). In experimentally co-infected groups, prevalence was greater in G. truncatula than in the other lymnaeid (6.3% instead of 3.0%). Significantly shorter patent periods and lower metacercarial production for each digenean were noted in L. glabra than in G. truncatula. However, in both lymnaeids, the two types of cercariae were released during the same shedding waves and several peaks during the patent period were synchronous. In spite of a greater shell height for L. glabra, metacercarial production of both digeneans in co-infected snails was lower than that in G. truncatula, thus indicating a still incomplete adaptation between these French L. glabra and both parasites.

  7. Tissue distribution and subcellular localization of trace metals in the pond snail Lymnaea stagnalis with special reference to the role of lysosomal granules in metal sequestration.

    PubMed

    Desouky, Mahmoud M A

    2006-05-01

    The present study was undertaken to elucidate the cellular mechanisms, which govern metal sequestration and detoxification in gastropods. For this purpose the pond snail, Lymnaea stagnalis was exposed to environmentally relevant concentrations of three species of metals (Al, Zn and Cd) for 30 days and the localization and fate of these metals were followed in different tissues of the snails. The measurement of relative distribution of metals between tissues revealed that the digestive gland and kidney account for most of the accumulated metals. Al and Cd (non-essential metals) were redistributed to the digestive gland, possibly because of the presence of specific binding entities in the digestive glands of the herein species. This study focuses on the role of intracellular metal-containing granules on metal sequestration. Three main types of granules were identified in the digestive gland cells namely small, green and yellow granules. The morphological examination and the progressive accumulation of elements within these granules revealed that they are developmental stages with the yellow granule being the mature one. The total number of these granules was found to be significantly increased upon exposure of the snails to Al only. This increase may be a response to the large amount of Al that is accumulated through feeding route of this grazing snail. X-ray microanalysis (XRMA) revealed that metals were localized in all three types of digestive gland granules. The increased amount of ligands (P and S) in the granules may give evidence for their role in metal sequestration. Levels of Al and P were positively correlated in the digestive gland granules. It is possible that aluminium is bound to phosphorus to render it insoluble and so to both immobilize it within the lysosome and to be excreted in a highly insoluble form. On the other hand, both Zn and Cd induced marked upregulation of S in mature (yellow) granules by 26- and 11-folds, respectively. The lysosomal

  8. Molecular characterisation of Galba truncatula, Lymnaea neotropica and L. schirazensis from Cajamarca, Peru and their potential role in transmission of human and animal fascioliasis

    PubMed Central

    2012-01-01

    Background Human and animal fascioliasis is emerging in many world regions, among which Andean countries constitute the largest regional hot spot and Peru the country presenting more human endemic areas. A survey was undertaken on the lymnaeid snails inhabiting the hyperendemic area of Cajamarca, where human prevalences are the highest known among the areas presenting a "valley transmission pattern", to establish which species are present, genetically characterise their populations by comparison with other human endemic areas, and discuss which ones have transmission capacity and their potential implications with human and animal infection. Methods Therefore, ribosomal DNA ITS-2 and ITS-1, and mitochondrial DNA 16S and cox1 were sequenced by the dideoxy chain-termination method. Results Results indicate the presence of three, morphologically similar, small lymnaeid species belonging to the Galba/Fossaria group: Galba truncatula, Lymnaea neotropica and L. schirazensis. Only one combined haplotype for each species was found. The ITS-1, 16S and cox1 haplotypes of G. truncatula are new. No new haplotypes were found in the other two species. This scenario changes previous knowledge, in which only L. viator (= L. viatrix) was mentioned. Galba truncatula appears to be the most abundant, with high population densities and evident anthropophyly including usual presence in human neighbourhood. Infection by Fasciola hepatica larval stages were molecularly confirmed in two populations of this species. The nearness between G. truncatula populations presenting liver fluke infection and both human settings and schools for children, together with the absence of populations of other lymnaeid species in the locality, suggest a direct relationship with human infection. Conclusions The geographical overlap of three lymnaeid species poses problems for epidemiological studies and control action. First, a problem in classifying lymnaeid specimens in both field and laboratory activities

  9. [Investigations on the pathogenesis of changes in somatic growth of Lymnaea stagnalis (Gastropoda: Pulmonata) experimentally infected with parthenites Opisthioglyphe ranae (Digenea: Plagiorchiida). I. Relative weight of accessory sex organs and synthetic activity of neurosecretory cells].

    PubMed

    Pokora, Z

    1996-01-01

    In the paper an attempt to define pathogenesis of changes in somatic growth of juvenile individuals of the popular freshwater snail Lymnaea stagnalis experimentally infected with parthenites of the trematode Opisthioglyphe ranae was undertaken. Significant enlargement of relative wet weight of examined accessory sex organs (albumen gland, oothecal gland, prostate, male copulatory organ) observed in infected snails permits to explain increase of their somatic growth basing on the hypothesis of disturbances in energetistic budget of the host-as a consequence of reduction by the parasite activity of the snail's reproductive system. Pathogenesis of this phenomenon has probably a complicated character, including also effect of parthenites on activity of the neurosecretory cells that control somatic growth in examined species of the snail. An argument for this standpoint is, observed in infected snails, increase of amount of neurosecretory material and RNA in cytoplasm of these cells (the light green cells of cerebral ganglia), as well as amount of the loose fraction of chromatine in their nuclei.

  10. Acute toxicity, critical body residues, Michaelis-Menten analysis of bioaccumulation, and ionoregulatory disturbance in response to waterborne nickel in four invertebrates: Chironomus riparius, Lymnaea stagnalis, Lumbriculus variegatus and Daphnia pulex.

    PubMed

    Leonard, Erin M; Wood, Chris M

    2013-06-01

    We investigated the bioaccumulation and acute toxicity (48 h or 96 h) of Ni in four freshwater invertebrate species in two waters with hardness of 40 (soft water) and 140 mg L(-1) as CaCO(3) (hard water). Sensitivity order (most to least) was Lymnaea stagnalis > Daphnia pulex > Lumbriculus variegatus > Chironomus riparius. In all cases water hardness was protective against acute Ni toxicity with LC(50) values 3-3.5× higher in the hard water vs. soft water. In addition, higher water hardness significantly reduced Ni bioaccumulation in these organisms suggesting that competition by Ca and Mg for uptake at the biotic ligand may contribute to higher metal resistance. CBR50 values (Critical Body Residues) were less dependent on water chemistry (i.e. more consistent) than LC(50) values within and across species by ~2 fold. These data support one of the main advantages of the Tissue Residue Approach (TRA) where tissue concentrations are generally less variable than exposure concentrations with respect to toxicity. Whole body Ni bioaccumulation followed Michaelis-Menten kinetics in all organisms, with greater hardness tending to decrease B(max) with no consistent effect on K(d). Across species, acute Ni LC(50) values tended to increase with both K(d) and B(max) values - i.e. more sensitive species exhibited higher binding affinity and lower binding capacity for Ni, but there was no correlation with body size. With respect to biotic ligand modeling, log K(NiBL) values derived from Ni bioaccumulation correlated well with log K(NiBL) values derived from toxicity testing. Both whole body Na and Mg levels were disturbed, suggesting that disruption of ionoregulatory homeostasis is a mechanism of acute Ni toxicity. In L. stagnalis, Na depletion was a more sensitive endpoint than mortality, however, the opposite was true for the other organisms. This is the first study to show the relationship between Na and Ni.

  11. Larval stages of trematodes in gastropods from Lake Chicnahuapan, State of Mexico, Mexico.

    PubMed

    Barragán-Sáenz, Francisco Adrián; Sánchez-Nava, Petra; Hernández-Gallegos, Oswaldo; Salgado-Maldonado, Guillermo

    2009-10-01

    During the period from January to December (2007), 1,095 freshwater molluscs of four species were captured (Lymnaea stagnalis, Stagnicola elodes, Physella cubensis and Physa acuta) in Lake Chicnahuapan, State of Mexico, Mexico. Two hundred seventy-two (24.84% prevalence) of these molluscs were parasitised by 11 trematode species (from which two were not identified at the species level) having six cercariae species and five metacercariae species represented in five families. The cercariae Telorchis corti (Plagiorchiidae) and the metacercariae Cotylurus cornutus (Strigeidae) were the species with the highest prevalence among the examined snails. The highest percentage of infection was observed in L. stagnalis (27.45% of prevalence, n = 572) and P. cubensis (23.96%, n = 455). Twenty-one of the examined snails had multiple infections with up to three trematode species.

  12. Molluscicidal steroid glycoalkaloids possessing stereoisomeric spirosolane structures.

    PubMed

    Alzérreca, A; Hart, G

    1982-07-01

    Both the steroidal glycoalkaloid mixture obtained from Solanum mammosum fruits (solasonine 1 and solamargine 2) and the stereoisomeric glycosidic alkaloid tomatine 3 are toxic at 10 and 25 ppm to Lymnaea cubensis and Biomphalaria glabratus, respectively. Their aglycones solasodine 4 and tomatidine 5 obtained by hydrolysis of the glycosides showed no toxicity to either mollusk under the test conditions used. Preliminary structure-activity correlations indicate that the molluscicidal properties depend on the type of aglycones and on the glycoside bond. A common molecular structure in molluscicidal spirosolane and spirostane glycosides is recognized.

  13. Fascioliasis of livestock and snail host for Fasciola in the Altiplano Region of Bolivia.

    PubMed

    Ueno, H; Arandia, R; Morales, G; Medina, G

    1975-01-01

    Fascioliasis caused by Fasciola hepatica was a serious problem for sheep and alpacas in the Altiplano Region of Bolivia. In some provinces close to Lake Titicaca, the raising of sheep was forced to discontinue, because infection with the fluke made it unprofitable and almost impossible. It was proved that in the Altiplano Region, two species of freshwater snails, Lymnaea viatrix and L. cubensis var., served as intermediate hosts for F. hepatica. In some subtropical areas of Bolivia, these snails could not be found, although other Lymnaea sp. was widely distributed there. As it is possible for Lymnaea sp. to be intermediate host for the fluke, further studies are required on the identification. Acute fascioliasis of sheep occurred in the Altiplano Region principally during a period from May to July, or the dry season. In some areas, the mortality rate of infected sheep was roughly estimated as 15 to 25% annually. Contamination with Fasciola metacercariae of herbage and semi-aquatic plants grown in a swamp in one of these areas was biologically assessed, using guinea pigs. Plants of Compositae and Eleocharis sp. were contaminated most intensely and those of Senicio sp. and Vallisneria sp. carried a fairly large number of cysts, while plants of Scirpus sp. and Ranunclaceae carried only a few cysts. No signs of Fasciola infection were observed in any animal given the plants of Liliaceae.

  14. Epidemiology and population biology of pseudoperonospora cubensis: a model system for management of downy mildews

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The resurgence of cucurbit downy mildew has dramatically influenced production of cucurbits and disease management systems at multiple scales. Long-distance dispersal is a fundamental aspect of epidemic development that influences the timing and extent of disease outbreaks. Dispersal potential of th...

  15. Spatial and Temporal Profiling of Griseofulvin Production in Xylaria cubensis Using Mass Spectrometry Mapping

    PubMed Central

    Sica, Vincent P.; Rees, Evan R.; Tchegnon, Edem; Bardsley, Robert H.; Raja, Huzefa A.; Oberlies, Nicholas H.

    2016-01-01

    A large portion of natural products research revolves around the discovery of new, bioactive chemical entities; however, studies to probe the biological purpose of such secondary metabolites for the host organism are often limited. Mass spectrometry mapping of secondary metabolite biosynthesis in situ can be used to probe a series of ecological questions about fungi that may be lost through traditional natural products chemistry extraction protocols. A griseofulvin-producing fungal culture of the Xylariaceae family, isolated as an endophyte of the tree Asimina triloba, was analyzed through a series of spatial and temporal mapping experiments. This fungus produced unique fungal characteristics, such as guttates and stroma, both of which were explored spatially. The distribution of griseofulvin on this culture in isolation was compared to its dispersal when grown in co-culture with a competing Penicillium species via a droplet–based surface sampling system. The fungistatic properties of griseofulvin were visualized, including the consequences for biosynthesis of polyhydroxyanthraquinones in a rival culture. PMID:27199902

  16. Spatial and Temporal Profiling of Griseofulvin Production in Xylaria cubensis Using Mass Spectrometry Mapping.

    PubMed

    Sica, Vincent P; Rees, Evan R; Tchegnon, Edem; Bardsley, Robert H; Raja, Huzefa A; Oberlies, Nicholas H

    2016-01-01

    A large portion of natural products research revolves around the discovery of new, bioactive chemical entities; however, studies to probe the biological purpose of such secondary metabolites for the host organism are often limited. Mass spectrometry mapping of secondary metabolite biosynthesis in situ can be used to probe a series of ecological questions about fungi that may be lost through traditional natural products chemistry extraction protocols. A griseofulvin-producing fungal culture of the Xylariaceae family, isolated as an endophyte of the tree Asimina triloba, was analyzed through a series of spatial and temporal mapping experiments. This fungus produced unique fungal characteristics, such as guttates and stroma, both of which were explored spatially. The distribution of griseofulvin on this culture in isolation was compared to its dispersal when grown in co-culture with a competing Penicillium species via a droplet-based surface sampling system. The fungistatic properties of griseofulvin were visualized, including the consequences for biosynthesis of polyhydroxyanthraquinones in a rival culture.

  17. A new baseline for fascioliasis in Venezuela: lymnaeid vectors ascertained by DNA sequencing and analysis of their relationships with human and animal infection

    PubMed Central

    2011-01-01

    Background Human and animal fascioliasis poses serious public health problems in South America. In Venezuela, livestock infection represents an important veterinary problem whereas there appear to be few human cases reported, most of which are passively detected in health centres. However, results of recent surveys suggest that the situation may be underestimated in particular areas. To obtain a baseline for future fascioliasis assessment, studies were undertaken by means of rDNA ITS-2 and ITS-1 and mtDNA cox1 sequencing to clarify the specific status of Venezuelan lymnaeids, their geographical distribution and fascioliasis transmission capacity, by comparison with other American countries and other continents. Results Results obtained completely change the lymnaeid scenario known so far. The relatively rich lymnaeid fauna of Venezuela has been proven to include (i) Lymnaea meridensis and L. neotropica as the only native members, (ii) L. cubensis and Pseudosuccinea columella introduced from the Caribbean area, and (iii) Galba truncatula and L. schirazensis introduced from the Old World. The absence of representatives of the stagnicoline and Radix groups is remarkable. Four species are fascioliasis vectors: G. truncatula, L. cubensis and L. neotropica, which have the capacity to give rise to human endemic areas, and P. columella, which is a source of animal infection and is responsible for the spread of disease. Vector capacity in the apparently highland endemic L. meridensis is to be confimed, although may be expected given its phylogenetic relationships. Similarly as elsewhere, the non-transmitting L. schirazensis has been confused with L. cubensis, also with G. truncatula and possibly with L. neotropica. Conclusions The new scenario leads to the re-opening of many disease aspects. In Venezuela, altitude appears to be the main factor influencing fascioliasis distribution. Human infection shows an altitude pattern similar to other Andean countries, although a

  18. Using next-generation sequencing to develop molecular diagnostics for Pseudoperonospora cubensis, the cucurbit downy mildew pathogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Advances in Next Generation Sequencing (NGS) allow for rapid development of genomics resources needed to generate molecular diagnostics assays for infectious agents. NGS approaches are particularly helpful for organisms that cannot be cultured, such as the downy mildew pathogens, a group of biotrop...

  19. SPATIAL DISTRIBUTION OF LYMNAEIDAE (MOLLUSCA, BASOMMATOPHORA), INTERMEDIATE HOST OF Fasciola hepatica LINNAEUS, 1758 (TREMATODA, DIGENEA) IN BRAZIL

    PubMed Central

    Medeiros, Camilla; Scholte, Ronaldo Guilherme Carvalho; D'ávila, Sthefane; Caldeira, Roberta Lima; Carvalho, Omar dos Santos

    2014-01-01

    Snails of the family Lymnaeidae act as intermediate hosts in the biological cycle of Fasciola hepatica, which is a biological agent of fasciolosis, a parasitic disease of medical importance for humans and animals. The present work aimed to update and map the spatial distribution of the intermediate host snails of F. hepatica in Brazil. Data on the distribution of lymnaeids species were compiled from the Collection of Medical Malacology (Fiocruz-CMM, CPqRR), Collection of Malacology (MZUSP), “SpeciesLink” (CRIA) network and through systematic surveys in the literature. Our maps of the distribution of lymnaeids show that Pseudosuccinea columella is the most common species and it is widespread in the South and Southeast with few records in the Midwest, North and Northeast regions. The distribution of the Galba viatrix, G. cubensis and G. truncatula showed a few records in the South and Southeast regions, they were not reported for the Midwest, North and Northeast. In addition, in the South region there are a few records for G. viatrix and one occurrence of Lymnaea rupestris. Our findings resulted in the first map of the spatial distribution of Lymnaeidae species in Brazil which might be useful to better understand the fasciolosis distribution and delineate priority areas for control interventions. PMID:24879003

  20. The participation of NMDA receptors, PKC, and MAPK in the formation of memory following operant conditioning in Lymnaea

    PubMed Central

    2010-01-01

    Background Memory is the ability to store, retain, and later retrieve information that has been learned. Intermediate term memory (ITM) that persists for up to 3 h requires new protein synthesis. Long term memory (LTM) that persists for at least 24 h requires: DNA transcription, RNA translation, and the trafficking of newly synthesized proteins. It has been shown in a number of different model systems that NMDA receptors, protein kinase C (PKC) and mitogen activated protein kinase (MAPK) are all involved in the memory formation process. Results Here we show that snails trained in control conditions are capable of forming, depending on the training procedure used, either ITM or LTM. However, blockage of NMDA receptors (MK 801), inhibition of PKC (GF109203X hydrochloride) and MAPK activity (UO126) prevent the formation of both ITM and LTM. Conclusions The injection of either U0126 or GF109203X, which inhibit MAPK and PKC activity respectively, 1 hour prior to training results in the inhibition of both ITM and LTM formation. We further found that NMDA receptor activity was necessary in order for both ITM and LTM formation. PMID:20807415

  1. Host specificity shapes population structure of pinworm parasites in Caribbean reptiles.

    PubMed

    Falk, Bryan G; Perkins, Susan L

    2013-09-01

    Host specificity is one of the potential factors affecting parasite diversification because gene flow may be facilitated or constrained by the number of host species that a parasite can exploit. We test this hypothesis using a costructure approach, comparing two sympatric pinworm parasites that differ in host specificity - Parapharyngodon cubensis and Spauligodon anolis - on the Puerto Rican Bank and St. Croix in the Caribbean. Spauligodon anolis specializes on Anolis lizards, whereas P. cubensis parasitizes Anolis lizards as well as many other species of lizards and snakes. We collected lizards from across the Puerto Rican Bank and St. Croix, sampled them for S. anolis and P. cubensis and generated nuclear and mitochondrial sequence data from the parasites. We used these data to show that P. cubensis is comprised of multiple cryptic species that exhibit limited population structure relative to S. anolis, which is consistent with our prediction based on their host specificity. We also provide evidence that the distribution of P. cubensis species is maintained by competitive exclusion, and in contrast to previous theoretical work, the parasites with the greatest number of host species also reach the highest prevalence rates. Overall, our results are consistent with the hypothesis that host specificity shapes parasite diversification, and suggest that even moderate differences in host specificity may contribute to substantial differences in diversification.

  2. PCR detection of Pseudoperonospora humuli in air samples from hop yards

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Downy mildew of hop, caused by Pseudoperonospora humuli, is an important disease in most regions of hop production and is managed largely with regular fungicide applications. A PCR assay specific to P. humuli and the related organism P. cubensis was developed and used to monitor airborne inoculum i...

  3. QTLs for downy mildew resistance and their association with LRR-RLK resistance gene analogs in cucumber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Downy mildew (DM) caused by the obligate oomycete Pseudoperonospora cubensis is an important disease of cucumber and other cucurbit crops. For more than 50 years, DM was effectively controlled in cucumber through host resistance. The dm-1 gene became less effective in 2004 due to the appearance of a...

  4. QTL mapping of downy mildew resistance in PI 197088 and PI 330628 cucumbers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Downy mildew (DM, Pseudoperonospora cubensis) is a devastating fungal disease of cucumber worldwide. Several plant introduction lines have been identified with high resistance to the post-2004 new DM strain found in the U.S. However, the inheritance of DM resistance is still not well defined. Molecu...

  5. The hyphomycete genus Piricauda, with the description of a new species.

    PubMed

    Mercado Sierra, Angel; Guarro, Josep; Heredia, Gabriela

    2005-06-01

    The genus Piricauda comprises eight species characterized by monotretic conidiogenous cells and micronematous arched conidiophores: P. cochinensis, P. cubensis, P. longispora, P. mexicana, P. paraguayensis, P. pseudarthriae, P. taiwanensis, and P. vulcanensis sp. nov. P. caribensis is transferred to Acrodictys. Species usually colonize dead petioles and rachides of palms in tropical and subtropical areas, but can also be found on branches and leaves of different trees. The species are illustrated and briefly described, and a key to them is provided.

  6. One-trial reward learning in the snail Lymnea stagnalis.

    PubMed

    Alexander, J; Audesirk, T E; Audesirk, G J

    1984-01-01

    We present evidence that the pond snail Lymnaea stagnalis is capable of aquisition and extensive retention of an appetitively reinforced feeding response after only a single training trial. Food-deprived snails presented with a single pairing of a phagostimulant (a mixture of sucrose and casein digest) and a novel, non-food chemostimulus (amyl acetate) subsequently made feeding responses to the amyl acetate and retained the association for at least 19 days. This demonstration of one-trial, non-aversive classical conditioning enhances the utility of Lymnaea stagnalis as a model system for the detailed analysis of neural mechanisms underlying plasticity.

  7. Morphological and molecular characterization of lymnaeid snails and their potential role in transmission of Fasciola spp. in Vietnam.

    PubMed

    Dung, Bui Thi; Doanh, Pham Ngoc; The, Dang Tat; Loan, Ho Thi; Losson, Bertrand; Caron, Yannick

    2013-12-01

    Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.

  8. First description of deep-water elasmobranch assemblages in the Exuma Sound, The Bahamas

    NASA Astrophysics Data System (ADS)

    Brooks, Edward J.; Brooks, Annabelle M. L.; Williams, Sean; Jordan, Lance K. B.; Abercrombie, Debra; Chapman, Demian D.; Howey-Jordan, Lucy A.; Grubbs, R. Dean

    2015-05-01

    Deep-sea chondrichthyans, like many deep-water fishes, are very poorly understood at the most fundamental biological, ecological and taxonomic levels. Our study represents the first ecological investigation of deep-water elasmobranch assemblages in The Bahamas, and the first assessment of species-specific resilience to capture for all of the species captured. Standardised deep-water longline surveys (n=69) were conducted September to December 2010 and 2011 between 472 m and 1024 m deep, resulting in the capture of 144 sharks from 8 different species. These included the Cuban dogfish, Squalus cubensis, the bigeye sixgill shark, Hexanchus nakamurai, the bluntnose sixgill shark, Hexanchus griseus, the smooth dogfish, Mustelus canis insularis, the roughskin dogfish, Centroscymnus owstoni, Springer's sawtail catshark, Galeus springeri and the false catshark, Pseudotriakis microdon. Preliminary genetic analysis indicated two or more species of gulper sharks, Centrophorus spp.; however, for the present study they were treated as a single species complex. Water depth and distance from the rocky structure of the Exuma Sound wall were inversely correlated with species richness, whereas seabed temperature was directly correlated with species richness. These variables also had a significant influence on the abundance and distribution of many species. Expanded depth ranges were established for S. cubensis and H. nakamurai, which, in the case of S. cubensis, is thought to be driven by thermal preferences. At-vessel mortality rates increased significantly with depth, and post-release mortality was thought to be high for some species, in part due to high post-release predation. This study highlights the importance of utilising strategic geographic locations that provide easy access to deep water, in combination with traditional expedition-based deep-ocean science, to accelerate the acquisition of fundamental ecological and biological insights into deep-sea elasmobranchs.

  9. A novel monoclonal antibody-based immunoenzymatic assay for epidemiological surveillance of the vector snails of Fasciola hepatica (Trematoda: Digenea).

    PubMed

    Alba, Annia; Hernández, Hilda M; Marcet, Ricardo; Vázquez, Antonio A; Figueredo, Mabel; Sánchez, Jorge; Otero, Oscar; Sarracent, Jorge

    2015-02-01

    Fasciolosis is a globally distributed snail-borne disease which requires economic consideration due to its enormous impact on veterinary medicine. During recent decades, this parasitosis has also shown increasing prevalence in human populations worldwide. The dissemination and successful transmission of fasciolosis ultimately depends on the existence of susceptible snails that act as intermediate hosts. Therefore, to accomplish effective control of this disease, surveillance and detection of the infected intermediate host would be essential. The screening of trematodes within snails using classical parasitological examination of the larvae can be unreliable (sensitivity and specificity vary depending on the time of infection and the experience of the observer) and relatively costly when using molecular biological methods during large-scale monitoring. Here we propose a novel monoclonal antibody-based immunoenzymatic assay to detect ongoing Fasciola hepatica infection in lymnaeid snails. Anti-F. hepatica rediae mouse monoclonal antibodies were generated and used to develop a double monoclonal antibody-based ELISA for parasite detection. Fasciola hepatica-infected and uninfected laboratory-reared Galba cubensis and Pseudosuccinea columella were used for assessment of the developed ELISA. Experimentally infected snails were dissected and examined for parasite larvae as the "gold standard" method. Sensitivity results were 100% for both snail species, while specificity was 98% for G. cubensis and 100% for P. columella. No cross-reactivity was detected in lymnaeids infected with Trichobilharzia sp. or Cotylophoron sp. The ELISA enabled detection of the infection from day 8 p.i. in G. cubensis while in P. columella it was noted as early as day 4. To our knowledge no previous immunoassays have been reported to detect helminth-infected snails and the developed sandwich ELISA method is therefore suggested for infection status validation in natural populations of lymnaeid

  10. Phylogeography of Pinus subsection Australes in the Caribbean Basin

    PubMed Central

    Jardón-Barbolla, Lev; Delgado-Valerio, Patricia; Geada-López, Gretel; Vázquez-Lobo, Alejandra; Piñero, Daniel

    2011-01-01

    Background and Aims Four species of Pinus subsection Australes occur in the Caribbean Basin: P. caribaea, P. cubensis, P. maestrensis and P. occidentalis. This study analyses the phylogeography of these species to assess possible colonization events from Central America to the islands and subsequent population expansions during glacial periods driven by both drier climate and larger emerged land areas. Methods Allele size data were obtained for plastid microsatellites for 314 individuals from 24 populations, covering the distribution range of subsection Australes in the Caribbean Basin. Key Results In total, 113 plastid haplotypes were identified. The highest genetic diversity was found in populations of P. caribaea. Overall, Caribbean Basin populations fit the isolation by distance model. Significant phylogeographical structure was found (RST = 0·671 > permuted RST = 0·101; P < 0·0001). The haplotype network and a Bayesian analysis of population structure (BAPS) indicated different Central American origins for P. caribaea var. bahamensis and P. caribaea var. caribaea plastids, with Central America populations in northern and south-eastern groups. Sudden expansion times for BAPS clusters were close to three glacial maxima. Conclusions Central America contains ancestral plastid haplotypes. Population expansion has played a major role in the distribution of genetic diversity in P. caribaea var. hondurensis. Two colonization events gave rise to the P. caribaea var. bahamensis and P. caribaea var. caribaea lineages. Plastid variation in the eastern species (P. cubensis, P. maestrensis and P. occidentalis) evolved independently from that in P. caribaea var. caribaea. Incomplete lineage sorting between P. cubensis and P. maestrensis is apparent. Inferred expansion times for P. caribaea var. bahamensis and for the eastern lineages correspond to glacial maxima, whereas those for P. caribaea var. hondurensis correspond to the beginning of the temperature decrease that

  11. [Ecology of river mollusks of medical and veterinary importance in 3 sites in La Habana province].

    PubMed

    Vázquez Perera, Antonio Alejandro; Gutiérrez Amador, Alfredo

    2007-01-01

    An ecological research study was carried out in freshwater mollusk populations of medical and veterinary importance, in order to determine the biotic and abiotic factors that affect their dynamics. It was observed that the principal abiotic factors influencing abundance of mollusks were total hardness, salinity, acidity, alkalinity and CO2 concentration. Both aquatic plants and specific relations among mollusk groups were the principal biotic factors that affected the molluskan fauna. Species like Fossaria cubensis and Tarebia granifera appeared affected when the site diversity increased whereas the tiarid Melanoides tuberculata prevailed in almost all the ecosystems.

  12. Aurapex penicillata gen. sp. nov. from native Miconia theaezans and Tibouchina spp. in Colombia.

    PubMed

    Gryzenhout, Marieka; Myburg, Henrietta; Rodas, Carlos A; Wingfield, Brenda D; Wingfield, Michael J

    2006-01-01

    Conidiomata of a fungus resembling Chrysoporthe cubensis, a serious canker pathogen of Eucalyptus spp. (Myrtaceae, Myrtales) in tropical and subtropical parts of the world, was found on Eucalyptus grandis in Colombia. Fruiting structures of the fungus could be distinguished from those of C. cubensis by their distinctly orange conidiomatal necks. This fungus also was found on several plant species native to Colombia including Tibouchina urvilleana, T. lepidota and Miconia theaezans (Melastomataceae, Myrtales). Morphological comparisons, as well as those based on sequences of the ITS1/ITS2 region of the ribosomal DNA repeat and the beta-tubulin gene, were used to characterize this fungus. Its pathogenicity was assessed on various plants from which it has been collected, either in field or greenhouse trials. Phylogenetic analyses showed that isolates reside in a clade distinct from the four clades accommodating Chrysoporthe, Cryphonectria, Endothia and Rostraureum. Members of this clade are distinguished by the presence of orange conidiomatal necks with black bases and a unique internal stromatal structure. No teleomorph has been found for this fungus, for which we have provided the name Aurapex penicillata gen. sp. nov. A. penicillata produced only small lesions after inoculation on young T. urvilleana, M. theaezans and E. grandis trees and appears not to be a serious pathogen.

  13. Spot light survey on fresh-water snails of medical importance in Al Fayoum Governorate, Egypt.

    PubMed

    Abo-Madyan, Ahmed A; Morsy, Tosson A; Motawea, Saad M; El Garhy, Manal F; Massoud, Ahmed M A

    2005-04-01

    In a survey carried out during Summer and Autumn of 2004, for snails of medical importance, nine species were recovered. These were Biomphalaria alexandrina, B. glabrata, B. pfeifferi, Bulinus truncatus, B. forskalii, Lymnaea natalensis, Bellamya (=Vivipara) unicolor, Physa acuta and Hydrobia musaensis. Parasitological examination revealed that B. alexandrina, B. glabrata and L. natalensis harboured immature stages of their concerned trematode parasites. Moreover, P. acuta harboured the immature stage of the nematode parasite Parastrongylus cantonensis.

  14. Gigantobilharzia, possible cause of cercarial dermatitis: Case report

    PubMed Central

    Omar, Hussein M.; Omer, Osama H.; Al-Dhubaibi, Mohammed S.

    2016-01-01

    Cercarial dermatitis (swimmer’s itch) is a worldwide, often neglected parasitic skin disease characterized by strong maculopapular skin eruption accompanied by intensive itching. A fisherman suffered from forearm dermatitis. Clinical history associated with the recovery of the avian schistosome; Gigantobilharzia from little green bee-eater (Merops orientalis najdanus) and collected Lymnaea snails supported the authors’ opinion that patient clinical signs are most likely due to the invasion of avian schistosome cercariae. PMID:27004065

  15. Coevolution between Hispaniolan crossbills and pine: does more time allow for greater phenotypic escalation at lower latitude?

    PubMed

    Parchman, Thomas L; Benkman, Craig W; Mezquida, Eduardo T

    2007-09-01

    Crossbills (Aves: Loxia) and several conifers have coevolved in predator-prey arms races over the last 10,000 years. However, the extent to which coevolutionary arms races have contributed to the adaptive radiation of crossbills or to any other adaptive radiation is largely unknown. Here we extend our previous studies of geographically structured coevolution by considering a crossbill-conifer interaction that has persisted for a much longer time period and involves a conifer with more variable annual seed production. We examined geographic variation in the cone and seed traits of two sister species of pines, Pinus occidentalis and P. cubensis, on the islands of Hispaniola and Cuba, respectively. We also compared the Hispaniolan crossbill (Loxia megaplaga) to its sister taxa the North American white-winged crossbill (Loxia leucoptera leucoptera). The Hispaniolan crossbill is endemic to Hispaniola whereas Cuba lacks crossbills. In addition and in contrast to previous studies, the variation in selection experienced by these pines due to crossbills is not confounded by the occurrence of selection by tree squirrels (Tamiasciurus and Sciurus). As predicted if P. occidentalis has evolved defenses in response to selection exerted by crossbills, cones of P. occidentalis have scales that are 53% thicker than those of P. cubensis. Cones of P. occidentalis, but not P. cubensis, also have well-developed spines, a known defense against vertebrate seed predators. Consistent with patterns of divergence seen in crossbills coevolving locally with other conifers, the Hispaniolan crossbill has evolved a bill that is 25% deeper than the white-winged crossbill. Together with phylogenetic analyses, our results suggest that predator-prey coevolution between Hispaniolan crossbills and P. occidentalis over approximately 600,000 years has caused substantial morphological evolution in both the crossbill and pine. This also indicates that cone crop fluctuations do not prevent crossbills and

  16. Spatiotemporal spread of cucurbit downy mildew in the eastern United States.

    PubMed

    Ojiambo, P S; Holmes, G J

    2011-04-01

    The dynamics of cucurbit downy mildew, caused by Pseudoperonospora cubensis, in the eastern United States in 2008 and 2009 were investigated based on disease records collected in 24 states as part of the Cucurbit downy mildew ipmPIPE monitoring program. The mean season-long rate of temporal disease progress across the 2 years was 1.4 new cases per day. Although cucurbit downy mildew was detected in mid-February and early March in southern Florida, the disease progressed slowly during the spring and early summer and did not enter its exponential phase until mid-June. The median nearest-neighbor distance of spread of new disease cases was ≈110 km in both years, with ≈15% of the distances being >240 km. Considering disease epidemics on all cucurbits, the epidemic expanded at a rate of 9.2 and 10.5 km per day in 2008 and 2009, respectively. These rates of spatial spread are at the lower range of those reported for the annual spread of tobacco blue mold in the southeastern United States, a disease that is also aerially dispersed over long distances. These results suggest that regional spread of cucurbit downy mildew may be limited by opportunities for establishment in the first half of the year, when fewer cucurbit hosts are available for infection. The O-ring statistic was used to determine the spatial pattern of cucurbit downy mildew outbreaks using complete spatial randomness as the null model for hypothesis testing. Disease outbreaks in both years were spatially aggregated and the extent of spatial dependence was up to 1,000 km. Results from the spatial analysis suggests that disease outbreaks in the Great Lakes and mid-Atlantic regions may be due to the spread of P. cubensis sporangia from outbreaks of the disease near the Georgia/South Carolina/North Carolina border rather than from overwintering sites in southern Florida. Space-time point pattern analysis indicated strong (P < 0.001) evidence for a space-time interaction and a space-time risk window of ≈3

  17. Presence of Fasciola hepatica in feral nutria (Myocastor coypus) living in a public park in Brazil.

    PubMed

    El-Kouba, Maysa M A N; Marques, Sandra M T; Pilati, Célso; Hamann, Waldir

    2009-03-01

    Sixteen nutria (Myocastor coypus) from a protected area in Curitiba, Brazil, were sampled to determine the prevalence of Fasciola hepatica eggs and intestinal parasites and the presence of snails in the habitat used by nutria. The overall prevalence rates were 56.25% (9) for F. hepatica eggs, 87.50% (14) for cestode eggs, 56.25% (9) for ascarid eggs, 50% (8) for coccidian (Eimeriidae) oocysts, and 56.25% (9) for Strongyloidea eggs. Fasciola hepatica eggs had an average size of 138 microm x 72 microm. The following mollusks were found: Physa cubensis, Physa marmorata, and Biomphalaria tenagophyla. These results suggest that the infected nutria could serve as a source of contamination for other animals, humans, and surface water. This is the first report about parasites in nutria in the state of Paraná, Brazil.

  18. Pulley reef: a deep photosynthetic coral reef on the West Florida Shelf, USA

    USGS Publications Warehouse

    Culter, J.K.; Ritchie, K.B.; Earle, S.A.; Guggenheim, D.E.; Halley, R.B.; Ciembronowicz, K.T.; Hine, A.C.; Jarrett, B.D.; Locker, S.D.; Jaap, W.C.

