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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.

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Mollusk hemocyanins, among the largest known proteins, are used as immunostimulants in biomedical and clinical applications. The hemocyanin of the Chilean gastropod Concholepas concholepas (CCH) exhibits unique properties, which makes it safe and effective for human immunotherapy, as observed in animal models of bladder cancer and melanoma, and dendritical cell vaccine trials. Despite its potential, the structure and amino acid sequence of CCH remain unknown. This study reports two sequence fragments of CCH, representing three complete functional units (FUs). We also determined the high-resolution (1.5 Å) X-ray crystal structure of an "FU-g type" from the CCHB subunit. This structure enables in-depth analysis of chemical interactions at the copper-binding center and unveils an unusual, truncated N-glycosylation pattern. These features are linked to eliciting more robust immunological responses in animals, offering insights into CCH's enhanced immunostimulatory properties and opening new avenues for its potential applications in biomedical research and therapies.

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This study investigated the prevalence and genetic diversity of gastroenteric viruses in mussels and oysters in Rio de Janeiro, Brazil. One hundred and thirty-four marketed bivalve samples were obtained between January and December 2022. The viral analysis was performed according to ISO/TS 15216, and the screening revealed the detection of norovirus GII/GI (40.3%), sapovirus (SaV; 12.7%), human mastadenovirus (7.5%), and rotavirus A (RVA; 5.9%). In total, 44.8% (60) of shellfish samples tested positive for one or more viruses, 46.7% (28/60) of the positive samples tested positive for a single viral agent, 26.7% (16) tested positive for two viral agents, 8.3% (5) for three viral agents, and 13.3% (8) for four viral agents. Additionally, three mussel samples were contaminated with the five investigated viruses (5%, 3/60). Norovirus GII showed the highest mean viral load (3.4 × 105 GC/g), followed by SaV (1.4 × 104 GC/g), RVA (1.1 × 104 GC/g), human mastadenovirus (3.9 × 103 GC/g), and norovirus GI (6.7 × 102 GC/g). Molecular characterization revealed that the recovered norovirus strains belonged to genotypes GII.2, GII.6, GII.9, GII.17, and GII.27; SaV belonged to genotypes GI.1 and GIV.1; RVA to genotypes G6, G8, P[8]-III, and human mastadenovirus to types F40 and F41. The GII.27 norovirus characterized in this study is the only strain of this genotype reported in Brazil. This study highlights the dissemination and diversity of gastroenteric viruses present in commercialized bivalves in a touristic area, indicating the potential risk to human health and the contribution of bivalves in the propagation of emerging pathogens.

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It is generally acknowledged that microplastic pollutants are prevalent in ocean waters and sediments across a range of tropical, temperate, subpolar, and polar regions. The waters surrounding King George Island are significantly impacted by human activities, particularly those related to scientific stations, fishing, and tourism. Organisms, such as Laternula elliptica, can be used as environmental monitors due to the likelihood that they will bioaccumulate pollutants. The goal of this study was to quantify and identify plastic and cellulosic micro-fragments and microfibers present in the soft body of clams (n = 21), collected from Fildes Bay near sewage and wastewater discharges. Plastic and cellulose microfragments and microfibers were counted, and their compositions were determined using FT-IR. All 21 individuals sampled contained fragments and fibers, with a total of 900 items detected (42.86 ± 25.36 mean ± SD items per individual), or 1.82 items g.wet mass-1. 58 % of items were cellulose and 22 % plastic. Considering the plastic polymer compositions, 28.57 % were polyethylene terephthalate (PET), 21.43 % acrylic, 14.29 % high-density polyethylene (HDPE), 14.29 % Polypropylene (PP), 7.14 % ultra-high drawn polyethylene filament (UHMWPE), 7.14 % polyester and 7.14 % Polyethylene. The quantities and prevalence of MP in L. elliptica were higher than those found in other Antarctic marine species, and even in bivalves from populated regions of the world. Our work assessed the pollution status of L. elliptica near an effluent of wastewater plants and found that 95 % of individuals displayed MP and 100 % microfibers that could impact their population.

