Mapping freshwater snails in Angola: distribution, identity and molecular diversity of medically important taxa

Introduction - This study was designed to determine the presence and identity of potential intermediate snail hosts of schistosomiasis in Bengo, Luanda, Kwanza Norte and Malanje Provinces in NW Angola. This is an area where infection with Schistosoma haematobium, causing urogenital schistosomiasis, is common but little is known about transmission of the disease. Angola has had a varied past with regards to disease control and is revitalising efforts to combat Neglected Tropical Diseases.

3D-ultrastructure, functions and stress responses of rhogocytes from the freshwater gastropods Biomphalaria glabrata and Lymnaea stagnalis

Rhogocytes, also termed ‘pore cells’, exist free in the hemolymph or embedded in the connective tissue of different body parts of molluscs, notably gastropods. These unique cells can be round, elongated or irregularly shaped, and up to 30 μm in diameter. Their hallmark is the so-called slit apparatus: i.e. pocket-like invaginations of the plasma membrane creating extracellular lacunae, bridged by cytoplasmic bars. These bars form distinctive slits of ca. 20 nm width. A slit diaphragm composed of proteins establishes a molecular sieve with holes of 20 x 20 nm. Different functions have been assigned to this special molluscan cell type, notably biosynthesis of the hemolymph respiratory protein hemocyanin. It has further been proposed, but not proven, that in the case of red-blooded snail species rhogocytes might synthesize the hemoglobin. However, the secretion pathway of these hemolymph proteins, and the functional role of the enigmatic slit apparatus remained unclear. Additionally proposed functions of rhogocytes, such as heavy metal detoxification or hemolymph protein degradation, are also not well studied. This work provides more detailed electron microscopical, histological and immunobiochemical information on the structure and function of rhogocytes of the freshwater snails Biomphalaria glabrata and Lymnaea stagnalis. By in situ hybridization on mantle tissues, it proves that B. glabrata rhogocytes synthesize hemoglobin and L. stagnalis rhogocytes synthesize hemocyanin. Hemocyanin is present, in endoplasmic reticulum lacunae and in vesicles, as individual molecules or pseudo-crystalline arrays. The first 3D reconstructions of rhogocytes are provided by means of electron tomography and show unprecedented details of the slit apparatus. A highly dense material in the cytoplasmic bars close to the diaphragmatic slits was shown, by immunogold labeling, to contain actin. By immunofluorescence microscopy, the protein nephrin was localized at the periphery of rhogocytes. The presence of both proteins in the slit apparatus supports the previous hypothesis, hitherto solely based on similarities of the ultrastructure, that the molluscan rhogocytes are phylogenetically related to mammalian podocytes and insect nephrocytes. A possible secretion pathway of respiratory proteins that includes a transfer mechanism of vesicles through the diaphragmatic slits is proposed and discussed. We also studied, by electron microscopy, the reaction of rhogocytes in situ to two forms of animal stress: deprivation of food and cadmium contamination of the tank water. Significant cellular reactions to both stressors were observed and documented. Notably, the slit apparatus surface and the number of electron-dense cytoplasmic vesicles increased in response to cadmium stress. Food deprivation led to an increase in hemocyanin production. These observations are also discussed in the framework of using such animals as potential environmental biomarkers.

Ancylidae (Mollusca, Heterobranchia, Pulmonata, Basommatophora) do Estado do Rio de Janeiro, Brasil: morfologia, sistemática e distribuição geográfica