    2006-01-01

    Pulley Reef (24°50′N, 83°40′W) lies on a submerged late Pleistocene shoreline feature that formed during a sea-level stillstand from 13.8 to 14.5 ka (Jarrett et al. 2005). The reef is currently 60–75 m deep, exhibits 10–60% coral cover, and extends over approximately 160 km2 of the sea floor. Zooxanthellate corals are primarily Agaricia lamarcki, A. fragilis, Leptoseris cucullata, and less common Madracis formosa, M. pharensis, M. decactis, Montastraea cavernosa, Porites divaricata, Scolymia cubensis and Oculina tenella. Coralline algae are comparable in abundance to stony corals. Other macroalgae include Halimeda tuna, Dictyota divaricata, Lobophora variegata, Ventricatri ventricosa, Verdigelas pelas, and Kallymenia sp. Anadyomene menziesii is abundant. The reef provides a habitat for organisms typically observed at much shallower depths, and is the deepest known photosynthetic coral reef on the North America continental shelf (Fig. 1).

  19. Glycyrrhiza glabra extract protects plants against important phytopathogenic fungi.

    PubMed

    Schuster, C; Konstantinidou-Doltsinis, S; Schmitt, A

    2010-01-01

    In previous investigations an ethanolic plant extract from Glycyrrhiza glabra (2.5% w/v) showed 100% efficacy against late blight (Phytophthora infestans) on detached tomato leaves. Based on these findings, the objective of this work was to investigate the effect of this extract against different important plant pathogenic fungi. Tests were carried out on potted plants. Against P. infestans, efficacies of 75% and 58% were achieved on tomato and potato plants with 5% extract concentration, respectively. Against another Oomycete, Pseudoperonospora cubensis, on cucumber, application of a 2.5% extract led to an efficacy of above 90%. The EC50-value was calculated to be 0.5% In a trial on beans against bean rust (Uromyces appendiculatus), G. glabra extract (5% concentration) showed 92% efficacy. In contrast, against powdery mildew on cucumber (Podosphaera xanthii), no disease reduction was found. Overall, the results indicate a high potential for the extract of G. glabra to control a number of important plant pathogens.

  20. Effects of a non-native cichlid fish (African jewelfish, Hemichromis letourneuxi Sauvage 1880) on a simulated Everglades aquatic community

    USGS Publications Warehouse

    Schofield, Pamela J.; Slone, Daniel H.; Gregoire, Denise R.; Loftus, William F.

    2014-01-01

    In an 8-month mesocosm experiment, we examined how a simulated Everglades aquatic community of small native fishes, snails, and shrimp changed with the addition of either a native predator (dollar sunfish Lepomis marginatus) or a non-native predator (African jewelfish Hemichromis letourneuxi) compared to a no-predator control. Two snail species (Planorbella duryi, Physella cubensis) and the shrimp (Palaemonetes paludosus) displayed the strongest predator-treatment effects, with significantly lower biomasses in tanks with Hemichromis. One small native fish (Heterandria formosa) was significantly less abundant in Hemichromis tanks, but there were no significant treatment effects for Gambusia holbrooki, Jordanella floridae, or Pomacea paludosa (applesnail). Overall, there were few treatment differences between native predator and no-predator control tanks. The results suggest that the potential of Hemichromis to affect basal food-web species that link primary producers with higher-level consumers in the aquatic food web, with unknown consequences for Florida waters.

  1. A revision of the genus Atelecrinus PH Carpenter (Echinodermata: Crinoidea).

    PubMed

    Messing, Charles G

    2013-01-01

    The unusual bathyal comatulid crinoid genus Atelecrinus is widespread in the Atlantic and tropical Pacific Oceans and currently includes three recognized species. A re-assessment based on examination of new and existing specimens requires establishment of two new genera and five new species, and returns three junior synonyms to species-level status. Paratelecrinus is erected to accommodate Atelecrinus wyvilli PH Carpenter, A. conifer AH Clark, A. cubensis PH Carpenter, P. orthotriremis, new species, P. amenouzume new species, P. laticonulus new species and P. telo new species. Adelatelecrinus is erected to accommodate Atelecrinus sulcatus AH Clark and Adelatelecrinus vallatus new species. Atelecrinus retains A. balanoides PH Carpenter and A. helgae AH Clark, which restricts the genus to the Atlantic. In both Paratelecrinus and Adelatelecrinus, the basals articulate with the centrodorsal via ligament bundles anchored in deep ringlike interradial pits that project into the centrodorsal cavity, whereas in Atelecrinus the centrodorsal rim has shallow interradial concavities and attaches to the basals via a tight junction with no obvious ligament bundles. The spoon-shaped aboral fossa in the basals of Paratelecrinus appears to be unique among articulate crinoids and differs from the smooth fossa found in both Atelecrinus and Adelatelecrinus. New material extends the range of the family to the Indian Ocean. A few species are now known from enough specimens to identify some ontogenetic and distributional variations. Proximal ray morphology varies substantially with size in P. cubensis and P. orthotriremis. A. balanoides generally occurs in deeper water in the Lesser Antilles than in the Bahamas and Strait of Florida, while P. orthotriremis occurs in shallower water in the Lesser Antilles and deeper in the Bahamas.

  2. Distribution and abundance of schistosomiasis and fascioliasis host snails along the Mara River in Kenya and Tanzania.

    PubMed

    Dida, Gabriel O; Gelder, Frank B; Anyona, Douglas N; Matano, Ally-Said; Abuom, Paul O; Adoka, Samson O; Ouma, Collins; Kanangire, Canisius K; Owuor, Phillip O; Ofulla, Ayub V O

    2014-01-01

    We purposively selected 39 sampling sites along the Mara River and its two perennial tributaries of Amala and Nyangores and sampled snails. In addition, water physicochemical parameters (temperature, turbidity, dissolved oxygen, conductivity, alkalinity, salinity and pH) were taken to establish their influence on the snail abundance and habitat preference. Out of the 39 sites sampled, 10 (25.6%) had snails. The snail species encountered included Biomphalaria pfeifferi Krauss - the intermediate host of Schistosoma mansoni Sambon, Bulinus africanus - the intermediate host of Schistosoma haematobium, and Lymnaea natalensis Krauss - the intermediate host of both Fasciola gigantica and F. hepatica Cobbold. Ceratophallus spp., a non-vector snail was also encountered. Most (61.0%) of the snails were encountered in streamside pools. Schistosomiasis-transmitting host snails, B. pfeifferi and B. africanus, were fewer than fascioliasis-transmitting Lymnaea species. All the four different snail species were found to be attached to different aquatic weeds, with B. pfeifferi accounting for over half (61.1%) of the snails attached to the sedge, followed by B. africanus and Lymnaea spp., accounting for 22.2 and 16.7%, respectively. Ceratophallus spp. were non-existent in sedge. The results from this preliminary study show that snails intermediate hosts of schistosomiasis and fascioliasis exists in different habitats, in few areas along the Mara River, though their densities are still low to have any noticeable impacts on disease transmission in case they are infected. The mere presence of the vector snails in these focal regions calls for their immediate control and institution of proper regulations, management, and education among the locals that can help curtail the spread of the snails and also schistosomiasis and fascioliasis within the Mara River basin.

  3. Synthesis, Spectroscopic and Toxicity Studies of Titanocene Chelates of Isatin-3-Thiosemicarbazones

    PubMed Central

    Vatsa, Garima; Pandey, O. P.; Sengupta, S. K.

    2005-01-01

    The reactions of bis(cyclopentadienyl)titanium(IV) dichloride with a new class of thiosemicarbazone (LH2), derived by condensing isatin with different N(4)-substituted thiosemicarbazides, have been studied and products of type [Cp2Ti(L)] have been isolated. On the basis of various physico-chemical and spectral studies, five coordinate structures have been assigned to these derivatives. Toxicity studies of titanocene complexes at tbur different concentrations have been carried out against snail Lymnaea acuminata. The effect of most potent compounds on the activity of acetylcholinesterase enzyme, which inhibits the activity of enzyme, possibly by the formation of enzyme-inhibitor complex, was also studied. PMID:18365096

  4. Lead uptake and lead loss in the fresh water field crab, Barytelphusa guerini, on exposure to organic and inorganic lead

    SciTech Connect

    Tulasi, S.J.; Yasmeen, R.; Reddy, C.P.; Rao, J.V.R.

    1987-07-01

    Lead is a heavy metal which is widely used in paint industry, pigments, dyes, electrical components and electronics, plastic chemicals and in various other things. Since some of the lead salts are soluble in water, lead presents a potential threat to aquatic organisms. Studies dealing with invertebrates include those on mortality, growth and lead uptake in Lymnaea palustris and bioaccumulation of heavy metals in oysters and mussels. Little information exists regarding the effect of lead on the fresh water crustaceans. Hence the present investigation has been undertaken to study the uptake and loss of lead on exposure to subtoxic levels or organic and inorganic lead.

  5. Peculiarities of studying an isolated neuron by the method of laser interference microscopy

    SciTech Connect

    Yusipovich, Alexander I; Kazakova, Tatiana A; Erokhova, Liudmila A; Brazhe, Nadezda A; Maksimov, Georgy V; Novikov, Sergey M; Lazarev, Grigory L

    2006-09-30

    Actual aspects of using a new method of laser interference microscopy (LIM) for studying nerve cells are discussed. The peculiarities of the LIM display of neurons are demonstrated by the example of isolated neurons of a pond snail Lymnaea stagnalis. A comparative analysis of the images of the cell and subcellular structures of a neuron obtained by the methods of interference microscopy, optical transmission microscopy, and confocal microscopy is performed. Various aspects of the application of LIM for studying the lateral dimensions and internal structure of the cytoplasm and organelles of a neuron in cytology and cell physiology are discussed. (laser biology)

  6. LASER BIOLOGY: Peculiarities of studying an isolated neuron by the method of laser interference microscopy

    NASA Astrophysics Data System (ADS)

    Yusipovich, Alexander I.; Novikov, Sergey M.; Kazakova, Tatiana A.; Erokhova, Liudmila A.; Brazhe, Nadezda A.; Lazarev, Grigory L.; Maksimov, Georgy V.

    2006-09-01

    Actual aspects of using a new method of laser interference microscopy (LIM) for studying nerve cells are discussed. The peculiarities of the LIM display of neurons are demonstrated by the example of isolated neurons of a pond snail Lymnaea stagnalis. A comparative analysis of the images of the cell and subcellular structures of a neuron obtained by the methods of interference microscopy, optical transmission microscopy, and confocal microscopy is performed. Various aspects of the application of LIM for studying the lateral dimensions and internal structure of the cytoplasm and organelles of a neuron in cytology and cell physiology are discussed.

  7. Chronic lead exposure reduces junctional resistance at an electrical synapse.

    PubMed

    Audesirk, G; Audesirk, T

    1984-01-01

    Both acute and chronic lead exposure have been found to inhibit transmission at chemical synapses, possibly by interfering with inward calcium current. We have found that chronic lead exposure slightly reduces input resistance and greatly reduces the junctional resistance between two strongly electrically coupled neurons in the pond snail Lymnaea stagnalis. The net effect is to increase the strength of electrical coupling. A reduction in gap junctional resistance would also be expected to increase the flow of small molecules between cells. However, Lucifer Yellow injections did not reveal dye-coupling between the cells. Lead exposure also increases the capacitance of the neurons.

  8. Microelectrode investigation of neuroneal ageing from a single identified neurone.

    PubMed

    Patel, Bhavik Anil; Arundell, Martin; Parker, Kim H; Yeoman, Mark S; O'Hare, Danny

    2010-09-14

    Microelectrode amperometry is uniquely suited for characterising the dynamics of neurotransmitter release, as it offers unparalleled spatial and temporal resolution. We have used carbon fibre microelectrodes to study release of the monoamine neurotransmitter serotonin (5-HT) and the gaseous transmitter nitric oxide (NO) in intact central nervous system of the water snail, Lymnaea stagnalis. Analysis of spontaneous vesicular release of 5-HT and depolarisation-induced release of NO reveals significant differences with ageing that may be associated with changes in protein structure and function.

  9. Effects of histochrom and emoxypin on biophysical properties of electroexitable cells.

    PubMed

    Mischenko, N P; Fedoreev, S A; Zapara, T A; Ratushnyak, A S

    2009-02-01

    Comparative investigation of effects of antioxidant agents histochrom and emoxypin on biophysical characteristics of isolated neurons from shell Lymnaea stagnalis under normal conditions and during oxidative stress was performed. Differently directed effects of these compounds on resting potential and transmembrane ion currents of neural cells under normal conditions were detected. Histochrom provides hyperpolarizing and emoxypin--depolarizing effect on neuronal membrane potential. Under conditions of oxidative stress both products possess antioxidant action. Obtained data allows coming closer to understanding of cellular-molecular mechanisms of protective action of compounds.

  10. Estimating Polygenic Models for Multivariate Data on Large Pedigrees

    PubMed Central

    Thompson, E. A.; Shaw, R. G.

    1992-01-01

    We have developed algorithms for the likelihood estimation of additive genetic models for quantitative traits on large pedigrees. The approach uses the expectation L-maximization (EM) algorithm, but avoids intensive computation. In this paper, we focus on extensions of previous work to the case of multivariate data. We exemplify the approach by analyses of bivariate data on a four-generation, 949-member pedigree of the snail Lymnaea elodes, and on a three-generation pedigree of the guppy Poecilia reticulata containing about 400 individuals. PMID:1516823

  11. Trophic factor-induced activity 'signature' regulates the functional expression of postsynaptic excitatory acetylcholine receptors required for synaptogenesis.

    PubMed

    Luk, Collin C; Lee, Arthur J; Wijdenes, Pierre; Zaidi, Wali; Leung, Andrew; Wong, Noelle Y; Andrews, Joseph; Syed, Naweed I

    2015-04-01

    Highly coordinated and coincidental patterns of activity-dependent mechanisms ("fire together wire together") are thought to serve as inductive signals during synaptogenesis, enabling neuronal pairing between specific sub-sets of excitatory partners. However, neither the nature of activity triggers, nor the "activity signature" of long-term neuronal firing in developing/regenerating neurons have yet been fully defined. Using a highly tractable model system comprising of identified cholinergic neurons from Lymnaea, we have discovered that intrinsic trophic factors present in the Lymnaea brain-conditioned medium (CM) act as a natural trigger for activity patterns in post- but not the presynaptic neuron. Using microelectrode array recordings, we demonstrate that trophic factors trigger stereotypical activity patterns that include changes in frequency, activity and variance. These parameters were reliable indicators of whether a neuron expressed functional excitatory or inhibitory nAChRs and synapse formation. Surprisingly, we found that the post- but not the presynaptic cell exhibits these changes in activity patterns, and that the functional expression of excitatory nAChRs required neuronal somata, de novo protein synthesis and voltage gated calcium channels. In summary, our data provides novel insights into trophic factor mediated actions on neuronal activity and its specific regulation of nAChR expression.

  12. Expression of a retinoic acid receptor (RAR)-like protein in the embryonic and adult nervous system of a protostome species.

    PubMed

    Carter, Christopher J; Rand, Christopher; Mohammad, Imtiaz; Lepp, Amanda; Vesprini, Nicholas; Wiebe, Olivia; Carlone, Robert; Spencer, Gaynor E

    2015-01-01

    The vitamin A metabolite, retinoic acid, is an important molecule in nervous system development and regeneration in vertebrates. Retinoic acid signaling in vertebrates is mediated by two classes of nuclear receptors, the retinoid X receptors (RXRs) and the retinoic acid receptors (RARs). Recently, evidence has emerged to suggest that many effects of retinoic acid are conserved between vertebrate and invertebrate nervous systems, even though the RARs were previously thought to be a vertebrate innovation and to not exist in non-chordates. We have cloned a full-length putative RAR from the CNS of the mollusc Lymnaea stagnalis (LymRAR). Immunoreactivity for the RAR protein was found in axons of adult neurons in the central nervous system and in growth cones of regenerating neurons in vitro. A vertebrate RAR antagonist blocked growth cone turning induced by exogenous all-trans retinoic acid, possibly suggesting a role for this receptor in axon guidance. We also provide immunostaining evidence for the presence of RAR protein in the developing, embryonic CNS, where it is also found in axonal processes. Using qPCR, we determined that LymRAR mRNA is detectable in the early veliger stage embryo and that mRNA levels increase significantly during embryonic development. Putative disruption of retinoid signaling in Lymnaea embryos using vertebrate RAR antagonists resulted in abnormal eye and shell development and in some instances completely halted development, resembling the effects of all-trans retinoic acid. This study provides evidence for RAR functioning in a protostome species.

  13. Background concentrations of heavy metals in benthos from transboundary rivers of the Transbaikalia region, Russia.

    PubMed

    Kuklin, Aleksei Petrovich; Matafonov, Petr Viktorovich

    2014-02-01

    The concentrations (mg/kg dry weight) of Cu, Zn, As, Cd, Hg, and Pb were measured in benthic macroalgae and invertebrates collected in the upper transboundary tributaries of the Onon River, Transbaikalia, Russia. The background concentration ranges in Cladophora fracta, Ulothrix zonata and Zygnemataceae were: 6.4-9.1 for Cu, 27.2-73.1 for Zn, 0.4-0.9 for Cd, 6.7-35.3 for As, 0.01-0.02 for Hg, and 1.9-4.3 for Pb. In Brachycentrus americanus and Lymnaea media the concentration ranges were: 9.0-25.5 for Cu, 21.4-96.0 for Zn, 0.1-0.3 for Cd, 1.7-5.6 for As, 0.004-0.02 for Hg, and 0.4-2.2 for Pb. The concentrations of Cu, Zn, Pb, and Hg were consistent with data for uncontaminated areas. Under contamination conditions the concentrations in C. fracta were: 938 for Zn, 513 for Pb, and 9.5 for Cd; in Lymnaea media were: 46.8 for Cu, 176 for Zn, 52.3 for Pb, and 3.0 for Cd. All the organisms showed a common response to contamination, and consequently can be used as biomonitors of contamination by heavy metals.

  14. Organic and inorganic lead inhibit neurite growth in vertebrate and invertebrate neurons in culture.

    PubMed

    Audesirk, G; Shugarts, D; Nelson, G; Przekwas, J

    1989-12-01

    Neurons from brains of chick embryos and pond snails (Lymnaea stagnalis) were cultured for 3 to 4 d in the presence of no toxins, inorganic lead (PbCl2), or organic lead (triethyl lead chloride). In chick neurons, inorganic lead reduced the percentage of cells that grew neurites (IC50 = 270 microM total lead, approximately 70 nM free Pb2+) but did not reduce the number of neurites per cell or the mean neurite length. Triethyl lead reduced the percentage of cells that grew neurites (IC50 = 0.24 microM) and the mean neurite length (extrapolated IC50 = 3.6 microM) but did not reduce the number of neurites per cell. In Lymnaea neurons, inorganic lead reduced the percentage of cells that grew neurites (IC50 = 13 microM total lead; approximately 10 nM free Pb2+). Triethyl lead reduced the percentage of cells that grew neurites (IC50 = 0.4 microM) and exerted significant toxicity at 0.2 microM. The two forms of lead affected neurite growth in qualitatively different ways, which suggests that their mechanisms of action are different.

  15. Fasciola hepatica in snails collected from water-dropwort fields using PCR.

    PubMed

    Kim, Hwang-Yong; Choi, In-Wook; Kim, Yeon-Rok; Quan, Juan-Hua; Ismail, Hassan Ahmed Hassan Ahmed; Cha, Guang-Ho; Hong, Sung-Jong; Lee, Young-Ha

    2014-12-01

    Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea.

  16. Bioanalytical Applications of Real-Time ATP Imaging Via Bioluminescence

    SciTech Connect

    Gruenhagen, Jason Alan

    2003-01-01

    The research discussed within involves the development of novel applications of real-time imaging of adenosine 5'-triphosphate (ATP). ATP was detected via bioluminescence and the firefly luciferase-catalyzed reaction of ATP and luciferin. The use of a microscope and an imaging detector allowed for spatially resolved quantitation of ATP release. Employing this method, applications in both biological and chemical systems were developed. First, the mechanism by which the compound 48/80 induces release of ATP from human umbilical vein endothelial cells (HUVECs) was investigated. Numerous enzyme activators and inhibitors were utilized to probe the second messenger systems involved in release. Compound 48/80 activated a G{sub q}-type protein to initiate ATP release from HUVECs. Ca2+ imaging along with ATP imaging revealed that activation of phospholipase C and induction of intracellular Ca2+ signaling were necessary for release of ATP. Furthermore, activation of protein kinase C inhibited the activity of phospholipase C and thus decreased the magnitude of ATP release. This novel release mechanism was compared to the existing theories of extracellular release of ATP. Bioluminescence imaging was also employed to examine the role of ATP in the field of neuroscience. The central nervous system (CNS) was dissected from the freshwater snail Lymnaea stagnalis. Electrophysiological experiments demonstrated that the neurons of the Lymnaea were not damaged by any of the components of the imaging solution. ATP was continuously released by the ganglia of the CNS for over eight hours and varied from ganglion to ganglion and within individual ganglia. Addition of the neurotransmitters K+ and serotonin increased release of ATP in certain regions of the Lymnaea CNS. Finally, the ATP imaging technique was investigated for the study of drug release systems. MCM-41-type mesoporous nanospheres were loaded with ATP and end-capped with mercaptoethanol

  17. Transmission patterns of Fasciola hepatica to ruminants in Sweden.

    PubMed

    Novobilský, Adam; Engström, Annie; Sollenberg, Sofia; Gustafsson, Katarina; Morrison, David A; Höglund, Johan

    2014-07-14

    Transmission patterns of Fasciola hepatica were investigated on beef cattle (n=3) and sheep (n=3) farms in Sweden between 2011 and 2012. The dynamics of fluke infection, particularly estimated time of infection, were screened each grazing season by ELISA detection of antibodies in lambs (n=94) and first grazing season calves (n=61). Colostral transfer of F. hepatica antibodies from seropositive ewes was detected in sheep up to 11 weeks of age. In sheep, the estimated time of infection differed significantly between herds and years. Typical 'winter infection' was observed on two sheep farms in 2012, but the most prevalent transmission pattern was found to be 'summer infection', characterised by infection of animals in late summer by F. hepatica originating from overwintered and/or spring-excreted eggs. In contrast, beef calves were infected mainly in September-October ('summer infection'). Furthermore, lymnaeid and succineid snails were collected on the pastures used by these animals both in spring and in the autumn each year. In total, 1726, 588, 138, 130, 93 and 42 specimens of Galba truncatula, Lymnaea palustris, Lymnaea glabra, Lymnaea fuscus, Radix peregra and Succinea putris, respectively, were collected and identified. These were subsequently examined for the presence of F. hepatica DNA by species-specific PCR and the findings compared against mean monthly rainfall and temperature data for each farm. The main intermediate host of the liver fluke was G. truncatula, with a prevalence range of F. hepatica infection from 0% to 82%. Only 1 out of 42 terrestrial S. putris tested positive for F. hepatica, casting doubt on the role of this species in transmission of F. hepatica in Sweden. In conclusion, two main peak periods of infection were observed: May-June (from overwintered infected snails='winter infection') and August-September (from metacercariae developed and produced by snails during summer='summer infection'). The occurrence and frequency of 'winter

  18. Isolation and characterization of an antifungal protein from Bacillus licheniformis HS10.

    PubMed

    Wang, Zhixin; Wang, Yunpeng; Zheng, Li; Yang, Xiaona; Liu, Hongxia; Guo, Jianhua

    2014-11-07

    Bacillus licheniformis HS10 is a good biocontrol agent against Pseudoperonospora cubensis which caused cucumber downy disease. To identify and characterize the antifungal proteins produced by B.licheniformis HS10, the proteins from HS10 were isolated by using 30-60% ammonium sulfate precipitation, and purified with column chromatography on DEAE Sepharose Fast Flow, RESOURCE Q and Sephadex G-75. And the SDS-PAGE and MALDI-TOF/TOF-MS analysis results demonstrated that the antifungal protein was a monomer with molecular weight of about 55 kDa, identified as carboxypeptidase. Our experiments also showed that the antifungal protein from B. licheniformis HS10 had significantly inhibition on eight different kinds of plant pathogenic fungi, and it was stable with good biological activity at as high as 100°C for 30 min and in pH value ranged from 6 to 10. The biological activity was negatively affected by protease K and 10mM metal cations except Ca(2+).

  19. Ultrasound and physical models shed light on the respiratory system of embryonic dogfishes.

    PubMed

    Tomita, Taketeru; Cotton, Charles F; Toda, Minoru

    2016-02-01

    Embryos of live-bearing elasmobranchs (sharks, skates, and rays) must acquire oxygen in the uterus for several months to more than a year, but the mechanisms of delivery and uptake are still largely unknown. Diagnostic sonography performed on a captive Japanese dogfish (Squalus japonicus) showed that a late-stage embryo used buccal movement to pump uterine fluid, suggesting that the embryo acquires oxygen from uterine fluid via gill ventilation. It has been assumed that embryonic respiration in aplacental sharks depends on oxygen supplied by the uterine wall. To test this hypothesis, the rate of oxygen diffusion was estimated by applying a physical model to the uterine wall of two dogfish species (Squalus cf. mitsukurii and Squalus cubensis). The model calculations indicate that the supply of oxygen via diffusion through the uterine villi contributes less than 15-30% of the total oxygen demand of late-stage embryos. Some previous authors have suggested that pregnant dogfish intermittently exchange uterine fluid with external seawater during late gestation. Thus, late-stage embryos may acquire oxygen primarily from uterine seawater introduced from the external environment.

  20. Cucumber disease diagnosis using multispectral images

    NASA Astrophysics Data System (ADS)

    Feng, Jie; Li, Hongning; Shi, Junsheng; Yang, Weiping; Liao, Ningfang

    2009-07-01

    In this paper, multispectral imaging technique for plant diseases diagnosis is presented. Firstly, multispectral imaging system is designed. This system utilizes 15 narrow-band filters, a panchromatic band, a monochrome CCD camera, and standard illumination observing environment. The spectral reflectance and color of 8 Macbeth color patches are reproduced between 400nm and 700nm in the process. In addition, spectral reflectance angle and color difference is obtained through measurements and analysis of color patches using spectrometer and multispectral imaging system. The result shows that 16 narrow-bands multispectral imaging system realizes good accuracy in spectral reflectance and color reproduction. Secondly, a horticultural plant, cucumber' familiar disease are the researching objects. 210 multispectral samples are obtained by multispectral and are classified by BP artificial neural network. The classification accuracies of Sphaerotheca fuliginea, Corynespora cassiicola, Pseudoperonospora cubensis are 100%. Trichothecium roseum and Cladosporium cucumerinum are 96.67% and 90.00%. It is confirmed that the multispectral imaging system realizes good accuracy in the cucumber diseases diagnosis.

  1. Taxonomic revision of Rochefortia Sw. (Ehretiaceae, Boraginales)

    PubMed Central

    Irimia, Ramona-Elena

    2016-01-01

    Abstract Background Rochefortia is a small taxon of woody plants in the Ehretiaceae (Boraginales) exhibiting coriaceous leaves with cystoliths, small whitish flowers and drupaceous fruits containing four pyrenes. It shares the dioecious sex distribution with its sister group Lepidocordia and can be delimited from the latter (and all other Ehretiaceae) by the presence of thorns. Neotropical Rochefortia is distributed over most Caribbean islands, Central America and northern South America. Twenty-eight validly published names (corresponding to twenty-one typified taxa at the species level and below) are available in Rochefortia, but the precise number of species to be accepted has been elusive before this revision. New information In the course of the present revision, 353 herbarium collections, comprising approximately 540 Rochefortia specimens, were entried into a BRAHMS data base providing information about protologues and types and retrospective georeferences if possible. Based on the combination of molecular and morphological data we propose to recognise nine species of Rochefortia, namely R. acanthophora, R. bahamensis, R. barloventensis, R. cubensis, R. cuneata, R. lundellii, R. oblongata, R. spinosa and R. stellata (the remaining nineteen validly published names are synonymised under such names). Morphological description of each species and an identification key are provided. PMID:27346952

  2. Intron Derived Size Polymorphism in the Mitochondrial Genomes of Closely Related Chrysoporthe Species

    PubMed Central

    Kanzi, Aquillah Mumo; Wingfield, Brenda Diana; Steenkamp, Emma Theodora; Naidoo, Sanushka; van der Merwe, Nicolaas Albertus

    2016-01-01

    In this study, the complete mitochondrial (mt) genomes of Chrysoporthe austroafricana (190,834 bp), C. cubensis (89,084 bp) and C. deuterocubensis (124,412 bp) were determined. Additionally, the mitochondrial genome of another member of the Cryphonectriaceae, namely Cryphonectria parasitica (158,902 bp), was retrieved and annotated for comparative purposes. These genomes showed high levels of synteny, especially in regions including genes involved in oxidative phosphorylation and electron transfer, unique open reading frames (uORFs), ribosomal RNAs (rRNAs) and transfer RNAs (tRNAs), as well as intron positions. Comparative analyses revealed signatures of duplication events, intron number and length variation, and varying intronic ORFs which highlighted the genetic diversity of mt genomes among the Cryphonectriaceae. These mt genomes showed remarkable size polymorphism. The size polymorphism in the mt genomes of these closely related Chrysoporthe species was attributed to the varying number and length of introns, coding sequences and to a lesser extent, intergenic sequences. Compared to publicly available fungal mt genomes, the C. austroafricana mt genome is the second largest in the Ascomycetes thus far. PMID:27272523

  3. Synthesis and biological evaluation of novel fluorine-containing stilbene derivatives as fungicidal agents against phytopathogenic fungi.

    PubMed

    Jian, Weilin; He, Daohang; Xi, Pinggen; Li, Xinwei

    2015-11-18

    The rising development of resistance to conventional fungicides is driving the search for new alternative candidates to control plant diseases. In this study, a series of new fluorine-containing stilbene derivatives was synthesized on the basis of our previous quantitative structure-activity relationship analysis results. Bioassays in vivo revealed that the title compounds exhibited potent fungicidal activities against phytopathogenic fungi (Colletotrichum lagenarium and Pseudoperonospora cubensis) from cucumber plants. In comparison to the previous results, the introduction of a fluorine moiety showed improved activities of some compounds against those fungi. Notably, compound 9 exhibited a control efficacy against C. lagenarium (83.4 ± 1.3%) comparable to that of commercial fungicide (82.7 ± 1.7%). For further understanding the possible mode of action of the stilbene against C. lagenarium, the effects on hyphal morphology, electrolyte leakage, and respiration of mycelial cell suspension were studied. Microscopic observation showed considerably deformed mycelial morphology. The conductivity of mycelial suspension increased in the presence of compound 9, whereas no significantly inhibitory effect on respiration was observed. Taken together, the fungicidal mechanism of this stilbene is associated with its membrane disruption effect, resulting in increased membrane permeability. These results provide important clues for mechanistic study and derivatization of stilbenes as alternative sources of fungicidal agents for plant disease control.

  4. Species diversity in the Antrodia crassa group (Polyporales, Basidiomycota).

    PubMed

    Spirin, Viacheslav; Runnel, Kadri; Vlasák, Josef; Miettinen, Otto; Põldmaa, Kadri

    2015-12-01

    Antrodia is a polyphyletic genus, comprising brown-rot polypores with annual or short-lived perennial resupinate, dimitic basidiocarps. Here we focus on species that are closely related to Antrodia crassa, and investigate their phylogeny and species delimitation using geographic, ecological, morphological and molecular data (ITS and LSU rDNA, tef1). Phylogenetic analyses distinguished four clades within the monophyletic group of eleven conifer-inhabiting species (five described herein): (1)A. crassa s. str. (boreal Eurasia), Antrodia cincta sp. nova (North America) and Antrodia cretacea sp. nova (holarctic), all three being characterized by inamyloid skeletal hyphae that dissolve quickly in KOH solution; (2) Antrodia ignobilis sp. nova, Antrodia sitchensis and Antrodia sordida from North America, and Antrodia piceata sp. nova (previously considered conspecific with A. sitchensis) from Eurasia, possessing amyloid skeletal hyphae; (3) Antrodia ladiana sp. nova from the southern part of the USA, Antrodia pinea from East Asia, and Antrodia ferox - so far known from subtropical North America, but here reported also from Eurasia. These three species have inamyloid hyphae and narrow basidiospores; (4) the North American Antrodia pini-cubensis, sharing similar morphological characters with A. pinea, forming a separate clade. The habitat data indicate that several species are threatened by intensive forestry.

  5. Synoptic revision of Blabericola (Apicomplexa: Eugregarinida: Blabericolidae) parasitizing blaberid cockroaches (Dictyoptera: Blaberidae), with comments on delineating gregarine species boundaries.

    PubMed

    Clopton, Richard E

    2012-06-01

    Complete synoptic redescriptions, including complete morphometric data for all life cycle stages, species recognition characters, and differential comparisons are presented for the 4 gregarine species comprising Blabericola . Blabericola cubensis ( Peregrine, 1970 ), Blabericola haasi (Geus, 1969), Blabericola migrator ( Clopton, 1995 ), and Blabericola princisi ( Peregrine, 1970 ) are redescribed from their type hosts, i.e., the discoid cockroach Blaberus discoidalis , the lobster cockroach Nauphoeta cinerea , the Madagascar hissing cockroach Gromphadorhina portentosa , and the Bolivian cockroach Blaberus boliviensis , respectively. These gregarine species descriptions are stabilized through deposition of extensive new voucher collections. Species of Blabericola are distinguished by differences in relative metric ratios, morphology of oocysts, and by relative metric ratios of mature gamonts in association. This work is discussed as a model for morphological species descriptions in the Eugregarinorida including the 6 principles for morphological gregarine species descriptions, i.e., a centroid and population variation approach, adequate sample size, partitioning developmental variation and sexual dimorphism, recognition and minimization of fixation and physiological artifacts to eliminate false morphotypes, and comparative data sets across multiple life cycle stages.

  6. Effects of Washing Produce Contaminated with the Snail and Slug Hosts of Angiostrongylus cantonensis with Three Common Household Solutions

    PubMed Central

    Yeung, Norine W; Hayes, Kenneth A

    2013-01-01

    The emerging infectious disease angiostrongyliasis (rat lungworm disease) is caused by ingesting snails and slugs infected by the nematode Angiostrongylus cantonensis. The definitive hosts of A. cantonensis are rats and the obligatory intermediate hosts are slugs and snails. Many cases result from accidentally ingesting infected snails or slugs on produce (eg, lettuce). This study assessed three readily available household products as washing solutions for removing snails and slugs from produce (romaine lettuce) to lower the probability of accidentally ingesting them. The solutions were acetic acid (vinegar), sodium hypochlorite (bleach), and sodium chloride (domestic salt). Snail and slug species known to be intermediate hosts and that are common in the Hawaiian Islands were used in the experiments: the alien snail Succinea tenella, the alien semi-slug Parmarion martensi, and the alien slugs Veronicella cubensis and Deroceras laeve. None of the products was any more effective than washing and rinsing with tap water alone. Most snails and slugs were removed after treatment but some remained on the lettuce even after washing and rinsing the produce. Only washing, rinsing, and then rinsing each leaf individually resulted in complete removal of all snails and slugs. The study did not address removal of any remaining slime left by the snails and slugs, nor did it address killing of worms. PMID:23901391

  7. Effects of washing produce contaminated with the snail and slug hosts of Angiostrongylus cantonensis with three common household solutions.

    PubMed

    Yeung, Norine W; Hayes, Kenneth A; Cowie, Robert H

    2013-06-01

    The emerging infectious disease angiostrongyliasis (rat lungworm disease) is caused by ingesting snails and slugs infected by the nematode Angiostrongylus cantonensis. The definitive hosts of A. cantonensis are rats and the obligatory intermediate hosts are slugs and snails. Many cases result from accidentally ingesting infected snails or slugs on produce (eg, lettuce). This study assessed three readily available household products as washing solutions for removing snails and slugs from produce (romaine lettuce) to lower the probability of accidentally ingesting them. The solutions were acetic acid (vinegar), sodium hypochlorite (bleach), and sodium chloride (domestic salt). Snail and slug species known to be intermediate hosts and that are common in the Hawaiian Islands were used in the experiments: the alien snail Succinea tenella, the alien semi-slug Parmarion martensi, and the alien slugs Veronicella cubensis and Deroceras laeve. None of the products was any more effective than washing and rinsing with tap water alone. Most snails and slugs were removed after treatment but some remained on the lettuce even after washing and rinsing the produce. Only washing, rinsing, and then rinsing each leaf individually resulted in complete removal of all snails and slugs. The study did not address removal of any remaining slime left by the snails and slugs, nor did it address killing of worms.

