Communities of metazoan parasites in seven sympatric skate species (Elasmobranchii, Rajidae) from the English Channel and Celtic Sea differing in conservation status.

Elasmobranch populations are in steep decline mainly due to overfishing bycatch, but parasites may accelerate the collapse of vulnerable and/or highly parasitized species. We therefore studied metazoan parasites of Rajidae from the northeast Atlantic: vulnerable Leucoraja fullonica, near-threatened Raja brachyura, Raja clavata, Raja microocellata and Raja undulata, and least-concerned Raja montagui and Leucoraja naevus. Overall prevalence varied from 19% for R. montagui to 100% for L. fullonica. Parasite communities differed between skate species, and prevalence and abundance were higher for L. fullonica, R. microocellata, and R. undulata. We recorded 11 parasite taxa in the study: three nematodes, six cestodes, one monogenean, and one myxosporean. Whatever the skate species, the parasite component community comprised at least two nematode taxa among Phocanema spp., Proleptus sp. and Anisakis simplex. DNA-sequencing revealed that Phocanema azarasi and Phocanema krabbei both occurred in R. microocellata and R. undulata. Phocanema spp. was first recorded in L. fullonica, L. naevus, R. microocellata, R. montagui, and R. undulata, as Proleptus sp. in L. fullonica, and A. simplex in L. fullonica and R. clavata, Rockacestus sp. and Nybelinia sp. in R. undulata, and gill-myxosporeans on L. fullonica, L. naevus, R. microocellata, and R. undulata. The occurrence of 16 new host-parasite associations suggests potential environmental changes. Information provided by trophically transmitted helminths confirmed an opportunistic skate diet based on crustaceans and fish. We discuss results in terms of host fitness loss, bioindicator role of parasites, and anisakiasis risk. We recommend incorporating parasitology in research to improve elasmobranch conservation.

Metazoan parasite communities in Alosa alosa (Linnaeus, 1758) and Alosa fallax (Lacépède, 1803) (Clupeidae) from North-East Atlantic coastal waters and connected rivers.

Metazoan parasites were studied in 96 Alosa alosa and 78 Alosa fallax from North-East Atlantic coastal waters and connected rivers (among them three sympatric sites) in order to increase knowledge on these anadromous endangered fish and measure the parasitic impact on host condition. All shads were infected by one to six metazoan parasite taxa among the 12 identified in the whole sampling, with a mean abundance of parasites higher for A. alosa (167 ± 10) than for A. fallax (112 ± 11). Helminths, mostly trophically transmitted, were the best represented (eight taxa, prevalence up to 99%) in contrast with crustaceans and Petromyzontidae that rarely occurred (four taxa, prevalence <6%). Despite some quantitative differences, metazoan parasite communities of A. alosa and A. fallax remained stable in composition whatever the host developmental stage, sex, sample site, and salinity. Among the nine parasite taxa harbored by each Alosa species, six were shared with some differences in distribution patterns including in sympatric conditions, suggesting increasing dissimilarities between A. alosa and A. fallax with the age. Information on feeding ecology provided by trophically transmitted helminths confirmed euryphagous opportunistic diet of immatures and adults of both shad species, and assessed feeding of adults during spawning migrations. Our study also revealed the significant negative impact of Hemiurus appendiculatus on A. alosa and Pronoprymna ventricosa on A. fallax. Because helminth parasites are omnipresent in the shads and decrease their fitness, parasitological data must be included in further investigations and management programs on A. alosa and A. fallax.

Petromyzon marinus (Petromyzontidae), an unusual host for helminth parasites in western Europe.

