Snail Biomphalaria Glabrata Research Articles

Molecular Characterization of Rotifers and Their Potential Use in the Biological Control of Biomphalaria.

BackgroundSchistosomiasis is one of the most important tropical parasitic diseases worldwide. Biomphalaria straminea, the intermediate host of Schistosoma mansoni, has invaded and spread to Southern China since 1974 and may pose enormous threats to public health. Controlling intermediate host snails is an effective strategy in schistosomiasis intervention. However, the only effective chemical molluscicide, niclosamide, currently recommended by WHO may cause environmental pollution, loss of biodiversity, and high costs. Thus, to counter intermediate hosts, a sustainable and environmentally friendly tool is urgently needed. Here, we conducted field investigations to collect and identify a potential snail competitor rotifer and evaluated its molluscicide effect.ResultsIn this study, we collected two samples of rotifers from Shenzhen. We found both red and black phenotypic B. straminea snails at the sampling sites. We identified the rotifer population as a species of the genus Philodina according to the amplification and phylogenetic analysis results of coxI gene. We found that rotifer exposure did not significantly affect the hatching rate of B. straminea eggs but promoted the killing of juvenile snails. Meanwhile, rotifer exposure did not significantly alter the fecundity of B. straminea quantified by the number of eggs per egg mass, the number of egg masses per snail, and the number of eggs per snail; but the snails exposed to rotifers showed lower fecundity performance than the control snails. Importantly, rotifer exposure could significantly affect the development of juvenile B. straminea, showing a smaller shell diameter of the exposed snails than that of the control snails. In addition, rotifer exposure affected the life span of B. straminea snails, showing a 16.61% decline in the average life span. After rotifer exposure, the S. mansoni-infected B. straminea snails died significantly faster than those without rotifer exposure. Similar findings were observed in S. mansoni-infected Biomphalaria glabrata snails. These results implied that rotifer exposure significantly promoted the mortality of S. mansoni-infected B. straminea and B. glabrata.ConclusionsOur study demonstrated the potential molluscicide effect of rotifers on intermediate hosts under laboratory conditions. Our findings may provide new insights into the development of biocontrol strategies for snail-borne disease transmission.

PIWI silencing mechanism involving the retrotransposon nimbus orchestrates resistance to infection with Schistosoma mansoni in the snail vector, Biomphalaria glabrata.

Schistosomiasis remains widespread in many regions despite efforts at its elimination. By examining changes in the transcriptome at the host-pathogen interface in the snail Biomphalaria glabrata and the blood fluke Schistosoma mansoni, we previously demonstrated that an early stress response in juvenile snails, manifested by induction of heat shock protein 70 (Hsp 70) and Hsp 90 and of the reverse transcriptase (RT) domain of the B. glabrata non-LTR- retrotransposon, nimbus, were critical for B. glabrata susceptibility to S. mansoni. Subsequently, juvenile B. glabrata BS-90 snails, resistant to S. mansoni at 25°C become susceptible by the F2 generation when maintained at 32°C, indicating an epigenetic response. To better understand this plasticity in susceptibility of the BS-90 snail, mRNA sequences were examined from S. mansoni exposed juvenile BS-90 snails cultured either at 25°C (non-permissive temperature) or 32°C (permissive). Comparative analysis of transcriptomes from snails cultured at the non-permissive and permissive temperatures revealed that whereas stress related transcripts dominated the transcriptome of susceptible BS-90 juvenile snails at 32°C, transcripts encoding proteins with a role in epigenetics, such as PIWI (BgPiwi), chromobox protein homolog 1 (BgCBx1), histone acetyltransferase (BgHAT), histone deacetylase (BgHDAC) and metallotransferase (BgMT) were highly expressed in those cultured at 25°C. To identify robust candidate transcripts that will underscore the anti-schistosome phenotype in B. glabrata, further validation of the differential expression of the above transcripts was performed by using the resistant BS-90 (25°C) and the BBO2 susceptible snail stock whose genome has now been sequenced and represents an invaluable resource for molecular studies in B. glabrata. A role for BgPiwi in B. glabrata susceptibility to S. mansoni, was further examined by using siRNA corresponding to the BgPiwi encoding transcript to suppress expression of BgPiwi, rendering the resistant BS-90 juvenile snail susceptible to infection at 25°C. Given transposon silencing activity of PIWI as a facet of its role as guardian of the integrity of the genome, we examined the expression of the nimbus RT encoding transcript at 120 min after infection of resistant BS90 piwi-siRNA treated snails. We observed that nimbus RT was upregulated, indicating that modulation of the transcription of the nimbus RT was associated with susceptibility to S. mansoni in BgPiwi-siRNA treated BS-90 snails. Furthermore, treatment of susceptible BBO2 snails with the RT inhibitor lamivudine, before exposure to S. mansoni, blocked S. mansoni infection concurrent with downregulation of the nimbus RT transcript and upregulation of the BgPiwi encoding transcript in the lamivudine-treated, schistosome-exposed susceptible snails. These findings support a role for the interplay of BgPiwi and nimbus in the epigenetic modulation of plasticity of resistance/susceptibility in the snail-schistosome relationship.