  8. Screening of some Nigerian plants for molluscicidal activity.

    PubMed

    Kela, S L; Ogunsusi, R A; Ogbogu, V C; Nwude, N

    1989-01-01

    Methanolic (MEOH), evaporated crude water (ECW) and unevaporated crude water (UECW) extracts of 25 Nigerian plants, used for different medicinal and domestic purposes were screened for molluscacidal activity on laboratory-reared Lymnaea natalensis Krauss. Seven of the plants were not active; extracts from 18 (72 per cent) of the plants, some of which are renowned fish poisons, had molluscicidal activity. These were Acacia nilotica, Aristolochia albida, Balanites aegyptiaca, Blighia sapida, Boswellia dalzielii, Detarium microcarpum, Gnidia kraussiana, Kigelia africana, Nauclea latifolia, Opilia celtidefolia, Parkia clappertoniana, Polygonum limbatum, Pseudocedrela kotschyi, Sclerocarya birrea, Securidaca longipedunculata, Ximenia americana, Vetiveria nigritana and Ziziphus abyssinica. The LC50 of these extracts were determined. It is strongly recommended that the toxic effects of these extracts against fish, cercariae, snail eggs and mammals be further investigated so as to determine the right concentration, especially for use in fish ponds.

  9. Commercial watercress as an emerging source of fascioliasis in Northern France in 2002: results from an outbreak investigation.

    PubMed

    Mailles, A; Capek, I; Ajana, F; Schepens, C; Ilef, D; Vaillant, V

    2006-10-01

    In April 2002, five cases of fascioliasis were diagnosed in Tourcoing. A case-finding and a case-control study were carried out to identify the source of the outbreak and take appropriate control measures. Eighteen cases were identified through the medical laboratories carrying out serology for fascioliasis. Fourteen cases and 23 controls, identified by the physicians of the cases, were interviewed on symptoms of the disease and their consumption of uncooked plants. Cases were more likely than controls to have eaten commercialized raw watercress (OR 86.7, P < 0.001) and 13 (93%) of the cases reported its consumption. A single producer common to all cases was identified. The inspection of his watercress beds showed a lack of protection against Lymnaea truncatula. This outbreak of fascioliasis due to commercialized watercress indicates that actual sanitary regulations do not allow for the efficient prevention of infestation of watercress production in France.

  10. [Epidemiological data on human distomatosis (Fasciola hepatica L.) in the Limousin region of France. The species of plants eaten and snail hosts].

    PubMed

    Rondelaud, D

    1980-01-01

    These epidemiological studies concern 187 cases of human fasciolasis which occurred in the region of Limousin, France on a period of 24 years. Four species of plants have been eaten: Nasturtium officinale, Roripa silvestris, Taraxacum gr. officinale, Valerianella olitoria. These plants have been eaten during all months of the year, but with higher numbers in July-August and from November to February. The cases of human fasciolasis almost take place on all years, but their numbers are high in years showing a very important infestation of cattle by Fasciola hepatica. In 2/3 of cases, the plants come from stations situated out of cattle pastures. The water-cress almost comes from natural stations or from familial plantations. Five species of snails have been observed in these stations: Lymnaea glabra, L. palustris, L. peregra, L. stagnalis, L. truncatula. The young snails of these species experimentally have been observed. The significance of these results is discussed.

  11. Interaction of neurons at the level of cell bodies in the snail CNS. Heterogeneity of the neuroactive environment.

    PubMed

    Chistopol'skii, I A

    2005-09-01

    Experiments on the CNS of snail Lymnaea stagnalis in which a cell isolated from the serotonin cluster PeA was used as a mobile sensor neuron demonstrated the presence of neuroactive factors at the surface of the cellular "cortex" of the pedal ganglion. Apart from the previously known factor serotonin, effective concentrations of a factor suppressing the electrical activity of PeA were found at this site, along with a depolarizing factor which, unlike serotonin, narrowed PeA action potentials. The ability of these factors to control the electrical activity of the sensor neuron demonstrates the possible involvement of chemical agents in the intercellular space of the "cortex" in neuronal signaling.

  12. A flavanoid component of chocolate quickly reverses an imposed memory deficit.

    PubMed

    Knezevic, Bogdan; Komatsuzaki, Yoshimasa; de Freitas, Emily; Lukowiak, Ken

    2016-03-01

    The ability to remember is influenced by environmental and lifestyle factors, such as stress and diet. A flavanol contained in chocolate, epicatechin (Epi), has been shown to enhance long-term memory (LTM) formation in Lymnaea. Combining two stressors (low-calcium pond water and crowding) blocks learning and all forms of memory; that is, this combination of environmentally relevant stressors creates a memory-unfriendly state. We tested the hypothesis that Epi will immediately reverse the memory-unfriendly state, i.e. that snails in the memory-deficit state when trained in Epi will immediately become competent to learn and form memory. We found that Epi not only reverses the memory-deficit state but also further enhances LTM formation. Thus, a naturally occurring bioactive plant compound can overcome a memory-unfriendly state. This supports the idea that bioactive substances may mitigate memory-making deficits that, for example, occur with ageing.

  13. Epicatechin, a component of dark chocolate, enhances memory formation if applied during the memory consolidation period

    PubMed Central

    Fernell, Maria; Swinton, Cayley; Lukowiak, Ken

    2016-01-01

    ABSTRACT Epicatechin (Epi), a flavanol found in foods such as dark chocolate has previously been shown to enhance memory formation in our model system, operant conditioning of aerial respiration in Lymnaea. In those experiments snails were trained in Epi. Here we ask whether snails exposed to Epi before training, during the consolidation period immediately following training, or 1 h after training would enhance memory formation. We report here that Epi is only able to enhance memory if snails are placed in Epi-containing pond water immediately after training. That is, Epi enhances memory formation if it is applied during the memory consolidation period as well as if snails are trained in Epi-containing pond water. PMID:27574544

  14. Epicatechin, a component of dark chocolate, enhances memory formation if applied during the memory consolidation period.

    PubMed

    Fernell, Maria; Swinton, Cayley; Lukowiak, Ken

    2016-01-01

    Epicatechin (Epi), a flavanol found in foods such as dark chocolate has previously been shown to enhance memory formation in our model system, operant conditioning of aerial respiration in Lymnaea. In those experiments snails were trained in Epi. Here we ask whether snails exposed to Epi before training, during the consolidation period immediately following training, or 1 h after training would enhance memory formation. We report here that Epi is only able to enhance memory if snails are placed in Epi-containing pond water immediately after training. That is, Epi enhances memory formation if it is applied during the memory consolidation period as well as if snails are trained in Epi-containing pond water.

  15. Receipt of seminal fluid proteins causes reduction of male investment in a simultaneous hermaphrodite.

    PubMed

    Nakadera, Yumi; Swart, Elferra M; Hoffer, Jeroen N A; den Boon, Onno; Ellers, Jacintha; Koene, Joris M

    2014-04-14

    Mating partners often have conflicting interests when copulating. One of the major agents affecting female mating partners is seminal fluid, transferred along with sperm. The role of seminal fluid proteins (SFPs) in reproductive success is well studied in separate-sexed animals but is much less so in simultaneous hermaphrodites. The latter potentially have a unique target to exploit for the sperm donor's own benefit: the male function of their mating partners. Here we show that, in the great pond snail Lymnaea stagnalis, receipt of specific SFPs reduces both sperm transfer and paternity success in a subsequent insemination event. Lowering investment in the mating partner's male function constitutes a novel role for SFPs. This demonstrates for the first time that hermaphrodites alter their mates' male as well as female reproductive output. Although it remains to be tested whether this represents mate manipulation or an adaptive response of recipients, our findings identify male investment as a new target for postcopulatory sexual selection.

  16. Molluscicidal activity of Lawsonia inermis and its binary and tertiary combinations with other plant derived molluscicides.

    PubMed

    Singh, A; Singh, D K

    2001-03-01

    Molluscicidal activity of leaf, bark and seed of Lawsonia inermis against Lymnaea acuminata and Indoplanorbis exustus was studied. Highest toxicity was observed in the seed of Lawsonia inermis. Toxicity of binary (1:1) and tertiary (1:1:1) combinations of the essential oil of cedar (Cedrus deodara Roxh) and neem (Azadirachta indica A. Juss), powder from bulb of garlic (Allium sativum Linn), and oleoresin extracted from rhizome of ginger (Zingiber officinale Rosc) with Lawsonia inermis and Embelia ribes fruit powder were studied against L. acuminata and I. exustus. L. inermis seed powder in combination with Cedrus deodara oil and Azadirachta indica oil was more toxic than their individual components and other combinations.

  17. Effect of different combinations of MGK-264 or piperonyl butoxide with plant-derived molluscicides on snail reproduction.

    PubMed

    Singh, K; Singh, D K

    2000-02-01

    Effect of sublethal treatment (20% and 60% of LC(50)/24 h) of plant-derived molluscicides, viz. Polianthes tuberosa, Trachyspermum ammi, Allium sativum powder; Azadirachta indica oil; oleoresin of Zingiber officinale; and their active molluscicidal component in combination (1:5) with MGK-264 or piperonyl butoxide on the reproduction of snail Lymnaea acuminata have been studied. It was observed that the combination of plant derived molluscicide and their active molluscicidal components, viz. tigogenin, hecogenin, azadirachtin, allicin, thymol, and [6]-gingerol combination with MGK-264 or piperonyl butoxide caused a significant reduction in fecundity, hatchability, and survival of young snails. Withdrawal of snails to fresh water after the above treatment caused a significant recovery in the fecundity of L. acuminata.

  18. Echinostoma friedi: the effect of age of adult worms on the infectivity of miracidia.

    PubMed

    Toledo, R; Espert, A; Carpena, I; Trelis, M; Muñoz-Antoli, C; Esteban, J G

    2004-03-01

    The effect of ageing of adults of Echinostoma friedi (Trematoda: Echinostomatidae) on the infectivity of miracidia yielded was analysed. Miracidia were obtained after hatching of eggs obtained from adult worms of E. friedi collected weekly during the course of experimental infections in golden hamsters. Miracidial infectivity, measured in terms of percentage of infection in Lymnaea peregra, was significantly influenced by the age of the adult worms from which the miracidia were derived. Infective miracidia only were obtained from adult worms in the age range from 4 to 9 weeks post-infection. Infectivity was maximal in those miracidia derived from adults collected 8 and 9 weeks post-infection. The results suggest that adult worms producing viable eggs require additional maturation to be able to yield eggs containing infective miracidia.

  19. The life cycle and transmission dynamics of the larval stages of Hypoderaeum conoideum.

    PubMed

    Muñoz-Antolí, C; Toledo, R; Esteban, J G

    2000-06-01

    The morphology of the different larval stages and life cycle of Hypoderaeum conoideum (Trematoda: Echinostomatidae) are described. The freshwater snail species Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural first intermediate host and this and L. corvus serve as experimental first intermediate hosts. These and other freshwater snails, such as Physella acuta and Gyraulus chinensis, in turn serve as second intermediate hosts. Adult worms were obtained from chicks and ducks, but not from rats, mice and golden hamsters. The morphology of the larval stages is compared with previous work on H. conoideum. Several aspects of the biology of the life history stages are described with emphasis on the transmission dynamics of the free-living stages. Differential suitability of the snail species that may act as first and/or second intermediate hosts is studied and discussed.

  20. Biosynthesis of galactogen: purification of a 1. -->. 6 D-galactosyltransferase from Helix pomatia

    SciTech Connect

    Goudsmit, E.M.

    1986-05-01

    A (1 ..-->.. 6) D-galactosyltransferase from a pellet fraction (8000xg) of Helix pomatia albumen gland has been purified over 2000-fold by affinity chromatography on UDP-p-amino-phenyl-Sepharose. The enzyme catalyzes transfer of D-galactose from UDP-galactose to a 1 ..-->.. 6 linkage on acceptor H. pomatia galactogen. Three other polymers served as acceptors; beef lung galactan, Lymnaea stagnalis galactogen and arabinogalactan from larch wood. To determine the linkage of added galactose termini, acceptor galactogen of H. pomatia that had been tritiated by treatment with galactose oxidase and (/sup 3/H)KBH/sub 4/ was incubated with UDP-D-galactose and purified enzyme extract. /sup 3/H-galactogen reaction product was recovered, methylated, hydrolyzed and acetylated; tritiated derivatives were identified by collection of effluent fractions from gas chromatography.

  1. Male Accessory Gland Protein Reduces Egg Laying in a Simultaneous Hermaphrodite

    PubMed Central

    Koene, Joris M.; Sloot, Wiebe; Montagne-Wajer, Kora; Cummins, Scott F.; Degnan, Bernard M.; Smith, John S.; Nagle, Gregg T.; ter Maat, Andries

    2010-01-01

    Seminal fluid is an important part of the ejaculate of internally fertilizing animals. This fluid contains substances that nourish and activate sperm for successful fertilization. Additionally, it contains components that influence female physiology to further enhance fertilization success of the sperm donor, possibly beyond the recipient's optimum. Although evidence for such substances abounds, few studies have unraveled their identities, and focus has been exclusively on separate-sex species. We present the first detailed study into the seminal fluid composition of a hermaphrodite (Lymnaea stagnalis). Eight novel peptides and proteins were identified from the seminal-fluid-producing prostate gland and tested for effects on oviposition, hatching and consumption. The gene for the protein found to suppress egg mass production, Ovipostatin, was sequenced, thereby providing the first fully-characterized seminal fluid substance in a simultaneous hermaphrodite. Thus, seminal fluid peptides and proteins have evolved and can play a crucial role in sexual selection even when the sexes are combined. PMID:20404934

  2. A novel, nongenomic mechanism underlies retinoic acid-induced growth cone turning.

    PubMed

    Farrar, Nathan R; Dmetrichuk, Jennifer M; Carlone, Robert L; Spencer, Gaynor E

    2009-11-11

    The vitamin A metabolite, retinoic acid (RA), is well known for its roles in neural development and regeneration. We have previously shown that RA can induce positive growth cone turning in regenerating neurons in vitro. In this study, we address the subcellular mechanisms underlying this chemo-attractive response, using identified central neurons from the adult mollusc, Lymnaea stagnalis. We show that the RA-induced positive growth cone turning was maintained in the presence of the transcriptional inhibitor, actinomycin D. We also physically transected the neurites from the cell body and showed that isolated growth cones retain the capacity to turn toward a gradient of RA. Moreover, this attractive turning is dependent on de novo local protein synthesis and Ca(2+) influx. Most of RA's actions during neurite outgrowth and regeneration require gene transcription, although these data show for the first time in any species, that the chemotropic action of RA in guiding neurite outgrowth, involves a novel, nongenomic mechanism.

  3. Prolonged modification of action potential shape by synaptic inputs in molluscan neurones.

    PubMed

    Winlow, W

    1985-01-01

    1. Somatic action potentials of Lymnaea neurons are modified by excitatory or inhibitory synaptic inputs and have been studied using phase-plane techniques and an action potential duration monitor. 2. Excitatory synaptic inputs increase the rate of neuronal discharge, cause action potential broadening, a decrease in the maximum rate of depolarization (Vd) and a decrease in the maximum rate of repolarization (Vr). 3. Inhibitory synaptic inputs decrease the discharge rate and cause narrowing of action potentials, an increase in Vd and an increase in Vr. 4. The effects reported above outlast the original synaptic inputs by many seconds and, if the somatic action potentials are similar to those in the axon terminals, they may have far-reaching effects on transmitter release.

  4. Sensitivity of midge larvae of Chironomus tentans Fabricius (Diptera Chironomidae) to heavy metals

    SciTech Connect

    Khangarot, B.S.; Ray, P.K.

    1989-03-01

    The discharge of heavy metals into the natural waters has numerous obvious impacts on physical, chemical and biological parameters of aquatic ecosystem. Bioassay tests are important steps in establishing appropriate water quality criteria and standards for diverse use of ponds, lakes, streams and river waters. Therefore, the acute toxicities of various heavy metals to water flea Daphnia magna, and snail Lymnaea acuminata, and toad tadpoles Bufo mentanostictus have been reported from the authors' laboratory. Chironomid larvae might be particularly useful as indicators of water quality because they are widely distributed in freshwater systems and often from diverse communities within particular habitat. The aim of this study was to determine the acute toxicity of ten heavy metals to the midge larvae Chironomus tentans Fabricius, which forms an important link in aquatic food chain(s).

  5. Comparative studies on ecotoxicology of synthetic detergents.

    PubMed

    Lal, H; Misra, V; Viswanathan, P N; Krishna Murti, C R

    1983-12-01

    To predict the comparative toxicological response of synthetic detergents on aquatic ecosystems, the effects of various concentrations of neutralized alkyl benzene sulfonate were studied. The median tolerance limit at 48 hr, 95% confidence limit, slope function, presumable harmless concentration, and rate of survival of different species of aquatic fauna such as water fleas (Daphnia magna), mosquito larvae (Culex pipiens), slug worms (Tubifex rivulorum), snails (Lymnaea vulgaris), tadpoles (Rana cyanophlyctis), and fish fingerlings (Cirrhina mrigala) were followed at 0, 24, 48, 72, and 96 hr. Any effect on quality of the water was also tested after the addition of various concentrations of detergents. The results showed that water fleas are more susceptible to detergent toxicity than fish fingerlings, tadpoles, slug worms, snails, and mosquito larvae. Behavioral changes were also observed as an index for detergent toxicity. The relative toxicity of the detergents to various species is discussed in relation to selective ecotoxicological response.

  6. Toxicity of anti-fouling biocides to encysted metacercariae of Echinoparyphium recurvatum (Digenea: Echinostomatidae) and their snail hosts.

    PubMed

    Morley, N J; Leung, K M Y; Morritt, D; Crane, M

    2004-07-01

    The toxicity of the anti-fouling biocides tributyltin (TBTO), copper, and Irgarol 1051 (irgarol) at a nominal concentration of 10 microg/l over a 30 day period were investigated against the viability of metacercarial cysts of the digenean parasite Echinoparyphium recurvatum resident within the body of two common freshwater snails, Lymnaea peregra and Physa fontinalis. Reduced parasite viability was found under most exposures in both snail species. However a greater effect of toxicant exposure was found in cysts within P. fontinalis compared to those in L. peregra. This was associated with an increased mortality of the host snail. Among all tested biocides, TBTO exposures induced the highest mortality to both the parasite and their hosts. These results suggest that parasite viability is interlinked with survival of the host snail. The mechanisms of differing toxicity between host species and its relevance to successful parasite transmission to the next host are discussed.

  7. Distribution of potential intermediate hosts for Fasciola hepatica and Fascioloides magna in Montana, USA.

    PubMed

    Dunkel, A M; Rognlie, M C; Johnson, G R; Knapp, S E

    1996-03-01

    A collection of lymnaeid snails in Montana was made over a 3 year period, in conjunction with a state-wide survey of the distribution of liver flukes in Montana. Collection areas were selected based on reports of infected cattle, sheep or wildlife, and with the intent of covering all geographic regions of the state. Snails were found at all 97 of the locations chosen for collections, with lymnaeids collected at 71 of the locations. The 97 sites were located in 28 of Montana's 56 counties. Nine lymnaeid species were collected, five of which have been reported either as natural or experimental intermediate hosts for Fasciola hepatica or Fascioloides magna. The two snail species most widely distributed over the areas enzootic for the flukes were Lymnaea modicella and Lymanaea caperata.

  8. Interactions among invaders: community and ecosystem effects of multiple invasive species in an experimental aquatic system.

    PubMed

    Johnson, Pieter T J; Olden, Julian D; Solomon, Christopher T; Vander Zanden, M Jake

    2009-02-01

    With ecosystems increasingly supporting multiple invasive species, interactions among invaders could magnify or ameliorate the undesired consequences for native communities and ecosystems. We evaluated the individual and combined effects of rusty crayfish (Orconectes rusticus) and Chinese mystery snails [Bellamya (=Cipangopaludina) chinensis] on native snail communities (Physa, Helisoma and Lymnaea sp.) and ecosystem attributes (algal chlorophyll a and nutrient concentrations). Both invaders are widespread in the USA and commonly co-occur within northern temperate lakes, underscoring the importance of understanding their singular and joint effects. An outdoor mesocosm experiment revealed that while the two invaders had only weakly negative effects upon one another, both negatively affected the abundance and biomass of native snails, and their combined presence drove one native species to extinction and reduced a second by >95%. Owing to its larger size and thicker shell, adult Bellamya were protected from crayfish attack relative to native species (especially Physa and Lymnaea), suggesting the co-occurrence of these invaders in nature could have elevated consequences for native communities. The per capita impacts of Orconectes (a snail predator) on native snails were substantially greater than those of Bellamya (a snail competitor). Crayfish predation also had a cascading effect by reducing native snail biomass, leading to increased periphyton growth. Bellamya, in contrast, reduced periphyton biomass, likely causing a reduction in growth by native lymnaeid snails. Bellamya also increased water column N:P ratio, possibly because of a low P excretion rate relative to native snail species. Together, these findings highlight the importance of understanding interactions among invasive species, which can have significant community- and ecosystem-level effects.

  9. Dose-dependent effects of the clinical anesthetic isoflurane on Octopus vulgaris: a contribution to cephalopod welfare.

    PubMed

    Polese, Gianluca; Winlow, William; Di Cosmo, Anna

    2014-12-01

    Recent progress in animal welfare legislation relating to invertebrates has provoked interest in methods for the anesthesia of cephalopods, for which different approaches to anesthesia have been tried but in most cases without truly anesthetizing the animals. For example, several workers have used muscle relaxants or hypothermia as forms of "anesthesia." Several inhalational anesthetics are known to act in a dose-dependent manner on the great pond snail Lymnaea stagnalis, a pulmonate mollusk. Here we report, for the first time, on the effects of clinical doses of the well-known inhalational clinical anesthetic isoflurane on the behavioral responses of the common octopus Octopus vulgaris. In each experiment, isoflurane was equilibrated into a well-aerated seawater bath containing a single adult O. vulgaris. Using a web camera, we recorded each animal's response to touch stimuli eliciting withdrawal of the arms and siphon and observed changes in the respiratory rate and the chromatophore pattern over time (before, during, and after application of the anesthetic). We found that different animals of the same size responded with similar behavioral changes as the isoflurane concentration was gradually increased. After gradual application of 2% isoflurane for a maximum of 5 min (at which time all the responses indicated deep anesthesia), the animals recovered within 45-60 min in fresh aerated seawater. Based on previous findings in gastropods, we believe that the process of anesthesia induced by isoflurane is similar to that previously observed in Lymnaea. In this study we showed that isoflurane is a good, reversible anesthetic for O. vulgaris, and we developed a method for its use.

  10. The inclusion of downy mildews in a multi-locus-dataset and its reanalysis reveals a high degree of paraphyly in Phytophthora.

    PubMed

    Runge, Fabian; Telle, Sabine; Ploch, Sebastian; Savory, Elizabeth; Day, Brad; Sharma, Rahul; Thines, Marco

    2011-12-01

    Pathogens belonging to the Oomycota, a group of heterokont, fungal-like organisms, are amongst the most notorious pathogens in agriculture. In particular, the obligate biotrophic downy mildews and the hemibiotrophic members of the genus Phytophthora are responsible for a huge variety of destructive diseases, including sudden oak death caused by P. ramorum, potato late blight caused by P. infestans, cucurbit downy mildew caused by Pseudoperonospora cubensis, and grape downy mildew caused by Plasmopara viticola. About 800 species of downy mildews and roughly 100 species of Phytophthora are currently accepted, and recent studies have revealed that these groups are closely related. However, the degree to which Phytophthora is paraphyletic and where exactly the downy mildews insert into this genus in relation to other clades could not be inferred with certainty to date. Here we present a molecular phylogeny encompassing all clades of Phytophthora as represented in a multi-locus dataset and two representatives of the monophyletic downy mildews from divergent genera. Our results demonstrate that Phytophthora is at least six times paraphyletic with respect to the downy mildews. The downy mildew representatives are consistently nested within clade 4 (contains Phytophthora palmivora), which is placed sister to clade 1 (contains Phytophthora infestans). This finding would either necessitate placing all downy mildews and Phytopthora species in a single genus, either under the oldest generic name Peronospora or by conservation the later name Phytophthora, or the description of at least six new genera within Phytophthora. The complications of both options are discussed, and it is concluded that the latter is preferable, as it warrants fewer name changes and is more practical.

  11. [An early warning method of cucumber downy mildew in solar greenhouse based on canopy temperature and humidity modeling].

    PubMed

    Wang, Hui; Li, Mei-lan; Xu, Jian-ping; Chen, Mei-xiang; Li, Wen-yong; Li, Ming

    2015-10-01

    The greenhouse environmental parameters can be used to establish greenhouse nirco-climate model, which can combine with disease model for early warning, with aim of ecological controlling diseases to reduce pesticide usage, and protecting greenhouse ecological environment to ensure the agricultural product quality safety. Greenhouse canopy leaf temperature and air relative humidity, models were established using energy balance and moisture balance principle inside the greenhouse. The leaf temperature model considered radiation heat transfer between the greenhouse crops, wall, soil and cover, plus the heat exchange caused by indoor net radiation and crop transpiration. Furthermore, the water dynamic balance in the greenhouse including leaf transpiration, soil evaporation, cover and leaf water vapor condensation, was considered to develop a relative humidity model. The primary infection and latent period warning models for cucumber downy mildew (Pseudoperonospora cubensis) were validated using the results of the leaf temperature and relative humidity model, and then the estimated disease occurrence date of cucumber downy mildew was compared with actual disease occurrence date of field observation. Finally, the results were verified by the measured temperature and humidity data of September and October, 2014. The results showed that the root mean square deviations (RMSDs) of the measured and estimated leaf temperature were 0.016 and 0.024 °C, and the RMSDs of the measured and estimated air relative humidity were 0.15% and 0.13%, respectively. Combining the result of estimated temperature and humidity models, a cucumber disease early warning system was established to forecast the date of disease occurrence, which met with the real date. Thus, this work could provide the micro-environment data for the early warning system of cucumber diseases in solar greenhouses.

  12. Thirty-seventh supplement to the American Ornithologists' Union checklist of North American birds

    USGS Publications Warehouse

    Monroe, Burt L.; Banks, Richard C.; Fitzpatrick, John W.; Howell, Thomas R.; Johnson, Ned K.; Ouellet, Henri; Remsen, J.V.; Storer, Robert W.

    1989-01-01

    This third supplement subsequent to the 6th edition (1983) of the A.O.U. "Check-list of North American Birds" consists of changes adopted by the Committee on Classification and Nomenclature as of 1 March 1989. The changes fall into nine categories: (1) six species are added to the main list (Pterodroma longirostris, Larus crassirostris, Streptopelia decaocto, Cocccyzus julieni, Chrysolampis mosquitus, Emberiza aureola) because of new distributional information; (2) five species (Ara cubensis, Chlorostilbon bracei, Empidonax occidentalis, Polioptila californica, Pipilo crissalis) are added to the main list because of the splitting of species already on the list; (3) one name (Anthus rubescens) is changed because of the splitting of a species from outside the Checklist area; (4) two names (Morus bassanus, Nyctanassa violacea) is removed from the main list to Appendix B because of re-evaluation of Northern Hemisphere records; (6) three species (Pterodrama rostrata, P. alba, P. solandri) are moved from Appendix A to Appendix B, and one (P. defilippiana) is added to Appendix B because of questionable sight records; (7)A.O.U. numbers are added to three species (Ciccaba virgata, Myiopagis viridicata, Molothrus bonariensis) on the basis on new distributional records or supporting data; (8) several corrections in spelling or citations are made; and (9) English names are changed for twelve species to accommodate worldwide usage of these names. No new distributional information is included except as indicated above (i.e. minor changes of distribution are not noted). These actions bring the number of species recognized as occurring in North America (main list) to 1,945.

  13. Conopressin affects excitability, firing, and action potential shape through stimulation of transient and persistent inward currents in mulluscan neurons.

    PubMed

    van Soest, P F; Kits, K S

    1998-04-01

    The molluscan vasopressin/oxytocin-related neuropeptide conopressin activates two persistent inward currents in neurons from the anterior lobe of the right cerebral ganglion of Lymnaea stagnalis that are involved in the control of male copulatory behavior. The low-voltage-activated (LVA) current is activated at a wide range of membrane potentials, its amplitude being only weakly voltage dependent. The high-voltage-activated (HVA) current is activated at potentials positive to -40 mV only and shows a steep voltage dependence. Occurrence of both currents varies from cell to cell, some expressing both and others only the HVA current. In most neurons that have the LVA current, a conopressin-independent persistent inward current (INSR) is found that resembles the HVA current in its voltage dependence. The functional importance of the LVA and HVA currents was studied under current-clamp conditions in isolated anterior lobe neurons. In cells exhibiting both current types, the effect of activation of the LVA current alone was investigated as follows: previously recorded LVA current profiles were injected into the neurons, and the effects were compared with responses induced by conopressin. Both treatments resulted in a strong depolarization and firing activity. No differences in firing frequency and burst duration were observed, indicating that activation of the LVA current is sufficient to evoke bursts. In cells exhibiting only the HVA current, the effect of conopressin on the response to a depolarizing stimulus was tested. Conopressin reversibly increased the number of action potentials generated by the stimulus, suggesting that the HVA current enhances excitability and counteracts accommodation. Conopressin enhanced action potential broadening during depolarizing stimuli in many neurons. Voltage-clamp experiments performed under ion-selective conditions revealed the presence of transient sodium and calcium currents. Using the action potential clamp technique, it was

  14. Failure of delayed nonsynaptic neuronal plasticity underlies age-associated long-term associative memory impairment

    PubMed Central

    2012-01-01

    Background Cognitive impairment associated with subtle changes in neuron and neuronal network function rather than widespread neuron death is a feature of the normal aging process in humans and animals. Despite its broad evolutionary conservation, the etiology of this aging process is not well understood. However, recent evidence suggests the existence of a link between oxidative stress in the form of progressive membrane lipid peroxidation, declining neuronal electrical excitability and functional decline of the normal aging brain. The current study applies a combination of behavioural and electrophysiological techniques and pharmacological interventions to explore this hypothesis in a gastropod model (Lymnaea stagnalis feeding system) that allows pinpointing the molecular and neurobiological foundations of age-associated long-term memory (LTM) failure at the level of individual identified neurons and synapses. Results Classical appetitive reward-conditioning induced robust LTM in mature animals in the first quartile of their lifespan but failed to do so in animals in the last quartile of their lifespan. LTM failure correlated with reduced electrical excitability of two identified serotonergic modulatory interneurons (CGCs) critical in chemosensory integration by the neural network controlling feeding behaviour. Moreover, while behavioural conditioning induced delayed-onset persistent depolarization of the CGCs known to underlie appetitive LTM formation in this model in the younger animals, it failed to do so in LTM-deficient senescent animals. Dietary supplementation of the lipophilic anti-oxidant α-tocopherol reversed the effect of age on CGCs electrophysiological characteristics but failed to restore appetitive LTM function. Treatment with the SSRI fluoxetine reversed both the neurophysiological and behavioural effects of age in senior animals. Conclusions The results identify the CGCs as cellular loci of age-associated appetitive learning and memory

  15. Laboratory maintenance of the bacterial endosymbiont, Neorickettsia sp., through the life cycle of a digenean, Plagiorchis elegans.

    PubMed

    Greiman, Stephen E; Tkach, Maksym; Vaughan, Jefferson A; Tkach, Vasyl V

    2015-10-01

    The Digenea (Platyhelminthes: Trematoda) are a diverse and complex group of internal metazoan parasites. These parasites can serve as hosts to obligate intracellular bacteria belonging to the genus Neorickettsia (Family: Anaplasmataceae). Neorickettsiae persist within all stages of the fluke life cycle and thus are maintained through vertical transmission. However, the low prevalence of Neorickettsia in nature limits study of their transmission biology at different steps of digenean life cycles. To resolve this dilemma, we have developed for the first time a laboratory model allowing to maintain Neorickettsia sp. through the whole life cycle of a digenean, Plagiorchis elegans. The laboratory life cycle of P. elegans consists of a snail first intermediate host, Lymnaea stagnalis, an aquatic arthropod second intermediate host, Culex pipiens (mosquito larva), and a vertebrate definitive host, Mesocricetus auratus (Syrian hamster). This paper focuses on the development of the laboratory life cycle, as well as outlines its potential uses in studying the transmission biology of Neorickettsia and its evolutionary relationship within its digenean host.

  16. The numbers game: quantitative analysis of Neorickettsia sp. propagation through complex life cycle of its digenean host using real-time qPCR.

    PubMed

    Greiman, Stephen E; Tkach, Vasyl V

    2016-07-01

    Bacteria of the genus Neorickettsia are obligate intracellular endosymbionts of parasitic flukes (Digenea) and are passed through the entire complex life cycle of the parasite by vertical transmission. Several species of Neorickettsia are known to cause diseases in domestic animals, wildlife, and humans. Quantitative data on the transmission of the bacteria through the digenean life cycle is almost completely lacking. This study quantified for the first time the abundance of Neorickettsia within multiple stages of the life cycle of the digenean Plagiorchis elegans. Snails Lymnaea stagnalis collected from a pond in North Dakota were screened for the presence of digenean cercariae, which were subsequently tested for the presence of Neorickettsia. Three L. stagnalis were found shedding P. elegans cercariae infected with Neorickettsia. These snails were used to initiate three separate laboratory life cycles and obtain all life cycle stages for bacterial quantification. A quantitative real-time PCR assay targeting the GroEL gene was developed to enumerate Neorickettsia sp. within different stages of the digenean life cycle. The number of bacteria significantly increased throughout all stages, from eggs to adults. The two largest increases in number of bacteria occurred during the period from eggs to cercariae and from 6-day metacercariae to 48-h juvenile worms. These two periods seem to be the most important for Neorickettsia propagation through the complex digenean life cycle and maturation in the definitive host.

  17. The definitive and intermediate hosts of Fasciola hepatica in the natural watercress beds in central France.

    PubMed

    Rondelaud, D; Vignoles, P; Abrous, M; Dreyfuss, G

    2001-06-01

    Field investigations were carried out over a two-year period in 52 natural watercress beds located in the Limousin region of central France to list the mammal and bird species that frequented these sites. This enabled detection of the definitive hosts of Fasciola hepatica and determination of the prevalence of natural infection in snails. A total of 13 mammal and five bird species were listed in these watercress beds. Adult flukes were found in Lepus capensis (39.2%), Oryctolagus cuniculus (42.0%), and Sylvilagus floridanus (25.0%). No infection with F. hepatica was noted in the five species of rodents studied. Snails infected with F. hepatica were found in 14 watercress beds. The global prevalence of natural infection was 1.1% in Lymnaea truncatula and 0.3% in L. glabra. Among the other trematode larval forms detected, the most frequent was Haplometra cylindracea (0.5%). In the Limousin region, the presence of hares and rabbits in watercress beds ensured the continuation of the F. hepatica life cycle and permitted the subsequent infection of humans when this wild watercress was eaten.

  18. Fasciola hepatica infections in livestock flock, guanacos and coypus in two wildlife reserves in Argentina.

    PubMed

    Issia, Laura; Pietrokovsky, Silvia; Sousa-Figueiredo, José; Stothard, J Russell; Wisnivesky-Colli, Cristina

    2009-11-12

    Between autumn and spring 2006, a coprological survey was performed in two wildlife reserves located in the north of Argentine Patagonia to determine the prevalence of Fasciola hepatica and the number of parasite eggs per gram (epg) of feces in wild guanacos (Lama guanicoe), coypus (Myocastor coypus), and locally born and raised goats and sheep. Snails of the Family Lymnaeidae were collected in freshwater habitats, identified taxonomically and analyzed parasitologically. Prevalence of patent infection was 100% in sheep (n=69) and coypus (n=9), 84% in goats (n=20) and 0.5% in guanacos (n=224). No significant differences in epg were found among animals, but the median epg of coypus (160) and sheep (160) was higher than that of goats (80). For guanacos and goats, a negative binomial model estimating the population egg-count frequency could be fitted, while for coypus and sheep parasite egg-count frequencies trended toward a normal distribution, indicative of a more even, and much less aggregated distribution across sampled hosts. All snails (n=175) were Lymnaea truncatula and none of them was found infected. This is the first report of fascioliasis in free-ranging guanacos in Argentina. Coypu appears to be a major wildlife reservoir of F. hepatica, which was presumably introduced locally by livestock.

  19. [Fascioliasis: diagnosis, epidemiology and treatment].

    PubMed

    Carrada-Bravo, Teodoro

    2003-01-01

    Fascioliasis is a trematode, disease of liver and bile ducts of sheep, cattle, and other ruminants throughout the world that is caused by the fluke, Fasciola hepatica. Human infection has been reported in Mexico, Cuba, Puerto Rico, Chile, Peru, Uruguay, Brazil, Argentina, the US, Europe, eastern Africa, Japan and Australia. The parasite's miracidium invades one of the various Lymnaea water snail hosts. Infection results from ingestion of encysted metacercariae attached to raw watercress (Nasturtium officinale). Symptoms recorded from human cases included irregular fever, epigastric pain and abdominal tenderness, obstructive jaundice and leucocytosis with eosinophilea up to 60%. Specific diagnosis is based on recovery of the eggs in the patient's stool or from biliary tract drainage. Treatment is with emetine hydrochloride given intramusculary. Bithionol is given orally at a dosage of 30-50 mg/kg but on alternate days from 10 to 15 doses. Praziquantel is probably effective. Preventive measures include education of the public on mode of transmission of life cycle of the parasite, and dipping fresh watercress into boiling water for a few sec, or drying suspected watercress.