The sea lamprey Petromyzon marinus, which is among the most phylogenetically ancient vertebrates, is a hematophagous ectoparasite that feeds on vertebrates and is considered vulnerable in Europe but is a pest in the North American Great Lakes. We conducted a literature review of helminth parasites of P. marinus and investigated postmetamorphic lampreys sampled in rivers and northeast Atlantic coastal waters (western France) during spawning migration. Based on the literature review, 16 helminth taxa have been recorded in P. marinus, among them 14 in North America but only 2 in Europe, with no species in common between these areas. Specific parasites are lacking, and helminth parasites recorded in P. marinus are mostly opportunistic and are trophically transmitted to fish hosts with both extremely low prevalence and mean intensity. Thus, P. marinus seems an unusual host that is probably infected through accidental ingestion of parasites by microphagous larvae (ammocoetes) and/or hematophagous postmetamorphs. Our field study supports this hypothesis, since only a single third-stage larva of Anisakis simplex sensu stricto was found in 2 postmetamorphic P. marinus among the 115 individuals dissected. This opportunistic, trophically transmitted, and cosmopolitan nematode species has never been recorded in North American sea lampreys and only once in Galician rivers (southern Europe). Infestation pathways of P. marinus by A. simplex are proposed vis-à-vis the feeding strategy of postmetamorphs and fish host species which potentially harbor anisakid larvae in their musculature. More generally, the complexity of biotic interactions is discussed considering P. marinus both as a host for helminth parasites and as a parasite for hosts such as fish and mammals, which are also potential predators of sea lamprey.

Land snails can trap trematode cercariae in their shell: Encapsulation as a general response against parasites?

Terrestrial gastropods are hosts of a wide variety of metazoan parasites and can respond to parasite exposure in various ways. One of these defence mechanisms, the ability to trap parasites in the host shell, was previously thought to apply only against nematodes. During a field survey along an urbanisation gradient, we found that the shell of Cornu aspersum and Cepaea nemoralis can contain encapsulated trematode cercariae, with prevalences of 7% and 1%, respectively over the entire sample, and up to 47% at the local population level. To our knowledge, this is the first case study unambiguously showing that land snails can trap non-nematode parasites in their shell at non-negligible prevalences. Shell-encapsulation could be a more general defence mechanism than previously described, and more studies are needed to understand its importance and variability.

Title: Les gastéropodes terrestres peuvent piéger les cercaires de trématodes dans leur coquille : l'encapsulation comme réponse générale contre les parasites ? Abstract: Les gastéropodes terrestres sont les hôtes d'une grande variété de métazoaires parasites, et peuvent répondre de plusieurs façons à l'exposition parasitaire. L'un de ces mécanismes de défense, la capacité à piéger le parasite dans la coquille, semblait ne concerner que les nématodes. Lors d'un suivi de terrain le long d'un gradient d'urbanisation, nous avons observé que la coquille de Cornu aspersum et de Cepaea nemoralis pouvait contenir des cercaires de trématodes encapsulées, à des prévalences respectives de 7 % et 1 % sur l'ensemble de l'échantillon, et pouvant atteindre 47 % au niveau d'une population locale. À notre connaissance, ceci est la première étude montrant sans ambiguïté que les escargots terrestres peuvent piéger de façon significative des parasites autres que les nématodes dans leur coquille. L'encapsulation des parasites dans la coquille pourrait donc être un mécanisme de défense plus général que précédemment décrit, plus d'études étant nécessaires pour comprendre son importance et sa variabilité.

Impact of microcystin-producing cyanobacteria on reproductive success of Lymnaea stagnalis (Gastropoda, Pulmonata) and predicted consequences at the population level.

Our previous studies showed that microcystin (MC)-accumulation in the gastropod Lymnaea stagnalis and effects on life-history traits (survival, growth, and fecundity) varied according to age, exposure pathway (MC-producing cyanobacteria or dissolved MC), and presence or not of additional non-toxic food. This study investigated effects of exposure to MC-producing cyanobacteria or to dissolved MC of parent and of parent and egg masses of L. stagnalis on hatching success, duration of embryonic development and neonate survival. Secondly, the potential impact of MC-producing cyanobacterial proliferations (blooms) on L. stagnalis population growth, depending on bloom frequencies and recovery duration of life traits after exposure, was evaluated using a modelling approach. Experimental results showed that embryonic development was shortened in case of parent exposure to toxic cyanobacteria. Parent and eggs exposure to dissolved MC extended embryonic development and reduced hatching percentage, suggesting a permeability of egg masses to MC. Whatever exposure, neonate survival was reduced. Neonates exposed to cyanobacteria accumulated MCs 24 h after hatching, suggesting very early cyanobacteria ingestion. Modelling results showed that L. stagnalis population growth was influenced by the recovery time of life-history traits after exposure. When setting the latest at 6 weeks according to previous experiments, a frequency of one to four blooms per year strongly affected population dynamics and induced up to a 80-weeks delay compared to control in time required for populations to grow from 1 to 1000 individuals. Results are discussed in terms of impact of intoxication pathways on parents, eggs and neonates, and on population dynamics of L. stagnalis.

