RESUMO
This work focused on assessing of the risk associated with the consumption of bivalve mollusks, potentially contaminated with phycotoxins. The studied phycotoxins are saxitoxin (STX), okadaic acid (OA), dinophysistoxins (DTXs), yessotoxins (YTXs), pectenotoxins (PTX), azaspiracids (AZAs), and domoic acid (DA). These toxins were investigated in three species of bivalve mollusks (Anadara senilis, Crassostrea gasar, and Perna perna), originating from the Ebrié lagoon. Chemical analyses were carried out by LC-MS/MS, HPLC-FLD, and HPLC-UV. The level of OA and DTXs, STX, and DA was 10.92 µg OA eq./kg, 9.6 µg STX eq./kg, and 0.17 mg DA eq./kg, respectively. The level of PTXs and AZAs was 3.3 µg PTX-2 eq./kg and 13.86 µg AZA-1 eq./kg; that of YTXs was 0.01 mg YTX eq./kg. The daily exposure dose (DED) was 0.019 µg OA eq./kg bw for OA and DTXs; 0.285 µg DA eq./kg bw for DA; 0.006 µg PTX-2 eq./kg bw for PTXs; 0.016 µg STX eq./kg bw for STX; 0.01 µg YTX eq./kg bw for YTXs; and 0.024 µg AZA-1 eq./kg bw for AZAs for the oyster Crassostrea gasar. These estimated values are lower than the acute reference dose (ARfD) of each phycotoxin recommended by the European Food Safety Agency (EFSA). The risk of harmful effects is acceptable. The absence of risk is valid only for the study period (11 months) and concerns coastal populations living near the sampling points.
Assuntos
Bivalves , Ecossistema , Furanos , Macrolídeos , Venenos de Moluscos , Oxocinas , Toxinas de Poliéter , Animais , Côte d'Ivoire , Cromatografia Líquida , Espectrometria de Massas em Tandem , Monitoramento Ambiental , Ácido OkadáicoRESUMO
Roundup (Rn), a glyphosate-based formulation, is one of the most commonly used herbicides in the world. It affects non-targeted organisms in several ways, including adhesive activity towards metal ions. Zinc (Zn) plays a crucial role in a number of biochemical processes. In this study, we aim to elucidate the direct impact of Rn on Zn accumulation and Zn-dependent activities in the ex vivo system. To this end, we exposed the samples of the digestive gland of a bivalve mollusk Unio tumidus to 3 µM of Rn (calculated as 3 µM of glyphosate), Zn, Zn chelator (N,N,N',N'-Tetrakis(2-pyridylmethyl)ethylenediamine) (TPEN, Tp), and their combinations ZnTp and ZnRn for 17 h. We determined the levels of Zn in the tissue (Zn t) and metallothioneins (Zn-MT), metallothioneins (MTSH), and glutathione (GSH & GSSG), total antioxidant capacity (TAC), lysosomal membrane integrity, and caspase-3 activity. Our study demonstrated that Rn and Tp had different effects on the accumulation and functionality of Zn. Rn did not affect the accumulation of Zn (Zn t, Zn-MT) in the Zn- and ZnRn-groups. On the contrary, Tp produced effects antagonistic to Zn on caspase-3 activity, lysosomal stability, and MTSH concentration. Rn caused particular pro-oxidative effect that decreased GSH level (Rn- and ZnRn-groups) and lysosomal stability (Rn-group). The shared affected index was the GSH/GSSG ratio, which decreased by 2-8 times in each exposure. As the first experience with the application of Tp to indicate Zn activity in mollusks, the study concluded that the ex vivo approach could be useful in the study of numeral aquatic pollutants.
Assuntos
Unio , Poluentes Químicos da Água , Animais , Metalotioneína , Moluscos , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Zinco/toxicidadeRESUMO
Even though bivalve molluscs are recognized as bioindicators of freshwater quality, their responses to multiple stressors are unpredictable. This study aims to elucidate the inter-population peculiarities of the effect in the sub-chronic environmentally relevant exposure to novel contaminants. The specimens of Unio tumidus from reference (Pr) and contaminated (Ct) areas were treated with ibuprofen (IBU, 0.8 µg L-1), microplastic (MP, 1.0 mg L-1, size 0.1-0.5 mm), or their combination (Mix) for 14 days. Untreated mussels (PrC- and CtC-groups) served as controls. The PrC-group had higher levels of antioxidants Mn-SOD, Cu,Zn-SOD, catalase, and cholinesterase (AChE) as well as lesser levels of oxidative lesions (TBARS and protein carbonyls) in digestive glands, indicating lower environmental impact than in the CtC-group. However, lysosomal stability was similar in both control groups. Among antioxidants, Mn-SOD activity was affected most prominently, increasing in all exposed Ct-groups. TBARS level was increased only in PrMP-group compared to responsive control. IBU and Mix enhanced protein carbonyl concentration in the Pr-groups, and decreased it in the Ct-groups. AChE was induced in the CtIBU- and PrMix-groups, and lysosomal integrity increased in the CtIBU and CtMix-groups. Discriminant analyses indicated lesser differences between Pr-groups, demonstrating lower cumulative stress compared to Ct-groups. Generally, the most remarkable response was revealed in the CtIBU-group, and distortion of individual effects was established in combined exposures. The qualification of stress-neutral and stress-positive populations was proposed for Pr- and Ct-populations correspondingly. Inter-site peculiarities must be taken into consideration when the environmental impact of MP and pharmaceuticals is evaluated.