  20. Western blotting of formaldehyde-fixed neuropeptides as small as 400 daltons on gelatin-coated nitrocellulose paper.

    PubMed

    Too, C K; Murphy, P R; Croll, R P

    1994-06-01

    A method is described for Western blotting of peptides as small as 400 daltons (Da). Peptides were separated by tricine-sodium dodecyl sulfate electrophoresis and electroblotted to gelatin-coated PH79 nitrocellulose paper (0.1 micron). The electroblotted peptides were fixed to the nitrocellulose paper for 5-10 min in 4% paraformaldehyde solution. Using anti-rabbit FMRF-amide (Phe-Met-Arg-Phe-NH2) as primary antibody, positive immunoreactivity was detected with an amplified alkaline phosphatase assay which was sensitive to at least 0.5 microgram FMRFamide/lane. When immunoreactivity was determined with 125I-protein A, it was possible to amplify and detect weak signals by increasing the autoradiography time. Therefore, using the 125I-protein A detection method, Western blot analysis of brain extracts from Lymnaea stagnalis (pond snail) and Poecilia reticulata (guppy) indicated the presence of four FMRFamide immunoreactive bands after a 7-day exposure to X-ray film. The most abundant peptide coelectrophoresed with the FMRFamide standard (M(r) 598.8 Da). In addition, this Western blotting procedure also detected APGWamide (Ala-Pro-Gly-Try-NH2; 428.5 Da) and [D-Ala2]-Leu-enkephalinamide (568.7 Da) with their respective specific antibodies.

  1. Morphology and life cycle of Apatemon hypseleotris species novum from Australia including metacercariae viability and excystment.

    PubMed

    Negm-Eldin, M; Davies, R W

    2002-07-01

    Experimental infection of pigeon squabs and rats with encysted metacercariae from the Western Carp gudgeon (Hypseleotris klunzingeri) showed them to be infected with a new strigeid trematode, Apatemon hypseleotris. Growth and development of A. hypseleotris in pigeons were significantly higher than in rats. Eggs appeared in pigeon faeces within 7-14 days; miracidia hatched within 15-21 days and in the snail Lymnaea tomentosa released within 21 days. Cercariae experimentally encysted in the leeches Helobdella papillornata (86.7%) and Alboglossiphonia australiensis (73.3%). In fish, encystation occurred in the abdominal cavity (100%) and muscles (40%) of Hypseleotris klunzingeri, in the abdominal cavity (80%) and head (30%) of Gambusia affinis and in the abdominal cavity (62.5%) of Oncorhynchus mykiss but no encystation occurred in Moenkhausia pittieri. Freezing (-7 degrees C for 3-7 days or -21 degrees C for 8-12 hours), chilling (6 degrees C for 12 days), boiling (3 minutes) or salting for 3-5 days of encysted metacercariae did not significantly reduce infectivity. In vitro excystation of metacercariae was achieved using pepsin followed by trypsin and/or bile salts.

  2. A biodynamic understanding of dietborne metal uptake by a freshwater invertebrate

    USGS Publications Warehouse

    Croteau, M.-N.; Luoma, S.N.

    2008-01-01

    Aquatic organisms accumulate metals from dissolved and particulate phases. Dietborne metal uptake likely prevails in nature, but the physiological processes governing metal bioaccumulation from diet are not fully understood. We characterize dietborne copper, cadmium, and nickel uptake by a freshwater gastropod (Lymnaea stagnalis) both in terms of biodynamics and membrane transport characteristics. We use enriched stable isotopes to trace newly accumulated metals from diet, determine food ingestion rate (IR) and estimate metal assimilation efficiency (AE). Upon 18-h exposure, dietborne metal influx was linear over a range encompassing most environmental concentrations. Dietary metal uptake rate constants (kuf) ranged from 0.104 to 0.162 g g -1 day-1, and appeared to be an expression of transmembrane transport characteristics. Although kuf values were 1000-times lower than uptake rate constants from solution, biodynamic modeling showed that diet is the major Cd, Cu, and Ni source in nature. AE varied slightly among metals and exposure concentrations (84-95%). Suppression of Cd and Cu influxes upon exposure to extreme concentrations coincided with a 10-fold decrease in food IR, suggesting that feeding inhibition could act as an end point for dietary metal toxicity in L. stagnalis.

  3. Predicting dietborne metal toxicity from metal influxes

    USGS Publications Warehouse

    Croteau, M.-N.; Luoma, S.N.

    2009-01-01

    Dietborne metal uptake prevails for many species in nature. However, the links between dietary metal exposure and toxicity are not well understood. Sources of uncertainty include the lack of suitable tracers to quantify exposure for metals such as copper, the difficulty to assess dietary processes such as food ingestion rate, and the complexity to link metal bioaccumulation and effects. We characterized dietborne copper, nickel, and cadmium influxes in a freshwater gastropod exposed to diatoms labeled with enriched stable metal isotopes. Metal influxes in Lymnaea stagnalis correlated linearly with dietborne metal concentrations over a range encompassing most environmental exposures. Dietary Cd and Ni uptake rate constants (kuf) were, respectively, 3.3 and 2.3 times higher than that for Cu. Detoxification rate constants (k detox) were similar among metals and appeared 100 times higher than efflux rate constants (ke). Extremely high Cu concentrations reduced feeding rates, causing the relationship between exposure and influx to deviate from linearity; i.e., Cu uptake rates leveled off between 1500 and 1800 nmol g-1 day-1. L. stagnalis rapidly takes up Cu, Cd, and Ni from food but detoxifies the accumulated metals, instead of reducing uptake or intensifying excretion. Above a threshold uptake rate, however, the detoxification capabilities of L. stagnalis are overwhelmed.

  4. Ecological studies on Al-Khadoud Spring, Al-Hassa, Saudi Arabia.

    PubMed

    Al-Kahtani, Mohammed A; Youssef, Ashraf M; Fathi, Adel A

    2007-11-15

    Al-Khadoud spring is one of the most important water resources in Al-Hassa Governorate, Saudi Arabia. However, much of its biotic information is still unknown. This study presented preliminary ecological information of this aquatic body. Regarding to macrophytes, a total of eight species were observed along the study sites. These species include two submerged aquatic plants (Potamogeton pectinatus L. and Ceratophyllum demersum L.). The common distributed species are Phragmites australis (Cav.) Trimex Steud and Cyperus rotundus (L.). On the other hand, a total of 20 algal genera were recorded with 7 genera of Chlorophyceae, 8 of Bacillariophyceae, 4 of Cyanophyceae and one of Euglenophyceae. The common phytoplankton occurred in all three investigated sites were Chlorella vulgaris, Mougeotia sp., Oscillatoria sp. and Actinastrum sp. Regarding to the biotic fauna, different forms of unicellular zooplankton such as Paramecium and Amoeba were recorded. Invertebrates such as freshwater insects and some freshwater snails were documented in the study sites including Melanodies tuberculata, Melanopsis praemorsa and Lymnaea auricularia. As regard to vertebrates, one species of fish, Aphanius dispar, dominate the spring basin and its extended channels.

  5. Species diversity of Plagiorchis Lühe, 1899 (Digenea: Plagiorchiidae) in lymnaeid snails from freshwater ecosystems in central Europe revealed by molecules and morphology.

    PubMed

    Zikmundová, Jana; Georgieva, Simona; Faltýnková, Anna; Soldánová, Miroslava; Kostadinova, Aneta

    2014-05-01

    Larval stages of Plagiorchis spp. are both ubiquitous and ecologically important parasites in snail populations of freshwater ecosystems in Europe. However, difficulties in distinguishing the morphologically similar cercariae used for species identification, may lead to underestimation of species diversity. In this study, 38 isolates of Plagiorchis spp. infecting two lymnaeid snails, Lymnaea stagnalis (L.) and Radix auricularia (L.), in five central European freshwater ecosystems were subjected to morphological and molecular assessment. Five morphologically homogeneous and genetically distinct lineages of Plagiorchis spp. were identified via matching molecular data for the mitochondrial cytochrome c oxidase subunit I (cox1) gene with detailed morphological and morphometric data of the cercariae. Comparative sequence analysis using partial 28S rDNA and ITS1-5.8S-ITS2 sequences revealed that three distinct cox1 lineages are conspecific with Plagiorchis elegans (Rudolphi, 1802), P. maculosus (Rudolphi, 1802) and P. koreanus Ogata, 1938, respectively, whereas the lineage identified based on cercarial morphology as P. neomidis Brendow, 1970 plus a single isolate that could not be assigned to a described species, did not match any of the available sequences for Plagiorchis spp. A key to the cercariae of Plagiorchis spp. parasitising lymnaeid populations in central Europe is provided to facilitate identification.

  6. Diaphanous gene mutation affects spiral cleavage and chirality in snails

    PubMed Central

    Kuroda, Reiko; Fujikura, Kohei; Abe, Masanori; Hosoiri, Yuji; Asakawa, Shuichi; Shimizu, Miho; Umeda, Shin; Ichikawa, Futaba; Takahashi, Hiromi

    2016-01-01

    L-R (left and right) symmetry breaking during embryogenesis and the establishment of asymmetric body plan are key issues in developmental biology, but the onset including the handedness-determining gene locus still remains unknown. Using pure dextral (DD) and sinistral (dd) strains of the pond snail Lymnaea stagnalis as well as its F2 through to F10 backcrossed lines, the single handedness-determining-gene locus was mapped by genetic linkage analysis, BAC cloning and chromosome walking. We have identified the actin-related diaphanous gene Lsdia1 as the strongest candidate. Although the cDNA and derived amino acid sequences of the tandemly duplicated Lsdia1 and Lsdia2 genes are very similar, we could discriminate the two genes/proteins in our molecular biology experiments. The Lsdia1 gene of the sinistral strain carries a frameshift mutation that abrogates full-length LsDia1 protein expression. In the dextral strain, it is already translated prior to oviposition. Expression of Lsdia1 (only in the dextral strain) and Lsdia2 (in both chirality) decreases after the 1-cell stage, with no asymmetric localization throughout. The evolutionary relationships among body handedness, SD/SI (spiral deformation/spindle inclination) at the third cleavage, and expression of diaphanous proteins are discussed in comparison with three other pond snails (L. peregra, Physa acuta and Indoplanorbis exustus). PMID:27708420

  7. The identification and characteristics of Echinoparyphium rubrum (Cort. 1914) new comb. (Trematoda, Echinostomatidae) based on experimental evidence of the life cycle

    USGS Publications Warehouse

    Kanev, I.; Sorensen, R.; Sterner, M.; Cole, R.; Fried, B.

    1998-01-01

    The life cycle of Echinoparyphium rubrum (Cort, 1914) comb. n. has been completed experimentally. All of the developmental stages egg, miracidium, sporocyst, mother and daughter rediae, cercaria, metacercaria, and adult were examined and described. The miracidia infected freshwater snails of the genus Physa , P. gyrina and P. occidentalis. Attempts to infect snails of the genera Lymnaea, L. auricularis, L. peregra, L. truncatula and Bulinus, B. truncatus failed. Cercariae infected various pulmonate and prosobranch freshwater snails, mussels, frogs, water turtles and planarians. The adults developed in the small intestine of birds and mammals. The identity and major characteristics of Echinoparyphium rubrum are discussed. Synonyms of E. rubrum are Cercaria rubra Cort, 1914; Cercaria biflexa Faust, 1917; Cercaria chisolenata Faust, 1918; Echinostoma callawayensis Barker et Noll, 1915; Echinostoma revolutum of Johnson (1920); Echinoparyphium elegans of Cannon (1938), of Bain and Trelfall (1977), of Mahoney and Trelfall (1977); and Echinoparyphium recurvatum of Jilek (1977), Harley (1972), Sankurathri and Holmes (1976). Comparisons are made between E. rubrum and its 43-collar-spined allies: E. flexum from North America, E. cinctum from Europe, E. dunni from Asia and E. elegans from Africa.

  8. Occurrence of Fasciola hepatica (Linnaeus, 1758) in capybara (Hydrochoerus hydrochaeris) (Linnaeus, 1766) in Minas Gerais, Brazil.

    PubMed

    Dracz, Ruth Massote; Ribeiro, Vinicius Marques Antunes; Pereira, Cintia Aparecida de Jesus; Lima, Walter Dos Santos

    2016-01-01

    Fasciola hepatica is a parasite that affects the hepatic ducts of several species of domestic and wild vertebrates, causing huge economic losses to livestock rearing worldwide. Reports on occurrences of F. hepatica in capybaras are an important epidemiological aspect of this disease, since these rodents can be a source of contamination for other animals and humans. In the present study, conducted in a rural area of the municipality of Confins, Minas Gerais, fresh feces from capybaras were collected from the ground near a lagoon at the edge of the Ribeirão da Mata river. These were examined using the technique of four metal sieves. F. hepatica eggs were recovered. This trematode species was confirmed by observing morphological characteristics and measuring the eggs recovered from the capybara feces, and through experimental infection of Lymnaea columella (Say, 1817) by miracidia from these eggs and subsequent infection of C57/BL06 mice with metacercariae originating from these infected mollusks. The data suggest the occurrence of natural cycle of F. hepatica in this region and provide a warning that expansion of the geographical distribution of this parasite by means of this rodent is possible. It is therefore important to adopting measures for epidemiological control of this helminthiasis.

  9. Mitochondrial Insult in a Parkinson's like symptoms model

    NASA Astrophysics Data System (ADS)

    Carrizales, Julio; Cantu, Manuel; Plas, Daniel; Daniel Plas Lab Team, Dr.

    2014-03-01

    Healthy cells require healthy mitochondria. If these organelles are damaged, many health consequences follow. For example, Parkinson's Disease (PD) is a major neurodegenerative disorder of unknown cause, but much evidence points to the mitochondrion as a key player in the onset of this disease. PD has been studied in animal models challenged with toxins that target the mitochondria. In our work, we have used the pesticide, Rotenone, a known inhibitor of protein Complex I in the mitochondrial electron transport chain. When this toxin is applied to the freshwater mollusk, Lymnaea stagnalis, or pond snail, severe motor deficits ensue. In this project, we are studying the direct effects of this toxin on mitochondrial structure and physiology. We expected that the morphology of the organelle may be altered. In addition, it is likely that the mitochondrial membrane potential necessary for normal function may decrease as the electron transport loses the ability to move protons from the matrix to the intermembrane space. we also are going to use Electrophysiology to compare and Identify the difference of the electrical signaling among healthy and unhealthy neurons. HHMI

  10. The Effect of Diet Mixing on a Nonselective Herbivore

    PubMed Central

    2016-01-01

    The balanced-diet hypothesis states that a diverse prey community is beneficial to consumers due to resource complementarity among the prey species. Nonselective consumer species cannot differentiate between prey items and are therefore not able to actively regulate their diet intake. We thus wanted to test whether the balanced-diet hypothesis is applicable to nonselective consumers. We conducted a laboratory experiment in which a nonselective model grazer, the freshwater gastropod Lymnaea stagnalis, was fed benthic green algae as single species or as a multi-species mixture and quantified the snails’ somatic growth rates and shell lengths over a seven-week period. Gastropods fed the mixed diet were found to exhibit a higher somatic growth rate than the average of the snails fed single prey species. However, growth on the multi-species mixture did not exceed the growth rate obtained on the best single prey species. Similar results were obtained regarding the animals’ shell height increase over time. The mixed diet did not provide the highest growth rate, which confirms our hypothesis. We thus suggest that the balanced-diet hypothesis is less relevant for non-selective generalist consumers, which needs to be considered in estimates of secondary production. PMID:27391787

  11. Brefeldin A or monensin inhibits the 3D organizer in gastropod, polyplacophoran, and scaphopod molluscs.

    PubMed

    Gonzales, Eric E; van der Zee, Maurijn; Dictus, Wim J A G; van den Biggelaar, Jo

    2007-02-01

    In molluscs, the 3D vegetal blastomere acts as a developmental signaling center, or organizer, and is required to establish bilateral symmetry in the embryo. 3D is similar to organizing centers in other metazoans, but detailed comparisons are difficult, in part because its organizing function is poorly understood. To elucidate 3D function in a standardized fashion, we used monensin and brefeldin A (BFA) to rapidly and reversibly interfere with protein processing and secretion, thereby inhibiting the signaling interactions that underlie its specification and patterning. In the gastropods, Patella vulgata and Lymnaea stagnalis, the polyplacophoran, Mopalia muscosa, and the scaphopod, Antalis entalis, treatments initiated before the organizer-dependent onset of bilateral cleavage resulted in radialization of subsequent development. In radialized P. vulgata, L. stagnalis, and M. muscosa, organizer specification was blocked, and embryos failed to make the transition to bilateral cleavage. In all four species, the subsequent body plan was radially symmetric and was similarly organized about a novel aboral-oral axis. Our results demonstrate that brefeldin A (BFA) and monensin can be used to inhibit 3D's organizing function in a comparative fashion and that, at least in M. muscosa, the organizer-dependent developmental architecture of the embryo predicts subsequent patterns of morphogenetic movements in gastrulation and, ultimately, the layout of the adult body plan.

  12. Ligand-binding domain of an α7-nicotinic receptor chimera and its complex with agonist.

    PubMed

    Li, Shu-Xing; Huang, Sun; Bren, Nina; Noridomi, Kaori; Dellisanti, Cosma D; Sine, Steven M; Chen, Lin

    2011-09-11

    The α(7) acetylcholine receptor (AChR) mediates pre- and postsynaptic neurotransmission in the central nervous system and is a potential therapeutic target in neurodegenerative, neuropsychiatric and inflammatory disorders. We determined the crystal structure of the extracellular domain of a receptor chimera constructed from the human α(7) AChR and Lymnaea stagnalis acetylcholine binding protein (AChBP), which shares 64% sequence identity and 71% similarity with native α(7). We also determined the structure with bound epibatidine, a potent AChR agonist. Comparison of the structures revealed molecular rearrangements and interactions that mediate agonist recognition and early steps in signal transduction in α(7) AChRs. The structures further revealed a ring of negative charge within the central vestibule, poised to contribute to cation selectivity. Structure-guided mutational studies disclosed distinctive contributions to agonist recognition and signal transduction in α(7) AChRs. The structures provide a realistic template for structure-aided drug design and for defining structure-function relationships of α(7) AChRs.

  13. Bioaccumulation dynamics and modeling in an estuarine invertebrate following aqueous exposure to nanosized and dissolved silver.

    PubMed

    Khan, Farhan R; Misra, Superb K; García-Alonso, Javier; Smith, Brian D; Strekopytov, Stanislav; Rainbow, Philip S; Luoma, Samuel N; Valsami-Jones, Eugenia

    2012-07-17

    Predicting the environmental impact of engineered nanomaterials (ENMs) is increasingly important owing to the prevalence of emerging nanotechnologies. We derived waterborne uptake and efflux rate constants for the estuarine snail, Peringia ulvae, exposed to dissolved Ag (AgNO(3)) and silver nanoparticles (Ag NPs), using biodynamic modeling. Uptake rates demonstrated that dissolved Ag is twice as bioavailable as Ag in nanoparticle form. Biphasic loss dynamics revealed the faster elimination of Ag from Ag NPs at the start of depuration, but similar slow efflux rate constants. The integration of biodynamic parameters into our model accurately predicted Ag tissue burdens during chronic exposure with 85% of predicted values within a factor of 2 of observed values. Zeta potentials for the Ag NPs were lower in estuarine waters than in waters of less salinity; and uptake rates in P. ulvae were slower than reported for the freshwater snail Lymnaea stagnalis in similar experiments. This suggests aggregation of Ag NPs occurs in estuarine waters and reduces, but does not eliminate, bioavailability of Ag from the Ag NPs. Biodynamic modeling provides an effective methodology to determine bioavailable metal concentrations (originating from metal and metal-oxide nanoparticles) in the environment and may aid future ENM risk assessment.

  14. Simultaneous analysis of multiple neurotransmitters by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry.

    PubMed

    Tufi, Sara; Lamoree, Marja; de Boer, Jacob; Leonards, Pim

    2015-05-22

    Neurotransmitters are endogenous metabolites that allow the signal transmission across neuronal synapses. Their biological role is crucial for many physiological functions and their levels can be changed by several diseases. Because of their high polarity, hydrophilic interaction liquid chromatography (HILIC) is a promising tool for neurotransmitter analysis. Due to the large number of HILIC stationary phases available, an evaluation of the column performances and retention behaviors has been performed on five different commercial HILIC packing materials (silica, amino, amide and two zwitterionic stationary phases). Several parameters like the linear correlation between retention and the distribution coefficient (logD), the separation factor k and the column resolution Rs have been investigated and the column performances have been visualized with a heat map and hierarchical clustering analysis. An optimized and validated HILIC-MS/MS method based on the ZIC-cHILIC column is proposed for the simultaneous detection and quantification of twenty compounds consisting of neurotransmitters, precursors and metabolites: 3-methoxytyramine (3-MT), 5-hydroxyindoleacetic acid (5-HIAA), 5-hydroxy-L-tripthophan, acetylcholine, choline, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, epinephrine, γ-aminobutyric acid (GABA), glutamate, glutamine, histamine, histidine, L-tryptophan, L-tyrosine, norepinephrine, normetanephrine, phenylalanine, serotonin and tyramine. The method was applied to neuronal metabolite profiling of the central nervous system of the freshwater snail Lymnaea stagnalis. This method is suitable to explore neuronal metabolism and its alteration in different biological matrices.

  15. Coolidge effect in pond snails: male motivation in a simultaneous hermaphrodite

    PubMed Central

    Koene, Joris M; Ter Maat, Andries

    2007-01-01

    Background The simultaneously hermaphroditic pond snail, Lymnaea stagnalis, can mate in the male and female role, but within one copulation only one sexual role is performed at a time. Previous work has shown that male motivation is determined by the availability of seminal fluid in the prostate gland, which is detected via a nervous connection by the brain area controlling male behaviour. Based on this knowledge, patterns of sexual role alternations within mating pairs can be explained. Results The data presented here reveal that these snails can donate and receive sperm several times within 24 hours, and that they have increased mating rates in larger groups (i.e. more mating opportunities). For mating pairs we show, by introducing novel mating partners after copulation, that animals do inseminate new partners, while they are no longer motivated to inseminate their original partners. Conclusion Our findings provide the first direct evidence for higher motivation in a hermaphrodite to copulate when a new partner is encountered. This Coolidge effect seems to be attenuated when mucus trails are excluded, which suggests that a chemical or textural cue may be responsible for mediating this response to sperm competition. PMID:17986351

  16. Structural and Population Polymorphism of RT-Like Sequences in Avian Schistosomes Trichobilharzia szidati (Platyhelminthes: Digenea: Schistosomatidae)

    PubMed Central

    Semyenova, S. K.; Chrisanfova, G. G.; Guliaev, A. S.; Yesakova, A. P.; Ryskov, A. P.

    2015-01-01

    Recently we developed the genus-specific markers of the avian schistosomes of the genus Trichobilharzia, the causative agents of human cercarial dermatitis. The 7 novel genome sequences of T. franki, T. regenti, and T. szidati revealed similarity with genome repeat region of African schistosome Schistosoma mansoni. In the present work we analyzed the 37 new T. szidati sequences to study intragenome variability and host specificity for the parasite from three localities of East Europe. DNAs were isolated from cercariae or single sporocysts obtained from 6 lymnaeid snails Lymnaea stagnalis and L. palustris from Belarus and Russia. All sequences formed three diverged groups, one of which consists of the sequences with multiple deletions; other groups involved two paralogous copies with stop codons and frameshift mutations. Strong association between geographical distribution and snail host specificity cannot be established. All studied sequences have homology with the reverse transcriptase domain (RT) of Penelope-like elements (PLE) of S. mansoni and S. japonicum and new members of RT family were identified. We proposed that three diverged groups RT sequences of T. szidati are results of duplication or transposition of PLE during parasite evolution. Implications of the retroelement dynamics in the life history of avian schistosomes are discussed. PMID:26114104

  17. Photomediated Larvicidal Activity of Pheophorbide a against Cercaria Larvae of Fasciola gigantica.

    PubMed

    Singh, Divya Jyoti; Singh, Vinay Kumar; Singh, D K

    2017-01-01

    Fasciolosis is a parasitic disease caused by Fasciola gigantica. The freshwater snail Lymnaea acuminata is the intermediate host of F. gigantica which cause endemic fasciolosis in the northern part of India. To investigate larvicidal activity of pure and laboratory extracted pheophorbide a (Pa) against cercaria larvae of F. gigantica, data were analyzed in different spectra of visible light, sunlight, and laboratory conditions. Photostimulation of chlorophyll derivative pheophorbide a (Pa) caused time and concentration dependent larvicidal activity against cercaria larvae of F. gigantica. Larvicidal activity of pure Pa under 650 nm and 400-650 nm (8 h LC50 0.006 mg/10 mL) was more pronounced than extracted Pa under same irradiations (650 nm LC50 0.12 mg/10 mL, 400-650 nm LC50 0.14 mg/10 mL). Lowest toxicity of pure (8 h LC50 0.14 mg/10 mL) and extracted Pa (8 h LC50 1.25 mg/10 mL) was noted under 400 nm. Pa was found to be toxic in laboratory conditions also. The results presented in this paper indicate that pheophorbide a possess potential larvicidal activity against Fasciola gigantica larvae in different wavelengths of visible light, sunlight, and laboratory conditions.

  18. Fascioliasis Control: In Vivo and In Vitro Phytotherapy of Vector Snail to Kill Fasciola Larva

    PubMed Central

    Sunita, Kumari; Singh, D. K.

    2011-01-01

    Snail is one of the important components of an aquatic ecosystem, it acts as intermediate host of Fasciola species. Control of snail population below a certain threshold level is one of the important methods in the campaign to reduce the incidence of fascioliasis. Life cycle of the parasite can be interrupted by killing the snail or Fasciola larva redia and cercaria in the snail body. In vivo and in vitro toxicity of the plant products and their active component such as citral, ferulic acid, umbelliferone, azadirachtin, and allicin against larva of Fasciola in infected snail Lymnaea acuminata were tested. Mortality of larvae were observed at 2 h, 4 h, 6 h, and 8 h, of treatment. In in vivo treatment, azadirachtin caused highest mortality in redia and cercaria larva (8 h, LC50 0.11, and 0.05 mg/L) whereas in in vitro condition allicin was highly toxic against redia and cercaria (8 h, LC50 0.01, and 0.009 mg/L). Toxicity of citral was lowest against redia and cercaria larva. PMID:22132306

  19. The life-cycle of Echinostoma friedi n. sp. (Trematoda: Echinostomatidae) in Spain and a discussion on the relationships within the 'revolutum' group based on cercarial chaetotaxy.

    PubMed

    Toledo, R; Muñoz-Antolí, C; Esteban, J G

    2000-03-01

    The morphology of the different stages and life-cycle of Echinostomna friedi n. sp. are described and figured. The freshwater snail Lymnaea peregra (Gastropoda: Lymnaeidae) serves as the natural and experimental first intermediate host and L. corvus and Gyraulus chinensis (Gastropoda: Planorbidae) as experimental first intermediate hosts. These, and Physella acuta (Gastropoda: Physidae), also serve as second intermediate hosts. Adult worms, possessing 37 collar spines, were obtained from naturally infected Rattus norvegicus and experimentally from albino rats, golden hamsters and chickens. Mice were not suitable experimental definitive hosts. E. friedi differs from the most closely related species in the 'revolutum' group mainly in terms of several morphological and biological features of the life-cycle stages and in its cercarial chaetotaxy. The chaetotaxy patterns of the species of the 'revolutum' group are analyzed and the results show that a taxonomic comparison of these species may be carried out on the basis of the number of sensilla in the clusters CIII VI, CIII V2 (or CIII V1 + CIII V2), CIV DL and UVb. These clusters appear adequate to establish taxonomic relationships between different species within the 'revolutum' group.

  20. In vitro studies of neuronal networks and synaptic plasticity in invertebrates and in mammals using multielectrode arrays.

    PubMed

    Massobrio, Paolo; Tessadori, Jacopo; Chiappalone, Michela; Ghirardi, Mirella

    2015-01-01

    Brain functions are strictly dependent on neural connections formed during development and modified during life. The cellular and molecular mechanisms underlying synaptogenesis and plastic changes involved in learning and memory have been analyzed in detail in simple animals such as invertebrates and in circuits of mammalian brains mainly by intracellular recordings of neuronal activity. In the last decades, the evolution of techniques such as microelectrode arrays (MEAs) that allow simultaneous, long-lasting, noninvasive, extracellular recordings from a large number of neurons has proven very useful to study long-term processes in neuronal networks in vivo and in vitro. In this work, we start off by briefly reviewing the microelectrode array technology and the optimization of the coupling between neurons and microtransducers to detect subthreshold synaptic signals. Then, we report MEA studies of circuit formation and activity in invertebrate models such as Lymnaea, Aplysia, and Helix. In the following sections, we analyze plasticity and connectivity in cultures of mammalian dissociated neurons, focusing on spontaneous activity and electrical stimulation. We conclude by discussing plasticity in closed-loop experiments.

  1. Invertebrate grazers affect metal/metalloid fixation during litter decomposition.

    PubMed

    Schaller, Jörg; Brackhage, Carsten

    2015-01-01

    Plant litter and organic sediments are main sinks for metals and metalloids in aquatic ecosystems. The effect of invertebrates as key species in aquatic litter decomposition on metal/metalloid fixation by organic matter is described only for shredders, but for grazers as another important animal group less is known. Consequently, a laboratory batch experiment was conducted to examine the effect of invertebrate grazers (Lymnaea stagnalis L.) on metal/metalloid fixation/remobilization during aquatic litter decomposition. It could be shown that invertebrate grazers facilitate significantly the formation of smaller sizes of particulate organic matter (POM), as shown previously for invertebrate shredders. The metal/metalloid binding capacity of these smaller particles of POM is higher compared to leaf litter residuals. But element enrichment is not as high as shown previously for the effect by invertebrate shredders. Invertebrate grazers enhance also the mobilization of selected elements to the water, in the range also proven for invertebrate shredders but different for the different elements. Nonetheless invertebrate grazers activity during aquatic litter decomposition leads to a metal/metalloid fixation into leaf litter as part of sediment organic matter. Hence, the effect of invertebrate grazers on metal/metalloid fixation/remobilization contrasts partly with former assessments revealing the possibility of an enhanced metal/metalloid fixation.

  2. In vitro PHYTOTHERAPY OF VECTOR SNAILS BY BINARY COMBINATIONS OF LARVICIDAL ACTIVE COMPONENTS IN EFFECTIVE CONTROL OF FASCIOLIASIS

    PubMed Central

    Sunita, Kumari; Kumar, Pradeep; Singh, Vinay Kumar; Singh, Dinesh Kumar

    2013-01-01

    SUMMARY A food-borne trematode infection fascioliasis is one among common public health problems worldwide. It caused a great economic loss for the human race. Control of snail population below a certain threshold level is one of the important methods in the campaign to reduce the incidence of fascioliasis. The life cycle of the parasite can be interrupted by killing the snail or Fasciola larva redia and cercaria inside of the snail Lymnaea acuminata. In vitro toxicity of different binary combinations (1:1 ratio) of plant-derived larvicidal active components such as citral, ferulic acid, umbelliferone, azadirachtin and allicin against Fasciola redia and cercaria were tested. The mortality of larvae was observed at 2h, 4h, 6h and 8h of treatment. In in vitro condition azadirachtin + allicin (1:1 ratio) was highly toxic against redia and cercaria (8h LC50 0.006 and 0.005 mg/L). Toxicity of citral + ferulic acid was lowest against redia and cercaria larvae. PMID:24037283

  3. The mechanism of extracellular stimulation of nerve cells on an electrolyte-oxide-semiconductor capacitor.

    PubMed

    Schoen, Ingmar; Fromherz, Peter

    2007-02-01

    Extracellular excitation of neurons is applied in studies of cultured networks and brain tissue, as well as in neuroprosthetics. We elucidate its mechanism in an electrophysiological approach by comparing voltage-clamp and current-clamp recordings of individual neurons on an insulated planar electrode. Noninvasive stimulation of neurons from pedal ganglia of Lymnaea stagnalis is achieved by defined voltage ramps applied to an electrolyte/HfO2/silicon capacitor. Effects on the smaller attached cell membrane and the larger free membrane are distinguished in a two-domain-stimulation model. Under current-clamp, we study the polarization that is induced for closed ion channels. Under voltage-clamp, we determine the capacitive gating of ion channels in the attached membrane by falling voltage ramps and for comparison also the gating of all channels by conventional variation of the intracellular voltage. Neuronal excitation is elicited under current-clamp by two mechanisms: Rising voltage ramps depolarize the free membrane such that an action potential is triggered. Falling voltage ramps depolarize the attached membrane such that local ion currents are activated that depolarize the free membrane and trigger an action potential. The electrophysiological analysis of extracellular stimulation in the simple model system is a basis for its systematic optimization in neuronal networks and brain tissue.

  4. pT305-CaMKII stabilizes a learning-induced increase in AMPA receptors for ongoing memory consolidation after classical conditioning

    PubMed Central

    Kemenes, György

    2014-01-01

    The role of CaMKII in learning-induced activation and trafficking of AMPA receptors (AMPARs) is well established. However, the link between the phosphorylation state of CaMKII and the agonist-triggered proteasomal degradation of AMPARs during memory consolidation remains unknown. Here we describe a novel CaMKII-dependent mechanism by which a learning-induced increase in AMPAR levels is stabilized for consolidation of associative long-term memory. Six hours after classical conditioning the levels of both autophosphorylated pT305-CaMKII and GluA1 type AMPAR subunits are significantly elevated in the ganglia containing the learning circuits of the snail Lymnaea stagnalis. CaMKIINtide treatment significantly reduces the learning-induced elevation of both pT305-CaMKII and GluA1 levels and impairs associative long-term memory. Inhibition of proteasomal activity offsets the deleterious effects of CaMKIINtide on both GluA1 levels and long-term memory. These findings suggest that increased levels of pT305-CaMKII play a role in AMPAR dependent memory consolidation by reducing proteasomal degradation of GluA1 receptor subunits. PMID:24875483

  5. The effects of the psychiatric drug carbamazepine on freshwater invertebrate communities and ecosystem dynamics.

    PubMed

    Jarvis, Amanda L; Bernot, Melody J; Bernot, Randall J

    2014-10-15

    Freshwater ecosystems are persistently exposed to pharmaceutical pollutants, including carbamazepine. Despite the ubiquity and recalcitrance of carbamazepine, the effects of this pharmaceutical on freshwater ecosystems and communities are unclear. To better understand how carbamazepine influences the invertebrate community and ecosystem dynamics in freshwaters, we conducted a mesocosm experiment utilizing environmentally relevant concentrations of carbamazepine (200 and 2000 ng/L). Mesocosms were populated with four gastropod taxa (Elimia, Physa, Lymnaea and Helisoma), zooplankton, filamentous algae and phytoplankton. After a 31 d experimental duration, structural equation modeling (SEM) was used to relate changes in the community structure and ecosystem dynamics to carbamazepine exposure. Invertebrate diversity increased in the presence of carbamazepine. Additionally, carbamazepine altered the biomass of Helisoma and Elimia, induced a decline in Daphnia pulex abundance and shifted the zooplankton community toward copepod dominance. Lastly, carbamazepine decreased the decomposition of organic matter and indirectly altered primary production and dissolved nutrient concentrations. Changes in the invertebrate community occurred through both direct (i.e., exposure to carbamazepine) and indirect pathways (i.e., changes in food resource availability). These data indicate that carbamazepine may alter freshwater community structure and ecosystem dynamics and could have profound effects on natural systems.

  6. Stimulus-dependent translocation of egg-laying hormone encoding mRNA into the axonal compartment of the neuroendocrine caudodorsal cells.

    PubMed

    van Minnen, J; Bergman, J J

    2003-11-01

    To get insight into the stimulus-dependent translocation of mRNA encoding neuropeptides to the axonal compartment of neurons, we investigated this process in the egg-laying hormone producing caudodorsal cells of the mollusk Lymnaea stagnalis. The axonal compartment including the nerve terminals of these neurons harbors high amounts of mRNA encoding the egg-laying hormone precursor. We determined how a sensory stimulus, that results in egg-laying, affected the amount of egg-laying hormone encoding transcripts in the axon endings. Four hours after stimulation high amounts of transcripts were detected in the axonal compartment and maximum values were reached after 8 h. Transcript levels in the somata were affected in a similar fashion, although the increase was not as pronounced as in the axons. Next, we investigated the ultrastructural localization of egg-laying hormone encoding transcripts in axons and axon terminals by means of electron microscopic in situ hybridization and showed that transcripts were localized in the axoplasm. By means of conventional electron microscopy we showed that axon terminals of egg-laying hormone producing neurons contained large amounts of polyribosomes. Together, these data support the notion that egg-laying hormone encoding transcripts are translated in the axonal compartment.

