Ramshorn Snail Research Articles

The Potential Use of Ultrasound to Control the TrematodeBolbophorus confususby Eliminating the Ram's Horn SnailPlanorbella trivolvisin Commercial Aquaculture Settings

Abstract One method in the investigation of acoustics to improve aquaculture production is the use of ultrasound to control the trematode Bolbophorus confusus in commercial catfish Ictalurus spp. ponds. The trematode population can be controlled by eliminating the host, ram's horn snail Planorbella trivolvis, which is typically accomplished with chemical treatments. The work presented here investigates snail elimination via exposure to high amplitude ultrasound. Initial laboratory tests indicated that a commercially available sonicator (operating at 20 kHz) is capable of quickly killing individual snails in test tanks. More thorough testing indicated efficacy rates of at least 35% (potentially 65%) on batches of 10 snails in a reverberant environment. The experimental setup of these initial laboratory tests provided nearly 20 decibels of gain in sound levels compared to what is expected in ponds. This is due to reverberation from the air surrounding the tank walls, as opposed to absorption at the bottom a...

Toxic Effects of Copper, Cadmium, and Methoxychlor Shown by Neutral Red Retention Assay in Two Species of Freshwater Molluscs

We used neutral red retention assay in lysosomes of digestive gland cells as an indicator for stress effects by the environmental contaminants Cu, Cd, and the pesticide methoxychlor in two freshwater molluscs, the unionid mussel, Elliptio complanata (Lightfoot) and the ramshorn snail Helisoma trivolvis (Say). Mussels and snails were exposed for 7 and 14 days to Cu and Cd each at nominal concentrations of 2.5 µg/L, 5.0 µg/L, and 10.0 µg/L, and to methoxychlor concentrations of 1.0 µg/L, 10.0µg/L, and 100.0 µg/L. Both mussels and snails exposed to Cu showed a significant increase in the percent of destabilized lysosomes compared with lab control and freshly-collected (field control) mussels at both 7 and 14 days exposure for all concentrations. Cd-exposed mussels did not show a significant difference with either of the controls at 7 days, but at 14 days exposure, Cd significantly increased the percent of destabilized lysosomes at all concentrations compared to field control mussels. Compared to laboratory controls, Cd increased lysosomal destabilization at 5.0 µg/L and 10.0 µg/L. Snails exposed to Cd for 7 days had a significantly higher percentage of lysosomal destabilization than both lab and field controls but at 14 days, significant differences were only seen at the two highest Cd concentrations. Methoxychlor-exposed mussels showed no significant lysosomal destabilization at 7 days compared to controls. But at 14 days exposure, the pesticide increased the percentage of lysosomal destabilization at 10.0µg/L compared to lab controls, and increased at both 10.0µg/L and 100.0µg/L compared to field control mussels. Methoxychlor-exposed snails had a higher percentage of lysosomal destabilization at 7 and 14 days at all concentrations compared to both controls with the exception of the 1.0 µg/L -7 day exposure group. Snails were more sensitive to Cd and to methoxychlor than were mussels possibly because they lack an operculum and are thus completely exposed to the environment. The lowest observed effect concentration (LOEC) for Cd was 2.5 µg/L (14 days) for Elliptio and 2.5 µg/L (7 and 14 days) for Helisoma. For methoxychlor, the LOEC was 10.0 µg/L (14 days) for Elliptio and 10.0 and 1.0 µg/L (7 and 14 days, respectively) for Helisoma. The LOEC for Cu was 2.5 µg/L (7 and 14 day exposure) for both Elliptio and Helisoma. These results show that lysosomal destabilization as indicated by neutral red retention is a reliable indicator of stress from heavy metals and a pesticide in freshwater molluscs, including a taxon that is endangered or threatened in North America.

Genetic Sequence Data Identifies the Cercaria of Drepanocephalus spathans (Digenea: Echinostomatidae), A Parasite of the Double-Crested Cormorant (Phalacrocorax auritus), with Notes on its Pathology in Juvenile Channel Catfish (Ictalurus punctatus)