Os moluscos pateliformes de água doce da região Neotropical são comumente atribuídos a família Ancylidae sensu latum, abrangendo sete gêneros com pelo menos 13 espécies válidas e sete com identificação duvidosa. Os ancilídeos possuem pequenas dimensões, alcançam no máximo 15 mm de comprimento. Sua concha é frágil, composta por duas regiões, a protoconcha e a teleoconcha, as quais apresentam caracteres relevantes para a sistemática. Na parte mole as impressões musculares, a pigmentação do manto, o sistema reprodutor e a rádula são importantes para o estudo da família. Apesar de existirem vários registros de ocorrência para ancílideos no Estado do Rio de Janeiro (ERJ), existem poucos dados morfológicos. O principal objetivo deste estudo foi fornecer e ampliar as informações sobre a morfologia e distribuição geográfica das espécies de Ancylidae encontradas no ERJ. Os materiais utilizados foram procedentes de coletas próprias, material depositado em Coleções Científicas e dados de revisão bibliográfica. O estudo da morfologia comparada das conchas foi realizado com o auxílio de imagens de microscópio óptico e de varredura. Para a comparação das partes moles, os espécimes foram corados e dissecados sob lupa. Através da conquiliometria analisamos as diferenças inter e intrapopulacionais. Com este trabalho, a riqueza conhecida para Ancylidae no ERJ, aumentou de cinco para sete espécies: Burnupia sp., Ferrissia sp., Gundlachia radiata (Guilding, 1828),G. ticaga (Marcus & Marcus, 1962), Gundlachia sp., Hebetancylus moricandi (d`Orbigny, 1837) e Uncancylus concentricus (d`Orbigny, 1837). Gundlachia radiata e U. concentricus constituem novos registros para o ERJ, e G. radiata para a Região Sudeste. As três espécies com o maior número de registros de ocorrência no ERJ foram: G. ticaga (66%), Ferrissia sp. (37%) e Gundlachia sp. (18%). A ampla distribuição de G. ticaga pode ser devido à capacidade de suportar ambientes impactados. Em relação à morfologia, Burnupia sp. difere de Burnupia ingae Lanzer, 1991, única espécie deste gênero descrita para o Brasil, por apresentar diferenças na microescultura da concha e também na forma das impressões musculares. Ferrissia sp. difere de F. gentilis Lanzer, 1991, devido a diferenças na microescultura apical e número de cúspides no dente central da rádula. Gundlachia sp. é diferente de G. ticaga e G. radiata, por apresentar a abertura da concha mais arredondada, ápice mais recurvado, ultrapassando a borda da concha, pontuações irregulares em toda a protoconcha e forma dos músculos adutores anterior direito e posterior mais elíptica. A morfologia interna também mostra diferenças entre Gundlachia sp. e G. ticaga, como o apêndice terminal do útero e o número de folículos do ovoteste. Através das análises conquiliométricas, constatamos para os gêneros Burnupia, Ferrrissia e Gundlachia, que os índices morfométricos se mostraram melhores que as medidas lineares para a discriminação das espécies, provavelmente porque esses índices diminuem o efeito da amplitude de tamanho das conchas, que são fortemente influenciadas pelas variações ecofenotípicas. Contudo, os caracteres diagnósticos das conchas e das partes moles (impressões musculares) são indispensáveis para a identificação dos gêneros e espécies de Ancylidae. Palavras-chave: Mollusca. Moluscos de água doce. Morfologia. Distribuição geográfica. Estado do Rio de Janeiro.uscos.

Estudo temporal de longo prazo da comunidade de moluscos límnicos em um riacho da Vila do Abraão, Ilha Grande, Angra dos Reis, RJ, Brasil