Molluscicidal activity of monoterpenes and their effects on inhibition of acetylcholinesterase activity on Biomphalaria glabrata, an intermediate host of Schistosoma mansoni

The molluscicidal action of essential oils have been attributed to the most prevalent terpene compounds. However, molluscicidal properties, mode of action, and toxicity to non-target organisms remain unclear. In this study, the molluscicidal potential of four monoterpenes (camphor, thymol, α-pinene, and 1,8-cineole) against the snail Biomphalaria glabrata, an intermediate host of Schistosoma mansoni, was analyzed. The molluscicide activity of each monoterpene was assessed by the standardized test of the World Health Organization (WHO) and the monoterpenes considered active against B. glabrata were analyzed as inhibitors of the enzymatic activity of acetylcholinesterase (AChE) extracted from snails. In addition, acute toxicity to non-target organisms was assessed against Danio rerio fish. The results show that camphor and 1,8-cineole monoterpenes did not induce snail mortality. Thymol and α-pinene were active against B. glabrata, inducing mortality in concentration-dependent patterns and showing a lethal effect in concentrations compatible with that recommended by the WHO (LC90 of 7.11 and LC90 10.34 μg ∙ mL−1, respectively). The toxic action of thymol and α-pinene on snails indicates that these monoterpenes may account for or largely contribute to the molluscicidal activity of essential oils that contain them as major compounds. Thymol and α-pinene inhibit the AChE of B. glabrata at concentrations higher than those used in the molluscicide test. These monoterpenes show low toxicity to non-target organisms compared to the commercial molluscicide niclosamide. Knowledge about monoterpene toxicity against B. glabrata contributes to its potential use in molluscicidal formulations and in alternatives to the control of snails that host intermediate S. mansoni, a crucial action in the prevention and transmission of schistosomiasis, a neglected tropical disease.

Gonadal histopathology and inflammatory response in the freshwater snail exposed to iron oxide nanoparticles and ferric chloride: Insights into reproductive nanotoxicity.

