Effects of municipal effluents on serotonin and dopamine levels in the freshwater mussel Elliptio complanata

Sex differentiation and gametogenesis represent critical steps in the reproductive process and are subject to hormonal control by serotonin, dopamine and steroids such as estradiol-17beta and testosterone. The purpose of this study sought to examine the endocrine-disrupting activity that a primary-treated municipal effluent might have on the metabolism of biogenic amine levels. First, serotonin receptors transfected in Chinese hamster ovary (CHO) cells were used to screen for the presence of serotonin receptor agonist or antagonist. Second, one group of Elliptio complanata mussels were exposed to single compounds likely to be found in municipal wastewaters and another group was exposed in situ to the municipal effluent plume for 90 days in experimental cages. Results showed that solid phase C-8 extracts of surface water downstream a municipal effluent could activate the transport of serotonin by receptors at a distance of at least 5 km from its outfall thereby indicating the presence of serotonin mimics in the effluent dispersion plume. Levels of serotonin and monoamine oxidase (MAO) activity in nerve ganglia of mussels exposed for 90 days to the municipal effluent were, respectively, reduced and increased at a distance 10-km downstream. Injections of estradiol-17beta and nonylphenol in mussels decreased the levels of serotonin and dopamine, but increased MAO activity in the gonad and nerve ganglia. Exposure to estrogenic chemicals present in municipal effluents may therefore alter the normal metabolism of serotonin and dopamine, both of which are involved in sexual differentiation in bivalves and fish. Chemicals acting through E2 receptor-mediated pathways and serotonin receptors are likely to cause the observed effects.     

Inflammatory properties of municipal effluents to Elliptio complanata mussels--lack of effects from anti-inflammatory drugs

The purpose of this study was to identify the pharmacological effects of anti-inflammatory drugs in freshwater mussels (Elliptio complanata) exposed to a primary-treated municipal effluent. Mussel specimens were injected with either increasing concentrations of ibuprofen or with a municipal effluent extract, and then left to stand for 24 h at 15 degrees C. They were also exposed to dilutions of a primary-treated effluent for 30 days at 15 degrees C under semi-static conditions. Gill and gonad cylcooxygenase (COX) were then determined after the incubation period. The influence of various drugs found in municipal effluents on serotonin and dopamine synaptosome transport was determined in visceral ganglia. The results show that injections of ibuprofen reduced COX activity nearly 4-fold in gills and 1.4-fold in gonads. However, COX activity was induced in both tissues after 24 h in mussels injected with a municipal effluent extract and after 30 days in those exposed to dilutions of the effluent. Moreover, synaptosomal dopamine transport activity was increased by ibuprofen, aspirin, caffeine and estradiol-17beta (E2), and decreased by loperamide and carbamazepine, suggesting increased and decreased turnover rates of this catecholamine, respectively. Serotonin transport activity was much less affected, decreasing with high doses of loperamide and increasing with ibuprofen, but with less intensity than with dopamine. The results suggest that although ibuprofen can effectively reduce COX activity in gill and gonadal tissues, exposure to both the municipal effluent and its organic extract increased COX activities, indicating the absence of NSAID (non-steroidal anti-inflammatory drugs)-related effects. Besides their known estrogenic and serotonergic properties, municipal effluents appear to elicit a state similar to inflammation in freshwater mussels.     

Evaluation of potential health risks to Eastern Elliptio (Elliptio complanata) (Mollusca: Bivalvia: Unionida: Unionidae) and implications for sympatric endangered freshwater mussel species