Estudos de longo prazo são essenciais para avaliar efeitos que em pouco tempo não seriam possíveis observá-los. Moluscos límnicos são parte importante dos ecossistemas aquáticos, além se serem vetores de parasitos de importância médica. A comunidade de moluscos do riacho da Vila do Abraão já havia sido estudada, havendo registro de sete espécies, incluindo a exótica Melanoides tuberculata. O objetivo geral do trabalho foi acompanhar a dinâmica das populações de moluscos límnicos, com ênfase em M. tuberculata durante sete anos. Avaliamos as variações na comunidade de moluscos límnicos em um riacho na Vila do Abraão Ilha Grande, Angra dos Reis, RJ, Brasil. Foram realizadas 42 coletas bimestrais no período de julho/2006 a novembro/2013. O trecho estudado foi dividido em oito pontos de coleta, com três coletores em cada, realizando buscas de 15 minutos, totalizando 45 minutos por ponto. Foram mensurados os fatores abióticos: luminosidade, temperatura do ar e da água, umidade, pH, condutividade, profundidade e oxigênio dissolvido. Para o acompanhamento da comunidade, foi calculada a abundância, abundância relativa, constância, índices de diversidade Simpson e Shannon, assim como equitabilidade e uniformidade, para cada espécie em cada expedição de coleta. Para investigar a associação dos moluscos com a helmintofauna, realizou-se o teste de exposição a luz. Para biomassa, de M. tuberculata foi calculado o peso seco da parte mole, e os valores de abundância foram convertidos para densidade. No total do estudo, foram coletados 90.718 espécimes: oito gastrópodes (M. tuberculata, Heleobia australis, Potamolithus sp., Physa acuta, Biomphalaria tenagophila, Gundlachia ticaga, Ferrissia fragilis e Omalonyx matheroni) e um bivalve (Pisidium punctiferum). As duas espécies mais abundantes foram: M. tuberculata, com 80% e P. acuta, com 8% do total dos indivíduos. Melanoides tuberculata foi a espécie mais constante do estudo, já que após Fevereiro/2009 foi encontrada em todos os pontos de coleta, exceto nas últimas expedições. Os índices de diversidade apresentaram valores diferentes entre Simpson e Shannon, mas com variações semelhantes. A equitabilidade e a uniformidade foram muito baixas, indicando uma dominância de M. tuberculata. Apenas M. tuberculata se apresentou parasitado por Centrocestus formosanus, com a maior prevalência em Abril/2013, quando mais de 50% da população estava parasitada. A biomassa foi calculada em 8155 g durante o estudo, e a produção secundária foi estimada em 423 g m-2year-1 no último ano estudado, sofrendo diminuição de acordo com a queda populacional de M. tuberculata que por sua vez, foi possivelmente influenciada pelo parasitismo. Podemos concluir que a riqueza da comunidade aumentou de sete para nove espécies, sendo seis exóticas e três nativas. A comunidade de moluscos da Vila do Abraão encontra-se dominada por M. tuberculata, este sobrepujando as demais, em abundância e biomassa. Tendo em vista o perigo das introduções, reforçamos a importância dos estudos de longo prazo para o acompanhamento de comunidades, sendo importantes para subsidiar estratégias de conservação principalmente em unidades de conservação.

Molluscicidal activity of some marine substances against the snail Biomphalaria glabrata (Mollusca, Planorbidae)

Freshwater snails of the genus Biomphalaria play a major role as intermediate hosts of Schistosoma mansoni, the etiologic agent of schistosomiasis. While Biomphalaria spp. control by molluscicides is one of the main strategies to reduce the snail population in infected areas, there are few effective molluscicides commercially available. Natural products may be considered as potentially useful and safe molluscicides. We have evaluated the molluscicidal activity of 12 extracts from ten marine organisms on adult and embryonic stages of Biomphalaria glabrata. Only extracts of the red algae Liagora farinosa and of the sponge Amphimedon viridis presented molluscicidal activity. Lethal concentration (LC)(50) values obtained were 120 mu g/mL for L. farinosa CH2Cl2 extract (apolar fraction) and 20 mu g/mL for A. viridis extract and halitoxin. The polar alga fraction and halitoxin had no effect on B. glabrata embryos. The algae apolar fraction was active on B. glabrata in all embryonic development stages, with LC50 values for blastulae at 42 mu g/mL, gastrulae at 124 mu g/mL, trochophore at 180 mu g/mL, and veliger at 222 mu g/mL. This is the first report of extracts from marine organisms which presented molluscicidal activity.

A marked effect of water snails on small crustaceans. [Translation from: Anz.Ost.Akad.Wiss 10, 95-99, 1952. ]

Cases of mutual exclusion of two species of organisms in nature are known in large numbers. In the majority, they make different demands on the environ- ment which makes co-occurrence impossible. Less frequent are those cases in which a definite activity of one species prevents the occurrence of the other in the same region. An experiment was carried out n order to establish if Chydorus sphaericus can co-occur with water-snails. It emerged that a substance soluble in water which is given off by snails is responsible for the negative effect on small crustacea.