Brachylaima spp. (Trematoda) parasitizing Cornu aspersum (Gastropoda) in France with potential risk of human consumption.

The edible land snail Cornu aspersum, native to the Mediterranean coastlines of North Africa, is widely distributed on most continents and often invasive in areas where introduction is recent. This species could contribute to the geographic spread of parasites as demonstrated for Brachylaima spp. These cosmopolitan trematodes may represent a threat to human health, like in Australia where Brachylaima cribbi infects humans. In this study, we demonstrate for the first time the occurrence of Brachylaima spp. in two French populations of C. aspersum, Thorigné-Fouillard (Ille-et-Vilaine), and Arçais (Deux-Sèvres), with an overall prevalence of 10.4% (Thorigné-Fouillard) and 73.3% (Arçais), respectively and a metacercarial intensity on average three times higher in Thorigné-Fouillard (37) than in Arçais (11). Cornu aspersum may act as a first and second intermediate host, as demonstrated in Arçais. The morphometrics of metacercariae, particularly the great body length about 2 mm, discriminate our Brachylaima species from those already described in C. aspersum (B. cribbi in Australia, and B. aspersae, B. llobregatensis and B. mascomai in Europe). Molecular analysis, based on 28S and COI, suggests the occurrence of two species in our study, one of which is probably Brachylaima mesostoma, an intestinal parasite of passeriform birds described in Central Europe. We underline the need for further research to identify species of Brachylaima in France and measure the health hazard of consuming field-collected snails.

TITLE: Brachylaima spp. (Trematoda) parasitant Cornu aspersum (Gastropoda) en France et risque potentiel pour la consommation humaine. ABSTRACT: L'escargot Cornu aspersum, originaire des côtes méditerranéennes d'Afrique du Nord, est largement répandu sur la plupart des continents et souvent invasif dans ses aires d'introduction récente. Cette espèce peut contribuer à l'expansion géographique de parasites comme démontré pour Brachylaima spp. Ces trématodes cosmopolites peuvent représenter une menace pour la santé comme en Australie où Brachylaima cribbi parasite l'espèce humaine. Dans cette étude, nous démontrons pour la première fois la présence de Brachylaima spp. dans deux populations françaises de C. aspersum, à Thorigné-Fouillard (Ille-et-Vilaine) et Arçais (Deux-Sèvres), avec une prévalence totale de 10.4 % (Thorigné-Fouillard) et 73.3 % (Arçais), et une intensité de métacercaires en moyenne trois fois plus importante à Thorigné-Fouillard (37) qu'à Arçais (11). Cornu aspersum peut jouer le rôle de premier et de second hôte intermédiaire, comme démontré à Arçais. La morphométrie des métacercaires, en particulier la grande taille corporelle d'environ 2 mm de long, suggèrent qu'il ne s'agit pas d'espèces de Brachylaima déjà décrites chez C. aspersum (B. cribbi en Australie et B. aspersae, B. llobregatensis et B. mascomai en Europe). L'analyse moléculaire, basée sur 28S et COI, suggère la présence de deux espèces dans notre étude, dont l'une est probablement Brachylaima mesostoma, un parasite intestinal d'oiseaux passeriformes décrit en Europe Centrale. Nous soulignons le besoin de recherche future pour identifier les espèces de Brachylaima présentes en France et mesurer le risque associé à la consommation d'escargots terrestres prélevés en milieu naturel.