Conservation efforts for freshwater musselspecies require identification and evaluationof potential health risks to populations. Sampling large numbers of individual mussels,however, could damage the small, fragile,extant populations of imperiled mussel species. In this study, a small sample size was designedto reduce lethal sampling, costs, andenvironmental disruption, while still allowing95% confidence in detecting health risks of25% or greater prevalence in the population. Health assessments were conducted on twentyspecimens of the Eastern Elliptio, Elliptio complanata, collected from two NorthCarolina sites as part of a survey to evaluatepotential disease threats to mussels in theregion. Bacteriological sampling of thegastrointestinal tracts yielded 18 aerobicbacterial species, of which Aeromonashydrophila (55.0%), Enterobacter spp.(40.0%), and Bacillus spp.(30.0%) were predominant. Histologicallesions of internal organs included mild tomoderate digestive gland atrophy andinflammation in one mussel, and mild tomoderate parasitism in several individuals. Adistinct difference in parasite prevalence wasevident between infections in E.complanata from the two collection sites. Thetrematode metacercaria of Homalometronarmatum and what appears to be three gillciliate species, the most abundant beingtentatively identified as the scyphidiidperitrich Mantoscyphidia sp., were foundin mussels from one site only. This studydemonstrates a comprehensive diagnosticapproach incorporating multiple modalities toassess the health status of mussel populations,while minimizing the sample size required toobtain valuable information. Furthermore, thisstudy provides baseline health data of E.complanata at two sites in south-central NorthCarolina and suggests the potential usefulnessof E. complanata as an environmentalbioindicator of health risks to sympatricthreatened freshwater mussel populations.     

Response of digestive gland cells of freshwater mussel Unio tumidus to phenolic compound exposure in vivo

The exposure of freshwater mussels Unio tumidus to phenolic compounds (tannic, ellagic and gallic acid) in vivo caused changes in proteins and DNA function of digestive gland cells. The mussels were exposed to various concentrations of tested polyphenols (60, 200 and 500 microM) for 24 and 48 h and their antioxidant and pro-oxidant effects were determined. The number of SH-groups was quantified spectrophotometrically using Ellman's reagent. Oxidative modification of proteins increased in the digestive gland cells in a dose- and time-dependent manner. The level of nuclear DNA damage was investigated using the comet assay. The results revealed that polyphenolic acids induce single and double-strand breaks in DNA. The highest changes were observed for tannic and gallic acids and the smallest ones for ellagic acid. 1h of DNA repair process was also studied using the same method. The data obtained in this experiment demonstrate that the most effective DNA repair occurs in the cells exposed to phenolic compounds for 24h. A longer incubation (up to 48 h) does not decrease the capacity of the repair mechanism. The antioxidant activity of the tested phenols was analyzed spectrofluorimetrically using a fluorescence probe DCFH-DA (dichlorofluorescein-diacetate). The experimental data showed that the tested acids can act as antioxidants when used at higher doses (200 and 500 microM) against the reactive oxygen species present in the digestive gland cells. The most effective was ellagic acid, also applied at the smallest dose of 60 microM, in comparison with tannic and gallic acids. In conclusion, our results demonstrate that chosen water-soluble polyphenols, which are located in various plant tissues and are also found in the aquatic environment, can influence organisms living in the water. They can be exposed to these chemicals that cause morphological alterations and changes in certain physiological processes in their organs (i.e. digestive gland cells of bivalve molluscs).     

Evaluation of genetic structure across freshwater mussel community (genus Elliptio) in the Altamaha River basin

Using mitochondrial DNA sequence data and genotypic data from four microsatellite loci, we evaluate the population genetic structure for three common species of the freshwater mussel genus Elliptio in the Altamaha River basin in Georgia, USA. These data are intended to evaluate whether distinct management strategies should be applied to upper and lower reaches of the Altamaha, which are separated by some discontinuities in appropriate habitat and may have been differentially influenced by heavier urbanization in the upstream reaches. Our results show no significant structure of mitochondrial data for E. dariensis, E. hopetonensis, or mussels in the E. icterina–complanata group. However, microsatellite data do suggest moderate genotypic structure of mussels in the E. icterina–complanata group, consistent with (but not necessarily caused by) the noted habitat disjunction. Application of these data may prove useful in management of rare endemic congeners such as the Altamaha spinymussel, E. spinosa, in showing that management resources can be applied uniformly throughout the Altamaha basin.     