Considering that most animals in an aquatic ecosystem are invertebrates, concerns about the ecotoxicological impact of emerging pollutants, such as nanomaterials, in these populations are relevant, which can lead to loss of aquatic biodiversity. However, knowledge concerning the effects of iron-based nanoparticles (IONPs) at cell and tissue-levels on freshwater gastropods remains limited. Thus, the present study aimed to analyse the histopathological changes and inflammatory response in the freshwater snail Biomphalaria glabrata after chronic exposure to gluconic-acid functionalized IONPs (GLA-IONPs) in comparison with their dissolved counterpart (FeCl3). Snails were exposed to both iron forms (1.0, 2.5, 6.25, and 15.62 mg L-1) for 28 days, and the qualitative and quantitative histopathological assessment on hermaphrodite gonads was conducted, following by analysis of histopathological indices and inflammatory responses. Results showed that both iron forms (GLA-IONPs and FeCl3) induced several gonadal histopathologies in the snails, mainly atresic acini, vacuolization of pre-vitellogenic oocytes, and atresic oocytes in a concentration-dependent pattern. GLA-IONPs induced a more intense inflammatory response and high frequency of vacuolized vitellogenic oocytes in comparison with FeCl3. Environmentally relevant concentration (2.5 mg L-1) of GLA-IONPs and FeCl3 induced high gonadal histopathological indices, indicating their potential reproductive toxicity. The current study showed that the chronic exposure of snails to GLA-IONPs and their dissolved counterpart (FeCl3) induced several gonadal histopathological changes and inflammatory responses in B. glabrata, confirming their potential risk to aquatic biodiversity.

Toxicity and genotoxicity of domestic sewage sludge in the freshwater snail Biomphalaria glabrata (Say, 1818).

Waste produced in homes is one of the main sources of pollutants in freshwater ecosystems. Therefore, it is imperative to implement methodologies that aid in environmental monitoring procedures. The use of organisms as biomonitors has grown increasingly prevalent as they are models that provide data that can be adequately evaluated. In this work, we investigated the genotoxic and cytotoxic effects caused by domestic sewage sludge through an analysis of biomarkers in the mollusk Biomphalaria glabrata. For the tests, increasing concentrations of 50, 100, 150, and 500 mg L-1 of domestic sewage sludge were standardized, in addition to control groups. Assays were performed after the mollusks were exposed to the domestic sewage sludge in acute (48 h) and chronic (15 d) manner. Toxicity tests were performed with embryonic and adult snails. The cytoplasmic and nuclear changes were analyzed in the hemocyte cells. Lastly, genotoxic damage was analyzed using the comet assay. Adult snails and embryos of B. glabrata showed no significant morphological changes. Domestic sludge caused deleterious effects on mollusks as confirmed after cell genotoxicity tests. Therefore, based on the results obtained from the analysis of B. glabrata hemocytes, we can affirm that domestic sewage sludge causes genotoxic and cytotoxic effects on mollusk cells. Therefore, it is possible to conclude that the mollusk Biomphalaria glabrata can be used as a good low-cost alternative to assist in the biomonitoring of freshwater environments. Graphical Abstract.

Multiple adverse effects of textile effluents and reactive Red 239 dye to aquatic organisms.

Textile dyeing consumes high volumes of water, generating proportional number of colored effluents which contain several hazardous chemical. These contaminants can implicate in significant changes in aquatic environmental, including several adverse effects to organisms in different trophic levels. The present study was developed to assess the ecotoxicological effects of textile effluent samples and reactive Red 239 dye (used in cotton dyeing) to aquatic organisms Vibrio fischeri bacteria, Daphnia similis crustacean, and Biomphalaria glabrata snail (adults and embryos). Chronic assays with lethal and sublethal effects for Daphnia similis were included and performed only for textile effluents samples. The mutagenicity was also evaluated with Salmonella/microsome assay (TA98, TA100, and YG1041 strains). V. fischeri bacteria was the most sensitive to reactive Red 239 dye (EC50 = 10.14 mg L-1) followed by mollusk embryos at all stages (EC50 = 116.41 to 124.14 mg L-1), D. similis (EC50= 389.42 mg L-1), and less sensitive to adult snails (LC50= 517.19 mg L-1). The textile effluent was toxic for all exposed organisms [E(L)C50 < 15%] and B. glabrata embryos showed different responses in the early stages of blastulae and gastrulae (EC50 = 7.60 and 7.08%) compared to advanced development stages trochophore and veliger (EC50 = 21.56 and 29.32%). Developmental and sublethal effects in B. glabrata embryos and D. similis were evidenced. In the chronic assay with effluent, the EC10/NOEC = 3% was obtained. Mutagenic effects were not detected for dye aqueous solutions neither for effluents samples. These data confirmed the importance of evaluating the effects in aquatic organisms from different trophic levels and reinforce the need for environmental aquatic protection.