An unidentified xiphidio-type cercaria, previously thought inconsequential to catfish health, was found to be released from marsh rams-horn snails (Planorbella trivolvis) inhabiting ponds on a commercial catfish operation in the Mississippi Delta. A preliminary challenge of cohabiting channel catfish ( Ictalurus punctatus ) with snails actively shedding the unidentified cercariae resulted in death of some fish. A second cohabitation trial yielded similar results, as did a third challenge of 250 cercariae/fish. Histopathology revealed developing metacercariae concentrated in the cranial region, especially within the branchial chamber, with several metacercariae at the base of the branchial arches within, or adjacent to, blood vessels, possibly the proximate cause of death. Genetic sequence analysis of the 18S small subunit ribosomal DNA (ssDNA), 28S large subunit rDNA (lsDNA), and cytochrome oxidase (Cox1) genes all matched the cercariae to Drepanocephalus spathans (Digenea: Echinostomatidae), a parasite of the double-crested cormorant (Phalacrocorax auritus), a piscivorous bird endemic on most catfish farms. This is the first commentary regarding pathology of D. spathans in juvenile channel catfish as well as the first report of the marsh rams-horn snail as an intermediate host in the D. spathans life cycle. The data presented here suggest this parasite could have limiting effects on catfish production, further supporting the need for adequate snail control programs to reduce trematode prevalence on commercial catfish operations.

Arresting mantle formation and redirecting embryonic shell gland tissue by platinum2+ leads to body plan modifications in Marisa cornuarietis (Gastropoda, Ampullariidae)

To evaluate the threat that anthropogenic substances pose to animals when they are emitted into the environment, tests like the invertebrate embryo toxicity test with the ramshorn snail Marisa cornuarietis have been developed. These tests are used to investigate substances like the heavy metal platinum (Pt) that is used in catalytic converters and is gradually released in car exhausts. In 2010, our group reported that high Pt concentrations cause body plan alterations in snails and prevent the formation of an external shell during M. cornuarietis embryogenesis. Now, this study presents scanning-electron micrographs and histological sections of platinum(2+) (Pt(2+))-treated and untreated M. cornuarietis embryos and compares "normally" developing and "shell-less" embryos during embryogenesis, to reveal the exact course of events that lead to this body plan shift. Both groups showed similar development until the onset of torsion 70- to 82-h postfertilization. In the Pt(2+)-exposed embryos, the rudimentary shell gland (=anlage of both shell gland and mantle, which usually evaginates, grows, and eventually covers the visceral sac) does not spread across the visceral sac but remains on its ventral side. Without the excessive growth of the shell gland, a horizontal rotation of the visceral sac relative to head and foot does not occur, as being normal during the process of torsion.

Does social facilitation affect responses to natural and anthropogenic stressors in the freshwater snail Planorbella trivolvis?

Social facilitation is the initiation or increase of a trait, such as stressor tolerance, when in the presence of conspecifics, members of the same species. It has been shown to alter the outcome of toxicity experiments in colonial organisms. We evaluated whether social facilitation would impact responses to stressors in the noncolonial New Mexico ramshorn snail (Planorbella trivolvis) by exposing snails to stressors either singly or in groups of three. Social facilitation did not impact snail responses to malathion but did affect responses to predator cues and temperature stress.

Histopathological effects of copper and lithium in the ramshorn snail, Marisa cornuarietis (Gastropoda, Prosobranchia)

The aim of this study was to determine and quantify effects of copper and lithium in tissues of early juveniles of the ramshorn snail, Marisa cornuarietis. For this purpose, hatchlings of M. cornuarietis were exposed for 7 days to a range of five different sublethal concentrations of copper (5, 10, 25, 50, and 75 μg Cu 2+ L −1) and lithium (50, 100, 200, 1000, and 5000 μg Li + L −1). Both metals changed the tissue structure of epidermis, hepatopancreas, and gills, varying between slight and strong reactions, depending on the copper and lithium concentration. The histopathological changes included alterations in epithelial and mucous cells of the epidermis, swelling of hepatopancreatic digestive cells, alterations in the number of basophilic cells, abnormal apices of digestive cells, irregularly shaped cilia and changes in the amount of mucus in the gills. The most sensible organ in M. cornuarietis indicating Cu or Li pollution is the hepatopancreas (LOECs were 10 μg Cu 2+ L −1, or 200 μg Li + L −1). In epidermis, mantle and gills relevant effects occurred with higher LOECs (50 μg Cu 2+ L −1, or 1000 μg Li + L −1). Base on LOECs, our results indicated that histopathological endpoints are high sensitivity to copper and lithium compared to endpoints for embryonic developmental toxicity.