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Massive mortalities in farmed larvae of the scallop Argopecten purpuratus have been associated with pathogenic Vibrio outbreaks. An energetic trade-off between development-associated demands and immune capacity has been observed. Given that highly unsaturated fatty acids (HUFAs) are essential nutrients for larval development, we evaluated the effect of diets based on microalgae low and high in HUFAs (LH and HH, respectively) on the energetic condition and the immune response of scallop larvae. The results showed that the HH diet increased cellular membrane fluidity in veliger larvae. The routine respiration rate was 64% higher in the HH-fed veligers than in the LH-fed veligers. Additionally, the metabolic capacity tended to be higher in the HH-fed veligers than in the LH-fed veligers after the Vibrio challenge. After the challenge, the HH-fed veligers presented higher transcript induction of ApTLR (immune receptor) and ApGlys (immune effector) genes, and the HH-fed pediveligers presented higher induction of ApLBP/BPI1 (antimicrobial immune effector) gene, than the LH-fed larvae. Furthermore, the HH-fed veligers controlled total Vibrio proliferation (maintaining near basal levels) after the bacterial challenge, while the LH-fed veligers were not able to control this proliferation, which increased three-fold. Finally, the HH-fed larvae showed 20-25% higher growth and survival rates than the LH-fed veligers. Overall, the results indicated that the administration of a HH diet increases cell membrane fluidity and energy metabolic capacity, which in turn enhances immunity and the ability to control Vibrio proliferation. The administration of microalgae high in HUFAs would be a promising strategy for improving scallop larval production efficiency.

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Mass mortality events caused by vibriosis have emerged in hatchery-reared scallop larvae from Chile, threatening scallop aquaculture. In an attempt to mitigate this emerging infectious disease and provide candidates for marker-assisted selective breeding, we tested here the existence of a genetic component of Argopecten purpuratus scallop resistance to the pathogen Vibrio bivalvicida. Through a dual RNA-seq approach we analyzed the basal transcriptome and the transcriptional response to infection in two resistant and two susceptible families as well as the pathogen transcriptomic response to host colonization. The results highlighted a genetic basis in the resistance of scallop larvae to the pathogen. The Vibrio response was characterized by a general metabolic adaptation to the host environment, along with several predicted virulence factors overexpressed in infected scallop larvae with no difference between resistant and susceptible host phenotypes. On the host side, several biological processes were enriched in uninfected resistant larvae. Within these enriched categories, immune-related processes were overexpressed, while morphogenesis, biomineral tissue development, and angiogenesis were under expressed. Particularly, genes involved in immune recognition and antimicrobial response, such as lipopolysaccharide-binding proteins (LBPs), lysozyme, and bactericidal permeability-increasing protein (BPI) were overexpressed in uninfected resistant larvae. As expected, immune-related biological processes were enriched in Vibrio-infected larvae, but they were more numerous in resistant larvae. Overexpressed immune genes in response to infection included several Toll-like receptors, TNF and NF-κB immune signaling genes, and the antimicrobial peptide Big defensin ApBD1. Results strongly suggest that both a front-loading of immune genes and an enhanced antimicrobial response to infection contribute to the resistance, while pathogen infective strategy does not discriminate between host phenotypes. Overall, early expression of host immune genes appears as a strong determinant of the disease outcome that could be used in marker-assisted selective breeding.

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Two Vibrio strains (VPAP36 and VPAP40) were isolated from moribund-settled larvae of the Chilean scallop Argopecten purpuratus during vibriosis outbreaks that occurred in two commercial scallop larvae hatcheries located in the Inglesa and Tongoy bays in Northern Chile. The strains were identified as Vibrio chagasii using phenotypic characterization and whole genome sequence analysis. Both strains exhibited the phenotypic properties associated with virulence, gelatin hydrolysis and ß-hemolysis, whereas only VPAP36 produced phospholipase and only VPAP40 produced caseinase. The whole genome analysis showed that the strains harbored genes encoding for the virulence factors, the EPS type II secretion system, and Quorum Sensing (auto-inductor 1 and auto-inductor 2), whereas genes encoding a metalloproteinase and a capsular polysaccharide were detected only in the VPAP40 genome. When challenge bioassays using healthy 11-day-old scallop larvae were performed, the V. chagasii VPAP36 and VPAP40 strains exhibited significant (p < 0.05) differences in their larval lethal activity, producing, after 48 h, larval mortalities of 65.51 ± 4.40% and 28.56 ± 5.35%, respectively. Otherwise, the cell-free extracellular products of the VPAP36 and VPAP40 strains produced larval mortalities of 20.86 ± 2.40% and 18.37 ± 2.40%, respectively, after 48 h of exposure. This study reports for the first time the isolation of V. chagasii from the massive larval mortalities of the farmed scallop (Argopecten purpuratus) in Chile, and demonstrates the pathogenic activity of V. chagasii towards the Chilean scallop, the second most important species for Chilean mariculture.