Uso do Sapajus apella na padronização de modelo experimental para Schistosoma mansoni em primatas neotropicais

ABSTRACT OBJECTIVES: To establish and evaluate an experimental model with non-human neotropical primates Sapajus apella to verify more effective diagnostic methods in the detection of schistosomiasis, supporting new studies that help in the cure and control of this disease. MATERIALS AND METHODS: Six adult primates of the Sapajus apella species were used, randomly distributed in two experimental groups of equal number. Of these, three animals were infected intradermally and three transcutaneously, with suspension of cercariae obtained from naturally infected Biomphalaria glabrata snails. To monitor the infection, urine samples were collected via probe or direct puncture, with the anesthetized animal, and stool samples were collected directly from the cages. The urine samples were examined by the rapid urine test (point-of-care cathodic circulating antigen - POC-CCA) and the stool samples by tests of spontaneous stool sedimentation (HPJ) and Kato-Katz. RESULTS: The use of transcutaneous and intradermal routes resulted in the infection of two of the three animals evaluated (66.6%) in each experimental group. The POC-CCA test was the first to detect positives (21st day), followed by HPJ (35th day), and Kato-Katz (42nd day) tests. CONCLUSION: The model analyzed is viable for reproduction in different studies on schistosomiasis mansoni, as the animals responded satisfactorily to the infection in both routes used, making it possible to verify the practicality of the intradermal route when compared to the transcutaneous one.

Genetic architecture of transmission stage production and virulence in schistosome parasites

ABSTRACT Both theory and experimental data from pathogens suggest that the production of transmission stages should be strongly associated with virulence, but the genetic bases of parasite transmission/virulence traits are poorly understood. The blood fluke Schistosoma mansoni shows extensive variation in numbers of cercariae larvae shed and in their virulence to infected snail hosts, consistent with expected trade-offs between parasite transmission and virulence. We crossed schistosomes from two populations that differ 8-fold in cercarial shedding and in their virulence to Biomphalaria glabrata snail hosts, and determined four-week cercarial shedding profiles in F0 parents, F1 parents and 376 F2 progeny from two independent crosses in inbred snails. Sequencing and linkage analysis revealed that cercarial production is polygenic and controlled by five QTLs (i.e. Quantitative Trait Loci). These QTLs act additively, explaining 28.56% of the phenotypic variation. These results demonstrate that the genetic architecture of key traits relevant to schistosome ecology can be dissected using classical linkage mapping approaches.

Biological activity survey of Pereskia aculeata Mill. and Pereskia grandifolia Haw. (Cactaceae)

Background: Pereskia aculeata and P. grandifolia are non-traditional Brazilian vegetables with high nutritional value used in traditional medicine. The antioxidant, anticholinesterase, molluscicidal, cytotoxic, and antiproliferative properties of hydroethanolic extracts of P. aculeata and P. grandifolia leaves (PAL, PGL) and fruits (PAF, PGF) are investigated in this study. Methods: All extracts were prepared by maceration with ethanol 70%. Their antioxidant properties were assessed through DPPH, ABTS, FRAP, and β-carotene bleaching inhibition assays. A TLC bioautography method was employed to evaluate the inhibiting capacity of the acetylcholinesterase enzyme. The molluscicidal activity was tested against the snail Biomphalaria glabrata, which serves as an intermediate host for Schistosoma mansoni. The cytotoxic activity was assessed by a Artemia salina lethality test and the antiproliferative properties against seven human carcinoma cell lines. Results: Compounds with anticholinesterase activity were found in all extracts. Polar compounds present in PAF and PGL extracts were the most active (IC50 &lt; 25 μg of dry mass) and had an adequate inhibition capacity of the AChE. PGF and PGL were classified as moderate (LC50 = 19.2 μg/ml) and modest molluscicidal agents (LC50 = 66.6 μg/ml), respectively. All extracts exhibited selective antiproliferative activity against human chronic myeloid leukemia cell lines (K562). PAL, PGL, and PGF presented potent antiproliferative activity (TGI ≤ 5 μg/ml). Conclusion: Both species exhibited anticholinesterase, cytotoxic and antiproliferative properties. This research supports the potential of these species as sustainable sources of nutraceutical compounds.