  7. Photomediated Larvicidal Activity of Pheophorbide a against Cercaria Larvae of Fasciola gigantica

    PubMed Central

    Singh, Divya Jyoti; Singh, Vinay Kumar

    2017-01-01

    Fasciolosis is a parasitic disease caused by Fasciola gigantica. The freshwater snail Lymnaea acuminata is the intermediate host of F. gigantica which cause endemic fasciolosis in the northern part of India. To investigate larvicidal activity of pure and laboratory extracted pheophorbide a (Pa) against cercaria larvae of F. gigantica, data were analyzed in different spectra of visible light, sunlight, and laboratory conditions. Photostimulation of chlorophyll derivative pheophorbide a (Pa) caused time and concentration dependent larvicidal activity against cercaria larvae of F. gigantica. Larvicidal activity of pure Pa under 650 nm and 400–650 nm (8 h LC50 0.006 mg/10 mL) was more pronounced than extracted Pa under same irradiations (650 nm LC50 0.12 mg/10 mL, 400–650 nm LC50 0.14 mg/10 mL). Lowest toxicity of pure (8 h LC50 0.14 mg/10 mL) and extracted Pa (8 h LC50 1.25 mg/10 mL) was noted under 400 nm. Pa was found to be toxic in laboratory conditions also. The results presented in this paper indicate that pheophorbide a possess potential larvicidal activity against Fasciola gigantica larvae in different wavelengths of visible light, sunlight, and laboratory conditions. PMID:28127494

  8. In Vitro Studies of Neuronal Networks and Synaptic Plasticity in Invertebrates and in Mammals Using Multielectrode Arrays

    PubMed Central

    Tessadori, Jacopo; Ghirardi, Mirella

    2015-01-01

    Brain functions are strictly dependent on neural connections formed during development and modified during life. The cellular and molecular mechanisms underlying synaptogenesis and plastic changes involved in learning and memory have been analyzed in detail in simple animals such as invertebrates and in circuits of mammalian brains mainly by intracellular recordings of neuronal activity. In the last decades, the evolution of techniques such as microelectrode arrays (MEAs) that allow simultaneous, long-lasting, noninvasive, extracellular recordings from a large number of neurons has proven very useful to study long-term processes in neuronal networks in vivo and in vitro. In this work, we start off by briefly reviewing the microelectrode array technology and the optimization of the coupling between neurons and microtransducers to detect subthreshold synaptic signals. Then, we report MEA studies of circuit formation and activity in invertebrate models such as Lymnaea, Aplysia, and Helix. In the following sections, we analyze plasticity and connectivity in cultures of mammalian dissociated neurons, focusing on spontaneous activity and electrical stimulation. We conclude by discussing plasticity in closed-loop experiments. PMID:25866681

  9. The conformation of acetylcholine at its target site in the membrane-embedded nicotinic acetylcholine receptor

    PubMed Central

    Williamson, P. T. F.; Verhoeven, A.; Miller, K. W.; Meier, B. H.; Watts, A.

    2007-01-01

    The conformation of the neurotransmitter acetylcholine bound to the fully functional nicotinic acetylcholine receptor embedded in its native membrane environment has been characterized by using frequency-selective recoupling solid-state NMR. Six dipolar couplings among five resolved 13C-labeled atoms of acetylcholine were measured. Bound acetylcholine adopts a bent conformation characterized with a quaternary ammonium-to-carbonyl distance of 5.1 Å. In this conformation, and with its orientation constrained to that previously determined by us, the acetylcholine could be docked satisfactorily in the agonist pocket of the agonist-bound, but not the agonist-free, crystal structure of a soluble acetylcholine-binding protein from Lymnaea stagnali. The quaternary ammonium group of the acetylcholine was determined to be within 3.9 Å of five aromatic residues and its acetyl group close to residues C187/188 of the principle and residue L112 of the complementary subunit. The observed >CO chemical shift is consistent with H bonding to the nicotinic acetylcholine receptor residues γY116 and δT119 that are homologous to L112 in the soluble acetylcholine-binding protein. PMID:17989232

  10. Left-right asymmetric expression of dpp in the mantle of gastropods correlates with asymmetric shell coiling

    PubMed Central

    2013-01-01

    Background Various shapes of gastropod shells have evolved ever since the Cambrian. Although theoretical analyses of morphogenesis exist, the molecular basis of shell development remains unclear. We compared expression patterns of the decapentaplegic (dpp) gene in the shell gland and mantle tissues at various developmental stages between coiled-shell and non-coiled-shell gastropods. Results We analyzed the expression patterns of dpp for the two limpets Patella vulgata and Nipponacmea fuscoviridis, and for the dextral wild-type and sinistral mutant lineage of the pond snail Lymnaea stagnalis. The limpets had symmetric expression patterns of dpp throughout ontogeny, whereas in the pond snail, the results indicated asymmetric and mirror image patterns between the dextral and sinistral lineages. Conclusion We hypothesize that Dpp induces mantle expansion, and the presence of a left/right asymmetric gradient of the Dpp protein causes the formation of a coiled shell. Our results provide a molecular explanation for shell, coiling including new insights into expression patterns in post-embryonic development, which should aid in understanding how various shell shapes are formed and have evolved in the gastropods. PMID:23711320

  11. Retinoic acid induces neurite outgrowth and growth cone turning in invertebrate neurons.

    PubMed

    Dmetrichuk, Jennifer M; Carlone, Robert L; Spencer, Gaynor E

    2006-06-01

    Identification of molecules involved in neurite outgrowth during development and/or regeneration is a major goal in the field of neuroscience. Retinoic acid (RA) is a biologically important metabolite of vitamin A that acts as a trophic factor and has been implicated in neurite outgrowth and regeneration in many vertebrate species. Although abundant in the CNS of many vertebrates, the precise role of RA in neural regeneration has yet to be determined. Moreover, very little information is available regarding the role of RA in invertebrate nervous systems. Here, we demonstrate for the first time that RA induces neurite outgrowth from invertebrate neurons. Using individually identified neurons isolated from the CNS of Lymnaea stagnalis, we demonstrated that a significantly greater proportion of cells produced neurite outgrowth in RA. RA also extended the duration of time that cells remained electrically excitable in vitro, and we showed that exogenously applied RA acted as a chemoattractive factor and induced growth cone turning toward the source of RA. This is the first demonstration that RA can induce turning of an individual growth cone. These data strongly suggest that the actions of RA on neurite outgrowth and cell survival are highly conserved across species.

  12. Different receptors mediate the electrophysiological and growth cone responses of an identified neuron to applied dopamine.

    PubMed

    Dobson, K S; Dmetrichuk, J M; Spencer, G E

    2006-09-15

    Neurotransmitters are among the many cues that may guide developing axons toward appropriate targets in the developing nervous system. We have previously shown in the mollusk Lymnaea stagnalis that dopamine, released from an identified pre-synaptic cell, differentially affects growth cone behavior of its target and non-target cells in vitro. Here, we describe a group of non-target cells that also produce an inhibitory electrophysiological response to applied dopamine. We first determined, using pharmacological blockers, which receptors mediate this physiological response. We demonstrated that the dopaminergic electrophysiological responses of non-target cells were sensitive to a D2 receptor antagonist, as are known target cell responses. However, the non-target cell receptors were linked to different G-proteins and intracellular signaling pathways than the target cell receptors. Despite the presence of a D2-like receptor at the soma, the growth cone collapse of these non-target cells was mediated by D1-like receptors. This study shows that different dopamine receptor sub-types mediated the inhibitory physiological and growth cone responses of an identified cell type. We therefore not only provide further evidence that D2- and D1-like receptors can be present on the same neuron in invertebrates, but also show that these receptors are likely involved in very different cellular functions.

  13. A role for host-parasite interactions in the horizontal transfer of transposons across phyla.

    PubMed

    Gilbert, Clément; Schaack, Sarah; Pace, John K; Brindley, Paul J; Feschotte, Cédric

    2010-04-29

    Horizontal transfer (HT), or the passage of genetic material between non-mating species, is increasingly recognized as an important force in the evolution of eukaryotic genomes. Transposons, with their inherent ability to mobilize and amplify within genomes, may be especially prone to HT. However, the means by which transposons can spread across widely diverged species remain elusive. Here we present evidence that host-parasite interactions have promoted the HT of four transposon families between invertebrates and vertebrates. We found that Rhodnius prolixus, a triatomine bug feeding on the blood of various tetrapods and vector of Chagas' disease in humans, carries in its genome four distinct transposon families that also invaded the genomes of a diverse, but overlapping, set of tetrapods. The bug transposons are approximately 98% identical and cluster phylogenetically with those of the opossum and squirrel monkey, two of its preferred mammalian hosts in South America. We also identified one of these transposon families in the pond snail Lymnaea stagnalis, a cosmopolitan vector of trematodes infecting diverse vertebrates, whose ancestral sequence is nearly identical and clusters with those found in Old World mammals. Together these data provide evidence for a previously hypothesized role of host-parasite interactions in facilitating HT among animals. Furthermore, the large amount of DNA generated by the amplification of the horizontally transferred transposons supports the idea that the exchange of genetic material between hosts and parasites influences their genomic evolution.

  14. Mercury remediation in wetland sediment using zero-valent iron and granular activated carbon.

    PubMed

    Lewis, Ariel S; Huntington, Thomas G; Marvin-DiPasquale, Mark C; Amirbahman, Aria

    2016-05-01

    Wetlands are hotspots for production of toxic methylmercury (MeHg) that can bioaccumulate in the food web. The objective of this study was to determine whether the application of zero-valent iron (ZVI) or granular activated carbon (GAC) to wetland sediment could reduce MeHg production and bioavailability to benthic organisms. Field mesocosms were installed in a wetland fringing Hodgdon Pond (Maine, USA), and ZVI and GAC were applied. Pore-water MeHg concentrations were lower in treated compared with untreated mesocosms; however, sediment MeHg, as well as total Hg (THg), concentrations were not significantly different between treated and untreated mesocosms, suggesting that smaller pore-water MeHg concentrations in treated sediment were likely due to adsorption to ZVI and GAC, rather than inhibition of MeHg production. In laboratory experiments with intact vegetated sediment clumps, amendments did not significantly change sediment THg and MeHg concentrations; however, the mean pore-water MeHg and MeHg:THg ratios were lower in the amended sediment than the control. In the laboratory microcosms, snails (Lymnaea stagnalis) accumulated less MeHg in sediment treated with ZVI or GAC. The study results suggest that both GAC and ZVI have potential for reducing MeHg bioaccumulation in wetland sediment.

  15. Avian schistosomes and human cercarial dermatitis in a wildlife refuge in Mazandaran Province, northern Iran.

    PubMed

    Gohardehi, S; Fakhar, M; Madjidaei, M

    2013-09-01

    Each year, hundreds of aquatic migratory birds migrate from northern hemisphere to the Mazandaran Province, northern Iran. Little information is available on prevalence and density of schistosomes in water birds in Iran and around the world. The objectives of this study were to determine definitive and intermediate hosts of avian schistosomes as well as to assess human cercarial dermatitis (HCD) in a wildlife refuge in Mazandaran Province. Of 1106 examined people, 589 (53.2%) had maculopapular rashes mainly on feet but also on hand. The majority of cases were adults and local residents. Of 260 ducks, 41 (15.8%) were found to be infected with Trichobilharzia spp. eggs or adult worms. Prevalence was highest in Anas clypeata and Anas platyrhynchos, 79% and 18.9%, respectively. A total of 1.2% snails, examined by both shedding and crushing methods, were infected with furcocercariae belonging to avian schistosomes. The most frequently infected snail was Lymnaea gedrosiana (5.9%). Our results showed that cercarial dermatitis and avian schistosomiasis is a common and yet neglected disease in this area. Anas clypeata played the most important role in exposing snails to miracidia in ponds and paddy fields. Moreover, because of the high prevalence in ducks and high prevalence of HCD in the region, it is considered as a new endemic focus in Iran.

  16. Assessing the Dietary Bioavailability of Metals Associated with Natural Particles: Extending the Use of the Reverse Labeling Approach to Zinc.

    PubMed

    Croteau, Marie-Noële; Cain, Daniel J; Fuller, Christopher C

    2017-02-22

    We extend the use of a novel tracing technique to quantify the bioavailability of zinc (Zn) associated with natural particles using snails enriched with a less common Zn stable isotope. Lymnaea stagnalis is a model species that has relatively fast Zn uptake rates from the dissolved phase, enabling their rapid enrichment in (67)Zn during the initial phase of labeling. Isotopically enriched snails were subsequently exposed to algae mixed with increasing amounts of metal-rich particles collected from two acid mine drainage impacted rivers. Zinc bioavailability from the natural particles was inferred from calculations of (66)Zn assimilation into the snail's soft tissues. Zinc assimilation efficiency (AE) varied from 28% for the Animas River particles to 45% for the Snake River particles, indicating that particle-bound, or sorbed Zn, was bioavailable from acid mine drainage wastes. The relative binding strength of Zn sorption to the natural particles was inversely related to Zn bioavailability; a finding that would not have been possible without using the reverse labeling approach. Differences in the chemical composition of the particles suggest that their geochemical properties may influence the extent of Zn bioavailability.

  17. Phylogenetic and individual variation in gastropod central pattern generators.

    PubMed

    Sakurai, Akira; Katz, Paul S

    2015-09-01

    Gastropod molluscs provide a unique opportunity to explore the neural basis of rhythmic behaviors because of the accessibility of their nervous systems and the number of species that have been examined. Detailed comparisons of the central pattern generators (CPGs) underlying rhythmic feeding and swimming behaviors highlight the presence and effects of variation in neural circuits both across and within species. The feeding motor pattern of the snail, Lymnaea, is stereotyped, whereas the feeding motor pattern in the sea hare, Aplysia, is variable. However, the Aplysia motor pattern is regularized with operant conditioning or by mimicking learning using the dynamic clamp to change properties of CPG neurons. Swimming evolved repeatedly in marine gastropods. Distinct neural mechanisms underlie dissimilar forms of swimming, with homologous neurons playing different roles. However, even similar swimming behaviors in different species can be produced by distinct neural mechanisms, resulting from different synaptic connectivity of homologous neurons. Within a species, there can be variation in the strength and even valence of synapses, which does not have functional relevance under normal conditions, but can cause some individuals to be more susceptible to lesion of the circuit. This inter- and intra-species variation provides novel insights into CPG function and plasticity.

  18. Biogeochemical controls of uranium bioavailability from the dissolved phase in natural freshwaters

    USGS Publications Warehouse

    Croteau, Marie-Noele; Fuller, Christopher C.; Cain, Daniel J.; Campbell, Kate M.; Aiken, George R.

    2016-01-01

    To gain insights into the risks associated with uranium (U) mining and processing, we investigated the biogeochemical controls of U bioavailability in the model freshwater speciesLymnaea stagnalis (Gastropoda). Bioavailability of dissolved U(VI) was characterized in controlled laboratory experiments over a range of water hardness, pH, and in the presence of complexing ligands in the form of dissolved natural organic matter (DOM). Results show that dissolved U is bioavailable under all the geochemical conditions tested. Uranium uptake rates follow first order kinetics over a range encompassing most environmental concentrations. Uranium uptake rates in L. stagnalis ultimately demonstrate saturation uptake kinetics when exposure concentrations exceed 100 nM, suggesting uptake via a finite number of carriers or ion channels. The lack of a relationship between U uptake rate constants and Ca uptake rates suggest that U does not exclusively use Ca membrane transporters. In general, U bioavailability decreases with increasing pH, increasing Ca and Mg concentrations, and when DOM is present. Competing ions did not affect U uptake rates. Speciation modeling that includes formation constants for U ternary complexes reveals that the aqueous concentration of dicarbonato U species (UO2(CO3)2–2) best predicts U bioavailability to L. stagnalis, challenging the free-ion activity model postulate.

  19. Effects of chronic low level lead exposure on the physiology of individually identifiable neurons.

    PubMed

    Audesirk, G; Audesirk, T

    1983-01-01

    Although chronic exposure to lead has been correlated with a variety of behavioral and neurochemical deficits in humans and other mammals, little is known of the mechanisms of action of chronic lead at the level of the individual nerve cell. We have used the individually identifiable neurons of the freshwater pond snail Lymnaea stagnalis as a model system to investigate the effects of chronic low level (5 microM) lead exposure on neuronal physiology. Thirteen neuronal parameters were measured with intracellular microelectrode recording in each of six different identifiable neurons or homogeneous neuron clusters. Results were analyzed by a multivariate analysis of variance (MANOVA). MANOVA analysis indicates that there is a significant overall effect of lead exposure (p = 0.0001) and a significant interaction between lead and neuron type (p = 0.01). In most neuron types, chronic lead causes an increase in the resting potential, a slowing of recovery of the membrane potential after the undershoot of a spike, a decrease in spontaneous spiking activity, and a decrease in the input resistance. Lead also has differential effects on identifiable neurons, depressing excitability in some neuron types while not altering excitability in others.

  20. About a Snail, a Toad, and Rodents: Animal Models for Adaptation Research

    PubMed Central

    Roubos, Eric W.; Jenks, Bruce G.; Xu, Lu; Kuribara, Miyuki; Scheenen, Wim J. J. M.; Kozicz, Tamás

    2010-01-01

    Neural adaptation mechanisms have many similarities throughout the animal kingdom, enabling to study fundamentals of human adaptation in selected animal models with experimental approaches that are impossible to apply in man. This will be illustrated by reviewing research on three of such animal models, viz. (1) the egg-laying behavior of a snail, Lymnaea stagnalis: how one neuron type controls behavior, (2) adaptation to the ambient light condition by a toad, Xenopus laevis: how a neuroendocrine cell integrates complex external and neural inputs, and (3) stress, feeding, and depression in rodents: how a neuronal network co-ordinates different but related complex behaviors. Special attention is being paid to the actions of neurochemical messengers, such as neuropeptide Y, urocortin 1, and brain-derived neurotrophic factor. While awaiting new technological developments to study the living human brain at the cellular and molecular levels, continuing progress in the insight in the functioning of human adaptation mechanisms may be expected from neuroendocrine research using invertebrate and vertebrate animal models. PMID:22649351

  1. Mapping of dihydropyridine binding residues in a less sensitive invertebrate L-type calcium channel (LCa v 1).

    PubMed

    Senatore, Adriano; Boone, Adrienne; Lam, Stanley; Dawson, Taylor F; Zhorov, Boris; Spafford, J David

    2011-01-01

    Invertebrate L-type calcium channel, LCa(v) 1, isolated from the pond snail Lymnaea stagnalis is nearly indistinguishable from mammalian Ca(v) 1.2 (α1C) calcium channel in biophysical characteristics observed in vitro. These L-type channels are likely constrained within a narrow range of biophysical parameters to perform similar functions in the snail and mammalian cardiovascular systems. What distinguishes snail and mammalian L-type channels is a difference in dihydropyridine sensitivity: 100 nM isradipine exhibits a significant block of mammalian Ca(v) 1.2 currents without effect on snail LCa(v)1 currents. The native snail channel serves as a valuable surrogate for validating key residue differences identified from previous experimental and molecular modeling work. As predicted, three residue changes in LCa(v)1 (N_3o18, F_3i10, and I_4i12) replaced with DHP-sensing residues in respective positions of Ca(v) 1.2, (Q_3o18, Y_3i10, and M_4i12) raises the potency of isradipine block of LCa(v)1 channels to that of mammalian Ca(v) 1.2. Interestingly, the single N_3o18_Q mutation in LCa(v) 1 channels lowers DHP sensitivity even further and the triple mutation bearing enhanced isradipine sensitivity, still retains a reduced potency of agonist, (S)-Bay K8644.

  2. Dietary uptake of Cu sorbed to hydrous iron oxide is linked to cellular toxicity and feeding inhibition in a benthic grazer

    USGS Publications Warehouse

    Cain, Daniel J.; Croteau, Marie-Noele; Fuller, Christopher C.; Ringwood, Amy H.

    2016-01-01

    Whereas feeding inhibition caused by exposure to contaminants has been extensively documented, the underlying mechanism(s) are less well understood. For this study, the behavior of several key feeding processes, including ingestion rate and assimilation efficiency, that affect the dietary uptake of Cu were evaluated in the benthic grazer Lymnaea stagnalis following 4–5 h exposures to Cu adsorbed to synthetic hydrous ferric oxide (Cu–HFO). The particles were mixed with a cultured alga to create algal mats with Cu exposures spanning nearly 3 orders of magnitude at variable or constant Fe concentrations, thereby allowing first order and interactive effects of Cu and Fe to be evaluated. Results showed that Cu influx rates and ingestion rates decreased as Cu exposures of the algal mat mixture exceeded 104 nmol/g. Ingestion rate appeared to exert primary control on the Cu influx rate. Lysosomal destabilization rates increased directly with Cu influx rates. At the highest Cu exposure where the incidence of lysosomal membrane damage was greatest (51%), the ingestion rate was suppressed 80%. The findings suggested that feeding inhibition was a stress response emanating from excessive uptake of dietary Cu and cellular toxicity.

  3. Survival and growth of freshwater pulmonate and nonpulmonate snails in 28-day exposures to copper, ammonia, and pentachlorophenol

    USGS Publications Warehouse

    Besser, John M.; Dorman, Rebecca A.; Hardesty, Douglas K.; Ingersoll, Christopher G.

    2016-01-01

    We performed toxicity tests with two species of pulmonate snails (Lymnaea stagnalis and Physa gyrina) and four taxa of nonpulmonate snails in the family Hydrobiidae (Pyrgulopsis robusta,Taylorconcha serpenticola, Fluminicola sp., and Fontigens aldrichi). Snails were maintained in static-renewal or recirculating culture systems with adults removed periodically to isolate cohorts of offspring for toxicity testing. This method successfully produced offspring for both species of pulmonate snails and for two hydrobiid species, P. robusta and Fluminicola sp. Toxicity tests were performed for 28 days with copper, ammonia, and pentachlorophenol in hard reconstituted water with endpoints of survival and growth. Tests were started with 1-week-old L. stagnalis, 2-week-old P. gyrina, 5- to 13-week-old P. robusta and Fluminicola sp., and older juveniles and adults of several hydrobiid species. For all three chemicals, chronic toxicity values for pulmonate snails were consistently greater than those for hydrobiid snails, and hydrobiids were among the most sensitive taxa in species sensitivity distributions for all three chemicals. These results suggest that the toxicant sensitivity of nonpulmonate snails in the family Hydrobiidae would not be adequately represented by results of toxicity testing with pulmonate snails.

  4. Efficient expression of acetylcholine-binding protein from Aplysia californica in Bac-to-Bac system.

    PubMed

    Lin, Bo; Meng, Hailing; Bing, Hui; Zhangsun, Dongting; Luo, Sulan

    2014-01-01

    The Bac-to-Bac baculovirus expression system can efficiently produce recombinant proteins, but the system may have to be optimized to achieve high-level expression for different candidate proteins. We reported here the efficient expression of acetylcholine-binding proteins from sea hares Aplysia californica (Ac-AChBP) and a convenient method to monitor protein expression level in this expression system. Three key factors affecting expression of Ac-AChBP were optimized for maximizing the yield, which included the cell density, volume of the infecting baculovirus inoculums, and the culturing time of postinfection. We have found it to reach a high yield of ∼5 mg/L, which needs 55 h incubation after infection at the cell density of 2 × 10(6) cells/mL with an inoculum volume ratio of 1 : 100. The optimized expression system in this study was also applied for expressing another protein Ls-AChBP from Lymnaea stagnalis successfully. Therefore, this established method is helpful to produce high yields of AChBP proteins for X-ray crystallographic structural and functional studies.

  5. Trials with N-tritylmorpholine (Shell WL 8008) as a molluscicide in Southern Rhodesia

    PubMed Central

    Shiff, C. J.

    1966-01-01

    In the course of research into the effectiveness of new molluscicides for bilharziasis control, a recently developed compound, Shell WL 8008 (N-tritylmorpholine), has been tested in Southern Rhodesia, both in the laboratory and in the field. It is highly toxic to Bulinus (Physopsis) globosus, Biomphalaria pfeifferi and Lymnaea natalensis, all of which succumbed to the compound at a concentration of 1.0 ppm in a contact time of only 15 minutes. In experiments in reservoirs and natural watercourses, a concentrated aqueous suspension was sprayed on or just under the water surface, forming a white ”blanket” that sank at a rate depending on temperature, amount of vegetation, and so on. The period of contact with snails is sufficiently long for the molluscicide to kill them as it sinks; this characteristic is of great importance in the economic treatment of large stagnant water bodies or slow-moving streams. The compound is highly piscicidal, but has little effect on a wide range of microfauna. PMID:5297004

  6. Inhibition of nicotinic acetylcholine receptors, a novel facet in the pleiotropic activities of snake venom phospholipases A2.

    PubMed

    Vulfius, Catherine A; Kasheverov, Igor E; Starkov, Vladislav G; Osipov, Alexey V; Andreeva, Tatyana V; Filkin, Sergey Yu; Gorbacheva, Elena V; Astashev, Maxim E; Tsetlin, Victor I; Utkin, Yuri N

    2014-01-01

    Phospholipases A2 represent the most abundant family of snake venom proteins. They manifest an array of biological activities, which is constantly expanding. We have recently shown that a protein bitanarin, isolated from the venom of the puff adder Bitis arietans and possessing high phospholipolytic activity, interacts with different types of nicotinic acetylcholine receptors and with the acetylcholine-binding protein. To check if this property is characteristic to all venom phospholipases A2, we have studied the capability of these enzymes from other snakes to block the responses of Lymnaea stagnalis neurons to acetylcholine or cytisine and to inhibit α-bungarotoxin binding to nicotinic acetylcholine receptors and acetylcholine-binding proteins. Here we present the evidence that phospholipases A2 from venoms of vipers Vipera ursinii and V. nikolskii, cobra Naja kaouthia, and krait Bungarus fasciatus from different snake families suppress the acetylcholine- or cytisine-elicited currents in L. stagnalis neurons and compete with α-bungarotoxin for binding to muscle- and neuronal α7-types of nicotinic acetylcholine receptor, as well as to acetylcholine-binding proteins. As the phospholipase A2 content in venoms is quite high, under some conditions the activity found may contribute to the deleterious venom effects. The results obtained suggest that the ability to interact with nicotinic acetylcholine receptors may be a general property of snake venom phospholipases A2, which add a new target to the numerous activities of these enzymes.

  7. Silver bioaccumulation dynamics in a freshwater invertebrate after aqueous and dietary exposures to nanosized and ionic Ag

    USGS Publications Warehouse

    le Croteau, Marie-Noe; Misra, Superb K.; Luoma, Samuel N.; Valsami-Jones, Eugenia

    2011-01-01

    We compared silver (Ag) bioavailability and toxicity to a freshwater gastropod after exposure to ionic silver (Ag+) and to Ag nanoparticles (Ag NPs) capped with citrate or with humic acid. Silver form, exposure route, and capping agent influence Ag bioaccumulation dynamics in Lymnaea stagnalis. Snails efficiently accumulated Ag from all forms after either aqueous or dietary exposure. For both exposure routes, uptake rates were faster for Ag+ than for Ag NPs. Snails efficiently assimilated Ag from Ag NPs mixed with diatoms (assimilation efficiency (AE) ranged from 49 to 58%) and from diatoms pre-exposed to Ag+ (AE of 73%). In the diet, Ag NPs damaged digestion. Snails ate less and inefficiently processed the ingested food, which adversely impacted their growth. Loss rates of Ag were faster after waterborne exposure to Ag NPs than after exposure to dissolved Ag+. Once Ag was taken up from diet, whether from Ag+ or Ag NPs, Ag was lost extremely slowly. Large Ag body concentrations are thus expected in L. stagnalis after dietborne exposures, especially to citrate-capped Ag NPs. Ingestion of Ag associated with particulate materials appears as the most important vector of uptake. Nanosilver exposure from food might trigger important environmental risks.

  8. Mercury remediation in wetland sediment using zero-valent iron and granular activated carbon

    USGS Publications Warehouse

    Lewis, Ariel S.; Huntington, Thomas G.; Marvin-DiPasquale, Mark C.; Amirbahman, Aria

    2016-01-01

    Wetlands are hotspots for production of toxic methylmercury (MeHg) that can bioaccumulate in the food web. The objective of this study was to determine whether the application of zero-valent iron (ZVI) or granular activated carbon (GAC) to wetland sediment could reduce MeHg production and bioavailability to benthic organisms. Field mesocosms were installed in a wetland fringing Hodgdon Pond (Maine, USA), and ZVI and GAC were applied. Pore-water MeHg concentrations were lower in treated compared with untreated mesocosms; however, sediment MeHg, as well as total Hg (THg), concentrations were not significantly different between treated and untreated mesocosms, suggesting that smaller pore-water MeHg concentrations in treated sediment were likely due to adsorption to ZVI and GAC, rather than inhibition of MeHg production. In laboratory experiments with intact vegetated sediment clumps, amendments did not significantly change sediment THg and MeHg concentrations; however, the mean pore-water MeHg and MeHg:THg ratios were lower in the amended sediment than the control. In the laboratory microcosms, snails (Lymnaea stagnalis) accumulated less MeHg in sediment treated with ZVI or GAC. The study results suggest that both GAC and ZVI have potential for reducing MeHg bioaccumulation in wetland sediment.

  9. Survival and Growth of Freshwater Pulmonate and Nonpulmonate Snails in 28-Day Exposures to Copper, Ammonia, and Pentachlorophenol.

    PubMed

    Besser, John M; Dorman, Rebecca A; Hardesty, Douglas L; Ingersoll, Christopher G

    2016-02-01

    We performed toxicity tests with two species of pulmonate snails (Lymnaea stagnalis and Physa gyrina) and four taxa of nonpulmonate snails in the family Hydrobiidae (Pyrgulopsis robusta, Taylorconcha serpenticola, Fluminicola sp., and Fontigens aldrichi). Snails were maintained in static-renewal or recirculating culture systems with adults removed periodically to isolate cohorts of offspring for toxicity testing. This method successfully produced offspring for both species of pulmonate snails and for two hydrobiid species, P. robusta and Fluminicola sp. Toxicity tests were performed for 28 days with copper, ammonia, and pentachlorophenol in hard reconstituted water with endpoints of survival and growth. Tests were started with 1-week-old L. stagnalis, 2-week-old P. gyrina, 5- to 13-week-old P. robusta and Fluminicola sp., and older juveniles and adults of several hydrobiid species. For all three chemicals, chronic toxicity values for pulmonate snails were consistently greater than those for hydrobiid snails, and hydrobiids were among the most sensitive taxa in species sensitivity distributions for all three chemicals. These results suggest that the toxicant sensitivity of nonpulmonate snails in the family Hydrobiidae would not be adequately represented by results of toxicity testing with pulmonate snails.

  10. Novel and non-traditional use of stable isotope tracers to study metal bioavailability from natural particles

    USGS Publications Warehouse

    Croteau, Marie-Noële; Cain, Daniel J.; Fuller, Christopher C.

    2013-01-01

    We devised a novel tracing approach that involves enriching test organisms with a stable metal isotope of low natural abundance prior to characterizing metal bioavailability from natural inorganic particles. In addition to circumventing uncertainties associated with labeling natural particles and distinguishing background metals, the proposed "reverse labeling" technique overcomes many drawbacks inherent to using radioisotope tracers. Specifically, we chronically exposed freshwater snails (Lymnaea stagnalis) to synthetic water spiked with Cu that was 99.4% 65Cu to increase the relative abundance of 65Cu in the snail’s tissues from 32% to >80%. The isotopically enriched snails were then exposed to benthic algae mixed with Cu-bearing Fe–Al particles collected from the Animas River (Colorado), an acid mine drainage impacted river. We used 63Cu to trace Cu uptake from the natural particles and inferred their bioavailability from calculation of Cu assimilation into tissues. Cu assimilation from these particles was 44%, indicating that 44% of the particulate Cu was absorbed by the invertebrate. This demonstrates that inorganic particulate Cu can be bioavailable. The reverse labeling approach shows great potential in various scientific areas such as environmental contamination and nutrition for addressing questions involving uptake of an element that naturally has multiple isotopes.

  11. Bioaccumulation and toxicity of CuO nanoparticles by a freshwater invertebrate after waterborne and dietborne exposures

    USGS Publications Warehouse

    Croteau, Marie-Noele; Misra, Superb K.; Luoma, Samuel N.; Valsami-Jones, Eugenia

    2014-01-01

    The incidental ingestion of engineered nanoparticles (NPs) can be an important route of uptake for aquatic organisms. Yet, knowledge of dietary bioavailability and toxicity of NPs is scarce. Here we used isotopically modified copper oxide (65CuO) NPs to characterize the processes governing their bioaccumulation in a freshwater snail after waterborne and dietborne exposures. Lymnaea stagnalis efficiently accumulated 65Cu after aqueous and dietary exposures to 65CuO NPs. Cu assimilation efficiency and feeding rates averaged 83% and 0.61 g g–1 d–1 at low exposure concentrations (–1), and declined by nearly 50% above this concentration. We estimated that 80–90% of the bioaccumulated 65Cu concentration in L. stagnalis originated from the 65CuO NPs, suggesting that dissolution had a negligible influence on Cu uptake from the NPs under our experimental conditions. The physiological loss of 65Cu incorporated into tissues after exposures to 65CuO NPs was rapid over the first days of depuration and not detectable thereafter. As a result, large Cu body concentrations are expected in L. stagnalis after exposure to CuO NPs. To the degree that there is a link between bioaccumulation and toxicity, dietborne exposures to CuO NPs are likely to elicit adverse effects more readily than waterborne exposures.

  12. Molecular diversity of avian schistosomes in Danish freshwater snails.

    PubMed

    Christiansen, Anne Ø; Olsen, Annette; Buchmann, Kurt; Kania, Per W; Nejsum, Peter; Vennervald, Birgitte J

    2016-03-01

    Avian schistosomes are widespread parasites of snails and waterfowl and may cause cercarial dermatitis (swimmer's itch) in humans, a disease that is frequently reported in European countries. These parasites are known to occur in Denmark, but here, we applied a new approach using molecular tools to identify the parasites at species level. In order to do that, 499 pulmonate freshwater snails (Radix sp., Lymnaea stagnalis, Stagnicola sp. and Planorbarius corneus) were sampled from 12 lakes, ponds, and marshes in the greater Copenhagen area. Avian schistosome cercariae were identified by microscopy and subjected to molecular investigation by sequencing and phylogenetic analysis of the 5.8S and ITS2 ribosomal DNA for species identification. Additionally, snail hosts belonging to the genus Radix were identified by sequencing and phylogenetic analysis of partial ITS2 ribosomal DNA. Three out of 499 snails shed different species of Trichobilharzia cercariae: Trichobilharzia szidati was isolated from L. stagnalis, Trichobilharzia franki from Radix auricularia and Trichobilharzia regenti from Radix peregra. In the light of the public health risk represented by bird schistosomes, these findings are of concern and, particularly, the presence of the potentially neuro-pathogenic species, T. regenti, in Danish freshwaters calls for attention.

  13. Associations between trematode infections in cattle and freshwater snails in highland and lowland areas of Iringa Rural District, Tanzania.

    PubMed

    Nzalawahe, Jahashi; Kassuku, Ayub A; Stothard, J Russell; Coles, Gerald C; Eisler, Mark C

    2015-09-01

    The epidemiology of trematode infections in cattle was investigated within highland and lowland areas of Iringa Rural District, in southern Tanzania. Fecal samples were collected from 450 cattle in 15 villages at altitudes ranging from 696 to 1800 m above the sea level. Freshwater snails were collected from selected water bodies and screened for emergence of cercariae. The infection rates in cattle were Fasciola gigantica 28·2%, paramphistomes 62·8% and Schistosoma bovis 4·8%. Notably, prevalence of trematode infections in cattle was much higher in highland (altitude > 1500 m) as compared with lowland (altitude < 1500 m) areas and was statistically significant (P-value = 0·000) for F. gigantica and paramphistomes but not for S. bovis. The snails collected included Lymnaea natalensis, Bulinus africanus, Bulinus tropicus, Bulinus forskali, Biomphalaria pfeifferi, Melanoides tuberculata and Bellamya constricta with a greater proportion of highland (75%) than lowland (36%) water bodies harbouring snails. Altitude is a major factor shaping the epidemiology of F. gigantica and paramphistomes infections in cattle in Iringa Rural District with greater emphasis upon control needed in highland areas.

  14. [A focus study from a case of human fascioliasis in Neuquén].