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PURPOSE: Eucotylid trematodes are parasites of the urinary system of birds with a cosmopolitan distribution. Despite the importance of these flukes, fundamental aspects of their biology, such as intermediate hosts and larval morphology, are poorly known. Herein, the potential involvement of aquatic mollusks in the transmission of a species of Tanaisia is reported for the first time. METHODS: During the search of non-emergent larval stages of trematodes in mollusks collected from an urban waterbody from Brazil in February of 2021, 1 out of 18 specimens (5.5%) of Biomphalaria straminea was found harboring sporocysts, cercariae and encysted metacercariae morphologically compatible with those described for eucotylid species. Sequences generated for 28S, ITS-2, and cox1 molecular markers were compared with sequences available in GenBank and subjected to phylogenetic analyses. RESULTS: Molecular analyses revealed parasite affiliation with members of the genus Tanaisia, given it groped in a strongly supported clade with species of this genus included in the 28S phylogenetic tree. The larvae tentatively identified as Tanaisia sp. can be conspecific with an unpublished isolate of Tanaisia valida found in birds in South Brazil (100% similarity in 28S and ITS-2). CONCLUSION: Biomphalaria straminea is reported as a natural host of a species of Tanaisia for the first time. This finding highlights the possibility, so far unknown, of transmission of species of the family Eucotylidae in aquatic environments.

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Abstract Estuaries are important ecosystems due to the ecological services they provide, acting as nurseries for many species of fish and invertebrates, and are also used as environments for the extraction and cultivation of mollusks. Oysters are animals that filter water to obtain oxygen and nutrients. In this process, they can bioaccumulate microorganisms and chemical substances in their tissues. The growth of mollusk culture in Northeastern Brazil requires the health identification of cultivated oysters through the quantification of the potentially harmful microbiota accumulated in the animals. Therefore, the present work aims to quantify and identify bacteria and possible pathogens found in the tissues of cultivated oysters and their culture waters. The Most Probable Number of Coliforms (MPN) in oysters and water were considered suitable according to the Brazilian current legislation, Vibrio sp. obtained low colonization and Salmonella sp. was not observed. The prevalence of microorganisms potentially pathogenic to oysters was 33.7%, highlighting metazoans and Nematopsis sp., however, the intensity of the infestation of these organisms was moderate. The low contamination of oysters demonstrates that this culture environment is promising for this activity. However, continuous environmental and sanitary monitoring is fundamental to guarantee the safety of the culture waters and the sustainability of aquaculture activities.

Resumo Os estuários são ecossistemas importantes devido a serviços ecológicos que fornecem, os quais conferem a função de berçário para muitas espécies de peixes e invertebrados, e também são utilizados como ambientes de extração e cultivo de moluscos. As ostras são animais que filtram a água para obtenção de oxigênio e nutrientes. Nesse processo podem bioacumular microorganismos e substâncias químicas em seus tecidos. O crescimento da malacocultura no Nordeste do Brasil fomenta a necessidade de identificar a sanidade das ostras cultivadas através da quantificação da microbiota potencialmente nociva acumulada nos animais. Portanto o presente trabalho visa quantificar e identificar bactérias e possíveis patógenos encontrados nos tecidos dos moluscos cultivados e nas suas águas de cultivo. O Número mais Provável de Coliformes (NMP) nas ostras e na água foram considerados próprios segundo as legislações vigentes, Vibrio sp. obteve baixa colonização e Salmonella sp. não foi observada. A prevalência de microorganismos potencialmente patógenos para as ostras foi de 33,7%, destacando como mais prevalentes os metazoários e Nematopsis sp., porém a intensidade da infestação desses organismos foi moderada. A baixa contaminação das ostras demonstra que este ambiente de cultivo é promissor para esta atividade. No entanto, o contínuo monitoramento ambiental e sanitário é fundamental para garantir a inocuidade das águas de cultivo e a sustentabilidade das atividades aquícolas.