East African Freshwater Fisheries Research Organization Annual Report 1972

Feeding habits of many animals have been used widely in animal classifications. This is so, because the type of diet an organism requires demands structural specialisation which will utilise the available resource. Many animals may however have many structural modifications to enable them to be described as omnivourous or generalised feeders such as H. empodisma and H. riponianus (GREENWOOD 1960) which may show varying degrees of structural and adaptational intermediacy between two trophic groups. Generally, however, the diet of many animals including fish changes as the animal grow larger. The change in structural modifications is usually correlated with changes in the diet. In fishes the change may involve change from tricuspid to biscuspid and finally to unicuspid type of teeth. The degree of modification in the structure depends on the diet, thus Haplochromis that feeds on soft tissues of snails only requires modifications in oral dentition while Haplochromis that feeds on both soft tissues and shells of snails require modification in the lower pharyngeal bone for grinding purposes. Other modifications connected with food utilisation may be located in the alimentary canal. (I) The fish species that are commercially exploited are Protopterus aethiopicus, Clarias mossambicus, Tilapia esculenta, Tilapia amphimelas and Tilapia hybrids. The other fish species present in the lake but not commercially exploited are: Gnathonemus sp. Alestes sp. Labeo sp., Barbus paludinoses, Barbus jacksoni, Barbus lineomaculatus, Barbus regersi, Leptogrlanis sp., Schilbe sp., Haplochromis spp. and Hemihaplochromis sp. (2) Protopterus sp. and Clarias sp. are mostly caught with hooks on long lines. There has been a steady increase in number of hooks on the lake. Since the stocks of Protopterus and C/arias in the lake have a limit, we should control the number of hooks used by each of the fishermen in order to avoid overharvesting. (3) All the previous studies on Lake Kitangiri fisheries suggested the use of gill nets with mesh size greater than 88.9 mm in order to avoid the capture of immature Ti/apia spp. But if the fishermen are to obtain economic gains from the fishery, the optimum mesh size for use is 88.9 -101.6 mm. (4) The gillnet is a passive gear with very beneficial selective characteristics. Unfortunately the drive-in fishery which exists on Lake Kitangiri more or less destroys the gillnet selectivity characteristics. It is therefore recommended that the beating of water with poles be discouraged and stopped. (5) There is need for provision of stable fishing canoes to replace the unstable bottle palm dug-out canoes which are currently being used and which are very risky to operate. (6) The fish processing facilities on Lake Kitangiri are still inadequate. Most of the fish is sun dried, Since sun drying is very difficult during the rainy season, most fishermen carry out intensive fishing during the dry season, Concentrating most of the fishing effort in anyone season instead of spreading evenly this effort over the whole year could damage the age structure of the exploitable stocks. (7) There are considerable fluctuations in the volume of water of the lake. The feasibility of regulating the water loss through the effluent Sibiti river should be investigated by the Water Development Department. (8) Damming the Sibiti river is an expensive undertaking and therefore, the Rural Development Bank of Tanzania should be asked to assess the economic feasibility of such a project.

Bioaccumulation of the hepatotoxic microcystins in various organs of a freshwater snail from a subtropical Chinese lake, Taihu lake, with dense toxic microcystis blooms

In this paper, we describe the seasonal dynamics of three common microcystins (MCs MC-RR, MC-YR, and MC-LR) in the whole body, hepatopancreas, intestine, gonad, foot, remaining tissue, and offspring of a freshwater snail, Bellamya aeruginosa, from Gonghu Bay of Lake Taihu, China, where dense toxic Microcystis blooms occur in the warm seasons. Microcystins were determined by liquid chromatography electrospray ionization mass spectrum. Microcystin (MC-RR + MC-YR + MC-LR) content of the offspring and gonad showed high positive correlation, indicating that microcystins could transfer from adult females to their young with physiological connection. This study is the first to report the presence of microcystins in the offspring of the adult snail. The majority of the toxins were present in the intestine (53.6%) and hepatopancreas (29.9%), whereas other tissues contained only 16.5%. If intestines are excluded, up to 64.3% of the toxin burden was allocated in the hepatopancreas. The microcystin content in the intestine, hepatopancreas, and gonad were correlated with the biomass of Microcystis and intracellular and extracellular toxins. Of the analyzed foot samples, 18.2% were above the tolerable daily microcystin intake recommended by the World Health Organization (WHO) for human consumption. This result indicates that public health warnings regarding human ingestion of snails from Taihu Lake are warranted. In addition, further studies are needed to evaluate the occurrence by Microcystis in relation to spatial and temporal changes in water quality.

Effects of selection and drift on G matrix evolution in a heterogeneous environment: a multivariate Qst-Fst Test with the freshwater snail Galba truncatula.