    PubMed

    Rubel, Diana; Prepelitchi, Lucila; Kleiman, Florencia; Carnevale, Silvana; Wisnivesky-Colli, Cristina

    2005-01-01

    An epidemiological focal study was performed in Loncopué, Neuquén, Argentina, in November 2002 to detect the origin of the infection in a human case of fascioliasis confirmed by an indirect-ELISA test, six months before the study. Thirty five individual fecal samples were taken from domestic livestock, and watercress plants and snails were collected from the irrigation ditches connected to a main canal in the surroundings of the patient's house. A new blood sample was taken from the already recovered patient. The patient was still seropositive to Fasciola hepatica antigens. No metacercariae were found in the 222 watercress leaves checked. All the snails collected (n=130) were identified as Lymnaea viatrix and two out of 101 (2%) were infected with F. hepatica larvae. Coprological analysis showed F. hepatica eggs in 100% of goats (10/10), 82% of sheep (9/11) and 86% of bovines (6/7). The number of eggs per gram shed by positive goats (median=20.7, Q1=6.2, Q3=34.5) and sheep (4, 1.8, 13) was significantly higher than in cows (0.3, 0.3, 1.7) (p < 0.01). Local veterinary control programs were apparently not effective in this case. Anthelmintics used and treatment schedule should be revised and small herds raised at households should also be included and treated.

  15. Are Sick Individuals Weak Competitors? Competitive Ability of Snails Parasitized by a Gigantism-Inducing Trematode

    PubMed Central

    Seppälä, Otto; Karvonen, Anssi; Kuosa, Marja; Haataja, Maarit; Jokela, Jukka

    2013-01-01

    Parasitized individuals are often expected to be poor competitors because they are weakened by infections. Many trematode species, however, although extensively exploiting their mollusc hosts, also induce gigantism (increased host size) by diverting host resources towards growth instead of reproduction. In such systems, alternatively to reduced competitive ability due to negative effects of parasitism on host performance, larger size could allow more efficient resource acquisition and thus increase the relative competitive ability of host individuals. We addressed this hypothesis by testing the effect of a trematode parasite Diplostomum pseudospathaceum on the competitive ability of its snail host Lymnaea stagnalis. We experimentally examined the growth of snails kept in pairs in relation to their infection status and intensity of resource competition (i.e. food availability). We found that parasitized snails grew faster and their reproduction was reduced compared to unparasitized individuals indicating parasite-induced gigantism. However, growth of the snails was faster when competing with parasitized individuals compared to unparasitized snails indicating reduced competitive ability due to parasitism. The latter effect, however, was relatively weak suggesting that the effects of the parasite on snail physiology may partly override each other in determining competitive ability. PMID:24205383

  16. Unraveling the complexities of circadian and sleep interactions with memory formation through invertebrate research

    PubMed Central

    Michel, Maximilian; Lyons, Lisa C.

    2014-01-01

    Across phylogeny, the endogenous biological clock has been recognized as providing adaptive advantages to organisms through coordination of physiological and behavioral processes. Recent research has emphasized the role of circadian modulation of memory in generating peaks and troughs in cognitive performance. The circadian clock along with homeostatic processes also regulates sleep, which itself impacts the formation and consolidation of memory. Thus, the circadian clock, sleep and memory form a triad with ongoing dynamic interactions. With technological advances and the development of a global 24/7 society, understanding the mechanisms underlying these connections becomes pivotal for development of therapeutic treatments for memory disorders and to address issues in cognitive performance arising from non-traditional work schedules. Invertebrate models, such as Drosophila melanogaster and the mollusks Aplysia and Lymnaea, have proven invaluable tools for identification of highly conserved molecular processes in memory. Recent research from invertebrate systems has outlined the influence of sleep and the circadian clock upon synaptic plasticity. In this review, we discuss the effects of the circadian clock and sleep on memory formation in invertebrates drawing attention to the potential of in vivo and in vitro approaches that harness the power of simple invertebrate systems to correlate individual cellular processes with complex behaviors. In conclusion, this review highlights how studies in invertebrates with relatively simple nervous systems can provide mechanistic insights into corresponding behaviors in higher organisms and can be used to outline possible therapeutic options to guide further targeted inquiry. PMID:25136297

  17. Laboratory maintenance of the bacterial endosymbiont, Neorickettsia sp., through the life cycle of a digenean, Plagiorchis elegans

    PubMed Central

    Greiman, Stephen E.; Tkach, Maksym; Vaughan, Jefferson A.; Tkach, Vasyl V.

    2016-01-01

    The Digenea (Platyhelminthes: Trematoda) are a diverse and complex group of internal metazoan parasites. These parasites can serve as hosts to obligate intracellular bacteria belonging to the genus Neorickettsia (Family: Anaplasmataceae). Neorickettsiae persist within all stages of the fluke life cycle and thus are maintained through vertical transmission. However, the low prevalence of Neorickettsia in nature limits study of their transmission biology at different steps of digenean life cycles. To resolve this dilemma, we have developed for the first time a laboratory model allowing to maintain Neorickettsia sp. through the whole life cycle of a digenean, Plagiorchis elegans. The laboratory life cycle of P. elegans consists of a snail first intermediate host, Lymnaea stagnalis, an aquatic arthropod second intermediate host, Culex pipiens (mosquito larva), and a vertebrate definitive host, Mesocricetus auratus (Syrian hamster). This paper focuses on the development of the laboratory life cycle, as well as outlines its potential uses in studying the transmission biology of Neorickettsia and its evolutionary relationship within its digenean host. PMID:26160679

  18. The role of synaptotagmin I C2A calcium-binding domain in synaptic vesicle clustering during synapse formation

    PubMed Central

    Gardzinski, Peter; Lee, David W K; Fei, Guang-He; Hui, Kwokyin; Huang, Guan J; Sun, Hong-Shuo; Feng, Zhong-Ping

    2007-01-01

    Synaptic vesicles aggregate at the presynaptic terminal during synapse formation via mechanisms that are poorly understood. Here we have investigated the role of the putative calcium sensor synaptotagmin I in vesicle aggregation during the formation of soma–soma synapses between identified partner cells using a simple in vitro synapse model in the mollusc Lymnaea stagnalis. Immunocytochemistry, optical imaging and electrophysiological recording techniques were used to monitor synapse formation and vesicle localization. Within 6 h, contact between appropriate synaptic partner cells up-regulated global synaptotagmin I expression, and induced a localized aggregation of synaptotagmin I at the contact site. Cell contacts between non-synaptic partner cells did not affect synaptotagmin I expression. Application of an human immunodeficiency virus type-1 transactivator (HIV-1 TAT)-tagged peptide corresponding to loop 3 of the synaptotagmin I C2A domain prevented synaptic vesicle aggregation and synapse formation. By contrast, a TAT-tagged peptide containing the calcium-binding motif of the C2B domain did not affect synaptic vesicle aggregation or synapse formation. Calcium imaging with Fura-2 demonstrated that TAT–C2 peptides did not alter either basal or evoked intracellular calcium levels. These results demonstrate that contact with an appropriate target cell is necessary to initiate synaptic vesicle aggregation during nascent synapse formation and that the initial aggregation of synaptic vesicles is dependent on loop 3 of the C2A domain of synaptotagmin I. PMID:17317745

  19. (Tri)butyltin biotic degradation rates and pathways in different compartments of a freshwater model ecosystem.

    PubMed

    Tessier, Emmanuel; Amouroux, David; Morin, Anne; Christian, Lehnhoff; Thybaud, Eric; Vindimian, Eric; Donard, Olivier F X

    2007-12-15

    Experiments were conducted in controlled temperate freshwater ecosystems (microcosms) to determine the persistence and biogeochemical dynamic of tributyltin (TBT) and its degradation products. TBT and its derivatives were monitored simultaneously for 23 days (552 h) in sediment-water systems, with or without macroorganisms (macrophytes: Elodea canadensis and gastropods: Lymnaea stagnalis). Biphasic TBT removal from the water column was significantly enhanced by the presence of biota. The persistence of TBT in biota was assessed by a kinetic approach of the different bioaccumulation pathways and associated metabolisms adopted by the snails and the macrophytes in response to the TBT contamination. Furthermore, sediment acted for the final sink for butyltins in both types of microcosms, with more than 70% of TBT and its metabolites recovered in this compartment after two weeks of exposure. Degradation pathways in sediments of both biotic and abiotic microcosms appeared to represent a key process in TBT cycle and were characterized by half-lives in the range of one month. Specific transformation and transfer pathways of TBT as reactional mechanisms are discussed and modelled assessing in detail the role of each compartment with regards to the fate of TBT in the model aquatic ecosystems.

  20. [The radioactive contamination dynamics of water body ecosystems of different types in the Chernobyl atomic station alienation zone in Belarus].

    PubMed

    Golubev, A P; Sikorskiĭ, V G; Kalinin, V N; Afonin, V Iu; Chekan, G S

    2007-01-01

    The long-term (1986-2005) gamma-activity dynamics in dominating zoobenthos species and the bottom sediments in the inlet of Pripyat river and the non-flowing Perstok lake within the Chernobyl alienation zone was determined. Immediately after the accident (1986-1987) zoonehthos y-activity achieved the maximal values (up to 300-1100 kBq/kg) and after that began to decline steadily due to natural decay of man-caused radionuclides of "Chernobyl origin". Up to summer 2005 gastropod mollusks gamma-activity (Lymnaea stagnalis, Viviparus viviparus) approached to the natural level (less than 6 Bq/kg) in the inlet of Pripyat river, but it remained at the very high level up to 979-1638 Bq/kg in the Perstok lake. The positive correlation between gamma-activity of mollusks and bottom sediments has been established. In turn, the long-term variations of atmospheric precipitate amounts which wash down radionuclides from surrounding territories to water bodies and the amounts of annual flow of the Pripyat river as well as shoreline position changes in water bodies within the Chernobyl alienation zone influence on these values too.

  1. α-Conotoxin OmIA Is a Potent Ligand for the Acetylcholine-binding Protein as Well as α3β2 and α7 Nicotinic Acetylcholine Receptors*

    PubMed Central

    Talley, Todd T.; Olivera, Baldomero M.; Han, Kyou-Hoon; Christensen, Sean B.; Dowell, Cheryl; Tsigelny, Igor; Ho, Kwok-Yiu; Taylor, Palmer; McIntosh, J. Michael

    2016-01-01

    The molluskan acetylcholine-binding protein (AChBP) is a homolog of the extracellular binding domain of the pentameric ligand-gated ion channel family. AChBP most closely resembles the α-subunit of nicotinic acetylcholine receptors and in particular the homomeric α7 nicotinic receptor. We report the isolation and characterization of an α-conotoxin that has the highest known affinity for the Lymnaea AChBP and also potently blocks the α7 nAChR subtype when expressed in Xenopus oocytes. Remarkably, the peptide also has high affinity for the α3β2 nAChR indicating that α-conotoxin OmIA in combination with the AChBP may serve as a model system for understanding the binding determinants of α3β2 nAChRs. α-Conotoxin OmIA was purified from the venom of Conus omaria. It is a 17-amino-acid, two-disulfide bridge peptide. The ligand is the first α-conotoxin with higher affinity for the closely related receptor subtypes, α3β2 versus α6β2, and selectively blocks these two subtypes when compared with α2β2, α4β2, and α1β1δε nAChRs. PMID:16803900

  2. Assessing the dietary bioavailability of metals associated with natural particles: Extending the use of the reverse labeling approach to zinc

    USGS Publications Warehouse

    Croteau, Marie-Noele; Cain, Daniel J.; Fuller, Christopher C.

    2017-01-01

    We extend the use of a novel tracing technique to quantify the bioavailability of zinc (Zn) associated with natural particles using snails enriched with a less common Zn stable isotope. Lymnaea stagnalis is a model species that has relatively fast Zn uptake rates from the dissolved phase, enabling their rapid enrichment in 67Zn during the initial phase of labeling. Isotopically enriched snails were subsequently exposed to algae mixed with increasing amounts of metal-rich particles collected from two acid mine drainage impacted rivers. Zinc bioavailability from the natural particles was inferred from calculations of 66Zn assimilation into the snail’s soft tissues. Zinc assimilation efficiency (AE) varied from 28% for the Animas River particles to 45% for the Snake River particles, indicating that particle-bound, or sorbed Zn, was bioavailable from acid mine drainage wastes. The relative binding strength of Zn sorption to the natural particles was inversely related to Zn bioavailability; a finding that would not have been possible without using the reverse labeling approach. Differences in the chemical composition of the particles suggest that their geochemical properties may influence the extent of Zn bioavailability.

  3. Phospholipase A2 – nexus of aging, oxidative stress, neuronal excitability, and functional decline of the aging nervous system? Insights from a snail model system of neuronal aging and age-associated memory impairment

    PubMed Central

    Hermann, Petra M.; Watson, Shawn N.; Wildering, Willem C.

    2014-01-01

    The aging brain undergoes a range of changes varying from subtle structural and physiological changes causing only minor functional decline under healthy normal aging conditions, to severe cognitive or neurological impairment associated with extensive loss of neurons and circuits due to age-associated neurodegenerative disease conditions. Understanding how biological aging processes affect the brain and how they contribute to the onset and progress of age-associated neurodegenerative diseases is a core research goal in contemporary neuroscience. This review focuses on the idea that changes in intrinsic neuronal electrical excitability associated with (per)oxidation of membrane lipids and activation of phospholipase A2 (PLA2) enzymes are an important mechanism of learning and memory failure under normal aging conditions. Specifically, in the context of this special issue on the biology of cognitive aging we portray the opportunities offered by the identifiable neurons and behaviorally characterized neural circuits of the freshwater snail Lymnaea stagnalis in neuronal aging research and recapitulate recent insights indicating a key role of lipid peroxidation-induced PLA2 as instruments of aging, oxidative stress and inflammation in age-associated neuronal and memory impairment in this model system. The findings are discussed in view of accumulating evidence suggesting involvement of analogous mechanisms in the etiology of age-associated dysfunction and disease of the human and mammalian brain. PMID:25538730

  4. Larval settlement and spat recovery rates of the oyster Crassostrea brasiliana (Lamarck, 1819) using different systems to induce metamorphosis.

    PubMed

    Silveira, R C; Silva, F C; Gomes, C H M; Ferreira, J F; Melo, C M R

    2011-05-01

    This study aimed at the assessment, in the laboratory, of the larval settlement and spat recovery rates of oysters of the species Crassostrea brasiliana using plastic collectors, epinephrine (C9H13NO3 C4H6O6) and shell powder in settlement tanks. Polypropylene was used attached to bamboo frames. The material was chosen due to its pliability--that favours the spat detachment. Two experiments were carried out; the first between February and April 2008, and the second between November and December 2008 at the Marine Mussel Laboratory of Santa Catarina Federal University (Laboratório de Moluscos Marinhos da Universidade Federal de Santa Catarina). In the first experiment, the scratched plastic collectors were tested consorting them with shell powder; on the second, the plastic collectors were tested consorted with shell powder, only shell powder and epinephrine as the metamorphosis stimulator. The quantification was carried out of the larvae settled in the plastic collectors, and of the recovery and integrity of the spats after their detachment. The first experiment has shown a recovery rate of 48.83% of the spats in comparison with the D larvae used. From this percentage, 4.9% settled in the plastic collectors and 43.93% in shell powder. The second experiment revealed 55.78% regarding the settled spats in comparison with the total of larvae used (using epinephrine), 78.62% in the treatment with the collector plus shell powder and 58.33% in the treatment only with shell powder. Thus, the use of the collector plus shell powder resulted in a greater spat recovery when compared to the other treatments.

  5. Mercury net methylation in five tropical flood plain regions of Brazil: high in the root zone of floating macrophyte mats but low in surface sediments and flooded soils.

    PubMed

    Guimarães, J R; Meili, M; Hylander, L D; de Castro e Silva, E; Roulet, M; Mauro, J B; de Lemos, R

    2000-10-16

    In aquatic systems, bottom sediments have often been considered as the main methylmercury (MeHg) production site. In tropical floodplain areas, however, floating meadows and flooded forests extend over large areas and can be important Hg methylating sites. We present here a cross-system comparison of the Hg net methylation capacity in surface sediments, flooded soils and roots of floating aquatic macrophytes, assayed by in situ incubation with 203Hg and extraction of formed Me203 Hg by acid leaching and toluene. The presence of mono-MeHg was confirmed by thin layer chromatography and other techniques. Study areas included floodplain lakes in the Amazon basin (Tapajós, Negro and Amazon rivers), the Pantanal floodplain (Paraguay river basin), freshwater coastal lagoons in Rio de Janeiro and oxbow lakes in the Mogi-Guaçú river, São Paulo state. Different Hg levels were added in assays performed in 1994-1998, but great care was taken to standardise all other test parameters, to allow data comparisons. Net MeHg production was one order of magnitude higher (mean 13.8%, range 0.28-35) in the living or decomposing roots of floating or rooted macrophyte mats (Eichhornia azurea, E. crassipes, Paspalum sp., Eleocharis sellowiana, Salvinia sp., S. rotundifolia and Scirpus cubensis) than in the surface layer of underlying lake sediments (mean 0.6%, range 0.022-2.5). Methylation in flooded soils presented a wide range and was in some cases similar to the one found in macrophyte roots but usually much lower. In a Tapajós floodplain lake, natural concentrations of MeHg in soil and sediment cores taken along a lake-forest transect agreed well with data on net methylation potentials in the same samples. E. azurea, E. crassipes and Salvinia presented the highest methylation potentials, up to 113 times higher than in sediments. Methylation in E. azurea from six lakes of the Paraguay and Cuiabá rivers, high Pantanal, was determined in the 1998 dry and wet seasons and ranged from

  6. Habitat characteristics for different freshwater snail species as determined biologically through macroinvertebrate information.

    PubMed

    El-Khayat, Hanaa M M; Mahmoud, Kadria M A; Mostafa, Bayomy B; Tantawy, Ahmad A; El-Deeb, Fatma A; Ragb, Fawzy M; Ismail, Nahed M; El-Said, Kalil M; Taleb, Hoda M Abu

    2011-12-01

    Macro-invertebrates including freshwater snails collected from 643 sites over 8 successive seasons among the River Nile, branches, main canals and certain drains in eight Egyptian Governorates. Thirteen snail species and one bivalve species were identified. The most distributed were Lanistus carinatus and Physa acuta while the most abundant were Cleopatra bulimoides and Physa acuta during the whole study. The sites that harbored each snail species in all the examined water-courses were grouped seasonally and their biological assessment was determined by their minimum and maximum total point similarity percentage to that of the corresponded reference site and mean of the total points. Habitats for most snail species attained minimum total point's similarity percentage less than 21% (very poor habitat) during autumn and winter then spring while during summer very poor habitat was harbored by only few snail species. P. acuta was the only survived snails in habitat which attained 0 as a minimum total point's similarity percentage during two seasons and L. carinatus and Succinea cleopatra during one season. With respect to medically important snails very poor sites constituted 23% of Biomphalaria alexandrina sites, 14% of Lymnaea natalensis and 9.4% of Bulinus truncatus sites. The studied macroinvertebrate matrices, total number of organisms, taxa richness, the Ephemeroptera, Plecoptera, and Trichoptera (EPT) index, ratio of EPT index to chironomidae, ratio of scraper to filtering collector, contribution of dominant macroinvertebrate major group, comparison revealed descending tolerances from B. alexanrina followed by L. natalensis then B. truncates, but Hilsenhoff Biotic Index (HBI) showed the same tolerance to organic pollution.

  7. Measuring nitric oxide in single neurons by capillary electrophoresis with laser-induced fluorescence: use of ascorbate oxidase in diaminofluorescein measurements.

    PubMed

    Kim, Won-Suk; Ye, Xiaoying; Rubakhin, Stanislav S; Sweedler, Jonathan V

    2006-03-15

    As a family of novel fluorescent indicators for nitric oxide (NO), the diaminofluoresceins (DAFs) have allowed real-time measurement of neuronal NO, an important gaseous neurotransmitter. However, the measurement of NO by the most commonly used NO sensor, 4,5-diaminofluorescein (DAF-2), is altered by two processes: the interaction of DAF-2 with intracellular dehydroascorbic acid (DHA) and the impact of ascorbic acid (AA) on the levels of N2O3, the intermediate product of the oxidation of NO that reacts with DAF-2. Similar AA/DHA effects are observed with other DAF probes, including DAF-FM and DAR-4M. To overcome these limitations, we use a specific enzymatic reaction to eliminate the confounding effect of AA on DAF quantitation of NO and then use capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection to distinguish the various reaction products. First, the enzyme ascorbate oxidase (AO) is used to catalyze the oxidation of AA to DHA. Next, CE-LIF separates the fluorescent products of the reaction of DAF-2 with NO and DHA. Control experiments, including standard mixtures and single neurons with added NO donor, successfully demonstrate the utility of this approach. This protocol is further tested with homogenates of the mouth area from the sea slug Aplysia californica, previously shown to be NO-positive, and individual nitric oxide synthase-containing buccal neurons from the freshwater snail, Lymnaea stagnalis. In each case, significant amounts of NO are detected. This AO DAF methodology is specific, effective, simple, and allows NO to be measured in single cells without detectable interference from other compounds.

  8. Triassic and Jurassic rocks at Currie, Nevada Preliminary paleontologic evidence

    SciTech Connect

    Johnson, E.A.; Dubiel, R.F.; Brouwers, E.M. ); Litwin, R.J. ); Ash, S.R. ); Good, S.C. )

    1993-04-01

    A sequence of continental rocks overlies the Lower Triassic Thaynes Formation in a poorly exposed syncline near Currie in northeastern NV. The authors recognize four lithostratigraphic units above the Thaynes near Currie and provide new paleontologic data. In ascending order, unit 1 (120 ft) consists of reddish-brown, very fine grained sandstone. Unit 2 (50 ft) consists of light-gray, trough cross-stratified, coarse-grained, conglomeratic sandstone. Unit 3 (at least 500 ft) consists of green, red, and brown sandstone and mudstone. Unit 4 occurs as isolated outcrops of reddish-orange, fine- to medium-grained sandstone. New fossil evidence, while not definitive, constrain the age of this sequence. Plant megafossils in unit 1 include (1) a specimen with narrow ovate leaves, possibly from an early Mesozoic conifer and (2) abundant fragments of probable Neocalamites. The presence of these fossils and the absence of any angiosperm leaves or wood fragments suggest an early Mesozoic age. Ostracodes in unit 3 are exclusively Darwinula sp., and their association with conchostracans in the absence of younger ostracodes suggests a Triassic age. Finally, two small outcrops, previously mapped as Triassic/Jurassic, contain the gastropods Pilidae indet. and Lymnaea sp., which resemble Late Cretaceous to Paleocene faunas. The sequence is similar to the nearest Lower Mesozoic section on the Colorado Plateau at Cove Fort, Utah, 165 miles to the southeast. The authors' new evidence supports the longstanding correlation of units 1--4 with the Lower Triassic Moenkopi Formation (part), the Shinarump and Petrified Forest Members of the Upper Triassic Chinle Formation, and the Lower Jurassic Navajo Sandstone of the Plateau. These rocks at Currie demonstrate that the Early Mesozoic depositional systems of the Colorado Plateau extended at least this far west and provide constraints on Early Mesozoic tectonism in the eastern Great Basin.

  9. Unusual snail species involved in the transmission of Fasciola hepatica in watercress beds in central France.

    PubMed

    Dreyfuss, G; Vignoles, P; Abrous, M; Rondelaud, D

    2002-06-01

    Four freshwater pulmonate species (Lymnaea ovata, L. stagnalis, Physa acuta, Planorbis leucostoma) were living in several watercress beds known for their relationships with human cases of fasciolosis, whereas L. truncatula was never found. The aims of these studies were to determine the prevalence of natural infections with Fasciola hepatica in snails and to verify if these species might ensure the full larval development of this trematode (with cercarial shedding) when they were experimentally subjected to F. hepatica only, or to co-infections with an other trematode species. Investigations were so carried out in six snail populations living in watercress beds (including three for P. acuta) and in four others originating from three brooks or a pond (as controls). Snails naturally infected with F. hepatica were found in two watercress beds inhabited by L. ovata (prevalence of infection: 1.4%) and P. leucostoma (0.1%), respectively. The L. ovata from the watercress bed could be infected at a higher size than those from the control population and the prevalence of this infection was greater in the bed population. Similar findings were noted for L. stagnalis. Despite single or dual infections, the results obtained with the four populations of P. acuta were unsuccessful. In contrast, the co-infections of young P. leucostoma with Paramphistomum daubneyi and F. hepatica resulted in the shedding of some F. hepatica cercariae. According to the authors, the occurrence of fasciolosis in these watercress beds would be the consequence of frequent natural encounters between parasite and snails (L. ovata, L. stagnalis), or of co-infections with P. daubneyi and F. hepatica (P. leucostoma). In watercress beds only colonized by P. acuta, a lymnaeid species would have ensured the larval development of F. hepatica but it would have been eliminated by P. acuta, as this last species was known to be invasive and could colonize open drainage ditches on siliceous soil.

  10. Fasciola hepatica and lymnaeid snails occurring at very high altitude in South America.

    PubMed

    Mas-Coma, S; Funatsu, I R; Bargues, M D

    2001-01-01

    Fascioliasis due to the digenean species Fasciola hepatica has recently proved to be an important public health problem, with human cases reported in countries of the five continents, including severe symptoms and pathology, with singular epidemiological characteristics, and presenting human endemic areas ranging from hypo- to hyperendemic. One of the singular epidemiological characteristics of human fascioliasis is the link of the hyperendemic areas to very high altitude regions, at least in South America. The Northern Bolivian Altiplano, located at very high altitude (3800-4100 m), presents the highest prevalences and intensities of human fascioliasis known. Sequences of the internal transcribed spacers ITS-1 and ITS-2 of the nuclear ribosomal DNA of Altiplanic Fasciola hepatica and the intermediate snail host Lymnaea truncatula suggest that both were recently introduced from Europe. Studies were undertaken to understand how the liver fluke and its lymnaeid snail host adapted to the extreme environmental conditions of the high altitude and succeeded in giving rise to high infection rates. In experimental infections of Altiplanic lymnaeids carried out with liver fluke isolates from Altiplanic sheep and cattle, the following aspects were studied: miracidium development inside the egg, infectivity of miracidia, prepatent period, shedding period, chronobiology of cercarial emergence, number of cercariae shed by individual snails, survival of molluscs at the beginning of the shedding process, survival of infected snails after the end of the shedding period and longevity of shedding and non-shedding snails. When comparing the development characteristics of European F. hepatica and L. truncatula, a longer cercarial shedding period and a higher cercarial production were observed, both aspects related to a greater survival capacity of the infected lymnaeid snails from the Altiplano. These differences would appear to favour transmission and may be interpreted as strategies

  11. Oxidative-stress induced increase in circulating fatty acids does not contribute to phospholipase A2-dependent appetitive long-term memory failure in the pond snail Lymnaeastagnalis

    PubMed Central

    2014-01-01

    Background Reactive oxygen species (ROS) are essential for normal physiological functioning of the brain. However, uncompensated increase in ROS levels may results in oxidative stress. Phospholipase A2 (PLA2) is one of the key players activated by elevated ROS levels resulting in the hydrolysis of various products from the plasmamembrane such as peroxidized fatty acids. Free fatty acids (FFAs) and fatty acid metabolites are often implicated to the genesis of cognitive impairment. Previously we have shown that age-, and experimentally induced oxidative stress causes PLA2-dependent long-term memory (LTM) failure in an aversive operant conditioning model in Lymnaea stagnalis. In the present study, we investigate the effects of experimentally induced oxidative stress and the role of elevated levels of circulating FFAs on LTM function using a non-aversive appetitive classical conditioning paradigm. Results We show that intracoelomic injection of exogenous PLA2 or pro-oxidant induced PLA2 activation negatively affects LTM performance in our learning paradigm. In addition, we show that experimental induction of oxidative stress causes significant temporal changes in circulating FFA levels. Importantly, the time of training coincides with the peak of this change in lipid metabolism. However, intracoelomic injection with exogenous arachidonic acid, one of the main FFAs released by PLA2, does not affect LTM function. Moreover, sequestrating circulating FFAs with the aid of bovine serum albumin does not rescue pro-oxidant induced appetitive LTM failure. Conclusions Our data substantiates previous evidence linking lipid peroxidation and PLA2 activation to age- and oxidative stress-related cognitive impairment, neuronal dysfunction and disease. In addition however, our data indicate that lipid peroxidation induced increased levels of circulating (per)oxidized FFAs are not a factor in oxidative stress induced LTM impairment. PMID:24886155

  12. Biotic variation in coastal water bodies in Sussex, England: Implications for saline lagoons

    NASA Astrophysics Data System (ADS)

    Joyce, Chris B.; Vina-Herbon, Cristina; Metcalfe, Daniel J.

    2005-12-01

    Coastal water bodies are a heterogeneous resource typified by high spatial and temporal variability and threatened by anthropogenic impacts. This includes saline lagoons, which support a specialist biota and are a priority habitat for nature conservation. This paper describes the biotic variation in coastal water bodies in Sussex, England, in order to characterise the distinctiveness of the saline lagoon community and elucidate environmental factors that determine its distribution. Twenty-eight coastal water bodies were surveyed for their aquatic flora and invertebrate fauna and a suite of exploratory environmental variables compiled. Ordination and cluster analyses were used to examine patterns in community composition and relate these to environmental parameters. Biotic variation in the coastal water body resource was high. Salinity was the main environmental parameter explaining the regional distribution of taxa; freshwater and saline assemblages were evident and related to sea water ingress. Freshwater sites were indicated by the plant Myriophyllum spicatum and gastropod mollusc Lymnaea peregra, while more saline communities supported marine and brackish water taxa, notably a range of chlorophytic algae and the bivalve mollusc Cerastoderma glaucum. Site community differences were also related to bank slope and parameters describing habitat heterogeneity. A saline lagoon community was discerned within the matrix of biotic variation consisting of specialist lagoonal species with associated typically euryhaline taxa. For fauna, the latter were the molluscs Abra tenuis and Hydrobia ulvae, and the crustaceans Corophium volutator and Palaemonetes varians, and for flora they were the algae Ulva lactuca, Chaetomorpha mediterranea, Cladophora spp. and Enteromorpha intestinalis. One non-native polychaete species, Ficopomatus enigmaticus, also strongly influenced community structure within the lagoonal resource. The community was not well defined as specialist and

  13. A flavonol present in cocoa [(-)epicatechin] enhances snail memory.

    PubMed

    Fruson, Lee; Dalesman, Sarah; Lukowiak, Ken

    2012-10-15

    Dietary consumption of flavonoids (plant phytochemicals) may improve memory and neuro-cognitive performance, though the mechanism is poorly understood. Previous work has assessed cognitive effects in vertebrates; here we assess the suitability of Lymnaea stagnalis as an invertebrate model to elucidate the effects of flavonoids on cognition. (-)Epicatechin (epi) is a flavonoid present in cocoa, green tea and red wine. We studied its effects on basic snail behaviours (aerial respiration and locomotion), long-term memory (LTM) formation and memory extinction of operantly conditioned aerial respiratory behaviour. We found no significant effect of epi exposure (15 mg l(-1)) on either locomotion or aerial respiration. However, when snails were operantly conditioned in epi for a single 0.5 h training session, which typically results in memory lasting ~3 h, they formed LTM lasting at least 24 h. Snails exposed to epi also showed significantly increased resistance to extinction, consistent with the hypothesis that epi induces a more persistent LTM. Thus training in epi facilitates LTM formation and results in a more persistent and stronger memory. Previous work has indicated that memory-enhancing stressors (predator kairomones and KCl) act via sensory input from the osphradium and are dependent on a serotonergic (5-HT) signalling pathway. Here we found that the effects of epi on LTM were independent of osphradial input and 5-HT, demonstrating that an alternative mechanism of memory enhancement exists in L. stagnalis. Our data are consistent with the notion that dietary sources of epi can improve cognitive abilities, and that L. stagnalis is a suitable model with which to elucidate neuronal mechanisms.

  14. From Polyplacophora to Cephalopoda: comparative analysis of nitric oxide signalling in mollusca.

    PubMed

    Moroz, L L; Gillette, R

    1995-01-01

    The distribution of putative nitric oxide synthase (NOS)-containing cells has been analysed using NADPH-d histochemistry in the CNS and peripheral tissues in more than 2D ecologically and systematically different molluscan genera representing 3 main classes of the phylum MOLLUSCA: Polyplacophora (Lepidopleurus, Lepidozona, Katharina), Gastropoda (Littorina, Lymnaea, Aplexa, Physa, Planorbarius, Planorbis, Helisoma, Biomphalaria, Helix, Limax, Cepaea, Bulla, Aplysia, Phyllaplysia, Philine, Pleurobranchea, Tritonia, Armina, Flabellina, Cadlina) and Cephalopoda (Octopus, Sepia, Rossia, Loligo). Several species were used for more detailed immunohistochemical, biochemical, biophysical and physiological studies to further assay of NOS activity and to analyse functional roles of nitric oxide (NO) in these animals. The main conclusions of our comparative analysis and literature survey can be summarised as following: (i) There is strong evidence for the presence of NO-dependent signalling pathways in different molluscan species. (ii) We hypothesise that a general tendency in the evolution of NADPH-d-reactive cells in Mollusca is a migration of nitrergic function from periphery to the CNS. Also, different isoforms of NOS appear to be present in any one species. (iii) One of the main functional targets of NO signalling is the feeding system. However, there are obvious differences between predators (many labelled central motoneurons) and herbivorous species (many labelled peripheral putative sensory cells) as well as between land/freshwater and marine animals. Nevertheless, in all species tested NO-activated feeding-like motor patterns in the buccal ganglia. Additional functional and cellular targets for NO in molluscs are also considered. We briefly review neuromodulatory mechanisms of NO action and we consider molluscs as useful model systems for investigations of the roles of NO.

  15. Development of a solid-phase receptor-based assay for the detection of cyclic imines using a microsphere-flow cytometry system.

    PubMed

    Rodríguez, Laura P; Vilariño, Natalia; Molgó, Jordi; Aráoz, Rómulo; Louzao, M Carmen; Taylor, Palmer; Talley, Todd; Botana, Luis M

    2013-02-19

    Biologically active macrocycles containing a cyclic imine were isolated for the first time from aquaculture sites in Nova Scotia, Canada, during the 1990s. These compounds display a "fast-acting" toxicity in the traditional mouse bioassay for lipophilic marine toxins. Our work aimed at developing a receptor-based detection method for spirolides using a microsphere/flow cytometry Luminex system. For the assay, two alternatives were considered as binding proteins, the Torpedo marmorata nicotinic acetylcholine receptor (nAChR) and the Lymnaea stagnalis acetylcholine binding protein (Ls-AChBP). A receptor-based inhibition assay was developed using the immobilization of nAChR or Ls-AChBP on the surface of carboxylated microspheres and the competition of cyclic imines with biotin-α-bungarotoxin (α-BTX) for binding to these proteins. The amount of biotin-α-BTX bound to the surface of the microspheres was quantified using phycoerythrin (PE)-labeled streptavidin, and the fluorescence was analyzed in a Luminex 200 system. AChBP and nAChR bound to 13-desmethyl spirolide C efficiently; however, the cross-reactivity profile of the nAChR for spirolides and gymnodimine more closely matched the relative toxic potencies reported for these toxins. The nAChR was selected for further assay development. A simple sample preparation protocol consisting of an extraction with acetone yielded a final extract with no matrix interference on the nAChR/microsphere-based assay for mussels, scallops, and clams. This cyclic imine detection method allowed the detection of 13-desmethyl spirolide C in the range of 10-6000 μg/kg of shellfish meat, displaying a higher sensitivity and wider dynamic range than other receptor-based assays previously published. This microsphere-based assay provides a rapid, sensitive, and easily performed screening method that could be multiplexed for the simultaneous detection of several marine toxins.