Longevity of Bolbophorus damnificus Infections in Channel Catfish

The digenean Bolbophorus damnificus infects commercial channel catfish Ictalurus punctatus, causing mortality, lower feed consumption, and reduced growth in surviving fish. The purpose of this study was to determine the length of time for which B. damnificus prodiplostomulum metacercariae (juvenile trematode stage that infects fish) would remain viable (parasite appearing to be intact or exhibiting movement) in channel catfish. Fish (n = 210) were infected with molecularly confirmed B. damnificus cercariae harvested from naturally infected marsh rams-horn snails Planorbella trivolvis. During the first sampling (at 20 d postinfection), 8.3 +/- 3.6 metacercariae/fish (mean +/- SD) were found in the host muscle and visceral organs. The channel catfish were then acclimated to a water temperature of either 18 degrees C or 28 degrees C. After 11 months, 6.8 +/- 3.5 and 5.9 +/- 3.0 metacercariae/fish were found in groups held at 18 degrees C and 28 degrees C, respectively. The mean number of parasites per fish did not significantly differ between fish held at the two temperatures and did not significantly decline over time at either temperature. Fish examined from 13 to 30 months postinfection all contained viable metacercariae that were morphologically and molecularly identified as B. damnificus. At 18 months, 12 metacercariae (of which 11 were intact and 10 displayed movement) were found in the one fish sampled; at 30 months, the last fish sampled contained three intact metacercariae (one displayed slight movement). Our results indicate that B. damnificus metacercariae can remain viable in channel catfish for at least an 18-30-month production cycle during which they have the potential to affect fish growth; in addition, infected fish may serve as intermediate hosts for these metacercariae for at least 2.5 years postinfection.

Hydroseral habitat requirement of the endangered Shining Ramshorn Snail Segmentina nitida

Before the 1950s, the Shining Ramshorn snail Segmentina nitida was widespread across marshes and drainage ditches in Britain, continental Europe and Scandinavia. However, the population has declined rapidly and the IUCN Red Data list highlights the species as seriously vulnerable to extinction. It is now one of the six freshwater molluscan species protected under the UK Biodiversity Action Plan. Most previous research has focused on species decline and conservation issues. The aim of the research reported here was to clarify the habitat requirements of S. nitida in East Kent within the context of a seral classification. Six hydroseral stages were defined according to plant species composition. Segmentina nitida was limited uniquely to the Carex-Juncus-Eleocharis-Oenanthe community, a late seral stage in ditch succession, which confirms the work of other authors. A general linear statistical model applied to data from these sites suggested that (i) the total number of molluscs within each species varied across samples; (ii) the number of molluscs varied between samples with or without S. nitida; (iii) there was no variation in the total number of molluscs across the locations and (iv) there was no relationship between number of molluscs and sample site within location, implying some degree of connectivity between molluscan communities. Also a two-way ANOVA with Factor 1 as the number of mollusc and Factor 2 as the presence/absence of S. nitida showed a significant interaction effect. These observations are novel for these sites in East Kent and have implications for the ecology, life history strategy and conservation management of S. nitida.

Genotoxicity of platinum in embryos of zebrafish ( Danio rerio) and ramshorn snail ( Marisa cornuarietis)

The metal platinum is inter alia used for industrial and medical purposes. Due to its application in automobile catalytic converters and as an anti-cancer drug, Pt enters the aquatic environment via road runoff and hospital sewage and raises concerns about its environmental impact and toxicity to organisms. Therefore, the genotoxicity of Pt at 0, 0.1, 1, 10, 50, 100 and 200 μg/l PtCl 2 was tested on two freshwater organisms, zebrafish ( Danio rerio) and ramshorn snail ( Marisa cornuarietis) using the single cell gel electrophoresis, also called comet assay. PtCl 2 did not show any genotoxicity for D. rerio at the tested concentrations, whereas significantly elevated DNA damage was observed in M. cornuarietis at 1 μg/l PtCl 2 and beyond. The results of the study suggest a high sensitivity of M. cornuarietis concerning the genotoxic impact of PtCl 2.