Interactions between cyanobacteria and gastropods II. Impact of toxic Planktothrix agardhii on the life-history traits of Lymnaea stagnalis.

Hepatotoxins are frequently produced by many cyanobacterial species. Microcystins (MCs) are the most frequent and widely studied hepatotoxins, with potentially hazardous repercussions on aquatic organisms. As a ubiquitous herbivore living in eutrophic freshwaters, the snail Lymnaea stagnalis (Gastropoda: Pulmonata) is particularly exposed to cyanobacteria. The toxic filamentous Planktothrix agardhii is common in temperate lakes and is therefore, a potential food resource for gastropods. In the first part of this study, we demonstrated the ingestion of toxic P. agardhii by L. stagnalis during a 5 weeks exposure, with concomitant accumulation of, on average, 60% of total MCs ingested. After 3 weeks of non-toxic food (lettuce), approximately 90% of MCs were eliminated from tissues. Here, we investigate the impact of toxic P. agardhii consumption on the life-history traits (survival, growth and fecundity), locomotion and the structure of digestive and genital glands of juvenile and adult L. stagnalis. We observed a decrease of growth regardless of age, although this was more marked in juveniles, and a reduction of fecundity in adults. Survival and locomotion were not affected. Reduction of growth and fecundity continued to be observed even after feeding of non-toxic food for 3 weeks. The structure of the digestive gland was altered during the intoxication period but not irreversibly as cells tended to recover a normal status after the 3-week detoxification period. No histopathological changes occurred in the genital gland and oocytes, and spermatozoids were present in the gonadic acini. The density of cyanobacterial suspensions used in this study was comparable to those regularly observed in lakes, particularly in eutrophic waters. These results are discussed in terms of the negative impact of toxic cyanobacteria on natural communities of freshwater gastropods, and potential cascading effects on the equilibrium and functioning of the ecosystem.

Interactions between cyanobacteria and gastropods I. Ingestion of toxic Planktothrix agardhii by Lymnaea stagnalis and the kinetics of microcystin bioaccumulation and detoxification.

The last two decades have been marked by an increasing occurrence of toxic cyanobacterial blooms in aquatic ecosystems. These pose an expanding threat to the environment and to human health. Among the intracellular toxins produced by cyanobacteria, microcystins (hepatotoxins) are the most frequent and widely studied. As an ubiquitous herbivore living in eutrophic freshwaters, the freshwater snail Lymnaea stagnalis (Gastropoda: Pulmonata) is particularly exposed to cyanobacteria. The toxic filamentous Planktothrix agardhii is common in temperate lakes and is, therefore, a potential food resource for gastropods. We have studied the consumption of P. agardhii by L. stagnalis juveniles and adults in the presence or absence of non-toxic food (lettuce) over a 5-weeks period. Intoxication was followed by a 3-week detoxification period when snails were fed only on lettuce. The kinetics of microcystin accumulation and detoxification in the gastropods were established using the ELISA analytical method. The results showed an ingestion of toxic P. agardhii by L. stagnalis, even in the presence of lettuce, and the absence of food selection regardless of the age of the snails. Juveniles and adults consumed the same number of cells per millilitre and consumption was proportional to food availability. On average, 63% of cyanobacteria available were taken up during the first 24h. After 5 weeks of intoxication, 61% of the toxins present in the ingested cyanobacterial cells had accumulated in snail tissues (95% in the digestive-genital gland complex) with a concentration up to 80.4+/-4.9microggDW(-1). Toxin accumulation was greater in the gastropods fed on P. agardhii alone than those fed on the mixed diet, and was also greater in juveniles than in adults. After the removal of toxic cyanobacteria, detoxification was rapid: 64% of the toxins disappeared from snail tissues during the first week, but microcystins were still detected after 3 weeks (on average, 3.5+/-0.9microggDW(-1)). These results are discussed in terms of potential contamination to the food web.

