A conserved gastropod withdrawal circuit in Biomphalaria glabrata, an intermediate host for schistosomiasis.

Neuronal signals mediated by the biogenic amine serotonin (5-HT) underlie critical survival strategies across the animal kingdom. This investigation examined serotonin-like immunoreactive neurons in the cerebral ganglion of the panpulmonate snail Biomphalaria glabrata, a major intermediate host for the trematode parasite Schistosoma mansoni. Five neurons comprising the cerebral serotonergic F (CeSF) cluster of B. glabrata shared morphological characteristics with neurons that contribute to withdrawal behaviors in numerous heterobranch species. The largest member of this group, designated CeSF-1, projected an axon to the tentacle, a major site of threat detection. Intracellular recordings demonstrated repetitive activity and electrical coupling between the bilateral CeSF-1 cells. In semi-intact preparations, the CeSF-1 cells were not responsive to cutaneous stimuli but did respond to photic stimuli. A large FMRF-NH2-like immunoreactive neuron, termed C2, was also located on the dorsal surface of each cerebral hemiganglion near the origin of the tentacular nerve. C2 and CeSF-1 received coincident bouts of inhibitory synaptic input. Moreover, in the presence of 5-HT they both fired rhythmically and in phase. As the CeSF and C2 cells of Biomphalaria share fundamental properties with neurons that participate in withdrawal responses in Nudipleura and Euopisthobranchia, our observations support the proposal that features of this circuit are conserved in the Panpulmonata.NEW & NOTEWORTHY Neuronal signals mediated by the biogenic amine serotonin underlie critical survival strategies across the animal kingdom. This investigation identified a group of serotonergic cells in the panpulmonate snail Biomphalaria glabrata that appear to be homologous to neurons that mediate withdrawal responses in other gastropod taxa. It is proposed that an ancient withdrawal circuit has been highly conserved in three major gastropod lineages.

Lethality and Acetylcholinesterase Inhibition in a Native Invertebrate Species Exposed to Water Samples of an Impacted Stream (Reconquista River Basin, Argentina).

The study of multiple biomarkers in bioindicator species is a useful tool to evaluate water quality in addition to physicochemical analysis. The aim of this work was to study the toxicity of water samples from two sites with different anthropogenic impacts (R: near a residential area and FP: close to horticultural farms and industrial waste treatment plants) from Las Catonas sub-basin (Reconquista River basin) in the native gastropod Biomphalaria straminea. Some physicochemical parameters and chlorpyrifos concentration were measured in water samples. Snails were exposed in laboratory conditions 48 h to the water samples and neurotoxicity, behavior, lethality and acetylcholinesterase, carboxylesterase, glutathione S-transferase, glutathione reductase and catalase activities were measured. In water from FP, chlorpyrifos was detected and conductivity and pH were higher than in R. Lethality (60%) and a decrease (30%) in acetylcholinesterase were observed in snails exposed to FP indicating that water contamination causes high toxicity in B. straminea.

Inhibition of alternative oxidase disrupts the development and oviposition of Biomphalaria glabrata snails.

BACKGROUND: Biomphalaria glabrata is one of the main intermediate hosts of Schistosoma mansoni, the most widespread species of Schistosoma. Our previous studies proved that alternative oxidase (AOX), the terminal oxidase in the mitochondrial respiratory chain, widely exists in several species of intermediate host snails of Schistosoma. Meanwhile, inhibition of AOX activity in Oncomelania hupensis snails could dramatically enhance the molluscicidal effect of niclosamide. As a hermaphroditic aquatic mollusc, the high fecundity and population density of B. glabrata increase the difficulty of snail control, which is one of the critical strategies for schistosomiasis elimination. The present study aimed to investigate the possible role of AOX in the development and fecundity of B. glabrata snail, which could be manipulated more manageable than other species of intermediate host snails of Schistosoma. METHODS: The dynamic expression of the AOX gene was investigated in different developmental stages and tissues of B. glabrata, with morphological change and oviposition behaviour observed from juvenile to adult snails. Furtherly, dsRNA-mediated knockdown of BgAOX mRNA and the AOX protein activity inhibiting was performed to investigate the effect of AOX on the development and oviposition of snails. RESULTS: The BgAOX gene expression profile is highly related to the development from late juveniles to adults, especially to the reproductive system of snails, with a positive correlation of 0.975 between egg production and BgAOX relative expression in ovotestis of snails. The inhibition of BgAOX at the transcriptional level and AOX activity could efficiently inhibit snail growth. However, the interference at the BgAOX protein activity level led to more severe tissue damage and more significant inhibition of oviposition than at the transcriptional level. This inhibition of growth and oviposition decreased gradually with the increase in the snail size. CONCLUSIONS: The inhibition of AOX could efficiently disrupt the development and oviposition of B. glabrata snails, and the intervention targeting AOX at the juvenile stage is more effective for snails. This investigation explored the role of AOX in the growth and development of snails. It would benefit snail control in the future by providing a potential target while using molluscicides more efficiently.