  16. Natural Compounds Interacting with Nicotinic Acetylcholine Receptors: From Low-Molecular Weight Ones to Peptides and Proteins

    PubMed Central

    Kudryavtsev, Denis; Shelukhina, Irina; Vulfius, Catherine; Makarieva, Tatyana; Stonik, Valentin; Zhmak, Maxim; Ivanov, Igor; Kasheverov, Igor; Utkin, Yuri; Tsetlin, Victor

    2015-01-01

    Nicotinic acetylcholine receptors (nAChRs) fulfill a variety of functions making identification and analysis of nAChR subtypes a challenging task. Traditional instruments for nAChR research are d-tubocurarine, snake venom protein α-bungarotoxin (α-Bgt), and α-conotoxins, neurotoxic peptides from Conus snails. Various new compounds of different structural classes also interacting with nAChRs have been recently identified. Among the low-molecular weight compounds are alkaloids pibocin, varacin and makaluvamines C and G. 6-Bromohypaphorine from the mollusk Hermissenda crassicornis does not bind to Torpedo nAChR but behaves as an agonist on human α7 nAChR. To get more selective α-conotoxins, computer modeling of their complexes with acetylcholine-binding proteins and distinct nAChRs was used. Several novel three-finger neurotoxins targeting nAChRs were described and α-Bgt inhibition of GABA-A receptors was discovered. Information on the mechanisms of nAChR interactions with the three-finger proteins of the Ly6 family was found. Snake venom phospholipases A2 were recently found to inhibit different nAChR subtypes. Blocking of nAChRs in Lymnaea stagnalis neurons was shown for venom C-type lectin-like proteins, appearing to be the largest molecules capable to interact with the receptor. A huge nAChR molecule sensible to conformational rearrangements accommodates diverse binding sites recognizable by structurally very different compounds. PMID:26008231

  17. Biodiversity of trematodes in their intermediate mollusc and fish hosts in the freshwater ecosystems of Europe.

    PubMed

    Faltýnková, Anna; Sures, Bernd; Kostadinova, Aneta

    2016-03-01

    We analysed two novel databases containing 2,380 and 8,202 host-parasite-locality records for trematode parasites of molluscs and fishes, respectively, to assess the biodiversity of trematodes in their intermediate mollusc and fish hosts in the freshwater environment in Europe. The "mollusc" dataset covers large numbers of pulmonate (29 spp.), "prosobranch" (15 spp.) and bivalve (11 spp.) molluscs acting as first intermediate hosts for 171 trematode species of 89 genera and 35 families. Of these, 23 and 40 species utilise freshwater fishes as definitive and second intermediate hosts, respectively. The most frequently recorded families are the Echinostomatidae Looss, 1899, Diplostomidae Poirier, 1886 and Schistosomatidae Stilles & Hassal, 1898, and the most frequently recorded species are Diplostomum spathaceum (Rudolphi, 1819), D. pseudospathaceum Niewiadomska, 1984 and Echinoparyphium recurvatum (von Linstow, 1873). Four snail species harbour extremely rich trematode faunas: Lymnaea stagnalis (L.) (41 spp.); Planorbis planorbis (L.) (39 spp.); Radix peregra (O.F. Müller) (33 spp.); and R. ovata (Draparnaud) (31 spp.). The "fish" dataset covers 99 fish species of 63 genera and 19 families acting as second intermediate hosts for 66 species of 33 genera and nine families. The most frequently recorded families are the Diplostomidae Poirier, 1886, Strigeidae Railliet, 1919 and Bucephalidae Poche, 1907, and the most frequently recorded species are Diplostomum spathaceum (Rudolphi, 1819), Tylodelphys clavata (von Nordmann, 1832) and Posthodiplostomum cuticola (von Nordmann, 1832). Four cyprinid fishes exhibit the highest species richness of larval trematodes: Rutilus rutilus (L.) (41 spp.); Abramis brama (L.) (34 spp.); Blicca bjoerkna (L.) (33 spp.); and Scardinius erythrophthalmus (L.) (33 spp.). Larval stages of 50 species reported in fish are also reported in freshwater molluscs, thus indicating a relatively good knowledge of the life-cycles of fish trematodes in

  18. Evidence of Fasciola hepatica infection in Radix peregra and a mollusc of the family Succineidae in Ireland.

    PubMed

    Relf, V; Good, B; McCarthy, E; de Waal, T

    2009-07-07

    Worldwide molluscs of the genera Lymnaea, Pseudosuccinea, Galba and Stagnicola act as intermediate hosts of the common liver fluke, Fasciola hepatica. Galba truncatula is the only recorded intermediate host of F. hepatica within Ireland and is the principle intermediate host within Europe. To investigate the transmission of liver fluke on the Teagasc hill sheep farm, Co. Mayo, Ireland, snail species other than G. truncatula, were collected from the lowland pasture (snail sp. 'A') in May (n=23) and from the hill area (snail sp. 'B') in March, April and May 2008 (n=78, 53 and 36 respectively). Morphological characteristics identified snail sp. 'A' as a Succinea sp. and snail sp. 'B' as Radix peregra. PCR and subsequent sequencing of the 18S gene and internal transcribed spacer 2 (ITS-2) region of the snail rDNA were used for molecular identification. The 18S ribosomal DNA (rDNA) sequences revealed a 99% similarity with Omalonyx matheroni (Succineidae) for snail sp. 'A', and for snail sp. 'B' a 99% and 98% similarity with Radix auricularia and R. peregra, respectively. The ITS-2 sequences of snail sp. 'B' revealed a 100% similarity with R. peregra. Using PCR F. hepatica DNA was identified in 73.9% (n=17) of snail sp. 'A' collected during May and 10.3% (n=8), 60.4% (n=32) and 61.1% (n=22) of snail sp. 'B' collected during March, April and May, respectively. A priori knowledge of habitat preference and environmental tolerances of G. truncatula has failed to explain the prevalence of infection recorded in the hill sheep flock on Teagasc hill sheep farm. These findings may explain the presence of infection in sheep populations grazing hill habitats considered too acidic to maintain G. truncatula populations.

  19. Exploration of the molecular architecture of the orthosteric binding site in the α4β2 nicotinic acetylcholine receptor with analogs of 3-(dimethylamino)butyl dimethylcarbamate (DMABC) and 1-(pyridin-3-yl)-1,4-diazepane.

    PubMed

    Bach, Tinna B; Jensen, Anders A; Petersen, Jette G; Sørensen, Troels E; Della Volpe, Serena; Liu, Jun; Blaazer, Antoni R; van Muijlwijk-Koezen, Jacqueline E; Balle, Thomas; Frølund, Bente

    2015-09-18

    X-ray crystal structures of acetylcholine binding proteins (AChBPs) have revealed two different possible extensions to the classical ligand binding pocket known to accommodate various nicotinic agonists. One of the pockets is limited in size while the other is of considerable dimensions and protrudes along the interfacial cleft between subunits. To probe these putative extensions in functional nicotinic acetylcholine receptors (nAChRs), elongated analogs of 3-(dimethylamino)butyl dimethylcarbamate (DMABC) and 1-(pyridine-3-yl)-1,4-diazepane were prepared and characterized pharmacologically at neuronal heteromeric nAChRs. Although the new analogs, relative to parent compounds, displayed lower binding affinities, functional characterization of selected compounds revealed that they had retained partial α4β2 nAChR agonist activity. The structure-activity relationship data did not indicate an upper limit to the size of substituents as would have been expected if the ligand was bound in the smaller pocket. The data were better in agreement with a binding mode in which substituents protrude along the interfacial cleft of the receptor. This was further supported by docking into a homology model of the α4-β2 nAChR interface and by surface plasmon resonance biosensor analysis of binding of the compounds to acetylcholine-binding proteins, where they exhibit preference for Lymnaea stagnalis ACh binding protein (Ls-AChBP) over the Aplysia california ACh binding protein (Ac-AChBP). These results suggest new opportunities for expanding chemical space in the development of partial agonist and may be of interest in relation to development of novel smoking cessation aids.

  20. Effect of cysteine and humic acids on bioavailability of Ag from Ag nanoparticles to a freshwater snail

    USGS Publications Warehouse

    Luoma, Samuel N.; Tasha Stoiber,; Croteau, Marie-Noele; Isabelle Romer,; Ruth Merrifeild,; Lead, Jamie

    2016-01-01

    Metal-based engineered nanoparticles (NPs) will undergo transformations that will affect their bioavailability, toxicity and ecological risk when released to the environment, including interactions with dissolved organic material. The purpose of this paper is to determine how interactions with two different types of organic material affect the bioavailability of silver nanoparticles (AgNPs). Silver uptake rates by the pond snail Lymnaea stagnalis were determined after exposure to 25 nmol l-1 of Ag as PVP AgNPs, PEG AgNPs or AgNO3, in the presence of either Suwannee River humic acid or cysteine, a high-affinity thiol-rich organic ligand. Total uptake rate of Ag from the two NPs was either increased or not strongly affected in the presence of 1 – 10 mg 1-1 humic acid. Humic substances contain relatively few strong ligands for Ag explaining their limited effects on Ag uptake rate. In contrast, Ag uptake rate was substantially reduced by cysteine. Three components of uptake from the AgNPs were quantified in the presence of cysteine using a biodynamic modeling approach: uptake of dissolved Ag released by the AgNPs, uptake of a polymer or large (>3kD) Ag-cysteine complex and uptake of the nanoparticle itself. Addition of 1:1 Ag:cysteine reduced concentrations of dissolved Ag, which contributed to, but did not fully explain the reductions in uptake. A bioavailable Ag-cysteine complex (> 3kD) appeared to be the dominant avenue of uptake from both PVP AgNPs and PEG AgNPs in the presence of cysteine. Quantifying the different avenues of uptake sets the stage for studies to assess toxicity unique to NPs.

  1. Costs of receipt and donation of ejaculates in a simultaneous hermaphrodite

    PubMed Central

    2010-01-01

    Background Sexual conflicts between mating partners can strongly impact the evolutionary trajectories of species. This impact is determined by the balance between the costs and benefits of mating. However, due to sex-specific costs it is unclear how costs compare between males and females. Simultaneous hermaphrodites offer a unique opportunity to determine such costs, since both genders are expressed concurrently. By limiting copulation of focal individuals in pairs of pond snails (Lymnaea stagnalis) to either the male role or the female role, we were able to compare the fecundity of single sex individuals with paired hermaphrodites and non-copulants. Additionally, we examined the investment in sperm and seminal fluid of donors towards feminized snails and hermaphrodites. Results Compared to non-mating focal snails, reciprocating individuals as well as male and female copulants experienced a significant fecundity reduction (~40%) after, on average, 3.07 ± 0.12 copulations in their allowed roles (for donors 2.98 ± 0.16 copulations and for recipients 3.14 ± 0.12 copulations). In a single copulation, significantly more sperm was donated to partners that were restricted to mating in the female role than to hermaphrodites, while seminal fluid transfer was unaffected by recipient type. Conclusions Our data indicate that the costs of mating in both sex functions are high in L. stagnalis. This conclusion is based on fecundity data collected separately for male and female copulants. Male mating costs result from investment in expensive ejaculates, composed of sperm and seminal fluid. For female copulants, fecundity reduction correlated with transferred sperm numbers in the first copulation, while differences in transferred quantities of seminal fluid were not detected. These findings may point toward a "sperm effect" as a novel feature of pond snail reproductive ecology. In conclusion, sex allocation and sexual conflict both contribute to decreased female fecundity in

  2. Infection success of Echinoparyphium aconiatum (Trematoda) in its snail host under high temperature: role of host resistance

    PubMed Central

    2014-01-01

    Background Extreme weather events such as summer heat waves become more frequent owing to global climate change and are predicted to alter disease dynamics. This is because high temperatures can reduce host immune function. Predicting the impact of climate change on host-parasite interactions is, however, difficult as temperature may also affect parasite infective stages and other host characteristics determining the outcome of interaction. Methods Two experiments were conducted to investigate these phenomena in a Lymnaea stagnalis–Echinoparyphium aconiatum (Trematoda) interaction. In the first experiment, the effects of exposure of snails to experimental heat waves [maintenance at 25°C vs. 15°C (control)] with different durations (3 days, 7 days) on the infection success of parasite cercariae was examined. In the second experiment, the infection success was examined under similar conditions, while controlling for the possible temperature effects on cercariae and at least partly also for host physiological changes that take place rapidly compared to alterations in immune function (exposure to cercariae at intermediate 20°C). Results In the first experiment, increased infection success at 25°C was found independently of the duration of the heat wave. In the second experiment, increased infection success was found only in snails maintained at 25°C for 7 days, a treatment in which snail immune defence is known to be impaired. Conclusions These results suggest that the effects of host resistance in determining overall parasite infection success can be overridden by effects of temperature on parasite transmission stages and/or alterations in other host traits than immune defence. PMID:24754889

  3. Ancient origin of somatic and visceral neurons

    PubMed Central

    2013-01-01

    Background A key to understanding the evolution of the nervous system on a large phylogenetic scale is the identification of homologous neuronal types. Here, we focus this search on the sensory and motor neurons of bilaterians, exploiting their well-defined molecular signatures in vertebrates. Sensorimotor circuits in vertebrates are of two types: somatic (that sense the environment and respond by shaping bodily motions) and visceral (that sense the interior milieu and respond by regulating vital functions). These circuits differ by a small set of largely dedicated transcriptional determinants: Brn3 is expressed in many somatic sensory neurons, first and second order (among which mechanoreceptors are uniquely marked by the Brn3+/Islet1+/Drgx+ signature), somatic motoneurons uniquely co-express Lhx3/4 and Mnx1, while the vast majority of neurons, sensory and motor, involved in respiration, blood circulation or digestion are molecularly defined by their expression and dependence on the pan-visceral determinant Phox2b. Results We explore the status of the sensorimotor transcriptional code of vertebrates in mollusks, a lophotrochozoa clade that provides a rich repertoire of physiologically identified neurons. In the gastropods Lymnaea stagnalis and Aplysia californica, we show that homologues of Brn3, Drgx, Islet1, Mnx1, Lhx3/4 and Phox2b differentially mark neurons with mechanoreceptive, locomotory and cardiorespiratory functions. Moreover, in the cephalopod Sepia officinalis, we show that Phox2 marks the stellate ganglion (in line with the respiratory — that is, visceral— ancestral role of the mantle, its target organ), while the anterior pedal ganglion, which controls the prehensile and locomotory arms, expresses Mnx. Conclusions Despite considerable divergence in overall neural architecture, a molecular underpinning for the functional allocation of neurons to interactions with the environment or to homeostasis was inherited from the urbilaterian ancestor by

  4. The Calmodulin-Binding, Short Linear Motif, NSCaTE Is Conserved in L-Type Channel Ancestors of Vertebrate Cav1.2 and Cav1.3 Channels

    PubMed Central

    Taiakina, Valentina; Boone, Adrienne N.; Fux, Julia; Senatore, Adriano; Weber-Adrian, Danielle

    2013-01-01

    NSCaTE is a short linear motif of (xWxxx(I or L)xxxx), composed of residues with a high helix-forming propensity within a mostly disordered N-terminus that is conserved in L-type calcium channels from protostome invertebrates to humans. NSCaTE is an optional, lower affinity and calcium-sensitive binding site for calmodulin (CaM) which competes for CaM binding with a more ancient, C-terminal IQ domain on L-type channels. CaM bound to N- and C- terminal tails serve as dual detectors to changing intracellular Ca2+ concentrations, promoting calcium-dependent inactivation of L-type calcium channels. NSCaTE is absent in some arthropod species, and is also lacking in vertebrate L-type isoforms, Cav1.1 and Cav1.4 channels. The pervasiveness of a methionine just downstream from NSCaTE suggests that L-type channels could generate alternative N-termini lacking NSCaTE through the choice of translational start sites. Long N-terminus with an NSCaTE motif in L-type calcium channel homolog LCav1 from pond snail Lymnaea stagnalis has a faster calcium-dependent inactivation than a shortened N-termini lacking NSCaTE. NSCaTE effects are present in low concentrations of internal buffer (0.5 mM EGTA), but disappears in high buffer conditions (10 mM EGTA). Snail and mammalian NSCaTE have an alpha-helical propensity upon binding Ca2+-CaM and can saturate both CaM N-terminal and C-terminal domains in the absence of a competing IQ motif. NSCaTE evolved in ancestors of the first animals with internal organs for promoting a more rapid, calcium-sensitive inactivation of L-type channels. PMID:23626724

  5. Structural basis for cooperative interactions of substituted 2-aminopyrimidines with the acetylcholine binding protein.

    PubMed

    Kaczanowska, Katarzyna; Harel, Michal; Radić, Zoran; Changeux, Jean-Pierre; Finn, M G; Taylor, Palmer

    2014-07-22

    The nicotinic acetylcholine receptor (nAChR) and the acetylcholine binding protein (AChBP) are pentameric oligomers in which binding sites for nicotinic agonists and competitive antagonists are found at selected subunit interfaces. The nAChR spontaneously exists in multiple conformations associated with its activation and desensitization steps, and conformations are selectively stabilized by binding of agonists and antagonists. In the nAChR, agonist binding and the associated conformational changes accompanying activation and desensitization are cooperative. AChBP, which lacks the transmembrane spanning and cytoplasmic domains, serves as a homology model of the extracellular domain of the nAChRs. We identified unique cooperative binding behavior of a number of 4,6-disubstituted 2-aminopyrimidines to Lymnaea AChBP, with different molecular variants exhibiting positive, nH > 1.0, and negative cooperativity, nH < 1.0. Therefore, for a distinctive set of ligands, the extracellular domain of a nAChR surrogate suffices to accommodate cooperative interactions. X-ray crystal structures of AChBP complexes with examples of each allowed the identification of structural features in the ligands that confer differences in cooperative behavior. Both sets of molecules bind at the agonist-antagonist site, as expected from their competition with epibatidine. An analysis of AChBP quaternary structure shows that cooperative ligand binding is associated with a blooming or flare conformation, a structural change not observed with the classical, noncooperative, nicotinic ligands. Positively and negatively cooperative ligands exhibited unique features in the detailed binding determinants and poses of the complexes.

  6. Role in the selectivity of neonicotinoids of insect-specific basic residues in loop D of the nicotinic acetylcholine receptor agonist binding site.

    PubMed

    Shimomura, Masaru; Yokota, Maiko; Ihara, Makoto; Akamatsu, Miki; Sattelle, David B; Matsuda, Kazuhiko

    2006-10-01

    The insecticide imidacloprid and structurally related neonicotinoids act selectively on insect nicotinic acetylcholine receptors (nAChRs). To investigate the mechanism of neonicotinoid selectivity, we have examined the effects of mutations to basic amino acid residues in loop D of the nAChR acetylcholine (ACh) binding site on the interactions with imidacloprid. The receptors investigated are the recombinant chicken alpha4beta2 nAChR and Drosophila melanogaster Dalpha2/chicken beta2 hybrid nAChR expressed in Xenopus laevis oocytes. Although mutations of Thr77 in loop D of the beta2 subunit resulted in a barely detectable effect on the imidacloprid concentration-response curve for the alpha4beta2 nAChR, T77R;E79V double mutations shifted the curve dramatically to higher affinity binding of imidacloprid. Likewise, T77K;E79R and T77N;E79R double mutations in the Dalpha2beta2 nAChR also resulted in a shift to a higher affinity for imidacloprid, which exceeded that observed for a single mutation of Thr77 to basic residues. By contrast, these double mutations scarcely influenced the ACh concentration-response curve, suggesting selective interactions with imidacloprid of the newly introduced basic residues. Computational, homology models of the agonist binding domain of the wild-type and mutant alpha4beta2 and Dalpha2beta2 nAChRs with imidacloprid bound were generated based on the crystal structures of acetylcholine binding proteins of Lymnaea stagnalis and Aplysia californica. The models indicate that the nitro group of imidacloprid interacts directly with the introduced basic residues at position 77, whereas those at position 79 either prevent or permit such interactions depending on their electrostatic properties, thereby explaining the observed functional changes resulting from site-directed mutagenesis.

  7. Multi-linear regression analysis, preliminary biotic ligand modeling, and cross species comparison of the effects of water chemistry on chronic lead toxicity in invertebrates.

    PubMed

    Esbaugh, A J; Brix, K V; Mager, E M; De Schamphelaere, K; Grosell, M

    2012-03-01

    The current study examined the chronic toxicity of lead (Pb) to three invertebrate species: the cladoceran Ceriodaphnia dubia, the snail Lymnaea stagnalis and the rotifer Philodina rapida. The test media consisted of natural waters from across North America, varying in pertinent water chemistry parameters including dissolved organic carbon (DOC), calcium, pH and total CO(2). Chronic toxicity was assessed using reproductive endpoints for C. dubia and P. rapida while growth was assessed for L. stagnalis, with chronic toxicity varying markedly according to water chemistry. A multi-linear regression (MLR) approach was used to identify the relative importance of individual water chemistry components in predicting chronic Pb toxicity for each species. DOC was an integral component of MLR models for C. dubia and L. stagnalis, but surprisingly had no predictive impact on chronic Pb toxicity for P. rapida. Furthermore, sodium and total CO(2) were also identified as important factors affecting C. dubia toxicity; no other factors were predictive for L. stagnalis. The Pb toxicity of P. rapida was predicted by calcium and pH. The predictive power of the C. dubia and L. stagnalis MLR models was generally similar to that of the current C. dubia BLM, with R(2) values of 0.55 and 0.82 for the respective MLR models, compared to 0.45 and 0.79 for the respective BLMs. In contrast the BLM poorly predicted P. rapida toxicity (R(2)=0.19), as compared to the MLR (R(2)=0.92). The cross species variability in the effects of water chemistry, especially with respect to rotifers, suggests that cross species modeling of invertebrate chronic Pb toxicity using a C. dubia model may not always be appropriate.

  8. Determining metal assimilation efficiency in aquatic invertebrates using enriched stable metal isotope tracers

    USGS Publications Warehouse

    Croteau, M.-N.; Luoma, S.N.; Pellet, B.

    2007-01-01

    We employ a novel approach that combines pulse-chase feeding and multi-labelled stable isotopes to determine gut passage time (GPT), gut retention time (GRT), food ingestion rate (IR) and assimilation efficiency (AE) of three trace elements for a freshwater gastropod. Lettuce isotopically enriched in 53Cr, 65Cu and 106Cd was fed for 2 h to Lymnaea stagnalis. The release of tracers in feces and water was monitored for 48 h, during which unlabelled lettuce was provided ad libidum. The first defecation of 53Cr occurred after 5 h of depuration (GPT), whereas 90% of the ingested 53Cr was recovered in the feces after 22.5 h of depuration (GRT). 53Chromium was not significantly accumulated in the soft tissues upon exposure. In contrast, 65Cu and 106Cd assimilation was detectable for most experimental snails, i.e., 65/63Cu and 106/114Cd ratios in exposed snails were higher than those for controls. Food IR during the labelled feeding phase was 0.16 ?? 0.07 g g-1 d-1. IR was inferred from the amount of 53Cr egested in the feces during depuration and the concentration of 53Cr in the labelled lettuce. Assimilation efficiencies (??95% CI) determined using mass balance calculations were 84 ?? 4% for Cu and 85 ?? 3% for Cd. The ratio method yields similar AE estimates. Expanding the application of this novel stable isotope tracer technique to other metals in a wide variety of species will provide unique opportunities to evaluate the interplay between digestive processes and dietary influx of metals. Understanding the biological processes that modulate dietborne metal uptake is crucial to assess the toxicity of dietborne metals. ?? 2007 Elsevier B.V. All rights reserved.

  9. Marine Natural Products Acting on the Acetylcholine-Binding Protein and Nicotinic Receptors: From Computer Modeling to Binding Studies and Electrophysiology

    PubMed Central

    Kudryavtsev, Denis; Makarieva, Tatyana; Utkina, Natalia; Santalova, Elena; Kryukova, Elena; Methfessel, Christoph; Tsetlin, Victor; Stonik, Valentin; Kasheverov, Igor

    2014-01-01

    For a small library of natural products from marine sponges and ascidians, in silico docking to the Lymnaea stagnalis acetylcholine-binding protein (AChBP), a model for the ligand-binding domains of nicotinic acetylcholine receptors (nAChRs), was carried out and the possibility of complex formation was revealed. It was further experimentally confirmed via competition with radioiodinated α-bungarotoxin ([125I]-αBgt) for binding to AChBP of the majority of analyzed compounds. Alkaloids pibocin, varacin and makaluvamines С and G had relatively high affinities (Ki 0.5–1.3 μM). With the muscle-type nAChR from Torpedo californica ray and human neuronal α7 nAChR, heterologously expressed in the GH4C1 cell line, no competition with [125I]-αBgt was detected in four compounds, while the rest showed an inhibition. Makaluvamines (Ki ~ 1.5 μM) were the most active compounds, but only makaluvamine G and crambescidine 359 revealed a weak selectivity towards muscle-type nAChR. Rhizochalin, aglycone of rhizochalin, pibocin, makaluvamine G, monanchocidin, crambescidine 359 and aaptamine showed inhibitory activities in electrophysiology experiments on the mouse muscle and human α7 nAChRs, expressed in Xenopus laevis oocytes. Thus, our results confirm the utility of the modeling studies on AChBPs in a search for natural compounds with cholinergic activity and demonstrate the presence of the latter in the analyzed marine biological sources. PMID:24686559

  10. Impact of toxic cyanobacteria on gastropods and microcystin accumulation in a eutrophic lake (Grand-Lieu, France) with special reference to Physa (= Physella) acuta.

    PubMed

    Lance, Emilie; Brient, Luc; Carpentier, Alexandre; Acou, Anthony; Marion, Loïc; Bormans, Myriam; Gérard, Claudia

    2010-08-01

    Hepatotoxic microcystins (MCs) produced by cyanobacteria are known to accumulate in gastropods following grazing of toxic cyanobacteria and/or absorption of MCs dissolved in water, with adverse effects on life history traits demonstrated in the laboratory. In the field, such effects may vary depending on species, according to their relative sensitivity and ecology. The aims of this study were to i) establish how various intensities of MC-producing cyanobacteria proliferations alter the structure of gastropod community and ii) compare MC tissue concentration in gastropods in the field with those obtained in our previous laboratory experiments on the prosobranch Potamopyrgus antipodarum and the pulmonate Lymnaea stagnalis. We explored these questions through a one-year field study at three stations at Grand-Lieu Lake (France) affected by different intensities of cyanobacteria proliferations. A survey of the community structure and MC content of both cyanobacteria and gastropods was associated with a caging experiment involving P. antipodarum and L. stagnalis. In total, 2592 gastropods belonging to 7 prosobranch and 16 pulmonate species were collected. However, distribution among the stations was unequal with 62% vs 2% of gastropods sampled respectively at the stations with the lowest vs highest concentrations of MC. Irrespective of the station, pulmonates were always more diverse, more abundant and occurred at higher frequencies than prosobranchs. Only the pulmonate Physa acuta occurred at all stations, with abundance and MC tissue concentration (< or = 4.32 microg g DW(-1)) depending on the degrees of MC-producing cyanobacteria proliferations in the stations; therefore, P. acuta is proposed as a potential sentinel species. The caging experiment demonstrated a higher MC accumulation in L. stagnalis (< or = 0.36 microg g DW(-1) for 71% of individuals) than in P. antipodarum (< or = 0.02 microg g DW(-1) for 12%), corroborating previous laboratory observations. Results

  11. Balanced Plasticity and Stability of the Electrical Properties of a Molluscan Modulatory Interneuron after Classical Conditioning: A Computational Study

    PubMed Central

    Vavoulis, Dimitris V.; Nikitin, Eugeny S.; Kemenes, Ildikó; Marra, Vincenzo; Feng, Jianfeng; Benjamin, Paul R.; Kemenes, György

    2010-01-01

    The Cerebral Giant Cells (CGCs) are a pair of identified modulatory interneurons in the Central Nervous System of the pond snail Lymnaea stagnalis with an important role in the expression of both unconditioned and conditioned feeding behavior. Following single-trial food-reward classical conditioning, the membrane potential of the CGCs becomes persistently depolarized. This depolarization contributes to the conditioned response by facilitating sensory cell to command neuron synapses, which results in the activation of the feeding network by the conditioned stimulus. Despite the depolarization of the membrane potential, which enables the CGGs to play a key role in learning-induced network plasticity, there is no persistent change in the tonic firing rate or shape of the action potentials, allowing these neurons to retain their normal network function in feeding. In order to understand the ionic mechanisms of this novel combination of plasticity and stability of intrinsic electrical properties, we first constructed and validated a Hodgkin-Huxley-type model of the CGCs. We then used this model to elucidate how learning-induced changes in a somal persistent sodium and a delayed rectifier potassium current lead to a persistent depolarization of the CGCs whilst maintaining their firing rate. Including in the model an additional increase in the conductance of a high-voltage-activated calcium current allowed the spike amplitude and spike duration also to be maintained after conditioning. We conclude therefore that a balanced increase in three identified conductances is sufficient to explain the electrophysiological changes found in the CGCs after classical conditioning. PMID:20485464

  12. Evidence for inflammation-mediated memory dysfunction in gastropods: putative PLA2 and COX inhibitors abolish long-term memory failure induced by systemic immune challenges

    PubMed Central

    2013-01-01

    Background Previous studies associate lipid peroxidation with long-term memory (LTM) failure in a gastropod model (Lymnaea stagnalis) of associative learning and memory. This process involves activation of Phospholipase A2 (PLA2), an enzyme mediating the release of fatty acids such as arachidonic acid that form the precursor for a variety of pro-inflammatory lipid metabolites. This study investigated the effect of biologically realistic challenges of L. stagnalis host defense response system on LTM function and potential involvement of PLA2, COX and LOX therein. Results Systemic immune challenges by means of β-glucan laminarin injections induced elevated H2O2 release from L. stagnalis circulatory immune cells within 3 hrs of treatment. This effect dissipated within 24 hrs after treatment. Laminarin exposure has no direct effect on neuronal activity. Laminarin injections disrupted LTM formation if training followed within 1 hr after injection but had no behavioural impact if training started 24 hrs after treatment. Intermediate term memory was not affected by laminarin injection. Chemosensory and motor functions underpinning the feeding response involved in this learning model were not affected by laminarin injection. Laminarin’s suppression of LTM induction was reversed by treatment with aristolochic acid, a PLA2 inhibitor, or indomethacin, a putative COX inhibitor, but not by treatment with nordihydro-guaiaretic acid, a putative LOX inhibitor. Conclusions A systemic immune challenge administered shortly before behavioural training impairs associative LTM function in our model that can be countered with putative inhibitors of PLA2 and COX, but not LOX. As such, this study establishes a mechanistic link between the state of activity of this gastropod’s innate immune system and higher order nervous system function. Our findings underwrite the rapidly expanding view of neuroinflammatory processes as a fundamental, evolutionary conserved cause of cognitive and

  13. Allelopathic potential and ecotoxicity evaluation of gallic and nonanoic acids to prevent cyanobacterial growth in lentic systems: A preliminary mesocosm study.

    PubMed

    Techer, Didier; Fontaine, Pascal; Personne, Aline; Viot, Sandrine; Thomas, Marielle

    2016-03-15

    The increase in anthropogenic nutrient loading affecting many freshwater ecosystems combined with global warming may lead to cyanobacterial blooms on an increasingly frequent basis. Among the various physicochemical and biological methods which have been proposed to rapidly control blue-green algae growth, the use of plant-derived substances such as allelochemicals has gained great interest as an environment-friendly approach. The primary aim of this work was to evaluate the efficiency of gallic and nonanoic acid application to preemptively inhibit cyanobacterial growth in lentic hydrosystems. In order to address the process feasibility under realistic exposure scenarios, thirteen outdoor freshwater mesocosms (unit volume: 3m(3)) were designed, each containing phytoplankton (including local blue-green algae species) and various non-target organisms from higher trophic levels (Physa, Lymnaea, Gammarus, and Scardinius erythrophthalmus). After an 8-week mesocosm stabilization period, a full factorial design based on the presence/absence of gallic acid (GA) and nonanoic acid (NA) (including a control group) was implemented into the exposure tanks. Regular monitoring of major phytoplankton taxa was conducted during a 28-day experiment using an on-line fluorometer. The main results suggested that gallic acid was more efficient than nonanoic acid at limiting cyanobacterial growth at concentrations as low as 1 mg L(-1). Successive gallic acid applications (at 1, 2 and 4 mg L(-1)) at the early stages of cyanobacterial growth did not allow the complete elimination of blue-green algae from the mesocosms. However, the specificity of the allelopathic effect of gallic acid towards cyanobacteria was compatible with the maintenance of a primary productivity in the treated tanks as indicated by the photoautotrophic growth of other algal taxa. Finally, no biomarker induction signal could be reported in non-target species. Further gallic acid application trials in lentic systems such

  14. Cloning and characterization of a novel G protein beta-subunit of pearl oyster (Pinctada fucata), and its interaction sites with calmodulin.

    PubMed

    Chen, Lei; Xie, Liping; Xiong, Xunhao; Dai, Yiping; Fan, Weimin; Zhang, Rongqing

    2005-10-01

    A cDNA clone encoding a novel G protein beta subunit of beta1 subclass, pfGbeta1 was isolated from the pearl oyster (Pinctada fucata). The deduced amino acid sequence of pfGbeta1 (341 amino acids) shares high homology to northern European squid (Loligo pealei) and great pond snail (Lymnaea stagnalis) pfGbeta1, while it has diverged from bovine (Bos taurus) and human. The well-conserved amino acid domains in G protein beta subunit, seven WD repeats, were founded in the deduced amino acid sequence. Alignment analysis showed that the beginning amino acid residues in variable fragment of the seventh WD motif are different from any other Gbeta. The prediction of 3D structure of pfGbeta showed that pfGbeta belongs to beta-propeller family proteins whose members contain 4-8 antiparallel beta-sheets resembling the blades of a propeller. In situ hybridization and Northern blotting analysis revealed that the pfGbeta mRNA hybridization signals were widely expressed in various tissues except muscle, with abundantly in epithelia of gill, gonad and outer fold of mantle. We also investigated the interactions between Gbetagamma and calmodulin (CaM), and specific amino acid residues that may be critical for the binding of Gbetagamma to CaM were also identified. Furthermore, the functional studies of the interaction showed that the binding of CaM and Gbetagamma increases the alkaline phosphatase (ALP) activity, an indicator for mineralization in MC3T3-E1 cells. The ALP activity of the mutants of pfGbetagamma that impaired the interactions of Gbetagamma with CaM is higher than the Control group; however, it is lower than the WTC group. Together, these results suggest that the Gbetagamma might interact with CaM and point to the important physiological function in modulating cellular functions.

  15. Metal-contaminated Sediment Effects on Biofilm Communities: Impairment of Multiple Stream Ecosystem Functions

    NASA Astrophysics Data System (ADS)

    Burton, G.; Costello, D.

    2012-12-01

    Photosynthetic biofilms are crucial drivers of many important stream ecosystem functions (e.g., primary and secondary production, N cycling), yet we have a limited understanding of how these critical communities respond to contaminated sediments. Divalent metals (e.g., Cu, Ni, Zn) are ubiquitous in urban streams and may be contributing to the decline in ecosystem function in urban waters. We exposed natural biofilm communities in five different streams to a common sediment amended with four concentrations of Ni and Cu. Contaminated sediments were placed into cups, covered with mesh disks for biofilm attachment, and secured to the streambed. After 6 weeks, biofilm-colonized disks were analyzed for net primary production (NPP), chlorophyll a, and metal content. Sediments below the biofilms were analyzed for total metals, acid volatile sulfide, and high-resolution vertical dissolved oxygen concentrations. Additional biofilm disks were separated from the sediment and fed to Lymnaea stagnalis to assess indirect effects of sediment metal on grazers. Among our five streams, we found variation in the biofilm response to metals with the most productive stream (Elm Creek) showing the strongest negative response to metal-contaminated sediment. Contaminated sediments in Elm Creek reduced biofilm growth, slowed primary production, and prevented penetration of oxygen into surface sediments. In the less productive streams, biofilms did not reduce NPP in the presence of sediment metal and there was still substantial penetration of oxygen into sediments; however, metals moved out of the sediment and accumulated in the biofilm. L. stagnalis exposed to metal-contaminated biofilms fed at a slower rate than those given clean biofilms. This study suggests that biofilms, and the biogeochemical cycles they drive, can potentially be impaired by contaminated sediment but the response is context dependent. Further, indirect dietary effects of contaminated sediment occur more widely than

  16. Structural mechanisms for α-conotoxin activity at the human α3β4 nicotinic acetylcholine receptor

    PubMed Central

    Abraham, Nikita; Healy, Michael; Ragnarsson, Lotten; Brust, Andreas; Alewood, Paul F.; Lewis, Richard J.

    2017-01-01

    Nicotinic acetylcholine receptors (nAChR) are therapeutic targets for a range of human diseases. α-Conotoxins are naturally occurring peptide antagonists of nAChRs that have been used as pharmacological probes and investigated as drug leads for nAChR related disorders. However, α-conotoxin interactions have been mostly characterised at the α7 and α3β2 nAChRs, with interactions at other subtypes poorly understood. This study provides novel structural insights into the molecular basis for α-conotoxin activity at α3β4 nAChR, a therapeutic target where subtype specific antagonists have potential to treat nicotine addiction and lung cancer. A co-crystal structure of α-conotoxin LsIA with Lymnaea stagnalis acetylcholine binding protein guided the design and functional characterisations of LsIA analogues that identified the minimum pharmacophore regulating α3β4 antagonism. Interactions of the LsIA R10F with β4 K57 and the conserved –NN– α-conotoxin motif with β4 I77 and I109 conferred α3β4 activity to the otherwise inactive LsIA. Using these structural insights, we designed LsIA analogues with α3β4 activity. This new understanding of the structural basis of protein-protein interactions between α-conotoxins and α3β4 may help rationally guide the development of α3β4 selective antagonists with therapeutic potential. PMID:28361878

  17. Isotopically modified silver nanoparticles to assess nanosilver bioavailability and toxicity at environmentally relevant exposures

    USGS Publications Warehouse

    Croteau, Marie-Noële; Dybowska, Agnieszka D.; Luoma, Samuel N.; Misra, Superb K.; Valsami-Jones, Eugenia

    2014-01-01

    A major challenge in understanding the environmental implications of nanotechnology lies in studying nanoparticle uptake in organisms at environmentally realistic exposure concentrations. Typically, high exposure concentrations are needed to trigger measurable effects and to detect accumulation above background. But application of tracer techniques can overcome these limitations. Here we synthesised, for the first time, citrate-coated Ag nanoparticles using Ag that was 99.7 % 109Ag. In addition to conducting reactivity and dissolution studies, we assessed the bioavailability and toxicity of these isotopically modified Ag nanoparticles (109Ag NPs) to a freshwater snail under conditions typical of nature. We showed that accumulation of 109Ag from 109Ag NPs is detectable in the tissues of Lymnaea stagnalis after 24-h exposure to aqueous concentrations as low as 6 ng L–1 as well as after 3 h of dietary exposure to concentrations as low as 0.07 μg g–1. Silver uptake from unlabelled Ag NPs would not have been detected under similar exposure conditions. Uptake rates of 109Ag from 109Ag NPs mixed with food or dispersed in water were largely linear over a wide range of concentrations. Particle dissolution was most important at low waterborne concentrations. We estimated that 70 % of the bioaccumulated 109Ag concentration in L. stagnalis at exposures –1 originated from the newly solubilised Ag. Above this concentration, we predicted that 80 % of the bioaccumulated 109Ag concentration originated from the 109Ag NPs. It was not clear if agglomeration had a major influence on uptake rates.