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Abstract Introduction: The snail Hexaplex princeps is the most important gastropod in the fishery of Guerrero, but capture is not legally regulated for lack of enough studies about its reproduction, growth and other fishing aspects. Objective: To assess fishery data for H. princeps in Guerrero. Methods: From October 2016 to December 2017, we analyzed histological samples of the gonads from 6 to 13 individuals per month. We also collected data on pre-reproductive organisms captured in Costa Chica (2013 and 2014), Costa Grande (2017) and Acapulco (2019 and 2020). Results: Both sexes have five stages of gonadal development; most reproductive activity was from December to July, related to sea surface temperatures of 26 to 28 °C. The LC M in females was 94.1 mm (males: 86.1 mm). There was a high percentage of juvenile organisms in the catches (88.6 %). Conclusions: To let these snails reproduce at least once, we recommend a fishing ban from December to July, and that the minimum size of commercial capture be set at 94.1 mm.

Resumen Introducción: El caracol Hexaplex princeps es el gasterópodo más importante en la pesquería de Guerrero, pero su captura no está regulada legalmente por falta de suficientes estudios sobre su reproducción, crecimiento y otros aspectos de la pesca. Objetivo: Evaluar los datos de pesca de H. princeps en Guerrero. Métodos: De octubre 2016 a diciembre 2017, analizamos muestras histológicas de las gónadas de 6 a 13 individuos por mes. También recolectamos datos sobre organismos pre-reproductivos capturados en Costa Chica (2013 y 2014), Costa Grande (2017) y Acapulco (2019 y 2020). Resultados: Ambos sexos tienen cinco etapas de desarrollo gonadal; la mayor actividad reproductiva fue de diciembre a julio, relacionada con las temperaturas superficiales del mar, que se encuentran entre 26 y 28 °C. El LC M en hembras fue de 94.1 mm (machos: 86.1 mm). Hubo un alto porcentaje de organismos juveniles en las capturas (88.6 %). Conclusiones: Para que estos caracoles se reproduzcan al menos una vez, se recomienda una veda de diciembre a julio y que la talla mínima de captura comercial se fije en 94.1 mm.

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Lead, like other trace elements, is incorporated in the growing bands of bivalve shells. The chemicals stored into the shells can provide valuable information about seawater conditions during the period of shell formation. In this study, we present a practical approach to determine Pb isotopic signatures in bivalve shells as a tool for evaluating lead pollution in coastal waters. To demonstrate the applicability of the method, Pb isotopic fingerprinting in bivalve shell layers were investigated using laser ablation multicollector inductively coupled plasma mass spectrometry (LA-MC-ICP-MS). Lead isotope ratios (208Pb/206Pb and 206Pb/207Pb) were measured along distinct sections of the maximum growth axis of the shells. Calibration and quantification of Pb isotopes were performed using NIST 612 as reference material. Our results demonstrated that Pb isotope ratios in the shells ranged from 1.143 to 1.201 for 206Pb/207Pb and from 2.061 to 2.161 for 208Pb/206Pb. The isotopic signatures recorded in the sample shells correspond to similar ranges of Pb signatures reported for marine sediments from the same study area. In general, this work shows that LA-MC-ICP-MS is a suitable technique for determining spatially resolved lead isotopic signatures in bivalve shells and that it can be used to estimate the origin of Pb pollution in aquatic environments.