New Insights Into Biomphalysin Gene Family Diversification in the Vector Snail Biomphalaria glabrata.

Aerolysins initially characterized as virulence factors in bacteria are increasingly found in massive genome and transcriptome sequencing data from metazoans. Horizontal gene transfer has been demonstrated as the main way of aerolysin-related toxins acquisition in metazoans. However, only few studies have focused on their potential biological functions in such organisms. Herein, we present an extensive characterization of a multigene family encoding aerolysins - named biomphalysin - in Biomphalaria glabrata snail, the intermediate host of the trematode Schistosoma mansoni. Our results highlight that duplication and domestication of an acquired bacterial toxin gene in the snail genome result in the acquisition of a novel and diversified toxin family. Twenty-three biomphalysin genes were identified. All are expressed and exhibited a tissue-specific expression pattern. An in silico structural analysis was performed to highlight the central role played by two distinct domains i) a large lobe involved in the lytic function of these snail toxins which constrained their evolution and ii) a small lobe which is structurally variable between biomphalysin toxins and that matched to various functional domains involved in moiety recognition of targets cells. A functional approach suggests that the repertoire of biomphalysins that bind to pathogens, depends on the type of pathogen encountered. These results underline a neo-and sub-functionalization of the biomphalysin toxins, which have the potential to increase the range of effectors in the snail’s immune arsenal.

Evaluation of original Roundup® effects on Biomphalaria glabrata (Say, 1818)

The objective of this work is to evaluate the action of original Roundup® on the snail Biomphalaria glabrata infected and not infected by Schistosoma mansoni by observing its mortality and reproductive biology. Snails were submitted for 30 days to aqueous solutions of original Roundup at concentrations of 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012 and 0.01 (%) to determine LC50. To analyze the reproductive activity of B. glabrata, against the pesticide, four B. glabrata groups were formed: infected and exposed to the pesticide; infected and not exposed (control); not infected and exposed; and not infected and not exposed(control). LC50 (0.012% aqueous solution of original Roundup) caused mortality of at most up to 95% for 24, 48 and 72 h during the experimental period (30 days), and lost effect totally after 27 days. The reproductive activity of the non-infected and exposed snails was 30.3 ± 20.5 eggs in 2.7 ± 1.2 egg masses, and there was a reduction of 51% in the average number of hatched snails (11.5 ± 7.5) in comparison to the control group (23.9 ± 18.0) for four weeks (p = 0.003). The results indicate that the lethal concentration (LC50) of the 0.012% aqueous solution of original Roundup has a significant action on Biomphalaria glabrata for up to 27 days. Roundup significantly diminished the fertility of infected and exposed snails in comparison with the control group.

Occurrence of BIOMPHALARIA GLABRATA and PHYSA ACUTA (gastropoda: mollusca) snails in Quinta da Boa Vista municipal park, Rio de Janeiro, RJ, Brazil