Unraveling the effect of selection vs. drift on the evolution of quantitative traits is commonly achieved by one of two methods. Either one contrasts population differentiation estimates for genetic markers and quantitative traits (the Q(st)-F(st) contrast) or multivariate methods are used to study the covariance between sets of traits. In particular, many studies have focused on the genetic variance-covariance matrix (the G matrix). However, both drift and selection can cause changes in G. To understand their joint effects, we recently combined the two methods into a single test (accompanying article by Martin et al.), which we apply here to a network of 16 natural populations of the freshwater snail Galba truncatula. Using this new neutrality test, extended to hierarchical population structures, we studied the multivariate equivalent of the Q(st)-F(st) contrast for several life-history traits of G. truncatula. We found strong evidence of selection acting on multivariate phenotypes. Selection was homogeneous among populations within each habitat and heterogeneous between habitats. We found that the G matrices were relatively stable within each habitat, with proportionality between the among-populations (D) and the within-populations (G) covariance matrices. The effect of habitat heterogeneity is to break this proportionality because of selection for habitat-dependent optima. Individual-based simulations mimicking our empirical system confirmed that these patterns are expected under the selective regime inferred. We show that homogenizing selection can mimic some effect of drift on the G matrix (G and D almost proportional), but that incorporating information from molecular markers (multivariate Q(st)-F(st)) allows disentangling the two effects.

Establishment of the comet assay in the freshwater snail Biomphalaria glabrata (Say, 1818)

The single cell gel eletrophoresis or the comet assay was established in the freshwater snail Biomphalaria glabrata. For detecting DNA damage in circulating hemocytes, adult snails were irradiated with single doses of 2.5. 5, 10 and 20 Gy of Co-60 gamma radiation. Genotoxic effect of ionizing radiation was detected at all doses as a dose-related increase in DNA migration. Comet assay in B. glabrata demonstrated to be a simple, fast and reliable tool in the evaluation of genotoxic effects of environmental mutagens. (c) 2008 Elsevier B.V. All rights reserved.

Feeding ecology, density and biomass of the freshwater turtle, Phrynops geoffroanus, inhabiting a polluted urban river in south-eastern Brazil

This study examines the ecology of a population of Geoffroy's side-necked turtle Phrynops geoffroanus inhabiting a polluted urban river in Ribeirão Preto city, São Paulo state, south-eastern Brazil. Adult turtles fed mainly on Chironomidae larvae and pupae (Chironomus cf. plumosus, 100% of occurrence frequency) and domestic waste, but they also consumed terrestrial items (cockroach, snails) and carrion. Juvenile turtles showed more feeding diversity than the adults and exhibited a trend for predation on Chironomidae pupae, but this is not reflected in resource partitioning. The elevated number of turtles (170-230 turtles/ha of river) and biomass (255-345 kg/ha of river) inhabiting this urban river is probably the result of the abundance of sewage and organic waste produced by humans, the absence of predators, and increased availability of nesting areas. Such factors convert this area into an environment highly advantageous for the survival of Geoffroy's side-necked turtle.

New fossil pulmonate snails from the Paleocene of Itaborai Basin, Brazil (Pulmonata: Cerionidae, Strophocheilidae, Orthalicidae)

The limestones of ltaborai Basin (Middle Paleocene), Rio de Janeiro, Brazil, harbor a rich fossil fauna of pulmonate snails. Here two new pulmonate species are described: Brasilennea guttula sp. nov. (Cerionidae) and Eoborus rotundus sp. nov. (Strophocheilidae). B. guttula is the third species of its genus endemic from Itaborai, characterized mainly by its conspicuous shell shaped like a "water drop", with an acuminated spire. E. rotundus is the second of its genus from ltaborai, characterized mainly by its rounded outline and its relative small size. Moreover, a record of Plagiodontes aff. dental us (WOOD 1828) (Orthalicidae) is presented here for the first time for Itaborai Basin.