Metal sensitivity of the embryonic development of the ramshorn snail Marisa cornuarietis (Prosobranchia)

We investigated the effects of metal ions on the embryonic development of the ramshorn snail, Marisa cornuarietis, by exposing embryos to varying concentrations of copper (0, 50, 100, and 250 μg Cu(2+)/L), lead (0, 5, 10, and 15 mg Pb(2+)/L), lithium (0, 1, 2.5, and 3 mg Li(+)/L), or palladium (0, 50, 100, and 500 μg Pd(2+)/L). Effects of these metals were examined by recording mortality, the rate of tentacles and eyes formation, heart rate, hatching success, and weight after hatching. Compared to the control, we found a significant delay in the formation of tentacles and eyes after treatment with 100 μg Cu(2+)/L, 15 mg Pb(2+)/L, 2.5 mg Li(+)/L or 500 μg Pd(2+)/L. The heart rate decreased significantly at 500 μg Pd(2+)/L. At 10 mg Pb(2+)/L, 2.5 mg Li(+)/L, or 500 μg Pd(2+)/L, hatching was delayed significantly; 50 μg Cu(2+)/L induced a significantly earlier hatching, and reduced body weight. The LC(50) values were calculated to be about 50 μg Cu(2+)/L, 500 μg Pd(2+)/L, 2500 μg Li(+)/L, and 10000 μg Pb(2+)/L. These results show that the embryonic development of M. cornuarietis is about as sensitive to copper and lithium, compared to the most sensitive fishes used in embryo toxicity testing. Even though the MariETT is a laboratory-based assay focusing on toxicological endpoints of a selected model species, future application is envisaged to include testing of "natural" samples such as stream water or sediment interstitial water.

A Duplex Real-Time Polymerase Chain Reaction Assay for Differentiation between Bolbophorus Damnificus and Bolbophorus Type II Species Cercariae

A duplex quantitative real-time polymerase chain reaction (qPCR) assay was developed to differentiate between Bolbophorus damnificus and Bolbophorus type II species cercariae. Both trematode species are prevalent throughout the commercial catfish industry, as both infect the ram's horn snail, Planorbella trivolvis, which is commonly found in catfish ponds. Identification of cercaria to species is important in catfish disease challenge experiments, as only B. damnificus has been shown to have negative impacts on channel catfish. Oligonucleotide primers and fluorescence resonance energy transfer hydrolysis probes were designed to amplify the 18S small subunit ribosomal DNA gene of each species. The quantification cycle indicative of the number of cercariae in the sample prep was determined, and standard curves correlating to cercaria numbers were established. For both species, the assay was found to be highly repeatable and reproducible, with a linear dynamic range covering 7 orders of magnitude. The sensitivity limit of the assay was approximately 1/256th of a cercaria, regardless of species, and there was no remarkable interference between the 2 assays when run simultaneously within the same reaction. In a field study, identification of cercaria by the duplex real-time qPCR assay was in complete agreement with previously established end-point PCR protocols, demonstrating the assay to be a more rapid, quantifiable means of parasite identification.

Coordination between locomotor and respiratory rhythms in the great ramshorn snail Planorbarius corneus: Transmitter-dependent modifications

In a reduced preparation of Planorbarius corneus consisting of the CNS and mantle complex, both the dopamine precursor L-DOPA and the serotonin precursor 5-HTP have been found to be able to induce and maintain rhythmic pneumostome (PN) movements phase-coupled to fictive cyclic locomotion in a neurotransmitter-specific manner. After the transection of pedal commissures, pharmacologically induced PN movements were coordinated with the locomotor activity rhythm generated by the left pedal ganglion, as in Lymnaea regardless of spatial inversion of its CNS. Nevertheless, in Planorbarius during the 5-HTP-induced fictive muscular locomotion, the PN was never opened, but cuddled up to the mantle at the same phase of the locomotor cycle corresponding to close down the PN in Lymnaea.

Histopathological alterations and induction of hsp70 in ramshorn snail ( Marisa cornuarietis) and zebrafish ( Danio rerio) embryos after exposure to PtCl 2

The platinum group metals (PGMs) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree during the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGMs on organisms are extremely rare. In the present study, effects of various concentrations of PtCl 2 (0.1, 1, 10, 50 and 100 μg/L) were investigated with respect to the induction of hsp70 and histopathological alterations in the zebrafish, Danio rerio and the ramshorn snail, Marisa cornuarietis. Histopathological investigations revealed effects of Pt on both species, which varied between slight and strong cellular reactions, depending on the PtCl 2 concentration. The hsp70 level in M. cornuarietis did not show an increase following Pt exposure whereas it was significantly elevated at 100 μg/L PtCl 2 in D. rerio.