A new accumulation assay of Schistosoma mansoni miracidia using square capillary glass tubes.

Current control measures for schistosomiasis have only been partially successful in endemic areas due to socioeconomic constraints. One possibility for controlling the disease is to aim at the miracidial stage of the trematode to avoid infecting intermediate snail hosts by introducing more attractive substances for miracidia in the environment. Here, we introduce an accumulation assay of Schistosoma mansoni miracidia using a square glass tube for analysis of the positive responses of miracidia toward several substances, including snail-conditioned water of Biomphalaria glabrata, Bulinus globosus and insusceptible snails collected in the Nagasaki area in Japan. The substances are not proteins because miracidia accumulated in boiled snail-conditioned water and the secretion or emission level of substances depended on the feeding conditions of Biomphalaria glabrata. The present study also showed that substances emitted from Biomphalaria glabrata with a molecular weight around 10 kDa accumulated Schistosoma mansoni miracidia. Further, we showed that Schistosoma mansoni miracidia did not accumulate in response to mono- or disaccharides tested in the study.

Opportunity or catastrophe? effect of sea salt on host-parasite survival and reproduction.

Seawater intrusion associated with decreasing groundwater levels and rising seawater levels may affect freshwater species and their parasites. While brackish water certainly impacts freshwater systems globally, its impact on disease transmission is largely unknown. This study examined the effect of artificial seawater on host-parasite interactions using a freshwater snail host, Biomphalaria alexandrina, and the human trematode parasite Schistosoma mansoni. To evaluate the impact of increasing salinity on disease transmission four variables were analyzed: snail survival, snail reproduction, infection prevalence, and the survival of the parasite infective stage (cercariae). We found a decrease in snail survival, snail egg mass production, and snail infection prevalence as salinity increases. However, cercarial survival peaked at an intermediate salinity value. Our results suggest that seawater intrusion into freshwaters has the potential to decrease schistosome transmission to humans.

Interventions can shift the thermal optimum for parasitic disease transmission.

Temperature constrains the transmission of many pathogens. Interventions that target temperature-sensitive life stages, such as vector control measures that kill intermediate hosts, could shift the thermal optimum of transmission, thereby altering seasonal disease dynamics and rendering interventions less effective at certain times of the year and with global climate change. To test these hypotheses, we integrated an epidemiological model of schistosomiasis with empirically determined temperature-dependent traits of the human parasite Schistosoma mansoni and its intermediate snail host (Biomphalaria spp.). We show that transmission risk peaks at 21.7 °C (Topt ), and simulated interventions targeting snails and free-living parasite larvae increased Topt by up to 1.3 °C because intervention-related mortality overrode thermal constraints on transmission. This Topt shift suggests that snail control is more effective at lower temperatures, and global climate change will increase schistosomiasis risk in regions that move closer to Topt Considering regional transmission phenologies and timing of interventions when local conditions approach Topt will maximize human health outcomes.

Comparative Vectorial Competence of Biomphalaria sudanica and Biomphalaria choanomphala, Snail Hosts of Schistosoma mansoni, From Transmission Hotspots In Lake Victoria, Western Kenya.