The human species has an emotional connection with nature called “biophilia” (or, love for life). This connection goes from generation to generation, being one of the reasons why green spaces are present in large cities. Quinta da Boa Vista is a municipal park in Rio de Janeiro located in the São Cristóvão neighborhood in the northern part of the city. Its main attraction is the artificial lake system, a leisure attraction for the local community. This article reports the presence of Biomphalaria glabrata and Physa acuta snails around the main lake in the Quinta da Boa Vista Municipal Park in Rio de Janeiro, RJ. Seventy-one B. glabrata and twenty-seven P. acuta snails were collected. The snails were measured and examined for the presence of trematode larvae according to standardized procedures. The prevalence rate of B. glabrata was estimated at 15.5%. Rediae and cercariae resembling those of the species Typhlocoelum cucumerinum (Typhlocoelidae, Digenea) which parasitizes aquatic birds of the order Anseriformes were found. This study reinforces the importance of monitoring park lakes and ponds to better understand the risk of helminth transmission by molluscan vectors.KEY WORDS: Snails; Schistosomiasis; Digenea; Quinta da Boa Vista; Rio de Janeiro.

Genomic and transcriptional analysis of genes containing fibrinogen and IgSF domains in the schistosome vector Biomphalaria glabrata, with emphasis on the differential responses of snails susceptible or resistant to Schistosoma mansoni.

Achieving a deeper understanding of the factors controlling the defense responses of invertebrate vectors to the human-infecting pathogens they transmit will provide needed new leads to pursue for control. Consequently, we provide new genomic and transcriptomic insights regarding FReDs (containing a fibrinogen domain) and FREPs (fibrinogen domain and one or two IgSF domains) from the planorbid snail Biomphalaria glabrata, a Neotropical vector of Schistosoma mansoni, causative agent of human intestinal schistosomiasis. Using new bioinformatics approaches to improve annotation applied to both genome and RNA-Seq data, we identify 73 FReD genes, 39 of which are FREPs. We provide details of domain structure and consider relationships and homologies of B. glabrata FBG and IgSF domains. We note that schistosome-resistant (BS-90) snails mount complex FREP responses following exposure to S. mansoni infection whereas schistosome-susceptible (M line) snails do not. We also identify several coding differences between BS-90 and M line snails in three FREPs (2, 3.1 and 3.2) repeatedly implicated in other studies of anti-schistosome responses. In combination with other results, our study provides a strong impetus to pursue particular FREPs (2, 3.1, 3.2 and 4) as candidate resistance factors to be considered more broadly with respect to schistosome control efforts, including involving other Biomphalaria species vectoring S. mansoni in endemic areas in Africa.

Essential oils of the leaves of Syzygium cumini (L.) Skeels and fruit peels of Hymenaea courbaril (L.) var. courbaril as molluscides against Biomphalaria glabrata

This study evaluated the molluscicidal activity of essential oils (EOs) of Syzygium cumini L. and Hymenaea courbaril L. Hydrodistillation was used to extract the EOs. The chemical characterization was performed by Gas Chromatography coupled to mass spectrometry. The bioassay of Artemia salina Leach was used for the toxicity test. The molluscicidal assay tested concentrations of 10-80 mg L-1 of the EOs against Biomphalaria glabrata. The major constituent of The EO of S. cumini was isokaryophyllene and H. courbaril the Germacreno-D. The toxicity assay classified The EOs as nontoxic with LC50 412.10 mg L-1 for S. cumini and LC50 354.80 mg L-1 for H. courbaril. The molluscicidal assay purchased LC50 44.76/ LC90 77.20 for S. cumini and LC50 37.34 mg L-1/ LC90 73.24 mg L-1 for H. courbaril. The results showed that both EOs can be used as natural molluscicides in combating the snail Biomphalaria glabrata.

Schistosoma mansoni Vector Snails in Antigua and Montserrat, with Snail-Related Considerations Pertinent to a Declaration of Elimination of Human Schistosomiasis.