Uptake of platinum by zebrafish ( Danio rerio) and ramshorn snail ( Marisa cornuarietis) and resulting effects on early embryogenesis

Platinum group elements (PGEs), platinum, palladium and rhodium are widely used in automobile catalytic converters. PGEs are emitted into the environment and enter the aquatic ecosystem via runoff rainwater. The present study investigated the bioavailability of platinum chloride for the zebrafish ( Danio rerio) and the ramshorn snail ( Marisa cornuarietis) and determined the bioaccumulation rate of platinum. Applying the fish early life stage assay for D. rerio (DarT) and the Marisa embryo toxicity test (“Mariett”) for M. cornuarietis, effects of platinum chloride on the embryonic development were investigated. Platinum concentrations tested in this study ranged from environmentally relevant concentrations of 38 ng L −1 up to a concentration of 74.2 μg L −1 for D. rerio and of 200 ng L −1 up to 98.7 μg L −1 for M. cornuarietis. Platinum was found to be accumulated in both organisms. Bioaccumulation factors ( BAFs) were in the range of 5–55 for D. rerio and of 218.4–723.9 for M. cornuarietis, depending on the tested Pt concentrations. During the embryonic development, platinum was shown to alter the heart rate of both organisms already at the lowest tested concentration. At higher concentrations, platinum decelerated the hatching rate of the embryos of both species. Additionally, a retardation of the general development and a loss of weight due to platinum exposure was observed in M. cornuarietis. Results of this study contribute important data on the ecotoxicity of a rarely studied element.

Embryo toxicity of pesticides and heavy metals to the ramshorn snail, Marisa cornuarietis (Prosobranchia)

An invertebrate embryo toxicity test with the ampullariid snail, Marisa cornuarietis, to assess the toxicity of pesticides and heavy metals recently was established. Snail embryos were treated with atrazine (100, 1000, 10 000, and 30 000 μg/L), imidacloprid (10 000, 25 000, and 50 000 μg/L), Ni 2+ (0.1, 1, 10, and 100 μg/L) or Zn 2+ (100, 200, 500, 1000, 2000, and 5000 μg/L). The effect of these substances was examined by monitoring the following endpoints: mortality, formation of tentacles and eyes, heart rate, hatching, and weight after hatching. Effects in term of a significant delay on the formation of both tentacles and eyes were found after treatment with 100 μg/L Ni 2+ or 200 μg/L Zn 2+. The heart rate was shown to significantly decrease at 25 000 μg/L imidacloprid or 1000 μg/L Zn 2+. At 100 μg/L atrazine, 10 μg/L Ni 2+, or 1000 μg/L Zn 2+ a significant delay in hatching became visible. No significant mortality was observed for the tested concentrations of atrazine, imidacloprid, or Ni 2+, while 5000 μg/L Zn 2+ resulted in 100% mortality after 10 d. The weight of freshly hatched individuals remained unaffected in all treatments. On the basis of the lowest observed effect concentrations (LOECs) recorded, we could show the M. cornuarietis embryo toxicity test (MariETT) to react up to three orders of magnitude more sensitive (for metals) and at least one order of magnitude more sensitive (for the tested organics) than the established Danio rerio embryo test.

Exposure to Treated Sewage Effluent Disrupts Reproduction and Development in the Seasonally Breeding Ramshorn Snail (Subclass: Pulmonata, Planorbarius corneus)

Experiments were conducted to assess the impacts of exposure to sewage treatment works (STW) effluent upon the growth, reproductive function, and sexual development of the European mollusc, Planorbarius corneus under seasonally varying temperatures and photoperiodic conditions. In river water, a clear seasonal change in the number and weight of egg masses (during both 2003 and 2004), and in the number of eggs produced, was found, providing evidence for profound effects of both changing temperature and photoperiod on reproduction. Exposure to STW effluent caused disturbances in this seasonal reproductive cycle at all concentrations tested. The effects included significant dose-dependent increases in fecundity and in the overall length of the reproductive cycle in adult snails exposed to both 50% and 100% effluent relative to river water for a period of up to 14 weeks. Disturbances in the development of both the male and female gametes of the both the adult snails and their developmentally exposed offspring were also seen. These effects were more evident in the offspring than in the adults.

Triphenyltin alters lipid homeostasis in females of the ramshorn snail Marisa cornuarietis

Molluscs are sensitive species to the toxic effects of organotin compounds, particularly to masculinisation. Both tributyltin (TBT) and triphenyltin (TPT) have been recently shown to bind to mollusc retinoid X receptor (RXR). If RXR is involved in lipid homeostasis, exposure to TPT would have an immediate effect on endogenous lipids. To test this hypothesis, the ramshorn snail Marisa cornuarietis was exposed to environmentally relevant concentrations of TPT (30, 125, 500 ng/L as Sn) in a semi-static water regime for 7 days. Percentage of lipids and total fatty acid content decreased significantly in TPT-exposed females while the activity of peroxisomal acyl-CoA oxidase, involved in fatty acid catabolism, increased. In addition, fatty acid profiles (carbon chain length and unsaturation degree) were significantly altered in exposed females but not in males. This work highlights the ability of TPT to disrupt lipid metabolism in M. cornuarietis at environmentally realistic concentrations and the higher susceptibility of females in comparison to males.