Schistosoma mansoni, which causes human intestinal schistosomiasis, continues to be a major public health concern in the Lake Victoria basin in western Kenya, with Biomphalaria sudanica (a shoreline inhabiting snail) and Biomphalaria choanomphala (a deep-water snail) playing roles in transmission. A recent study showed that B. sudanica was abundantly present near all study villages on the lakeshore, but B. choanomphala was significantly more abundant near villages known to be persistent transmission hotspots. The present study investigated the relative compatibility of B. sudanica and B. choanomphala with S. mansoni. A reciprocal cross-infection experiment used young adult F1 generation B. sudanica and B. choanomphala that were exposed to either 1, 5, or 10 sympatric or allopatric human-derived S. mansoni miracidia. Three weeks post-exposure (PE) and weekly thereafter, the snails were counted and screened for schistosome cercariae, and at 7 wk PE, total cercariae shed during a 2 hr period by each infected snail was determined. Pre-patent periods for S. mansoni in both B. sudanica and B. choanomphala were similar, and most snails in all exposure combinations started shedding cercariae 5 wk PE. Prevalences were significantly higher in B. choanomphala (12.2-80.9%) than in B. sudanica (5.2-18.6%) at each dose, regardless of whether miracidia were of an allopatric or a sympatric source (P < 0.0001). Overall, the odds of a snail becoming infected with 5 or 10 miracidia were significantly higher than the odds of being infected with 1 miracidium, (P < 0.0001), and fewer cercariae were produced by snails exposed to single as compared to 5 or 10 miracidia. On average, B. choanomphala produced more cercariae ( = 458, SD = 414) than B. sudanica ( = 238, SD = 208) (P < 0.0001). These results suggest that B. choanomphala is more compatible with S. mansoni than B. sudanica. Though B. choanomphala can be found in shallow shoreline waters, it is, for the most part, a deeper-water taxon. Because dredging is a relatively inefficient means of sampling, B. choanomphala is likely underestimated with respect to its population size, the number of S. mansoni-positive snails, and its role in maintaining transmission.

Modeling schistosomiasis transmission: the importance of snail population structure.

BACKGROUND: Schistosomiasis is a neglected tropical disease endemic in 54 countries. A major Schistosoma species, Schistosoma mansoni, is sustained via a life cycle that includes both human and snail hosts. Mathematical models of S. mansoni transmission, used to elucidate the complexities of the transmission cycle and estimate the impact of intervention efforts, often focus primarily on the human host. However, S. mansoni incurs physiological costs in snails that vary with the age of the snail when first infected. Snail demography and the age of snail infection could thus affect the force of infection experienced by humans, which is frequently used to predict the impact of various control strategies. METHODS: To address how these snail host and parasite interactions influence model predictions, we developed deterministic models of schistosomiasis transmission that include varying complexity in the snail population age structure. Specifically, we examined how model outputs, such as schistosome prevalence in human and snail populations, respond to the inclusion of snail age structure. RESULTS: Our models suggest that snail population age structure modifies the force of infection experienced by humans and the relationship between snail infection prevalence and corresponding human infection prevalence. There are significant differences in estimated snail infection, cercarial density and mean worm burden between models without snail population dynamics and those with snail populations, and between models with a homogeneous snail population and those with age stratification. The variation between finely age-stratified snail populations and those grouped into only juvenile and adult life stages is, however, minimal. CONCLUSIONS: These results indicate that including snails and snail age structure in a schistosomiasis transmission model alters the relationship between snail and human infection prevalence. This highlights the importance of accounting for a heterogeneous intermediate host population in models of schistosomiasis transmission where the impact of proposed control measures is being considered.

Modelling the present and future distribution of Biomphalaria species along the watershed of the Middle Paranapanema region, São Paulo, Brazil.

The Middle Paranapanema region in the state of São Paulo, Brazil, is an area with high diversity for Biomphalaria species, with municipalities historically marked by cases of schistosomiasis transmission. The objectives of the study were to evaluate the current distribuition and predict the future distribution of habitats of Biomphalaria species at a high spatial resolution along 114 freshwater sites in the Middle Paranapanema watershed. The modelling encompassed 55 municipalities of the Middle Paranapanema region, which were analyzed through the maximum entropy algorithm. All geographic coordinates of the Biomphalaria species collected from 2015-2018 and environmental data were obtained through WorldClim, HydroSHEDS, TOPODATA and Secretaria do Meio Ambiente for the 1970-2017 period. For the 2041-2060 period we used the HadGEM2-ES climate model. Due to climate change, MaxEnt showed that there was a high probability for the maintenance of B. glabrata habitats near Ourinhos and Assis, an expansion of scattered spots, and a 50% probability that the species will spread throughout new suitable areas. The results showed that the geographical range of B. straminea will most likely expand in the future along the Middle Paranapanema hydrographic basin, especially in the municipalities near Ourinhos. For B. glabrata and B. straminea, the geographic expansion was related to the predicted increase in the annual temperature range. The habitats suitable for B. tenagophila and B. peregrina seemed to slightly expand around the west border of the Middle Paranapanema region. Biomphalaria occidentalis may have a small reduction in its distribution due to climate change. The variables that contributed the most to the future modelling for these three species were precipitation and temperature. Identifying the sites with intermediate hosts for schistosomiasis may guide public health measures to avoid or reduce future transmissions in this region.