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In marine ecosystems, most invertebrates possess diverse microbiomes on their external surfaces, such as those found in the pedal mucus of grazing gastropods and chitons that aids displacement on different surfaces. The microbes are then transported around and placed in contact with free-living microbial communities of micro and other macro-organisms, potentially exchanging species and homogenizing microbial composition and structure among grazer hosts. Here, we characterize the microbiota of the pedal mucus of five distantly related mollusk grazers, quantify differences in microbial community structure, mucus protein and carbohydrate content, and, through a simple laboratory experiment, assess their effects on integrated measures of biofilm abundance. Over 665 Amplicon Sequence Variants (ASVs) were found across grazers, with significant differences in abundance and composition among grazer species and epilithic biofilms. The pulmonate limpet Siphonaria lessonii and the periwinkle Echinolittorina peruviana shared similar microbiota. The microbiota of the chiton Chiton granosus, keyhole limpet Fissurella crassa, and scurrinid limpet Scurria araucana differed markedly from one another, and form those of the pulmonate limpet and periwinkle. Flavobacteriaceae (Bacteroidia) and Colwelliaceae (Gammaproteobacteria) were the most common among microbial taxa. Microbial strict specialists were found in only one grazer species. The pedal mucus pH was similar among grazers, but carbohydrate and protein concentrations differed significantly. Yet, differences in mucus composition were not reflected in microbial community structure. Only the pedal mucus of F. crassa and S. lessonii negatively affected the abundance of photosynthetic microorganisms in the biofilm, demonstrating the specificity of the pedal mucus effects on biofilm communities. Thus, the pedal mucus microbiota are distinct among grazer hosts and can affect and interact non-trophically with the epilithic biofilms on which grazers feed, potentially leading to microbial community coalescence mediated by grazer movement. Further studies are needed to unravel the myriad of non-trophic interactions and their reciprocal impacts between macro- and microbial communities.

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The introduction of the African snail Lissachatina fulica worldwide poses a risk to the environment, agriculture and human health. Considered a socio-environmental pest it causes problems to plantations and gardens, in addition to threatening native species. Also, it can be an intermediate host of worms that affect humans and mammals. This work aims to evaluate the socioenvironmental impact caused by L. fulica in Alto Paraíso de Goiás, Goiás, Brazil. Specifically, it aimed to analyze the residents' knowledge about L. fulica, if they were able to recognize the invasive snail and differentiate it from the giant native snail Megalobulimus sp., in addition to the community's ability to carry out actions to combat the invasive species. Semi-structured interviews were conducted, guided by a questionnaire, with the urban population, and an unstructured interview with a health surveillance agent in the municipality. The area of the city was divided into five zones and ten residences were visited in each zone. It was found that the residents most affected by L. fulica are those who live in the most urbanized region of the city, with 86% of respondents reporting seeing the animal in the city center. The presence of the Megalobulimus sp. was reported by four residents in the city's border regions with the native Cerrado. Most of the reports of the African snail in the houses occurred by residents of Alto Paraíso de Goiás who had plants in their backyard and personally took care of them.(AU)

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We present herein a short-term impact on marketed fisheries and human health safety in the first three months following the 2019 oil spill in Brazil. Total PAHs in the edible tissues of 34 finfish and shellfish species ranged from 8.71 to 418 ng g-1 wet weight, with robust evidence supporting crude oil contamination. A prevalence of low molecular weight PAHs was observed, mainly naphthalenes. A decreasing trend in mean total PAHs from mollusks (134 ng g-1) to crustaceans (73.9 ng g-1) and to fishes (45.3 ng g-1) was noted. The spilled oil caused immediate negative impacts on the local seafood market, despite less than 3% of samples exhibited concentrations above levels of concern, revealing a low probability for human health risks. These findings demonstrate that governments must be prepared to provide not only science-based quick responses but also effective science communication for society upon environmental disasters.

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This study was developed to examine the effect of using charru mussel-, maçunim- and oyster-shell meals as calcium sources on the productive performance, carcass yield and bone parameters of meat quail. A total of 250 one-day-old non-sexed European quail were used. The birds were distributed into five treatments (calcium sources: calcitic lime, calcium carbonate, charru mussel shell meal, maçunim shell meal or oyster shell meal) in a completely randomized design with five replicates and ten birds per experimental unit. Feed intake, weight gain and feed conversion were evaluated from one to 35 days. At 35 days, two birds with the average weight of the plot were slaughtered per plot and used for analysis of carcass yield and bone parameters. There were no significant differences (P > 0.05) in feed intake, weight gain, feed conversion, carcass yield or tibia dry matter content, ash content and strength. The charru mussel-, maçunim- and oyster-shell meals can be used as calcium sources in the diet of European quail without affecting their productive performance, carcass yield or bone mineralization.