Biomphalaria havanensis Identified as a Potential Intermediate Host for the Digenetic Trematode Bolbophorus damnificus

AbstractThe digenetic trematode Bolbophorus damnificus has been associated with mortalities in commercial channel catfish Ictalurus punctatus in the Mississippi Delta. In the life cycle of B. damnificus, the only confirmed first intermediate host is the ram's horn snail Planorbella trivolvis. Recently, the exotic snail Biomphalaria havanensis has been isolated in several channel catfish ponds in the Mississippi Delta. The aim of this study was to determine whether this invasive snail species could also serve as a first intermediate host in the B. damnificus life cycle. Bolbophorus damnificus ova were collected from an American white pelican Pelecanus erythrorhynchos that had been artificially infected with B. damnificus metacercariae; the ova were then introduced into an aquarium with parasite‐negative B. havanensis. Bolbophorus damnificus cercariae (confirmed by polymerase chain reaction) were detected in B. havanensis 45 d after exposure to B. damnificus ova. The snails continued to shed B. damnificus cercariae throughout the 80‐d postexposure examination period. Before this research, the only reported natural B. damnificus infection of snails involved P. trivolvis. This study indicates that the range of snail hosts is broader than previously suspected. A survey should be implemented to identify additional snail species that can serve as hosts for B. damnificus. Further research should determine whether B. havanensis is a natural host for B. damnificus, and if so the prevalence of B. havanensis in commercial channel catfish ponds should be assessed.

Introduction, distribution, spread, and impacts of exotic freshwater gastropods in Texas

We examined the patterns of distribution, vectors of introduction, and potential ecological impacts of freshwater exotic species in Texas over the last 45 years. Currently, five species of exotic gastropods are established: channeled-type applesnail (Pomacea insularum), red-rim melania (Melanoides tuberculatus), quilted melania (Tarebia granifera), giant rams-horn snail (Marisa cornuarietis), and Chinese mysterysnail (Cipangopaludinachinensis). In contrast to the northern part of the US, where shipping appears to be the most important vector for the introduction of aquatic invasive species, aquarium and ornamental trade dominated among unintentional vectors of introduction of all freshwater exotics in Texas, resulting in different patterns of distribution, spread, and ecological impacts. The rate of spread of exotic gastropods in Texas varied from 39 waterbodies colonized over 18 years for P. insularum to only three waterbodies during last 45 years for C.chinensis. Four of five exotic gastropods were found in highly vulnerable aquifer-fed springs and rivers, which contain numerous endemic and endangered species. The fifth species, Pomacea insularum, is an agricultural pest. Potential negative ecological effects of exotic gastropods include impacts on wetlands and wetland restoration, competitive exclusion of native snails, and the introduction of exotic parasites, trematodes, which could infect fish and waterfowl, including federally protected species. Aquifer springs with stable temperature regimes are refuges for both cold and warm intolerant species.

Short-term exposure to the organotin compound triphenyltin modulates esterified steroid levels in females of Marisa cornuarietis

Long-term exposures to organotin compounds have shown alterations on endogenous steroid levels in gastropods together with the development of imposex. However, information regarding short-term effects of these compounds on the endocrine system of gastropods is lacking. This work aimed at investigating those responses in the ramshorn snail Marisa cornuarietis by looking at both endogenous levels of free and esterified steroids and the metabolism of the androgen precursor androstenedione by digestive gland/gonad microsomal fractions. One-week exposure to the organotin compound triphenyltin (TPT) led to a significant increase in esterified testosterone (60–85%) and a decrease in esterified estradiol (50–84%) in females, but had no effect on males. The observed alterations in esterified steroids were not directly related to changes in P450 aromatase activity that remained unchanged in exposed females. The enzymes involved in the metabolism of the androgen precursor androstenedione, namely 17β-hydroxysteroid dehydrogenases and 5α-reductases, were not significantly altered by TPT exposure, suggesting that such enzymes are not primary targets of TPT in M. cornuarietis. Additional studies are needed to fully understand the significance of the observed alterations in females and their potential relationship with the development of imposex.