A comparative study of enzymatic and immunological parameters in Planorbarius corneus and Biomphalaria glabrata exposed to the organophosphate chlorpyrifos.

This study aimed to investigate the acute effects of chlorpyrifos on biomarkers related to neurotoxicity and immunotoxicity in two allopatric freshwater gastropod species belonging to the family Planorbidae. For this purpose, Planorbarius corneus and Biomphalaria glabrata were exposed to chlorpyrifos (active ingredient or commercial formulation) for 48 h at environmentally realistic concentrations (1 and 7.5 µg L-1). Basal acetylcholinesterase activity in soft tissues and hemolymph was almost one order of magnitude higher in P. corneus than in B. glabrata. However, upon chlorpyrifos exposure, statistically significant inhibition of enzymatic activity was registered in both species. Acetylcholinesterase was more sensitive to inhibition in soft tissues than in hemolymph. The highest inhibition was observed in the B. glabrata soft tissues exposed to the commercial formulation (88 % at 1 µg L-1 and 93 % at 7.5 µg L-1). Hemocyte number and lysosomal membrane stability did not show significant changes with respect to controls in any of the exposed groups. Superoxide anion generation was diminished (21-46 %) in P. corneus hemocytes exposed to the active ingredient and in B. glabrata hemocytes exposed to the active ingredient or the formulation. In contrast, hemocyte phagocytic activity increased in all exposed groups. Phagocytosis was most stimulated (89 %) in hemocytes sampled from B. glabrata treated with 7.5 µg L-1 chlorpyrifos. Altogether the results suggest that the freshwater gastropods P. corneus and B. glabrata are suitable model animals for environmental monitoring studies in the Northern Hemisphere and Latin America, respectively. Furthermore, these results add information on the relevance of testing pesticide formulations and on the usefulness of acetylcholinesterase inhibition and immunological parameters as biomarkers of the acute effects of chlorpyrifos in these species.

Behavioral response of Biomphalaria glabrataexposed to a sublethal concentration of Euphorbia miliivar. hislopii latex.

The Euphobia milii var. hislopii latex has been tested in the control of schistosomiasis but its action in the locomotor activity of Biomphalaria glabrata is unknown. The objective of this work was to study the locomotor and reproductive behaviors of B. glabrata exposed to E. milii var. hislopii latex. For this, 96 snails were individually exposed to the latex (LC50 - 0.53 mg / L) for 24 hours. The specimens were submitted to biomonitoring for image analysis to record the locomotor parameters at different times: before exposure (control), one day post exposure (group 1 d-p-e) and 30 days post exposure (group 30 d-p-e). The reproductive parameters were recorded weekly for 10 weeks. All locomotor activities of group 1 d-p-e decreased significantly. The egg/egg mass ratio decreased the week after exposure while there was an increase in the hatching rate. After 30 days, these reproductive parameters were similar to those observed in these same snails before exposure. However, the number of hatched snails declined after exposure until the end of the experiment. The influence of the latex in the parameters of B. glabrata added further evidence that this natural water-soluble product can be an important tool for the control of schistosomiasis.

Study of genotoxic and cytotoxic effects after acute and chronic exposures to industrial sewage sludge on Biomphalaria glabrata hemocytes.