.Investigations leading to a WHO-validated declaration of elimination of schistosomiasis transmission are contemplated for several countries, including Caribbean island nations. With assistance from the Pan American Health Organization, we undertook freshwater snail surveys in two such nations, Antigua and Barbuda, and Montserrat in September and October 2017. Historically, the transmission of Schistosoma mansoni supported by the Neotropical vector snail Biomphalaria glabrata occurred in both countries. Transmission on the islands is thought to have been interrupted by the treatment of infected people, improved sanitation, introduction of competitor snails, and on Montserrat with the eruption of the Soufrière volcano which decimated known B. glabrata habitats. Guided by the available literature and local expertise, we found Biomphalaria snails in seven of 15 and one of 14 localities on Antigua and Montserrat, respectively, most of which were identified anatomically and molecularly as Biomphalaria kuhniana. Two localities on Antigua harbored B. glabrata, but no schistosome infections in snails were found. For snail-related aspects of validation of elimination, there are needs to undertake basic local training in medical malacology, be guided by historical literature and recent human schistosomiasis surveys, improve and validate sampling protocols for aquatic habitats, enlist local expertise to efficiently find potential transmission sites, use both anatomical and molecular identifications of schistosomes or putative vector snail species found, if possible determine the susceptibility of recovered Biomphalaria spp. to S. mansoni, publish survey results, and provide museum vouchers of collected snails and parasites as part of the historical record.

Clusters of polymorphic transmembrane genes control resistance to schistosomes in snail vectors.

Schistosomiasis is a debilitating parasitic disease infecting hundreds of millions of people. Schistosomes use aquatic snails as intermediate hosts. A promising avenue for disease control involves leveraging innate host mechanisms to reduce snail vectorial capacity. In a genome-wide association study of Biomphalaria glabrata snails, we identify genomic region PTC2 which exhibits the largest known correlation with susceptibility to parasite infection (>15 fold effect). Using new genome assemblies with substantially higher contiguity than the Biomphalaria reference genome, we show that PTC2 haplotypes are exceptionally divergent in structure and sequence. This variation includes multi-kilobase indels containing entire genes, and orthologs for which most amino acid residues are polymorphic. RNA-Seq annotation reveals that most of these genes encode single-pass transmembrane proteins, as seen in another resistance region in the same species. Such groups of hyperdiverse snail proteins may mediate host-parasite interaction at the cell surface, offering promising targets for blocking the transmission of schistosomiasis.

Heat shock increases hydrogen peroxide release from circulating hemocytes of the snail Biomphalaria glabrata

Planorbid freshwater snails are important intermediate hosts for parasitic diseases caused by parasitic worms, most notably schistosomiasis. There are numerous reports of snails, specifically Biomphalaria glabrata, having compromised defences against schistosomes after being exposed to thermal stress. Environmental modifications to the defenses of schistosome transmitting snails could have negative ramifications for human disease risk in the context of climate change. Here the effects of heat shock on the production of hydrogen peroxide, a primary anti-microbial effector in many molluscs, were examined. The present findings show that heat shock increases NADPH oxidase 2 mRNA levels and hydrogen peroxide produced by snail hemocytes, and that both of these phenotypes could be reversed by an HSP-90 inhibitor. These findings indicate that snail defense systems are altered by heat shock at a molecular level in B. glabrata, and that snail immunity to many pathogens may be altered by the rapid variations in temperature that are associated with global climate change.

Molluscicidal activity of polyvinylpyrrolidone (PVP)-functionalized silver nanoparticles to Biomphalaria glabrata: Implications for control of intermediate host snail of Schistosoma mansoni

Silver nanoparticles (Ag NPs) have been applied in several commercial products due to their antimicrobial properties, while their molluscicide properties, mode of action and toxicity to snail species remain unclear. In this study, the comparative toxicity of polyvinylpyrrolidone (PVP)-functionalized Ag NPs and their dissolved counterpart (Ag ions) was analyzed during the early developmental stages of the freshwater snail Biomphalaria glabrata, intermediate host of Schistosoma mansoni. Ag NPs were synthesized and characterized by multiple techniques, and the snail embryotoxicity was analyzed in terms of mortality, hatching, developmental stages and morphological alterations, while the acute toxicity to newly-hatched snails was analyzed by mortality and behavioral impairments. Results showed that both Ag forms induced mortality, hatching delay and morphological alterations (especially hydropic abnormalities) in snail embryos in a concentration and exposure time dependent patterns. Ag NPs showed low embryotoxic effects and similar toxicity for newly-hatched snails when compared to their dissolved counterparts, indicating that the nanotoxicity was dependent of snail developmental stages. The knowledge about the Ag NP toxicity to different early development stages of B. glabrata contributes to its potential use as molluscicide and control of neglected tropical diseases, including schistosomiasis.