  18. Influence of hardness on the bioavailability of silver to a freshwater snail after waterborne exposure to silver nitrate and silver nanoparticles

    USGS Publications Warehouse

    Stoiber, Tasha L.; Croteau, Marie-Noele; Romer, Isabella; Tejamaya, Mila; Lead, Jamie R.; Luoma, Samuel N.

    2015-01-01

    The release of Ag nanoparticles (AgNPs) into the aquatic environment is likely, but the influence of water chemistry on their impacts and fate remains unclear. Here, we characterize the bioavailability of Ag from AgNO3 and from AgNPs capped with polyvinylpyrrolidone (PVP AgNP) and thiolated polyethylene glycol (PEG AgNP) in the freshwater snail, Lymnaea stagnalis, after short waterborne exposures. Results showed that water hardness, AgNP capping agents, and metal speciation affected the uptake rate of Ag from AgNPs. Comparison of the results from organisms of similar weight showed that water hardness affected the uptake of Ag from AgNPs, but not that from AgNO3. Transformation (dissolution and aggregation) of the AgNPs was also influenced by water hardness and the capping agent. Bioavailability of Ag from AgNPs was, in turn, correlated to these physical changes. Water hardness increased the aggregation of AgNPs, especially for PEG AgNPs, reducing the bioavailability of Ag from PEG AgNPs to a greater degree than from PVP AgNPs. Higher dissolved Ag concentrations were measured for the PVP AgNPs (15%) compared to PEG AgNPs (3%) in moderately hard water, enhancing Ag bioavailability of the former. Multiple drivers of bioavailability yielded differences in Ag influx between very hard and deionized water where the uptake rate constants (kuw, l g-1 d-1 ± SE) varied from 3.1 ± 0.7 to 0.2 ± 0.01 for PEG AgNPs and from 2.3 ± 0.02 to 1.3 ± 0.01 for PVP AgNPs. Modeling bioavailability of Ag from NPs revealed that Ag influx into L. stagnalis comprised uptake from the NPs themselves and from newly dissolved Ag.

  19. Predicting impacts of climate change on Fasciola hepatica risk.

    PubMed

    Fox, Naomi J; White, Piran C L; McClean, Colin J; Marion, Glenn; Evans, Andy; Hutchings, Michael R

    2011-01-10

    Fasciola hepatica (liver fluke) is a physically and economically devastating parasitic trematode whose rise in recent years has been attributed to climate change. Climate has an impact on the free-living stages of the parasite and its intermediate host Lymnaea truncatula, with the interactions between rainfall and temperature having the greatest influence on transmission efficacy. There have been a number of short term climate driven forecasts developed to predict the following season's infection risk, with the Ollerenshaw index being the most widely used. Through the synthesis of a modified Ollerenshaw index with the UKCP09 fine scale climate projection data we have developed long term seasonal risk forecasts up to 2070 at a 25 km square resolution. Additionally UKCIP gridded datasets at 5 km square resolution from 1970-2006 were used to highlight the climate-driven increase to date. The maps show unprecedented levels of future fasciolosis risk in parts of the UK, with risk of serious epidemics in Wales by 2050. The seasonal risk maps demonstrate the possible change in the timing of disease outbreaks due to increased risk from overwintering larvae. Despite an overall long term increase in all regions of the UK, spatio-temporal variation in risk levels is expected. Infection risk will reduce in some areas and fluctuate greatly in others with a predicted decrease in summer infection for parts of the UK due to restricted water availability. This forecast is the first approximation of the potential impacts of climate change on fasciolosis risk in the UK. It can be used as a basis for indicating where active disease surveillance should be targeted and where the development of improved mitigation or adaptation measures is likely to bring the greatest benefits.

  20. Structural and functional studies of the modulator NS9283 reveal agonist-like mechanism of action at α4β2 nicotinic acetylcholine receptors.

    PubMed

    Olsen, Jeppe A; Ahring, Philip K; Kastrup, Jette S; Gajhede, Michael; Balle, Thomas

    2014-09-05

    Modulation of Cys loop receptor ion channels is a proven drug discovery strategy, but many underlying mechanisms of the mode of action are poorly understood. We report the x-ray structure of the acetylcholine-binding protein from Lymnaea stagnalis with NS9283, a stoichiometry selective positive modulator that targets the α4-α4 interface of α4β2 nicotinic acetylcholine receptors (nAChRs). Together with homology modeling, mutational data, quantum mechanical calculations, and pharmacological studies on α4β2 nAChRs, the structure reveals a modulator binding mode that overlaps the α4-α4 interface agonist (acetylcholine)-binding site. Analysis of contacts to residues known to govern agonist binding and function suggests that modulation occurs by an agonist-like mechanism. Selectivity for α4-α4 over α4-β2 interfaces is determined mainly by steric restrictions from Val-136 on the β2-subunit and favorable interactions between NS9283 and His-142 at the complementary side of α4. In the concentration ranges where modulation is observed, its selectivity prevents NS9283 from directly activating nAChRs because activation requires coordinated action from more than one interface. However, we demonstrate that in a mutant receptor with one natural and two engineered α4-α4 interfaces, NS9283 is an agonist. Modulation via extracellular binding sites is well known for benzodiazepines acting at γ-aminobutyric acid type A receptors. Like NS9283, benzodiazepines increase the apparent agonist potency with a minimal effect on efficacy. The shared modulatory profile along with a binding site located in an extracellular subunit interface suggest that modulation via an agonist-like mechanism may be a common mechanism of action that potentially could apply to Cys loop receptors beyond the α4β2 nAChRs.

  1. Molecular Characterization and Functional Analysis of a Putative Octopamine/Tyramine Receptor during the Developmental Stages of the Pacific Oyster, Crassostrea gigas

    PubMed Central

    Ji, Peng; Xu, Fei; Huang, Baoyu; Li, Yingxiang; Li, Li; Zhang, Guofan

    2016-01-01

    Octopamine (OA) and its precursor, tyramine (TA), participate in invertebrate development such as growth, maturation, and reproduction by activating their corresponding G protein-coupled receptors (GPCRs). Although OA was first discovered in mollusks (octopus), subsequent studies on OA, TA and related receptors have primarily been conducted in Ecdysozoa, especially in insects. Accordingly, only limited reports on OA/TA receptors in mollusks are available and their physiological roles remain unclear. Here, a full-length cDNA encoding a putative 524 amino acid OA/TA receptor (CgGPR1) was isolated from the Pacific oyster Crassostrea gigas. CgGPR1 was most closely related to the Lymnaea stagnalis OA receptor OAR2 in sequence. Phylogenetic analysis showed that CgGPR1 belongs to a poorly studied subfamily of invertebrate OA/TA receptors. The spatio-temporal expression of CgGPR1 in C. gigas larvae was examined by quantitative real-time PCR and Western blot analysis. CgGPR1 was expressed during all developmental stages of C. gigas with higher levels at mid-developmental stages, indicating its potential role in embryogenesis and tissue differentiation. Immunoreactive fluorescence of CgGPR1 was mainly observed in the velum, foot, gill and mantle of C. gigas larvae. CgGPR1 transcripts were detected in all the tested organs of adult C. gigas, with highest level in the mantle. Pharmacological analysis showed that cAMP and Ca2+ concentrations remained unchanged in HEK293 cells expressing CgGPR1 upon addition of OA, TA or related amines, suggesting that CgGPR1 modulates other unknown molecules rather than cAMP and Ca2+. Our study sheds light on CgGPR1 function in oysters. PMID:27992549

  2. Deep mRNA Sequencing of the Tritonia diomedea Brain Transcriptome Provides Access to Gene Homologues for Neuronal Excitability, Synaptic Transmission and Peptidergic Signalling

    PubMed Central

    Senatore, Adriano; Edirisinghe, Neranjan; Katz, Paul S.

    2015-01-01

    Background The sea slug Tritonia diomedea (Mollusca, Gastropoda, Nudibranchia), has a simple and highly accessible nervous system, making it useful for studying neuronal and synaptic mechanisms underlying behavior. Although many important contributions have been made using Tritonia, until now, a lack of genetic information has impeded exploration at the molecular level. Results We performed Illumina sequencing of central nervous system mRNAs from Tritonia, generating 133.1 million 100 base pair, paired-end reads. De novo reconstruction of the RNA-Seq data yielded a total of 185,546 contigs, which partitioned into 123,154 non-redundant gene clusters (unigenes). BLAST comparison with RefSeq and Swiss-Prot protein databases, as well as mRNA data from other invertebrates (gastropod molluscs: Aplysia californica, Lymnaea stagnalis and Biomphalaria glabrata; cnidarian: Nematostella vectensis) revealed that up to 76,292 unigenes in the Tritonia transcriptome have putative homologues in other databases, 18,246 of which are below a more stringent E-value cut-off of 1x10-6. In silico prediction of secreted proteins from the Tritonia transcriptome shotgun assembly (TSA) produced a database of 579 unique sequences of secreted proteins, which also exhibited markedly higher expression levels compared to other genes in the TSA. Conclusions Our efforts greatly expand the availability of gene sequences available for Tritonia diomedea. We were able to extract full length protein sequences for most queried genes, including those involved in electrical excitability, synaptic vesicle release and neurotransmission, thus confirming that the transcriptome will serve as a useful tool for probing the molecular correlates of behavior in this species. We also generated a neurosecretome database that will serve as a useful tool for probing peptidergic signalling systems in the Tritonia brain. PMID:25719197

  3. The effect of pH on chronic aquatic nickel toxicity is dependent on the pH itself: Extending the chronic nickel bioavailability models.

    PubMed

    Nys, Charlotte; Janssen, Colin R; Van Sprang, Patrick; De Schamphelaere, Karel A C

    2016-05-01

    The environmental quality standard for Ni in the European Commission's Water Framework Directive is bioavailability based. Although some of the available chronic Ni bioavailability models are validated only for pH ≤ 8.2, a considerable fraction of European surface waters has a pH > 8.2. Therefore, the authors investigated the effect of a change in pH from 8.2 to 8.7 on chronic Ni toxicity in 3 invertebrate (Daphnia magna, Lymnaea stagnalis, and Brachionus calyciflorus) and 2 plant species (Pseudokirchneriella subcapitata and Lemna minor). Nickel toxicity was almost always significantly higher at pH 8.7 than at pH 8.2. To test whether the existing chronic Ni bioavailability models developed for pH ≤ 8.2 can be used at higher pH levels, Ni toxicity at pH 8.7 was predicted based on Ni toxicity observed at pH 8.2. This resulted in a consistent underestimation of toxicity. The results suggest that the effect of pH on Ni(2+) toxicity is dependent on the pH itself: the slope of the pH effect is steeper above than below pH 8.2 for species for which a species-specific bioavailability model exists. Therefore, the existing chronic Ni bioavailability models were modified to allow predictions of chronic Ni toxicity to invertebrates and plants in the pH range of 8.2 to 8.7 by applying a pH slope (SpH ) dependent on the pH of the target water. These modified Ni bioavailability models resulted in more accurate predictions of Ni toxicity to all 5 species (within 2-fold error), without the bias observed using the bioavailability models developed for pH ≤ 8.2. The results of the present study can decrease the uncertainty in implementing the bioavailability-based environmental quality standard under the Water Framework Directive for high-pH regions in Europe.

  4. Fasciolosis in cattle in Louisiana. II. Development of a system to use soil maps in a geographic information system to estimate disease risk on Louisiana coastal marsh rangeland.

    PubMed

    Zukowski, S H; Wilkerson, G W; Malone, J B

    1993-03-01

    A geographic information system (GIS) model of habitat for Lymnaea bulimoides, the snail intermediate host of Fasciola hepatica and the rumen fluke, Calicophoron microbothrioides, on the Chenier Plain of southwest Louisiana was revised to incorporate broad (greater than 100 m) chenier (relict beaches) along with adjacent marsh and transitional soils associated with spatial distribution of L. bulimoides habitat. The proportion of farmland comprised of soils of the GIS model coincided with actual habitat from a previous survey with a sensitivity of 91.3% and a specificity of 80.1%, and regressed significantly against the proportion of farmland comprised of L. bulimoides habitat (n = 12, P = 0.01, r2 = 0.50, slope = 0.015). A soil model index was calculated to incorporate (1) the proportion of farmland comprised of soils of the GIS model and (2) stocking rate. Fluke egg shedding indices (mean egg count multiplied by prevalence) were calculated for each fluke species on nine farms at four sampling times over a 2-year period. The maximum egg shedding indices for each farm, taken to indicate potential fluke transmission intensity, were correlated for the two fluke species on herds not recently treated for fasciolosis (n = 9, P = 0.004, r2 = 0.72, slope = 0.2), although at no one sampling period were shedding indices of the two fluke species significantly correlated. Egg shedding of C. microbothrioides by cattle correlated with the proportion of farmland comprised of soils included in the GIS model (n = 9, Spearman's rank coefficient was 0.7, P = 0.05). We conclude that (1) the maximum of several observations of the C. microbothrioides egg shedding index may be useful as a surrogate for F. hepatica in estimating risk from snail habitat on a farm when regular flukicide treatment interferes with F. hepatica egg shedding, and (2) the GIS model may estimate site-specific differences in fasciolosis risk to cattle operations in the Chenier Plain based on the association of

  5. The effect of organic and inorganic aqueous uranium speciation on U(VI) bioavailability to an aquatic invertebrate

    NASA Astrophysics Data System (ADS)

    Fuller, C.; Croteau, M. N.; Campbell, K. M.; Cain, D.; Aiken, G.

    2015-12-01

    Growing world-wide demand for uranium (U) as an energy source has raised concerns of the human and ecological risks of U extraction and processing in the United States. Because of limited information on the relationship between U speciation and bioavailability, particularly in aquatic animals, we are characterizing U uptake by a model freshwater invertebrate (the snail Lymnaea stagnalis). This species grazes on biofilms and is thus key in the trophic transfer of contaminants through aquatic food webs. We determined the bioavailability of dissolved U(VI) over a range of water hardness, pH (6 to 8), and the presence of dissolved natural organic matter (NOM) as a competing ligand, to test the effect of aqueous speciation on uptake. Bioavailability was assessed using U uptake rate constants (kuw) derived from a kinetic bioaccumulation model. Dissolved U (1 to 1000 nM) was bioavailable over the range of geochemical conditions tested with kuw (L/g/d) decreasing with increasing dissolved Ca and with increasing pH. For example, kuw decreased from 1.6 to 0.3 as dissolved Ca was increased from 0.04 to 1.5 mM, suggesting competition between bioavailable U(VI) species and strong ternary calcium uranyl carbonato complexes. At pH 7.5 in synthetic moderately hard freshwater, kuw decreased from 0.22 in the absence of NOM to 0.07 in the presence of a hydrophobic acid NOM isolate of high aromaticity (SUVA = 5) consistent with strong aqueous complexation of U(VI) by the NOM. The co-variance of U uptake and aqueous U species distribution is being analyzed to determine which U species are bioavailable. U speciation in systems with NOM is calculated using conditional U-NOM binding constants derived by equilibrium dialysis ligand exchange methodology. The bioavailability of dietborne U is being tested since dietary metal uptake prevails for many aquatic species. These experiments include addition of ferrihydrite with U sorbed, both in the presence and absence of NOM, and mixed with diet.

  6. Gene Splicing of an Invertebrate Beta Subunit (LCavβ) in the N-Terminal and HOOK Domains and Its Regulation of LCav1 and LCav2 Calcium Channels

    PubMed Central

    Dawson, Taylor F.; Boone, Adrienne N.; Senatore, Adriano; Piticaru, Joshua; Thiyagalingam, Shano; Jackson, Daniel; Davison, Angus; Spafford, J. David

    2014-01-01

    The accessory beta subunit (Cavβ) of calcium channels first appear in the same genome as Cav1 L-type calcium channels in single-celled coanoflagellates. The complexity of this relationship expanded in vertebrates to include four different possible Cavβ subunits (β1, β2, β3, β4) which associate with four Cav1 channel isoforms (Cav1.1 to Cav1.4) and three Cav2 channel isoforms (Cav2.1 to Cav2.3). Here we assess the fundamentally-shared features of the Cavβ subunit in an invertebrate model (pond snail Lymnaea stagnalis) that bears only three homologous genes: (LCav1, LCav2, and LCavβ). Invertebrate Cavβ subunits (in flatworms, snails, squid and honeybees) slow the inactivation kinetics of Cav2 channels, and they do so with variable N-termini and lacking the canonical palmitoylation residues of the vertebrate β2a subunit. Alternative splicing of exon 7 of the HOOK domain is a primary determinant of a slow inactivation kinetics imparted by the invertebrate LCavβ subunit. LCavβ will also slow the inactivation kinetics of LCav3 T-type channels, but this is likely not physiologically relevant in vivo. Variable N-termini have little influence on the voltage-dependent inactivation kinetics of differing invertebrate Cavβ subunits, but the expression pattern of N-terminal splice isoforms appears to be highly tissue specific. Molluscan LCavβ subunits have an N-terminal “A” isoform (coded by exons: 1a and 1b) that structurally resembles the muscle specific variant of vertebrate β1a subunit, and has a broad mRNA expression profile in brain, heart, muscle and glands. A more variable “B” N-terminus (exon 2) in the exon position of mammalian β3 and has a more brain-centric mRNA expression pattern. Lastly, we suggest that the facilitation of closed-state inactivation (e.g. observed in Cav2.2 and Cavβ3 subunit combinations) is a specialization in vertebrates, because neither snail subunit (LCav2 nor LCavβ) appears to be compatible with this observed property

  7. Influence of hardness on the bioavailability of silver to a freshwater snail after waterborne exposure to silver nitrate and silver nanoparticles.

    PubMed

    Stoiber, Tasha; Croteau, Marie-Noële; Römer, Isabella; Tejamaya, Mila; Lead, Jamie R; Luoma, Samuel N

    2015-01-01

    The release of Ag nanoparticles (AgNPs) into the aquatic environment is likely, but the influence of water chemistry on their impacts and fate remains unclear. Here, we characterize the bioavailability of Ag from AgNO(3) and from AgNPs capped with polyvinylpyrrolidone (PVP AgNP) and thiolated polyethylene glycol (PEG AgNP) in the freshwater snail, Lymnaea stagnalis, after short waterborne exposures. Results showed that water hardness, AgNP capping agents, and metal speciation affected the uptake rate of Ag from AgNPs. Comparison of the results from organisms of similar weight showed that water hardness affected the uptake of Ag from AgNPs, but not that from AgNO(3). Transformation (dissolution and aggregation) of the AgNPs was also influenced by water hardness and the capping agent. Bioavailability of Ag from AgNPs was, in turn, correlated to these physical changes. Water hardness increased the aggregation of AgNPs, especially for PEG AgNPs, reducing the bioavailability of Ag from PEG AgNPs to a greater degree than from PVP AgNPs. Higher dissolved Ag concentrations were measured for the PVP AgNPs (15%) compared to PEG AgNPs (3%) in moderately hard water, enhancing Ag bioavailability of the former. Multiple drivers of bioavailability yielded differences in Ag influx between very hard and deionized water where the uptake rate constants (k(uw), l g(-1) d(-1) ± SE) varied from 3.1 ± 0.7 to 0.2 ± 0.01 for PEG AgNPs and from 2.3 ± 0.02 to 1.3 ± 0.01 for PVP AgNPs. Modeling bioavailability of Ag from NPs revealed that Ag influx into L. stagnalis comprised uptake from the NPs themselves and from newly dissolved Ag.

  8. Discovery, synthesis, biological evaluation and structure-based optimization of novel piperidine derivatives as acetylcholine-binding protein ligands

    PubMed Central

    Shen, Jian; Yang, Xi-cheng; Yu, Ming-cheng; Xiao, Li; Zhang, Xun-jie; Sun, Hui-jiao; Chen, Hao; Pan, Guan-xin; Yan, Yu-rong; Wang, Si-chen; Li, Wei; Zhou, Lu; Xie, Qiong; Yu, Lin-qian; Wang, Yong-hui; Shao, Li-ming

    2017-01-01

    The homomeric α7 nicotinic receptor (α7 nAChR) is widely expressed in the human brain that could be activated to suppress neuroinflammation, oxidative stress and neuropathic pain. Consequently, a number of α7 nAChR agonists have entered clinical trials as anti-Alzheimer's or anti-psychotic therapies. However, high-resolution crystal structure of the full-length α7 receptor is thus far unavailable. Since acetylcholine-binding protein (AChBP) from Lymnaea stagnalis is most closely related to the α-subunit of nAChRs, it has been used as a template for the N-terminal domain of α-subunit of nAChR to study the molecular recognition process of nAChR-ligand interactions, and to identify ligands with potential nAChR-like activities. Here we report the discovery and optimization of novel acetylcholine-binding protein ligands through screening, structure-activity relationships and structure-based design. We manually screened in-house CNS-biased compound library in vitro and identified compound 1, a piperidine derivative, as an initial hit with moderate binding affinity against AChBP (17.2% inhibition at 100 nmol/L). During the 1st round of optimization, with compound 2 (21.5% inhibition at 100 nmol/L) as the starting point, 13 piperidine derivatives with different aryl substitutions were synthesized and assayed in vitro. No apparent correlation was demonstrated between the binding affinities and the steric or electrostatic effects of aryl substitutions for most compounds, but compound 14 showed a higher affinity (Ki=105.6 nmol/L) than nicotine (Ki=777 nmol/L). During the 2nd round of optimization, we performed molecular modeling of the putative complex of compound 14 with AChBP, and compared it with the epibatidine-AChBP complex. The results suggested that a different piperidinyl substitution might confer a better fit for epibatidine as the reference compound. Thus, compound 15 was designed and identified as a highly affinitive acetylcholine-binding protein ligand. In

  9. The chronic toxicity of molybdate to freshwater organisms. I. Generating reliable effects data.

    PubMed

    De Schamphelaere, K A C; Stubblefield, W; Rodriguez, P; Vleminckx, K; Janssen, C R

    2010-10-15

    The European Union regulation on Registration, Evaluation, Authorization and Restriction of Chemical substances (REACH) (EC, 2006) requires the characterization of the chronic toxicity of many chemicals in the aquatic environment, including molybdate (MoO(4)(2-)). Our literature review on the ecotoxicity of molybdate revealed that a limited amount of reliable chronic no observed effect concentrations (NOECs) for the derivation of a predicted no-effect concentration (PNEC) existed. This paper presents the results of additional ecotoxicity experiments that were conducted in order to fulfill the requirements for the derivation of a PNEC by means of the scientifically most robust species sensitivity distribution (SSD) approach (also called the statistical extrapolation approach). Ten test species were chronically exposed to molybdate (added as sodium molybdate dihydrate, Na(2)MoO(4)·2H(2)O) according to internationally accepted standard testing guidelines or equivalent. The 10% effective concentrations (EC10, expressed as measured dissolved molybdenum) for the most sensitive endpoint per species were 62.8-105.6 (mg Mo)/L for Daphnia magna (21day-reproduction), 78.2 (mg Mo)/L for Ceriodaphnia dubia (7day-reproduction), 61.2-366.2 (mg Mo)/L for the green alga Pseudokirchneriella subcapitata (72h-growth rate), 193.6 (mg Mo)/L for the rotifer Brachionus calyciflorus (48h-population growth rate), 121.4 (mg Mo)/L for the midge Chironomus riparius (14day-growth), 211.3 (mg Mo)/L for the snail Lymnaea stagnalis (28day-growth rate), 115.9 (mg Mo)/L for the frog Xenopus laevis (4day-larval development), 241.5 (mg Mo)/L for the higher plant Lemna minor (7day-growth rate), 39.3 (mg Mo)/L for the fathead minnow Pimephales promelas (34day-dry weight/biomass), and 43.2 (mg Mo)/L for the rainbow trout Oncorhynchus mykiss (78day-biomass). These effect concentrations are in line with the few reliable data currently available in the open literature. The data presented in this study can

  10. Laboratory assessment of the molluscicidal and cercariacidal activities of Balanites aegyptiaca

    PubMed Central

    Molla, Eshetu; Giday, Mirutse; Erko, Berhanu

    2013-01-01

    Objective To assess the molluscicidal and cercariacidal activities of aqueous extracts of Balanites aegyptiaca (B. aegyptiaca) against Ethiopian Biomphalaria pfeifferi (B. pfeifferi), Lymnaea natalensis (L. natalensis) and Schistosoma mansoni (S. mansoni) cercariae. Methods Extracts of seeds, endocarp, mesocarp, and fruit of B. aegyptiaca were tested for their activities against adult B. pfeifferi and L. natalensis. The cercariacidal activity of the seeds of the plant was also evaluated against S. mansoni. Bioassays were carried out following the methods recommended by WHO. Snail mortalities were compared between each plant part and snail species, and LC50 and LC90 values for the plant parts tested were computed. The cercariacidal activity of the plant was assessed by exposing the mice to the cercariae pre-exposed to aqueous extract of B. aegyptiaca seeds. Results For the molluscicidal activities of seeds, endocarp, mesocarp and whole fruit, the LC50 values against B. pfeifferi were 56.32, 77.53, 65.51 and 66.63 mg/L, respectively, while the respective LC90 values were 77.70, 120.04, 89.50 and 97.55 mg/L. Similarly, the LC50 values for the seeds, endocarp, mesocarp and whole fruit against L. natalensis were 80.33, 92.61, 83.52 and 87.84 mg/L, respectively, while the respective LC90 values were 102.30, 138.21, 115.42 and 127.69 mg/L. B. pfeifferi were found to be more susceptible to B. aegyptiaca than L. natalensis. S. mansoni cercariae exposed to 15 mg/L of extract of seeds were incapable of infecting mice. The mean egg load of tissue was reduced in mice infected with the cercariae exposed to 5 and 10 mg/L of the extract. Conclusions The aqueous extracts of different parts of B. aegyptiaca exhibited reasonable molluscicidal activity against B. pfeifferi and L. natalensis, as well as cercariacidal activity against S. mansoni cercariae. However, comprehensive laboratory evaluation is recommended prior to field tests of the plant parts since their impact on other

  11. [Latin American malacology. Freshwater mollusks from Argentina].

    PubMed

    Rumi, Alejandra; Gregoric, Diego E Gutiérrez; Núñez, Verónica; Darrigran, Gustavo A

    2008-03-01

    . Species of sanitary interest are the propagators of: schistosomiasis -Biomphalaria peregrina, B. straminea y B. tenagophila, Planorbidae-, fasciolasis -Lymnaea viatrix and L. columnella, Lymnaeidae- and dermatitis -Chilina gibbosa and C. fluminea, Chilinidae. Invasive species are: Corbicula fluminea (Corbiculidae) and Limnoperna fortunei (Mytilidae). The construction of new areas for the protection and conservation of the high risk endemic species of freshwater molluscs is a priority. It is necessary to give special attention to the species of the patagonic mountain range and of the mesopotamic area of the Del Plata Basin.

  12. Radiation-induced cytogenetic and hematologic effects on aquatic biota within the Chernobyl exclusion zone.

    PubMed

    Gudkov, D I; Shevtsova, N L; Pomortseva, N A; Dzyubenko, E V; Kaglyan, A E; Nazarov, A B

    2016-01-01

    During 1998-2014 the rate of chromosomal aberrations in embryo tissues of the pond snail (Lymnaea stagnalis) and root meristems of higher aquatic plants, and also hematologic indexes of mantle liquid of the adult snails and peripheral blood of fishes in water bodies within the Chernobyl exclusion zone (EZ) was studied. The absorbed dose rate for hydrobionts from water bodies of the EZ registered in a range from 0.25 to 420 μGy h(-1) and in the reference ones - up to 0.09 μGy h(-1). The level of chromosomal aberrations in the molluscs from the most contaminated water bodies of the EZ was registered within range of 18-27% and for the molluscs from the reference lakes this index was on the average 1.5% with the maximal values 2.3%. The rate of chromosomal aberrations in root meristematic cells of higher aquatic plants from the contaminated lakes of the EZ was in range of 7-17% and in the plants from reference water bodies was not exceed 2.1%. The positive correlation between chromosomal aberration rate and absorbed dose rate in the pond snail's embryos and root meristems of higher aquatic plants in water bodies of the EZ was registered. Analysis of hemolymph structure of snails from the most contaminated water bodies showed a high rate of dead and phagocytic cells as well as decrease of the young amoebocytes quantity. Hematologic research of fish allows to determine on the one hand an insignificant changes of leukogram structure, and from the other hand a high level of red cells with different abnormalities in the peripheral blood of fishes from the water bodies with high levels of radioactive contamination. It is suppose that qualitative indexes of red cells in peripheral blood of fish are more sensitive to long-term radiation impact in comparison with elements of white blood, which can be used for conducting of the hematologic monitoring of radioactive contaminated water bodies.

  13. Acute sensitivity of the vernal pool fairy shrimp, Branchinecta lynchi (Anostraca; Branchinectidae), and surrogate species to 10 chemicals.

    PubMed

    Ivey, Chris D; Besser, John M; Ingersoll, Chris G; Wang, Ning; Rogers, D Christopher; Raimondo, Sandy; Bauer, Candice R; Hammer, Edward J

    2017-03-01

    Vernal pool fairy shrimp, Branchinecta lynchi, (Branchiopoda; Anostraca) and other fairy shrimp species have been listed as threatened or endangered under the US Endangered Species Act. Because few data exist about the sensitivity of Branchinecta spp. to toxic effects of contaminants, it is difficult to determine whether they are adequately protected by water quality criteria. A series of acute (24-h) lethality/immobilization tests was conducted with 3 species of fairy shrimp (B. lynchi, Branchinecta lindahli, and Thamnocephalus platyurus) and 10 chemicals with varying modes of toxic action: ammonia, potassium, chloride, sulfate, chromium(VI), copper, nickel, zinc, alachlor, and metolachlor. The same chemicals were tested in 48-h tests with other branchiopods (the cladocerans Daphnia magna and Ceriodaphnia dubia) and an amphipod (Hyalella azteca), and in 96-h tests with snails (Physa gyrina and Lymnaea stagnalis). Median effect concentrations (EC50s) for B. lynchi were strongly correlated (r(2 ) = 0.975) with EC50s for the commercially available fairy shrimp species T. platyurus for most chemicals tested. Comparison of EC50s for fairy shrimp and EC50s for invertebrate taxa tested concurrently and with other published toxicity data indicated that fairy shrimp were relatively sensitive to potassium and several trace metals compared with other invertebrate taxa, although cladocerans, amphipods, and mussels had similar broad toxicant sensitivity. Interspecies correlation estimation models for predicting toxicity to fairy shrimp from surrogate species indicated that models with cladocerans and freshwater mussels as surrogates produced the best predictions of the sensitivity of fairy shrimp to contaminants. The results of these studies indicate that fairy shrimp are relatively sensitive to a range of toxicants, but Endangered Species Act-listed fairy shrimp of the genus Branchinecta were not consistently more sensitive than other fairy shrimp taxa. Environ Toxicol

  14. Laboratory evaluation of molluscicidal & mosquito larvicidal activities of leaves of Solanum nigrum L.

    PubMed Central

    Rawani, Anjali; Ghosh, Anupam; Chandra, Goutam

    2014-01-01

    Background & objectives: Indiscriminate use of synthetic pesticides has created serious problem for the aquatic flora and fauna, and also resulted in appearance of pesticide resistance in vector population. This study was designed to evaluate the biocontrol efficacy of aqueous and solvent extracts of mature leaves of Solanum nigrum L., against fresh water snail Lymnaea acuminata f. rufescens (Gray) (an intermediate host of parasites causing fasciolopsiasis) and larvae of Culex vishnui group (Reuben) (vector of Japanese encephalitis). Methods: Aqueous and solvent extracts of fresh, mature, green/shed dried leaves of S. nigrum were tested against adult L. acuminata and larvae of Cx. vishnui group. The lethal concentration was determined and the appropriate lethal concentration at 24 h of benzene extract was also studied on non target organisms such as Daphnia sp, Diplonychus annulatum and Chironomus circumdatus. A qualitative phytochemical analysis was carried out in search of active ingredient and the chemical nature of the active substance was also evaluated by infrared (IR) analysis. Results: In a 72 h bioassay experiment with the aqueous extract, the highest mortality was recorded in 0.5 and 3 per cent extract against larvae of Cx. vishnui group and L. acuminata, respectivela. In the benzene solvent extract, the maximum mortality was recorded at a concentration of 150 ppm against L. acuminata and at 50 ppm against larvae of Cx. vishnui group with LC50 values of 55.45 and 11.59 ppm, respectively at 72 h. The log probit analysis (95% confidence level) recorded lowest value at 72 h of exposure. Qualitative phytochemical analysis reported the presence of some biochemical compounds, such as saponin, flavonoids, steroid and tannin. Among these, the toxic compound was detected by IR analysis having Rf = 0.87 (showed 66.70% and 76.70% mortality of L. acuminata and larvae of Cx. vishnui group, respectively). IR analysis provided preliminary information about the aliphatic

  15. Acute sensitivity of the vernal pool fairy shrimp, Branchinecta lynchi (Anostraca; Branchinectidae), and surrogate species to 10 chemicals

    USGS Publications Warehouse

    Ivey, Chris D.; Besser, John M.; Ingersoll, Christopher G.; Wang, Ning; Rogers, Christopher; Raimondo, Sandy; Bauer, Candice R.; Hammer, Edward J.

    2017-01-01

    Vernal pool fairy shrimp, Branchinecta lynchi, (Branchiopoda; Anostraca) and other fairy shrimp species have been listed as threatened or endangered under the US Endangered Species Act. Because few data exist about the sensitivity of Branchinecta spp. to toxic effects of contaminants, it is difficult to determine whether they are adequately protected by water quality criteria. A series of acute (24-h) lethality/immobilization tests was conducted with 3 species of fairy shrimp (B. lynchi, Branchinecta lindahli, and Thamnocephalus platyurus) and 10 chemicals with varying modes of toxic action: ammonia, potassium, chloride, sulfate, chromium(VI), copper, nickel, zinc, alachlor, and metolachlor. The same chemicals were tested in 48-h tests with other branchiopods (the cladocerans Daphnia magna and Ceriodaphnia dubia) and an amphipod (Hyalella azteca), and in 96-h tests with snails (Physa gyrina and Lymnaea stagnalis). Median effect concentrations (EC50s) for B. lynchi were strongly correlated (r2 = 0.975) with EC50s for the commercially available fairy shrimp species T. platyurus for most chemicals tested. Comparison of EC50s for fairy shrimp and EC50s for invertebrate taxa tested concurrently and with other published toxicity data indicated that fairy shrimp were relatively sensitive to potassium and several trace metals compared with other invertebrate taxa, although cladocerans, amphipods, and mussels had similar broad toxicant sensitivity. Interspecies correlation estimation models for predicting toxicity to fairy shrimp from surrogate species indicated that models with cladocerans and freshwater mussels as surrogates produced the best predictions of the sensitivity of fairy shrimp to contaminants. The results of these studies indicate that fairy shrimp are relatively sensitive to a range of toxicants, but Endangered Species Act-listed fairy shrimp of the genus Branchinecta were not consistently more sensitive than other fairy shrimp taxa. Environ Toxicol Chem

  16. Toxicity of lead (Pb) to freshwater green algae: development and validation of a bioavailability model and inter-species sensitivity comparison.

    PubMed

    De Schamphelaere, K A C; Nys, C; Janssen, C R

    2014-10-01

    model to reduce uncertainty in site-specific risk assessment. A model-based comparison with other species indicated that the sensitivity difference between P. subcapitata and two of the most chronically Pb-sensitive aquatic invertebrates (the crustacean Ceriodaphnia dubia and the snail Lymnaea stagnalis) is strongly pH dependent, with P. subcapitata becoming the most sensitive of the three at pH > 7.4. This indicates that inter-species differences in Pb bioavailability relationships should be accounted for in risk assessment and in the derivation of water quality criteria or environmental quality standards for Pb. The chronic toxicity data with three algae species and the bioavailability model presented here will help to provide a stronger scientific basis for evaluating ecological effects of Pb in the freshwater environment.