Endocrine disruption and health effects of caged mussels, Elliptio complanata, placed downstream from a primary-treated municipal effluent plume for 1 year

Freshwater mussels, Elliptio complanata, were caged in special benthic pens and were immersed at one upstream (Ups) site and two downstream sites (8 and 11 km) of a primary-treated municipal effluent plume for 1 year. The levels of metallothionein-like proteins (MT), lipid peroxidation, protein-free DNA strands and glutathione S-transferase (GST) activity were assayed in digestive gland, gill and gonad tissues to evaluate biological effects and damage. The levels of monoamines (serotonin and dopamine) in nerve ganglia, ATP-dependent transport activity and monoamine oxidase (MAO) activity were also investigated in the homogenates, synaptosomes and mitochondria, respectively. Results showed that significant amounts of sediment accumulated in cages and 82% of mussels survived the yearlong exposure period at the downstream sites. MT-like proteins were induced in all tissues with the following response intensity: gill (3-fold), digestive gland (1.4-fold) and gonad tissues (1.3-fold). Lipid peroxidation decreased (2.5-fold) in digestive gland but increased in gill (1.6-fold) and in gonad tissues (1.5-fold). GST activity was readily increased in digestive gland (2.5-fold), suggesting the presence of organic contaminants in the plume. Levels of protein-free DNA strands did not vary significantly in digestive gland and gill tissues but were significantly reduced in gonad tissues (2.5-fold) relative to the upstream site. In visceral nerve ganglia, both serotonin and ATP-dependent serotonin transport decreased 1.7-fold with a 4-fold increase of 5-hydroxyindole acetate (5-HIAA, a serotonin metabolite) level relative to the upstream site. However, MAO activity was somewhat reduced at downstream sites (0.7- to 0.9-fold of the activity at the upstream site). Dopamine levels were found to be decreased (1.5-fold), but dopamine ATP-dependent transport activity was increased 1.8-fold, suggesting reduced dopaminergic activity. These results indicate that estrogenic chemicals are likely at play, and the increased dopamine and decreased serotonin ATP-dependent transport suggest that the municipal plume was serotonergic for mussels located at the downstream sites. Mussels exposed for 1 year display a complex but characteristic pattern of responses that could lead to harmful health effects including neuroendocrine disruption of reproduction.     

Sediment accumulation predicts the distribution of a unionid mussel (Elliptio complanata) in nearshore areas of a Canadian Shield lake

1. The importance of native freshwater mussels for ecosystem processes depends on their density, size distribution and activity. In lakes, many of the factors that affect mussels (fish hosts, habitat, food) could be directly or indirectly related to wind‐driven physical processes. 2. We tested whether the abundance and size of Elliptio complanata in the shallow, nearshore areas of a medium‐sized lake were related to site exposure, substratum type and fish distribution. To disentangle some of the correlated variables known to affect mussel distribution, we used paired exposed and sheltered sampling sites along the 7‐km fetch of the lake basin. 3. The distribution of sediment characteristics in nearshore areas was highly predictable. The mean depth of accumulated soft sediments decreased with increasing fetch at wind‐exposed sites, but increased with increasing fetch at sheltered sites. Sediments were deeper along the main shoreline than around islands. Deeper sediments tended to be finer and higher in silt content and organic fraction. 4. The density and proportion of juvenile mussels along the main shoreline varied in a unimodal way with sediment depth. These results suggest that wind‐driven physical forces affect the transport of young juveniles to sediment depositional areas and create sediment conditions that influence their growth and survival. In contrast, the proportion of juvenile mussels around islands was not related to sediment characteristics, but decreased with remoteness of the island, suggesting that the distribution of juvenile mussels may be limited by fish movements. These results are tentative since they do not include buried juvenile mussels. 5. We also found a unimodal relationship between total mussel density (juveniles and adults) and sediment depth but, in contrast to the relationship for juveniles only, it applied to all sites with soft sediments, including islands. We conclude that factors related to sediment depth affect the growth and survival of adult mussels around islands and that these factors are strong enough to modify the pattern of distribution established via dispersal during earlier life stages. 6. The mean shell length of adults at different sites within the lake basin ranged from 60 to 85 mm. Mussel shell length decreased with increasing fetch at sites exposed to the prevalent winds, but was relatively constant on the sheltered side of peninsulas and islands. The size of unionid mussels in different parts of the lake seems to be determined both by their exposure to physical forces and by sediments. 7. The local distribution of E. complanata is determined, directly and indirectly, by wind‐driven forces. These processes are likely to be important for other benthic organisms affected by similar habitat conditions (e.g. sediment characteristics, physical disturbance).     