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

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.

Variation in the response of the invasive species Potamopyrgus antipodarum (Smith) to natural (cyanobacterial toxin) and anthropogenic (herbicide atrazine) stressors.

In the context of increasing freshwater pollution, the impact on life-traits (survival, growth and fecundity) and locomotion of Potamopyrgus antipodarum of a 5-week field-concentration exposure to the cyanobacterial toxin microcystin-LR and the triazine herbicide, atrazine was studied. Whatever the age of exposed snails (juveniles, subadults, adults), microcystin-LR induced a decrease in survival, growth and fecundity but had no effect on locomotion. Atrazine induced a decrease in locomotory activity but had no significant effect on the life-traits. These results are discussed in terms of consequences to field populations.

The Clinical Learning Dyad Model: An Innovation in Midwifery Education.

There is a national shortage of women's health and primary care providers in the United States, including certified nurse-midwives and certified midwives. This shortage is directly related to how many students can be trained within the existing system. The current model of midwifery clinical training is based on apprenticeship, with one-on-one interaction between a student and preceptor. Thus, the number of newly trained midwifery providers is limited by the number of available and willing preceptors. The clinical learning dyad model (CLDM), which pairs 2 beginning midwifery students with one preceptor in a busy practice, addresses this problem. In addition, this model brings in a senior midwife student as a near-peer mentor when the students are first oriented into outpatient clinical practice. The model began as a pilot project to improve the quality of training and increase available student spots in clinical education. This article discusses the origins of the model, the specifics of its design, and the results of a midterm and one-year postintervention survey. Students and preceptors involved in this model identified several advantages to the program, including increased student accountability, enhanced socialization into the profession, improved learning, and reduced teaching burden on preceptors. An additional benefit of the CLDM is that students form a learning community and collaborate with preceptors to care for women in busy clinical settings. Challenges of the model will also be discussed. Further research is needed to evaluate the effectiveness of the CLDM. This article is part of a special series of articles that address midwifery innovations in clinical practice, education, interprofessional collaboration, health policy, and global health.

Abiotic stressors and stress responses: What commonalities appear between species across biological organization levels?

Organisms are regularly subjected to abiotic stressors related to increasing anthropogenic activities, including chemicals and climatic changes that induce major stresses. Based on various key taxa involved in ecosystem functioning (photosynthetic microorganisms, plants, invertebrates), we review how organisms respond and adapt to chemical- and temperature-induced stresses from molecular to population level. Using field-realistic studies, our integrative analysis aims to compare i) how molecular and physiological mechanisms related to protection, repair and energy allocation can impact life history traits of stressed organisms, and ii) to what extent trait responses influence individual and population responses. Common response mechanisms are evident at molecular and cellular scales but become rather difficult to define at higher levels due to evolutionary distance and environmental complexity. We provide new insights into the understanding of the impact of molecular and cellular responses on individual and population dynamics and assess the potential related effects on communities and ecosystem functioning.

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

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-1DW) 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.

Influence of introduced vs. native parasites on the body condition of migrant silver eels.

Because parasitism is among the reasons invoked to explain the collapse of Anguilla anguilla, we evaluated the parasitic constraint on body condition (BC) of migrant silver eels as a proxy of fitness with inter-site comparisons. Metazoan parasites were studied in 149 silver eels from five sites (northern Europe). In total, 89% were infected by 13 species including Myxozoa, Monogenea, Cestoda, Nematoda, and Acanthocephala. Anguillicoloides crassus was most common (56%), then Acanthocephalus clavula (30%), and Pseudodactylogyrus sp. (17%). BC, calculated for 58 females, was negatively correlated by abundance of the introduced Pseudodactylogyrus sp. but not by other parasite taxa. Nevertheless, the introduced A. crassus was considered as a severe pathogen based on previous data, whereas the native A. clavula was supposed to have limited impact. Parasite component communities and BC were different between sites. Silver eels from Stockholm Archipelago (Sweden) were the least parasitized (40% vs. 90-95% for other sites) with no parasites on the gills. Burrishoole (Ireland) differed by the absence of A. crassus and high prevalence of A. clavula (84%) but without consequences on BC. Gudenaa (Denmark), Corrib (Ireland), and Frémur (France) were close due to high prevalence of A. crassus (89-93%). Gudenaa and Corrib were the most similar because Pseudodactylogyrus sp. was also highly prevalent (respectively 71% and 60%) whereas absent in Frémur. Our results suggest that the fitness loss induced by the introduced parasites could affect the spawning success of migrant silver eels from Gudenaa and Corrib, and to a lesser extent from Frémur, but probably not those from Stockholm Archipelago and Burrishoole.