Industrial development has provided numerous benefits to improve quality of life in modern times, however, it has also led to the development and use of a large number of toxic chemicals which have caused damage to various ecosystems. Consequently, knowledge of techniques and organisms that can be used to monitor, identify and quantify environmental pollutants has become increasingly relevant. Therefore, the objective of this study was to use the mollusk Biomphalaria glabrata to analyze biomarker and biomonitoring parameters of industrial sewage sludge. To perform the tests, concentrations of 50, 100, 150 and 500 mg L-1 of industrial sewage sludge were standardized. All the tests were performed after the animals were exposed to the sludge in acute and chronic forms. Embryos exposure to sludge did not show a significant percentage of the animals non-viable when compared to the control group. Subsequently, hemocytes were analyzed for the presence of cytoplasmic and nuclear alterations. Finally, the comet test was performed to quantify the genotoxic damage caused by exposure to industrial sludge. Analysis hemocytes showed a significant number of cellular alterations was observed, mainly due to the high frequency of apoptosis. Moreover, during the analysis of nucleoids several degrees of nuclear damage were identified, with the groups exposed to the highest concentrations presenting the greatest genotoxic damage. Thus, we can conclude that the parameters evaluated in the mollusk Biomphalaria glabrata have proven to be a good tool, along with other techniques and complementary organisms, to assist aspects related to biomonitoring of freshwater ecosystems.

Striking differences in virulence, transmission and sporocyst growth dynamics between two schistosome populations.

BACKGROUND: Parasite traits associated with transmission success, such as the number of infective stages released from the host, are expected to be optimized by natural selection. However, in the trematode parasite Schistosoma mansoni, a key transmission trait, i.e. the number of cercariae larvae shed from infected Biomphalaria spp. snails, varies significantly within and between different parasite populations and selection experiments demonstrate that this variation has a strong genetic basis. In this study, we compared the transmission strategies of two laboratory schistosome population and their consequences for their snail host. METHODS: We infected inbred Biomphalaria glabrata snails using two S. mansoni parasite populations (SmBRE and SmLE), both isolated from Brazil and maintained in the laboratory for decades. We compared life history traits of these two parasite populations by quantifying sporocyst growth within infected snails (assayed using qPCR), output of cercaria larvae and impact on snail host physiological response (i.e. hemoglobin rate, laccase-like activity) and survival. RESULTS: We identified striking differences in virulence and transmission between the two studied parasite populations. SmBRE (low shedder (LS) parasite population) sheds very low numbers of cercariae and causes minimal impact on the snail physiological response (i.e. laccase-like activity, hemoglobin rate and snail survival). In contrast, SmLE (high shedder (HS) parasite population) sheds 8-fold more cercariae (mean ± SE cercariae per shedding: 284 ± 19 vs 2352 ± 113), causes high snail mortality and has strong impact on snail physiology. We found that HS sporocysts grow more rapidly inside the snail host, comprising up to 60% of cells within infected snails, compared to LS sporocysts, which comprised up to 31%. Cercarial production is strongly correlated to the number of S. mansoni sporocyst cells present within the snail host tissue, although the proportion of sporocyst cells alone does not explain the low cercarial shedding of SmBRE. CONCLUSIONS: We demonstrated the existence of alternative transmission strategies in the S. mansoni parasite consistent with trade-offs between parasite transmission and host survival: a "boom-bust" strategy characterized by high virulence, high transmission and short duration infections and a "slow and steady" strategy with low virulence, low transmission but long duration of snail host infections.

A Search for Snail-Related Answers to Explain Differences in Response of Schistosoma mansoni to Praziquantel Treatment among Responding and Persistent Hotspot Villages along the Kenyan Shore of Lake Victoria.

Following a 4-year annual praziquantel (PZQ) treatment campaign, the resulting prevalence of Schistosoma mansoni was seen to differ among individual villages along the Kenyan shore of Lake Victoria. We have investigated possible inherent differences in snail-related aspects of transmission among such 10 villages, including six persistent hotspot (PHS) villages (≤ 30% reduction in prevalence following repeated treatments) located along the west-facing shore of the lake and four PZQ-responding (RESP) villages (> 30% prevalence reduction following repeated treatment) along the Winam Gulf. When taking into account all sampling sites, times, and water hyacinth presence/absence, shoreline-associated Biomphalaria sudanica from PHS and RESP villages did not differ in relative abundance or prevalence of S. mansoni infection. Water hyacinth intrusions were associated with increased B. sudanica abundance. The deeper water snail Biomphalaria choanomphala was significantly more abundant in the PHS villages, and prevalence of S. mansoni among villages both before and after control was positively correlated with B. choanomphala abundance. Worm recoveries from sentinel mice did not differ between PHS and RESP villages, and abundance of non-schistosome trematode species was not associated with S. mansoni abundance. Biomphalaria choanomphala provides an alternative, deepwater mode of transmission that may favor greater persistence of S. mansoni in PHS villages. As we found evidence for ongoing S. mansoni transmission in all 10 villages, we conclude that conditions conducive for transmission and reinfection occur ubiquitously. This argues for an integrated, basin-wide plan for schistosomiasis control to counteract rapid reinfections facilitated by large snail populations and movements of infected people around the lake.