Objetivou-se avaliar o efeito da utilização das farinhas de conchas de sururu, maçunim e ostra como fontes de cálcio sobre o desempenho produtivo, rendimento de carcaça e parâmetros ósseos de codornas de corte. Foram utilizadas 250 codornas europeias, não sexadas, com um dia de idade. As aves foram distribuídas em delineamento inteiramente casualizado, constituído por cinco fontes de cálcio (calcário calcítico, carbonato de cálcio, farinha de conchas de sururu, farinha de conchas de maçunim e farinha de conchas de ostras), com cinco repetições e dez aves por unidade experimental. Foram avaliados o consumo de ração, ganho de peso e conversão alimentar no período de um a 35 dias. Aos 35 dias, duas aves de peso médio de cada parcela foram abatidas e utilizadas para as análises de rendimento de carcaça e parâmetros ósseos. Não foram observadas diferenças significativas (P > 0,05) para os dados de consumo de ração, ganho de peso, conversão alimentar, rendimento de carcaça, matéria seca, cinzas e resistência óssea de tíbias. As farinhas de conchas de sururu, maçunim e ostra podem ser utilizadas na alimentação de codornas europeias, como fontes de cálcio, sem afetar o desempenho produtivo, rendimento de carcaça e a mineralização óssea das aves.

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Big defensins is a large family of antimicrobial peptides found in restricted groups of invertebrates, in particular mollusks where they have highly diversified. Big defensins are composed of a highly hydrophobic N-terminal region and a C-terminal region containing six cysteine residues whose arrangement is identical to that of vertebrate ß-defensins. They have been shown to be active against both Gram-positive and Gram-negative bacteria and fungi. Antimicrobial aggregates called nanonets entrapping and killing bacteria have been recently described for the hydrophobic N-terminal region of the Cg-BigDef1 from the oyster Crassostrea gigas. To determine whether nanonets formation is a conserved trait of mollusk big defensins, we assessed the potential entrapping of bacteria through nanonets of the big defensin from the scallop Argopecten purpuratus, ApBD1. Recombinant ApBD1 was produced with a thrombin-cleavable N-terminal His6 tag, followed by the mature peptide carrying a mutation of the last cysteine residue of the C-terminal region by and arginine, named rApBD1(C87R). This mutation did not apparently affect the three-dimensional structure and the biological properties of rApBD1(C87R), as evidenced by in silico modeling and in vitro antimicrobial assays. Strong immune staining of rApBD1(C87R) in numerous areas surrounding bacteria was observed by confocal microscopy, suggesting that rApBD1(C87R) entraps bacteria in peptide aggregates similar to those reported to the oyster big defensin. This study suggests the conservation of bactericidal activity and nanonet formation across big defensins from bivalve mollusks.

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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.

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Freshwater snails of the genus Biomphalaria serve as intermediate hosts for the digenetic trematode Schistosoma mansoni, the etiological agent for the most widespread form of intestinal schistosomiasis. As neuropeptide signaling in host snails can be altered by trematode infection, a neural transcriptomics approach was undertaken to identify peptide precursors in Biomphalaria glabrata, the major intermediate host for S. mansoni in the Western Hemisphere. Three transcripts that encode peptides belonging to the FMRF-NH2 -related peptide (FaRP) family were identified in B. glabrata. One transcript encoded a precursor polypeptide (Bgl-FaRP1; 292 amino acids) that included eight copies of the tetrapeptide FMRF-NH2 and single copies of FIRF-NH2 , FLRF-NH2 , and pQFYRI-NH2 . The second transcript encoded a precursor (Bgl-FaRP2; 347 amino acids) that comprised 14 copies of the heptapeptide GDPFLRF-NH2 and 1 copy of SKPYMRF-NH2 . The precursor encoded by the third transcript (Bgl-FaRP3; 287 amino acids) recapitulated Bgl-FaRP2 but lacked the full SKPYMRF-NH2 peptide. The three precursors shared a common signal peptide, suggesting a genomic organization described previously in gastropods. Immunohistochemical studies were performed on the nervous systems of B. glabrata and B. alexandrina, a major intermediate host for S. mansoni in Egypt. FMRF-NH2 -like immunoreactive (FMRF-NH2 -li) neurons were located in regions of the central nervous system associated with reproduction, feeding, and cardiorespiration. Antisera raised against non-FMRF-NH2 peptides present in the tetrapeptide and heptapeptide precursors labeled independent subsets of the FMRF-NH2 -li neurons. This study supports the participation of FMRF-NH2 -related neuropeptides in the regulation of vital physiological and behavioral systems that are altered by parasitism in Biomphalaria.