Climate effects on inter‐annual variation in growth of the freshwater mussel (Anodonta beringiana) in an Alaskan lake

1. Warming trends are evident in many parts of the globe but are especially marked at higher latitudes, with complex effects on the biota that include direct effects on growth potential and indirect effects through food webs. 2. Air temperatures have been increasing over the past 50 years in southwestern Alaska, affecting the growth and population dynamics of many organisms, including a variety of aquatic species such as the freshwater mussel Anodonta beringiana. 3. We collected freshwater mussels from Iliamna Lake, in the Bristol Bay region of Alaska, and measured their shells to examine climatic effects on growth patterns. 4. Linear mixed effects models and ordinary least square linear regressions revealed strong positive correlations between local air temperatures (especially in May, October and the summer months) and inter‐annual variation in mussel growth. Slower mussel growth was also significantly correlated with later date of ice break‐up, which was linked to air temperatures in late spring.     

Accumulation of a peptide toxin from the cyanobacterium Oscillatoria agardhii in the freshwater mussel Anadonta cygnea

Swan mussels (Anodonta cygnea) were exposed to a toxic strain of the cyanobacterium Oscillatoria agardhii. Mussels accumulated large amounts of the peptide Oscillatoria toxin which was present in low concentrations within the cyanobacterial cells in the test aquaria (40–60 µg Oscillatoria toxin/1). The toxin concentration in the mussels increased during the experiment and after 15 days of exposure the concentration was 70 ± 2 µg/g freeze dried tissue (mean ± range of values). The highest concentration of the toxin (130 µg/g of freeze dried tissue) was found in the hepatopancreatic tissue. The toxin did not seem to be metabolized in the mussels and they were not killed by the high toxin concentrations within them. After two months in clean water still detectable amounts of toxin were present in the mussels.     

Range-wide population genetic analysis of the endangered northern riffleshell mussel, <Emphasis Type="Italic">Epioblasma torulosa rangiana</Emphasis> (Bivalvia: Unionoida)

The northern riffleshell (Epioblasma torulosa rangiana) is a critically endangered unionoid species in need of conservation throughout its range. It is the only unionoid to be federally protected in both Canada and the U.S. We use sequences from two mtDNA genes and 15 microsatellite loci to assess genetic variation among 86 individuals from the four populations in the three remaining drainages in which E. t. rangiana is known to be reproducing. All of these populations are in formerly glaciated landscapes that emerged <10 kya. The mtDNA sequence data do not indicate significant geographic structure among the populations. However, allelic data from the microsatellite loci reveal highly significant population structuring. Individuals of E. t. rangiana can be assigned to their own river of origin with 98.8% accuracy. Significant isolation-by-distance occurs. This analysis will be useful to conservation managers in documenting the genetic structure, patterns of isolation, and genetic variability within and among populations of E. t. rangiana.     

Genetic diversity and divergence among freshwater mussel (Anodonta) populations in the Bonneville Basin of Utah

Populations of the freshwater mussel genus Anodonta appear to be in a state of rapid decline in western North America, following a trend that unfortunately seems to be prevalent among these animals (Mollusca: Unionoida). Here we describe the patterns of molecular divergence and diversity among Anodonta populations in the Bonneville Basin, a large sub-basin of the Great Basin in western North America. Using amplified fragment length polymorphism (AFLP) analysis, we found a striking lack of nuclear diversity within some of these populations, along with a high degree of structuring among populations (FST = 0.61), suggesting post-Pleistocene isolation, due either to a long-term loss of hydrologic connectivity among populations or to more recent fish introductions. We also found evidence of recent hybridization in one of these populations, possibly mediated by fish-stocking practices. Using mitochondrial sequence data, we compared the Bonneville Basin populations to Anodonta in several other drainages in western North America. We found a general lack of resolution in these phylogenetic reconstructions, although there was a tendency for the Bonneville Basin Anodonta (tentatively A. californiensis) to cluster with A. oregonensis from the adjacent Lahontan Basin in Nevada. We recommend further investigation of anthropogenic factors that may be contributing to the decline of western Anodonta and a broad-scale analysis and synthesis of genetic and morphological variation among Anodonta in western North America.     