Reprotoxicity of 4-nonylphenol to Biomphalaria alexandrina snails at certain temperatures.

One of the most common compounds in pesticide formulations, plastics, and papers is 4-nonylphenol (4-NP). It is contained in agricultural, industrial, and wastewater effluents, which when discharged into surface waters affect aquatic fauna. Therefore, the present study aimed to use Biomphalaria alexandrina snails to evaluate the chronic toxicity of 4-NP. Its concentrations in collected water samples from Giza Governorate ranged from 400 to 1600 µg/l. Based on these environmentally relevant concentrations, laboratory experiments were carried out using standard 4-NP to investigate the effect of three concentrations; namely 400, 750, and 1600 µg/l. Survival rate of the exposed snails to 4-NP concentrations was affected after 4 weeks. Reproduction of the exposed snails to 4-NP concentrations was lower than that of the control at 30 °C, while the exposed snails to 400 µg/l of 4-NP showed maximum reproduction at 15 °C. The lowest hatchability percentage was recorded with egg masses laid by the exposed snails to 400 and 1600 µg/l of 4-NP at 15 and 30 °C, respectively. Furthermore, the results showed fluctuated levels of progesterone, estradiol, and testosterone depending upon the concentration and the temperature, which played a key role in determining the degree of 4-NP toxicity.

Host-parasite life-histories of the diurnal vs. nocturnal chronotypes of Schistosoma mansoni: adaptive significance.

OBJECTIVES: To optimise host-to-host transmission, digenean trematodes (parasites) synchronise their cercarial emission patterns with the aquatic activities of their vertebrate hosts. Schistosoma mansoni has two different circadian chronotypes: a diurnal shedding pattern with a mean peak occurring at 11:00 h, and a nocturnal shedding pattern with a mean peak occurring at 20:00 h. We analysed the life-history variations between these two chronotypes at the levels of the parasite and its hosts. METHODS: For each chronotype, we quantified three life-history traits related to the parasite (prepatent period, infection rate and cercarial production) and analysed the morphometry and the morphology of the parasite eggs; we also quantified three life-history traits related to the snail intermediate host (shell diameter, fecundity and survival rate) and one life-history trait related to the experimental definitive host (survival rate). A phylogeny based on the mitochondrial cytochrome-oxidase gene was made on samples of both chronotypes. RESULTS: Life-history analysis revealed significant variations between the two chronotypes. Life-history traits were optimal for both the parasite and the snail host for the diurnal chronotype compared to the nocturnal one. The new chronotype behaved like an allopatric population towards its snail host. Phylogenetic analysis supports the hypothesis of a lateral transfer of S. mansoni from humans to Rattus rattus. These results were interpreted in terms of an ongoing sympatric speciation. CONCLUSION: The nocturnal chronotype of S. mansoni showed non-adapted life-history traits in its relation with the snail intermediate host Biomphalaria pfeifferi. The emergence of this new phenotype is probably linked to divergent natural selection.