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Programmed cell death (PCD) is an essential process for the immune system's development and homeostasis, enabling the remotion of infected or unnecessary cells. There are several PCD's types, depending on the molecular mechanisms, such as non-inflammatory or pro-inflammatory. Hemocytes are the main component of cellular immunity in bivalve mollusks. Numerous infectious microorganisms produce toxins that impair hemocytes functions, but there is little knowledge on the role of PCD in these cells. This study aims to evaluate in vitro whether marine toxins induce a particular type of PCD in hemocytes of the bivalve mollusk Crassostrea gigas during 4 h at 25°C. Hemocytes were incubated with two types of marine toxins: non-proteinaceous toxins from microalgae (saxitoxin, STX; gonyautoxins 2 and 3, GTX2/3; okadaic acid/dynophysistoxin-1, OA/DTX-1; brevetoxins 2 and 3, PbTx-2,-3; brevetoxin 2, PbTx-2), and proteinaceous extracts from bacteria (Vibrio parahaemolyticus, Vp; V. campbellii, Vc). Also, we used the apoptosis inducers, staurosporine (STP), and camptothecin (CPT). STP, CPT, STX, and GTX 2/3, provoked high hemocyte mortality characterized by apoptosis hallmarks such as phosphatidylserine translocation into the outer leaflet of the cell membrane, exacerbated chromatin condensation, DNA oligonucleosomal fragments, and variation in gene expression levels of apoptotic caspases 2, 3, 7, and 8. The mixture of PbTx-2,-3 also showed many apoptosis features; however, they did not show apoptotic DNA oligonucleosomal fragments. Likewise, PbTx-2, OA/DTX-1, and proteinaceous extracts from bacteria Vp, and Vc, induced a minor degree of cell death with high gene expression of the pro-inflammatory initiator caspase-1, which could indicate a process of pyroptosis-like PCD. Hemocytes could carry out both PCD types simultaneously. Therefore, marine toxins trigger PCD's signaling pathways in C. gigas hemocytes, depending on the toxin's nature, which appears to be highly conserved both structurally and functionally.

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Near Infrared Spectroscopy (NIRS) is a spectroscopic technique that evaluates the vibrational energy levels of the chemical bonds of molecules within a wavelength range of 750-2,500 nm. This simple method acquires spectra that provide qualitative and quantitative data on the chemical components of the biomass of living organisms through the interaction between the electromagnetic waves and the sample. NIRS is an innovative, rapid, and non-destructive technique that can contribute to the differentiation of species based on their chemical phenotypes. Chemical profiles were obtained by NIRS from three snail species (Biomphalaria glabrata, Biomphalaria straminea, and Biomphalaria tenagophila) that are intermediate hosts of Schistosoma mansoni in Brazil. The correct identification of these species is important from an epidemiological viewpoint, given that each species has distinct biological and physiological characteristics. The present study aimed to develop a chemometric model for the interspecific and intra-specific classification of the three species, focusing on laboratory and field populations. The data were obtained from 271 live animals, including 150 snails recently collected from the field, with the remainder being raised in the laboratory. Populations were sampled at three localities in the Brazilian state of Rio de Janeiro, in the municipalities of Sumidouro (B. glabrata) and Paracambi (B. straminea), and the borough of Jacarepaguá in the Rio de Janeiro city (B. tenagophila). The chemometric analysis was run in the Unscrambler® software. The intra-specific classification of the field and laboratory populations obtained accuracy rates of 72.5% (B. tenagophila), 77.5% (B. straminea), and 85.0% (B. glabrata). The interspecific differentiation had a hit rate of 75% for the field populations and 80% for the laboratory populations. The results indicate chemical and metabolic differences between populations of the same species from the field and the laboratory. The chemical phenotype, which is closely related to the metabolic profile of the snails, varied between environments. Overall, the NIRS technique proved to be a potentially valuable tool for medical malacology, enabling the systematic discrimination of the Biomphalaria snails that are the intermediate hosts of S. mansoni in Brazil.

Assuntos