Accumulation of free and covalently bound microcystins in tissues of Lymnaea stagnalis (Gastropoda) following toxic cyanobacteria or dissolved microcystin-LR exposure.

Accumulation of free microcystins (MCs) in freshwater gastropods has been demonstrated but accumulation of MCs covalently bound to tissues has never been considered so far. Here, we follow the accumulation of total (free and bound) MCs in Lymnaea stagnalis exposed to i) dissolved MC-LR (33 and 100 microg L(-1)) and ii) Planktothrix agardhii suspensions producing 5 and 33 microg MC-LR equivalents L(-1) over a 5-week period, and after a 3-week depuration period. Snails exposed to dissolved MC-LR accumulated up to 0.26 microg total MCs g(-1) dry weight (DW), with no detection of bound MCs. Snails exposed to MCs producing P. agardhii accumulated up to 69.9 microg total MCs g(-1) DW, of which from 17.7 to 66.7% were bound. After depuration, up to 15.3 microg g(-1) DW of bound MCs were detected in snails previously exposed to toxic cyanobacteria, representing a potential source of MCs transfer through the food web.

Histopathology and microcystin distribution in Lymnaea stagnalis (Gastropoda) following toxic cyanobacterial or dissolved microcystin-LR exposure.

The accumulation of hepatotoxic microcystins (MCs) in gastropods has been demonstrated to be higher following grazing of toxic cyanobacteria than from MCs dissolved in ambient water. Previous studies, however, did not adequately consider MCs covalently bound to protein phosphatases, which may represent a considerably part of the MC body burden. Thus, using an immunohistochemical method, we examined and compared the histopathology and organ distribution of covalently bound MCs in Lymnaea stagnalis following a 5-week exposure to (i) dmMC-LR, dmMC-RR, and MC-YR-producing Planktothrix agardhii (5 microg MC-LReqL(-1)) and (ii) dissolved MC-LR (33 and 100 microgL(-1)). A subsequent 3-week depuration investigated potential MC elimination and tissue regeneration. Following both exposures, bound MCs were primarily observed in the digestive gland and tract of L. stagnalis. Snails exposed to toxic cyanobacteria showed severe and widespread necrotic changes in the digestive gland co-occurring with a pronounced cytoplasmic presence of MCs in digestive cells and in the lumen of digestive lobules. Snails exposed to dissolved MC-LR showed moderate and negligible pathological changes of the digestive gland co-occurring with a restrained presence of MCs in the apical membrane of digestive cells and in the lumen of digestive lobules. These results confirm lower uptake of dissolved MC-LR and correspondingly lower cytotoxicity in the digestive gland of L. stagnalis. In contrast, after ingestion of MC-containing cyanobacterial filaments, the most likely longer residual time within the digestive gland and/or the MC variant involved (e.g., MC-YR) allowed for increased MC uptake, consequently a higher MC burden in situ and thus a more pronounced ensuing pathology. While no pathological changes were observed in kidney, foot and the genital gland, MCs were detected in spermatozoids and oocytes of all exposed snails, most likely involving a hemolymph transport from the digestive system to the genital gland. The latter results indicate the potential for adverse impact of MCs on gastropod health and reproduction as well as the possible transfer of MCs to higher trophic levels of the food web.

Impact of toxic cyanobacteria on gastropods and microcystin accumulation in a eutrophic lake (Grand-Lieu, France) with special reference to Physa (= Physella) acuta.