OBJECTIFS: Afin d'optimiser la transmission d'hôte à hôte, les trématodes digènes (parasites) synchronisent leurs schémas d'émission cercarienne avec les activités aquatiques de leurs hôtes vertébrés. Schistosoma mansoni a deux chronotypes circadiens différents: un schéma de libérations diurnes avec un pic moyen survenant à 11h00 et un schéma nocturne avec un pic moyen à 20h00. Nous avons analysé les variations de l'histoire de vie entre ces deux chronotypes aux niveaux du parasite et de ses hôtes. MÉTHODES: Pour chaque chronotype, nous avons quantifié trois traits d'histoire de vie liés au parasite (période prépatente, taux d'infection et production cercarienne) et avons analysé la morphométrie et la morphologie des œufs du parasite; nous avons également quantifié trois traits d'histoire de vie liés à l'hôte intermédiaire escargot (diamètre de la coquille, fécondité et taux de survie) et un trait d'histoire de vie lié à l'hôte définitif expérimental (taux de survie). Une phylogénie basée sur le gène mitochondrial de la cytochrome oxydase a été réalisée sur des échantillons des deux chronotypes. RÉSULTATS: L'analyse de l'histoire de vie a révélé des variations significatives entre les deux chronotypes. Les traits d'histoire de vie étaient optimaux à la fois pour le parasite et pour l'hôte escargot pour le chronotype diurne par rapport au chronotype nocturne. Le nouveau chronotype se comportait comme une population allopatrique vis-à-vis de son hôte escargot. L'analyse phylogénétique soutient l'hypothèse d'un transfert latéral de S. mansoni de l'homme à Rattus rattus. Ces résultats ont été interprétés en termes de spéciation sympatrique en cours. CONCLUSION: Le chronotype nocturne de S. mansoni montre des traits d'histoire de vie non adaptés dans sa relation avec l'hôte intermédiaire escargot, Biomphalaria pfeifferi. L'émergence de ce nouveau phénotype est probablement liée à une sélection naturelle divergente.

Alterations in the Mitochondrial Physiology of Biomphalaria glabrata (Mollusca: Gastropoda) After Experimental Infection by Angiostrongylus cantonensis (Nematoda: Metastrongylidae).

BACKGROUND: Angiostrongylus cantonensis is a metastrongylid nematode that has a heteroxenous cycle, where snails act as intermediate hosts and the rodents Rattus rattus and Rattus novergicus are the definitive hosts. However, humans may act as accidental hosts presenting an atypical form of parasitism. This fact has motivated research to better understand systems of relationships involving A. cantonensis, targeting the control of species of gastropods that act as intermediary hosts. METHODS: For this, six groups were formed: three control groups (uninfected) and three infected groups, exposed to approximately 1200 L1 larvae of A. cantonensis. At the end of each week (1, 2, and 3 weeks), snails were dissected without anesthesia and the gonad-digestive gland (DGG) complex was separated for determination of oxygen consumption through high-resolution titration-injection respirometer (Oroboros, Oxygraph; Innsbruck, Austria). RESULTS: The results indicate suppression of mitochondrial oxidative metabolism of the host and compromised in different mitochondrial respiratory states. This effect, mainly observed in the group exposed to 1 week of infection, showed a decrease of approximately 38% (2.78 ± 0.37 pmol O2/mg of tissue; P < 0.05), 41% (2.76 ± 0.34 pmol O2/mg of tissue; P < 0.05) e 46% (2.91 ± 0.36 pmol O2/mg of tissue; P < 0.05) in the basal oxygen consumption after sequential addition (P + M), succinate and (ADP) in the respiratory medium, differing significantly from the control group. CONCLUSION: The results presented indicate that the prepatent infection by this metastrongylid impairs the aerobic oxidative metabolism of its host, causing a reduction in basal oxygen consumption. This effect, observed at the start of development of the parasites, indicates that this stage is the most critical for the success of the infection, and can be explained by a reduction of the mitochondrial density of the tissue analyzed, or also by suppression of enzyme centers related to the oxidative reactions.

An immunohistochemical analysis of peptidergic neurons apparently associated with reproduction and growth in Biomphalaria alexandrina.

Peptide hormones and neurotransmitters involved in reproduction and growth have been studied extensively in certain gastropod molluscs, such as Lymnaea stagnalis and Aplysia californica. The present study employs antisera that have been used to study peptidergic neurons in those species to probe the central nervous system of another gastropod, Biomphalaria alexandrina, an intermediate host of the parasitic trematode that causes schistosomiasis in humans. Whole mount preparations of central ganglia were stained immunohistochemically, and several populations of neurons appeared to be homologous to those forming the neuroendocrine axis that has been previously described in L. stagnalis. These cells include the caudodorsal cells and the light green and canopy cells, which produce hormones that regulate ovulation and growth, respectively. Other populations of cells containing APGWamide, FMRFamide and/or related peptides are consistent with ones that innervate the penis in L. stagnalis and other gastropods. Identification of neurons that might be responsible for the control of reproduction and growth in Biomphalaria provides an important initial step toward the development of novel methods of disease control and pest management directed toward reducing snail populations.