Risk assessment of iron oxide nanoparticles in an aquatic ecosystem: A case study on Biomphalaria glabrata

Iron oxide nanoparticles (IONPs) have been applied in several sectors in the environmental field, such as aquatic nanoremediation, due to their unique superparamagnetic and nanospecific properties. However, the knowledge of chronic toxicity of IONPs on aquatic invertebrate remains limited. Thus, the present study aimed to analyze the chronic toxicity of gluconic acid-functionalized IONPs (GLA-IONPs) and their dissolved counterpart (FeCl3) to freshwater snail Biomphalaria glabrata. GLA-IONPs were synthesized and characterized by multiple techniques, and the snails were exposed to both Fe forms at environmentally relevant concentrations (1.0–15.6 mg L−1) for 28 days. The bioaccumulation, mortality rate, behavior impairments, morphological alterations, fecundity and fertility of snails were analyzed. Results showed that GLA-IONPs induced high iron bioaccumulation in the entire soft tissue portion. Chronic exposure to GLA-IONP increased the behavioral impairments of snails compared to iron ions and control groups. Both Fe forms reduced the fecundity, while the mortality and reduced fertility were observed only after the exposure to GLA-IONPs at 15.6 mg L−1. Overall results indicated the behavioral impairments and reproductive toxicity associated, possibly, to bioaccumulation of GLA-IONPs in the B. glabrata. These results can be useful for the development of eco-friendly nanotechnologies.

Mollusk microbiota shift during Angiostrongylus cantonensis infection in the freshwater snail Biomphalaria glabrata and the terrestrial slug Phillocaulis soleiformis.

In the present work, we reported for the first time the microbiome from Phyllocaulis soleiformis and Biomphalaria glabrata assessed using high-throughput DNA sequencing pre- and post-infection with the helminth parasite Angiostrongylus cantonensis. B. glabrata and P. soleiformis were experimentally infected with A. cantonensis. Fecal DNAs from control and infected groups were extracted and subjected to 16S rRNA high-throughput sequencing survey. No significant differences were found in the alpha diversity indexes in Phyllocaulis and Biomphalaria experiments independently. PCoA analysis using the unweighted UniFrac measures showed that both microbiotas behaved differently depending on the host. In Biomphalaria microbiota, control and infected groups were significantly different (p = 0.0219), while Phyllocaulis samples were not (p = 0.5190). The microbiome of P. soleiformis infected with A. cantonensis showed a significant decrease of Sphingobacterium and a substantial increase of Cellvibrio when compared to a control group. The microbiome of B. glabrata infected with A. cantonensis showed a significant decline in the abundance of Flavobacterium, Fluviicola, Nitrospira, Vogesella and an OTU belonging to the family Comamonadaceae, and a significant increase of Uliginosibacterium and an OTU belonging to the family Weeksellaceae when compared to a control group. Overall, the microbiome data reported here provided valuable information with regard to the diversity of bacterial communities that comprise the gut microbiome of gastropods. Furthermore, we report here the effect of the infection of the helminth A. cantonensis in the ratio and distribution of the fecal microbiome of the snails. Further studies are highly valuable in order to better understand those interactions by comparing different microbiome profiles and mollusk models. By now, we anticipate that ecological studies will take significant advantage of these advances, particularly concerning improving our understanding of helminth-microbiome-host interactions.