Within- and among-lake variation in shell morphology of the freshwater clam Elliptio complanata (Bivalvia: Unionidae) from south-central Ontario lakes

To compare the relative magnitude of variation in shell morphology within and among lakes, Elliptio complanata were collected from low and high exposure areas in each of four small lakes in south-central Ontario. Nested ANOVA's on shell length, height, width and weight revealed that shell morphology varied much more between sites of differing exposure within a lake than among lakes of differing alkalinity. Canonical variates analysis showed that clams from high exposure areas had larger and proportionately taller and heavier shells than those from low exposure areas. There was no relationship between alkalinity of lakes and shell morphology. These results suggest that the use of unionid shell morphology to predict long-term whole lake water chemistry (e.g. alkalinity) requires sampling designs which take into account within-lake variation in shell morphology.     

Sediment particles as a cause of nacre staining in the freshwater mussel,Amblema plicata (Say) (Bivalvia: Unionidae)

Brown stains sometimes appear in the inner shell layers (nacre) of freshwater mussels. An electron microprobe was used to analyze the stained nacre of the unionid Amblema plicata (Say, 1817) from selected localities on the Mississippi River in the vicinity of LaCrosse and Prairie du Chien, Wisconsin.Several elements such as Na, Mg, Al, Si, P, S, Cl, K, and Fe are more highly concentrated in stained than in unstained nacre. Concentrations of these elements relative to Ca were found to vary significantly among the localities from which the specimens were obtained. Ratios have significantly higher variances downstream of the confluence of the Yellow and Mississippi Rivers, downstream of a barge fleeting area, near the town of Marquette, Iowa, near the site of sewage effluent for Prairie du Chien, Wisconsin and downstream of a scrap metal yard near LaCrosse, Wisconsin in contrast to control localities.Suspended silt, the result of runoff and river activity (barge traffic, dredging, pleasure boating) may be the stimulus for stain formation. Clay minerals adsorb accessory elements and in turbid water are trapped within the pallial space of A. plicata. The mussel secretes an organic-rich, periostracum-like layer over the entrapped sediment, and later reverts to prismatic and finally nacreous shell deposition. Some of the elements found in the stain could directly disturb Ca metabolism by competing with Ca for binding sites in shell aragonite.     

An integrative approach underscores the taxonomic status of Lamellidens exolescens,a freshwater mussel from the Oriental tropics (Bivalvia: Unionidae)

The Oriental Region harbours the second richest fauna of freshwater bivalves in the world, including many endangered endemic taxa. However, the Oriental fauna of the Unionidae have been very poorly studied using an integrative taxonomic approach, which may provide reasonable revisions of complicated (cryptic) taxa based on morphological, molecular, biogeographic and ecological evidence. Here, we present the first example of an integrative taxonomic revision concerning the status of Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]), a nominal mussel taxon that was accepted as a valid species within the genus Trapezoideus Simpson (1900). Currently, Trapezoideus exolescens is considered the type of the genus as far as the originally designated type species, U. foliaceus Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]), was considered to be a synonym of T. exolescens. Using nucleotide sequences obtained from mitochondrial (COI and 16S rRNA) and nuclear (28S rDNA) genes, we found that the topotypes of Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) cluster together with representatives of another mussel genus, Lamellidens Simpson (1900). Based on these results and on morphological data, we transfer Unio exolescens Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) from Trapezoideus to Lamellidens and propose Lamellidens exolescens (Gould, 1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) comb. nov. In addition, we revisited the status of Unio foliaceus Gould (1843 Gould, A.A. (1843). Dr. Gould had examined the shells not long since announced as having been received from the Rev. Francis Mason, missionary at Tavoy, in British Burmah. Proceedings of the Boston Society of Natural History, 1, 139–141. [Google Scholar]) as a valid species and the type of the genus Trapezoideus based on the morphological study of the type specimen, although a question concerning the true position of this taxon is still open because its molecular sequences are not available. Our findings highlight that an integrative taxonomic approach is an important tool, particularly when dealing with such species-rich Unionidae fauna as those of the Oriental Realm.     