Toxicological impact of oxyfluorfen 24% herbicide on the reproductive system, antioxidant enzymes, and endocrine disruption of Biomphalaria alexandrina (Ehrenberg, 1831) snails.

Oxyfluorfen (Goal 24%EC) herbicide is widely used in agriculture for weed control. Biomphalaria alexandrina snails can be used as bioindicator of the chemical pollution in the aquatic environment. The objective of this study was to evaluate the molluscicidal activity of this herbicide on Biomphalaria alexandrina snails and how it affected its biological system. The present study revealed a molluscicidal effect of oxyfluorfen 24%EC on these snails at LC50 5.9 mg/l. After exposure of snails to the sub-lethal concentrations (LC0, LC10, or LC25) of this herbicide, the survival rates, reproductive rate (R0), and fecundity (Mx) of adult B. alexandrina snails were significantly decreased in comparison with the control group. Also, levels of testosterone and estradiol were decreased significantly. It caused alterations in the antioxidant system, where exposure to sub-lethal concentration of this herbicide caused significant increases in levels of lipid peroxide malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD), while it significantly decreased glutathione transferase (GST). Histopathological changes in the digestive gland included severe damage in the digestive cells, where, they lost their tips and some were degenerated, while the secretory cells increased in number. Regarding the hermaphrodite gland, there were losses of the connective tissues, irregular sperms, and the eggs degenerated. These findings concluded that B. alexandrina snails can be used as a bioindicator for pollution with pesticide in the aquatic environment.

Morphological and molecular characterization of invasive Biomphalaria straminea in southern China.

BACKGROUND: Schistosomiasis is a common parasitic disease designated as a neglected tropical disease by the World Health Organization. Schistosomiasis mansoni is a form of the disease that is caused by the digenean trematode Schistosoma mansoni, transmitted through Biomphalaria spp. as an intermediate host. Biomphalaria was introduced to Hong Kong, China in aquatic plants shipments coming from Brazil and the snail rapidly established its habitats in southern China. Earlier studies of Biomphalaria spp. introduced to southern China identified the snails as Biomphalaria straminea, one of the susceptible species implicated in S. mansoni transmission in South America. However, recent molecular investigations also indicated the presence of another South American species, B. kuhniana, which is refractory to infection. As such, it is important to identify accurately the species currently distributed in southern China, especially with emerging reports of active S. mansoni infections in Chinese workers returning from Africa. METHODS: We combined morphological and molecular taxonomy tools to precisely identify Biomphalaria spp. distributed in Guangdong Province, southern China. In order to clearly understand the molecular profile of the species, we constructed a phylogeny using mtDNA data (COI and 16S rRNA sequences) from six populations of Biomphalaria spp. from Shenzhen City in Guangdong Province. In addition, we examined the external morphology of the shell and internal anatomy of the reproductive organs. RESULTS: Both morphological and molecular evidences indicated a close affinity between Biomphalaria spp. populations from Guangdong and B. straminea from Brazil. The shell morphology was roughly identical in all the populations collected with rounded whorls on one side and subangulated on the other, a smooth periphery, an egg-shaped aperture bowed to one side, and a deep umbilicus. The shape and number of prostate diverticula (ranged from 11.67 to 17.67) in Guangdong populations supports its close affinity to B. straminea rather than B. kuhniana. Molecular analysis did not conflict with morphological analysis. Little genetic differentiation was observed within Biomphalaria populations collected. Phylogenetic analysis of COI and 16S rRNA haplotypes from snails collected and B. straminea sequences from Brazil and China using Bayesian inference revealed that Guangdong populations were clustered in one clade with B. straminea from Hong Kong of China and B. straminea from Brazil indicating their close affinity to each other. CONCLUSIONS: Data obtained in the current study clearly show that the populations of Biomphalaria spp. investigated are B. straminea, and we assume that those snails were either introduced via passive dispersal from Hong Kong of China or as a result of multiple introduction routes from Brazil.