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See detailInvestigating apical adverse effects of four endocrine active substances in the freshwater gastropod Lymnaea stagnalis
Giusti, Arnaud ULg; Lagadic, Laurent; Barsi, Alpar et al

in Science of the Total Environment (2014), 493

The hermaphroditic gastropod Lymnaea stagnalis is proposed as a candidate species for the development of OECD guidelines for testing of the reprotoxicity of chemicals, including endocrine active ... [more ▼]

The hermaphroditic gastropod Lymnaea stagnalis is proposed as a candidate species for the development of OECD guidelines for testing of the reprotoxicity of chemicals, including endocrine active substances (EASs). Up to now, only a few putative EASs have been tested for their reproductive toxicity in this species. In this study, we investigate the effects of four EASs with different affinities to the vertebrate estrogen and androgen receptors (chlordecone as an estrogen; cyproterone acetate, fenitrothion and vinclozolin as anti-androgens) on the reproduction of L. stagnalis in a 21-day semi-static test. Testosterone and 17α-ethinylestradiol (EE2) were used as the reference compounds. The tested EASs had no significant effect on growth and survival at the tested concentration ranges (ng to μg/L). Classical reproduction endpoints (i.e., oviposition and fecundity) were not responsive to the tested chemicals, except for chlordecone and 17α-ethinylestradiol, which hampered reproduction from 19.6 μg/L and 17.6 μg/L, respectively. The frequency of polyembryonic eggs, used as an additional endpoint, demonstrated the effects of all compounds except EE2. The molecular pathways, which are involved in such reproduction impairments, remain unknown. Our results suggest that egg quality is a more sensitive endpoint as compared to other reproductive endpoints commonly assessed in mollusk toxicity tests. [less ▲]

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See detailProteomic Analysis of the Reproductive Organs of the Hermaphroditic Gastropod Lymnea stagnalis Exposed to Different Endocrine Disrupting Chemicals
Giusti, Arnaud ULg; Leprince, Pierre ULg; Mazzucchelli, Gabriel ULg et al

in PLoS ONE (2013), 8(11), 81086

Many studies have reported perturbations of mollusc reproduction following exposure to low concentrations (ng/L range) of endocrine disrupting chemicals (EDCs). However, the mechanisms of action of these ... [more ▼]

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

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See detailTestosterone levels and fecundity in the hermaphroditic aquatic snail Lymnaea stagnalis exposed to testosterone and endocrine disruptors
Giusti, Arnaud ULg; Ducrot, Virginie; Joaquim-Justo, Célia ULg et al

in Environmental Toxicology & Chemistry (2013), 32(8), 1740-1745

Endocrine disruptors are known to alter endogenous free and/or esterified levels of androgenic and estrogenic steroid hormones in aquatic molluscs. However, there is still a controversy on the origin of ... [more ▼]

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

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See detailReproductive impacts of tributyltin (TBT) and triphenyltin (TPT) in the hermaphroditic freshwater gastropod Lymnaea stagnalis
Giusti, Arnaud ULg; Barsi, Alpar; Dugué, Maël et al

in Environmental Toxicology & Chemistry (2013), 32(7), 1552-1560

Tributyltin (TBT) and triphenyltin (TPT) are emblematic endocrine disruptors, which have been mostly studied in gonochoric prosobranchs. Although both compounds can simultaneously occur in the environment ... [more ▼]

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

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See detailEffects of tributyltin (TBT) and triphenyltin (TPT) on the reproduction of the freshwater snail Lymnaea stagnalis
Lagadic, Laurent; Giusti, Arnaud ULg; Barsi, Alpar et al

Conference (2012, December 10)

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See detailReproductive impact and proteomic analysis of androgenic and anti-androgenic disruptors on the hermaphroditic freshwater gastropod Lymnaea stagnalis
Giusti, Arnaud ULg; Ducrot, Virginie; Joaquim-Justo, Célia ULg et al

Poster (2011, November)

In this study, e ects of two androgens (tributyltin and testosterone), one antiandrogen (cyproterone acetate) and one estrogen (chlordecone) on growth and reproduction were investigated in the ... [more ▼]

In this study, e ects of two androgens (tributyltin and testosterone), one antiandrogen (cyproterone acetate) and one estrogen (chlordecone) on growth and reproduction were investigated in the hermaphrodite gastropod Lymnaea stagnalis. In this study, exposure to a range of concentrations (ng/l to μg/l) of each chemical was performed during 21 days. e number of clutches and the number of eggs per clutch were monitored. A decrease in clutches laid per snail was observed after exposure to tributyltin (540, 1180 and 2600 ng/l) and chlordecone (10, 22, 50 and 110 μg/l). A signi cant decrease in egg laid per snail was observed after exposure to tributyltin (540, 1180 and 2600 ng/l) and chlordecone (50 and 110 μg/l). An increase of egg abnormalities ratio in exposed snails (atrophied albumen, polyembryonicity,...) was observed in L. stagnalis following exposure to testosterone (2, 22, 50 and 110 ng/l), cyproterone acetate (4,5 and 50 μg/l), tributyltin (110 and 244 ng/l) and chlordecone (4,5 and 10 μg/l). Investigation of alteration in protein expression in exposed snails was performed using proteomic analysis such as 2D-DIGE. Mass spectrometry identi cation was performed on proteins with altered expression. We could establish correlation between reproductive endpoints and changes in proteins involved in egg formation and in egg laying were underlined. Egg yolk ferritin, the main protein of egg yolk, was shown to be reduced signi cantly in relationship with a decrease of egg yolk quality after exposure to tributyltin 540 ng/l and cyproterone acetate 4,5 μg/l. Ovipostatin, a protein proved to reduce egg masses, was signi cantly over expressed in snails exposed to 50 μg/l of chlordecone and were in relationship with a reduction of clutches laid by individuals. Further western blot analysis on those proteins involved in the reproduction are underway. ese analysis will enable us to con rm and re ne with more speci city the 2D-DIGE results for the selected proteins. e results of this study can help to establish new biomarkers of exposure of endocrine disruptors in freshwater environment and can provide new insight on mode of action of endocrine disruptors in L. stagnalis. [less ▲]

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See detailEffects of androgenic and anti-androgenic substances on the freshwater gastropod Lymnaea stagnalis
Giusti, Arnaud ULg; Ducrot, Virginie; Joaquim-Justo, Célia ULg et al

Poster (2010, May 25)

Knowledge on the impacts of endocrine disrupting chemicals (EDCs) on gastropods is scarce and their mechanisms of action are still poorly understood. In this study, effects of 3 androgens (tributyltin ... [more ▼]

Knowledge on the impacts of endocrine disrupting chemicals (EDCs) on gastropods is scarce and their mechanisms of action are still poorly understood. In this study, effects of 3 androgens (tributyltin, testosterone and fenitrothion), 2 anti-androgens (cyproterone acetate and vinclozolin) and 1 estrogen (chlordecone) on growth and reproduction were investigated in the hermaphrodite gastropod Lymnaea stagnalis. [less ▲]

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