Ecological resistance to the invasion of a freshwater clam,Corbicula fluminea: fish predation effects

Summary Fish predation is shown to have a twenty nine fold effect on the abundance of the invasive freshwater clam, Corbicula fluminea, in a Texas reservoir. This predation has prevented the clam from establishing the high densities commonly reported for it elsewhere. The high magnitude of the fish effect is attributed to Corbicula being an invader to this reservoir and not being able to cope well with the mix of resident fish species. In the absence of fish, colonization of the reservoir by Corbicula is spatially patchy. When fish interact with these clams, they remove sufficient numbers of individuals from dense patches to create the appearance of a spatially uniform distribution.     

Studies on fouling by the freshwater mussel limnoperna fortunei and the antifouling effects of low energy surfaces

Attachment of the freshwater mussel, Limnoperna fortunei, was tested using non‐treated surfaces, viz. glass, nylon, rubber, silicone and Teflon, together with glass surfaces modified with nine kinds of silane coupling agents. Among the surfaces tested, the mussel avoided attaching to Teflon, silicone, and glass modified with 3‐bromopropyltrimethoxysilane or 3,3,3‐(trifluo‐ropropyl)‐trimethoxysilane. With respect to the relationship between the percentage attachment and the surface free energy (sfe) of the substrates, it was found that attachment was considerably reduced on the substrates which exhibited relatively low sfe, as above. The mean number of secreted byssuses per attaching mussel also decreased with decreasing substrate sfe. Furthermore, when the sfe was divided into the dispersion and polar components, the percentage mussel attachment was related to the polar component. These results suggest that effective antifouling towards L. fortunei is achieved on substrates with a low sfe polar component.     

Three deeply divided lineages of the freshwater mussel genus <Emphasis Type="Italic">Anodonta</Emphasis> in western North America

The surprising diversity and recent dramatic decline of freshwater mussels in North America have been well documented, although inventory efforts to date have been concentrated in the eastern United States. Unlike their eastern counterparts, western freshwater mussels have received comparatively little attention. The accurate identity of western lineages is a necessary component for future inventory, monitoring, and ecological work involving these taxa. Here we initiate a study involving the most speciose genus (Anodonta) in western North America, incorporating information about type localities and type specimen morphology and describing the discovery of three highly divergent lineages among four western Anodonta species. In a limited phylogenetic analysis, we find (1) that A. californiensis/nuttalliana and A.oregonensis/kennerlyi are distinct, highly divergent clades, and (2) that A. beringiana is more closely allied with A. woodiana, an Asian species, than either of the other two western North American clades. We were largely unable to resolve the placement of these three clades with respect to other anodontines, and suggest the need for a broader phylogenetic framework. We recommend, however, that the existence of these three deeply divergent groups be considered in the development of regional monitoring, conservation and research plans despite the taxonomic uncertainty.     

A model assessing the conservation threats to freshwater turtles of Sub-Saharan Africa predicts urgent need for continental conservation planning

Global assessment of conservation threats to several taxonomic groups are urgently needed for species living in regions of the world where field research is curtailed by logistic or economic constraints. For instance, it is now widely recognized that freshwater turtles represent one of the most endangered groups of vertebrates in the world. This situation has been particularly evident with Asian species. Although in Sub-Saharan Africa species are exposed to very similar threats (e.g., intense hunting for food and traditional medicine, habitat loss, collection for the pet trade, etc.), there is no comparable concern in the international conservation community simply because the African species are still too poorly known. Here, I propose a model for analyzing the threat levels to freshwater turtles of Sub-Saharan Africa and I test this model by taking into consideration a series of six risk variables of distribution and ecology. Each variable was categorized into five ranks (0–4) of increasing survival risk. The results, although preliminary, show a great sensitivity of most species. This strongly contrasts with the fact that apart from Malagasy endemics very few species are currently considered as threatened according to IUCN lists. There were clear differences among-families in the major threats. Softshell turtles are especially threatened because their large body size makes them a very good food source for native populations. The single podocnemidid species is also vulnerable, and is particularly sensitive to human hunting for domestic consumption. Concerning the Pelomedusidae, there are several species that are considered by the model as sensitive or highly sensitive to rarity. They are affected by a combination of threats (Extent of distribution in Sub-Saharan Africa, Habitat breadth, Habits, Likeliness of human direct/indirect persecution, and Habitat vulnerability). The conservation implications of these results are discussed.