The interactive effects of predators, resources, and disturbance on freshwater snail populations from the Everglades

The origins of population dynamics depend on interplay between abiotic and biotic factors; the relative importance of each changing across space and time. Predation is a central feature of ecological communities that removes individuals (consumption) and alters prey traits (non-consumptive). Resource quality mitigates non-consumptive predator effects by stimulating growth and reproduction. Disturbance resets predator-prey interactions by removing both. I integrate experiments, time-series analysis, and performance trials to examine the relative importance of these on the population dynamics of a snail species by studying a variety of their traits. A review of ninety-three published articles revealed that snail abundance was much less in the Everglades and similar ecosystems compared to all other freshwater ecosystems considered. Separating consumptive from non-consumptive (cues) predator effects at different phosphorous levels with an experiment determined that phosphorous stimulated, but predator cues inhibited snail growth (34% vs. 23%), activity (38% vs. 53%), and reproductive effort (99% vs. 90%) compared to controls. Cues induced taller shells and smaller openings and moved to refugia where they reduced periphyton by 8%. Consumptive predator effects were minor in comparison. In a reciprocal transplant cage experiment along a predator cue and phosphorous gradient created by a canal, snails grew 10% faster and produced 37% more eggs far from the canal (fewer cues) when fed phosphorous-enriched periphyton from near the canal. Time-series analysis at four sites and predator performance trials reveal that phosphorous-enriched regions support larger snail populations, seasonal drying removes snails at all sites, crayfish negatively affect populations in enriched regions, and molluscivorous fish consume snails in the wet season. Combining these studies reveals interplay between resources, predators, and seasonality that limit snail populations in the Everglades and lead to their low abundance compared to other freshwater ecosystems. Resource quality is emerging as the critical factor because improving resources profoundly improved growth and reproduction; seasonal drying and predation become important at times and places. This work contributes to the general understanding in ecology of the relative importance of different factors that structure populations and provides evidence that bolsters monitoring efforts to assess the Comprehensive Everglades Restoration Plan that show phosphorous enrichment is a major driver of ecosystem change.

A molecular and stable isotopic approach to investigate the importance of algal and detrital energy pathways in a freshwater marsh

The relative importance of algal and detrital energy pathways remains a central question in wetlands ecology. We used bulk stable isotope analysis and fatty acid composition to investigate the relative contributions of periphyton (algae) and floc (detritus) in a freshwater wetland with the goal of determining the inputs of these resource pools to lower trophic-level consumers. All animal samples revealed fatty acid markers indicative of both microbial (detrital) and algal origins, though the relative contributions varied among species. Vascular plant markers were in low abundance in most consumers. Detritivory is important for chironomids and amphipods, as demonstrated by the enhanced bacterial fatty acids present in both consumers, while algal resources, in the form of periphyton, likely support ephemeropteran larvae. Invertebrates such as amphipods and grass shrimp appear to be important resources for small omnivorous fish, while Poecilia latipinna appear to strongly use periphyton and Ephemeroptera larvae as food sources. Both P. latipinna and Lepomis spp. assimilated small amounts of vascular plant debris, possibly due to unintentional ingestion of floc while foraging for invertebrates and insect larvae. Physid snails, Haitia spp., were characterized by considerably different fatty acid compositions than other taxa examined, and likely play a unique role in Everglades’ food webs.

Comparison of snail density, standing stock, and body size between Caribbean karst wetlands and other freshwater ecosystems

Synthesizing data from multiple studies generates hypotheses about factors that affect the distribution and abundance of species among ecosystems. Snails are dominant herbivores in many freshwater ecosystems, but there is no comprehensive review of snail density, standing stock, or body size among freshwater ecosystems. We compile data on snail density and standing stock, estimate body size with their quotient, and discuss the major pattern that emerges. We report data from 215 freshwater ecosystems taken from 88 studies that we placed into nine categories. Sixty-five studies reported density, seven reported standing stock, and 16 reported both. Despite the breadth of studies, spatial and temporal sampling scales were limited. Researchers used 25 different sampling devices ranging in area from 0.0015 to 2.5 m2. Most ecosystem categories had similar snail densities, standing stocks, and body sizes suggesting snails shared a similar function among ecosystems. Caribbean karst wetlands were a striking exception with much lower density and standing stock, but large body size. Disparity in body size results from the presence of ampullariids in Caribbean karst wetlands suggesting that biogeography affects the distribution of taxa, and in this case size, among aquatic ecosystems. We propose that resource quality explains the disparity in density and standing stock between Caribbean karst wetlands and other categories. Periphyton in Caribbean karst wetlands has high carbon-to-phosphorous ratios and defensive characteristics that inhibit grazers. Unlike many freshwater ecosystems where snails are key grazers, we hypothesize that a microbial loop captures much of the primary production in Caribbean karst wetlands.