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 are discussed in terms of differential gastropod sensitivity and MC transfer through the food web.

Influence of toxic cyanobacteria on community structure and microcystin accumulation of freshwater molluscs.

Community structure and microcystin accumulation of freshwater molluscs were studied before and after cyanobacterial proliferations, in order to assess the impact of toxic blooms on molluscs and the risk of microcystin transfer in food web. Observed decrease in mollusc abundance and changes in species richness in highly contaminated waters were not significant; however, relative abundances of taxa (prosobranchs, pulmonates, bivalves) were significantly different before and after cyanobacterial bloom. Pulmonates constituted the dominant taxon, and bivalves never occurred after bloom. Microcystin accumulation was significantly higher in molluscs from highly (versus lowly) contaminated waters, in adults (versus juveniles) and in pulmonates (versus prosobranchs and bivalves). Results are discussed according to the ecology of molluscs, their sensitivity and their ability to detoxify.

Variation in the response of juvenile and adult gastropods (Lymnaea stagnalis) to cyanobacterial toxin (microcystin-LR).

Owing to the increasing public health problem related to the proliferation of toxin-producing cyanobacteria in aquatic ecosystems, we have investigated the response of the pond snail Lymnaea stagnalis exposed to 33 microg/L microcystin-LR for 6 weeks, through its life traits (survival, growth, fecundity) and locomotion; uptake of microcystin-LR was also quantified in the snail body tissues. To study the potential plasticity of the response related to the development stage, snails were exposed to the toxin as sexually immature and mature. According to our results, microcystin-LR accumulated in snail tissues at a higher level in juveniles (7.96 ng/g fresh weight) versus adults (2.17 ng/g fresh weight). Whatever the age, survival, growth, and locomotion were not affected by the toxin, and fecundity of polluted adults was reduced by half. These results are discussed in terms of negative effects of aqueous microcystin-LR occurrence on the dynamics of natural populations of gastropods.

Spontaneous arousals in supine infants while swaddled and unswaddled during rapid eye movement and quiet sleep.

OBJECTIVE: Supine sleep is recommended for infants to decrease the risk of sudden infant death syndrome, but many parents report that their infants seem uncomfortable supine. Many cultures swaddle infants for sleep in the supine position. Swaddled infants are said to "sleep better"; presumably they sleep longer or with fewer arousals. However, there have been no studies of the effect of swaddling on spontaneous arousals during sleep. Arousal is initiated in brainstem centers and manifests as a sequence of reflexes: from sighs to startles and then to thrashing movements. Such "brainstem arousals" may progress to full arousal, but most do not. METHODS: Twenty-six healthy infants, aged 80 +/- 7 days, were studied during normal nap times. Swaddled (cotton spandex swaddle) and unswaddled trials were alternated for each infant. Sleep state (rapid eye movement [REM] or quiet sleep [QS]) was determined by behavioral criteria (breathing pattern, eye movements) and electroencephalogram/electrooculogram (10 infants). Respitrace, submental and biceps electromyogram, and video recording were used to detect startles and sighs (augmented breaths). Full arousals were classified by eye opening and/or crying. Frequencies of sighs, startles, and full arousals per hour were calculated. Progression of events was calculated as percentages in each sleep state, as was duration of sleep state. RESULTS: Swaddling decreased startles in QS and REM, full arousal in QS, and progression of startle to arousal in QS. It resulted in shorter arousal duration during REM sleep and more REM sleep. CONCLUSIONS: Swaddling has a significant inhibitory effect on progression of arousals from brainstem to full arousals involving the cortex in QS. Swaddling decreases spontaneous arousals in QS and increases the duration of REM sleep, perhaps by helping infants return to sleep spontaneously, which may limit parental intervention. For these reasons, a safe form of swaddling that allows hip flexion/abduction and chest wall excursion may help parents keep their infants in the supine sleep position and thereby prevent the sudden infant death syndrome risks associated with the prone sleep position.