Assuntos
Bivalves , Unio , Poluentes Químicos da Água , Animais , Unio/metabolismo , Microplásticos , Ibuprofeno/toxicidade , Plásticos/metabolismo , Antioxidantes/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Poluentes Químicos da Água/toxicidade , Poluentes Químicos da Água/metabolismo , Bivalves/metabolismo , Superóxido Dismutase/metabolismo , Estresse OxidativoRESUMO
Scrobicularia plana is a coastal and estuarine bivalve widely used in ecotoxicological studies. However, the underlying molecular mechanisms for S. plana pollutant responses are hardly known due to the lack of molecular databases. Thus, in this study we present a holistic approach to assess a robust reference transcriptome and proteome of this clam. A mixture of control and metal-exposed individuals was used for mRNA isolation. Four sets of high quality filtered preprocessed reads were generated (two quality scores and two sequenced lengths) and assembled with Mira, Ray and Trinity algorithms. The sixty-four generated assemblies were refined, filtered and evaluated for their proteomic quality. Eight assemblies presented top Detonate scores but one was selected due to its compactness and biological representation, which was generated: (i) from the highest quality dataset (Q20L100), (ii) using Trinity algorithm with all k-mers (AtKa), (iii) removing redundancy by CD-HIT (RR80), and (iv) filtering out poor contigs (F), that was subsequently named Q20L100AtKaRR80F. S. plana proteomic analysis revealed 10,017 peptide groups that corresponded to 2066 proteins with a wide coverage of molecular functions and biological processes, confirming the strength of the database generated.
Assuntos
Bivalves , Proteoma , Animais , Bivalves/genética , Sequenciamento de Nucleotídeos em Larga Escala , Proteômica , TranscriptomaRESUMO
C1q domain-containing (C1qDC) proteins are a group of biopolymers involved in immune response as pattern recognition receptors (PRRs) in a lectin-like manner. A new protein MkC1qDC from the hemolymph plasma of Modiolus kurilensis bivalve mollusk widespread in the Northwest Pacific was purified. The isolation procedure included ammonium sulfate precipitation followed by affinity chromatography on pectin-Sepharose. The full-length MkC1qDC sequence was assembled using de novo mass-spectrometry peptide sequencing complemented with N-terminal Edman's degradation, and included 176 amino acid residues with molecular mass of 19 kDa displaying high homology to bivalve C1qDC proteins. MkC1qDC demonstrated antibacterial properties against Gram-negative and Gram-positive strains. MkC1qDC binds to a number of saccharides in Ca2+-dependent manner which characterized by structural meta-similarity in acidic group enrichment of galactose and mannose derivatives incorporated in diversified molecular species of glycans. Alginate, κ-carrageenan, fucoidan, and pectin were found to be highly effective inhibitors of MkC1qDC activity. Yeast mannan, lipopolysaccharide (LPS), peptidoglycan (PGN) and mucin showed an inhibitory effect at concentrations three orders of magnitude greater than for the most effective saccharides. MkC1qDC localized to the mussel hemal system and interstitial compartment. Intriguingly, MkC1qDC was found to suppress proliferation of human adenocarcinoma HeLa cells in a dose-dependent manner, indicating to the biomedical potential of MkC1qDC protein.
Assuntos
Glicoproteínas de Membrana/genética , Moluscos , Proteínas/genética , Receptores de Complemento/genética , Receptores de Reconhecimento de Padrão/genética , Animais , Organismos Aquáticos , Humanos , Glicoproteínas de Membrana/química , Oceano Pacífico , Proteínas/química , Receptores de Complemento/química , Receptores de Reconhecimento de Padrão/químicaRESUMO
Hydropower plants (HPPs) can affect the hydrological regime. However, biochemical responses of aquatic animals for the evaluation of this disturbing are not applied yet. The specimens of Unio tumidus were sampled in a reservoir (R) of a small HPP as well as downstream from the dam (DS). Biochemical indexes in the digestive gland and alkali labile phosphates (ALP) in the gonads were examined. The R-mollusks showed low cholinesterase, catalase and caspase-3 activities, and metallothionein concentration, but elevated levels of zinc and copper, oxidized glutathione and protein carbonyls. Concentrations of lactate, pyruvate and ALP, activity of superoxide dismutase and glutathione S-transferase, and lipid peroxidation level were similar in both groups. Integrated biomarker response (IBR/n) index (n = 13) was 2.17 and 0.29 in the R- and DS-groups correspondingly. We suggest that using integrative biological response based on the biochemical markers of bivalve mollusks can be a valid early warning step in assessing 'environmental flow' impact.
Assuntos
Monitoramento Ambiental , Centrais Elétricas , Unio/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Biomarcadores/metabolismo , Catalase/metabolismo , Glutationa Transferase/metabolismo , Peroxidação de Lipídeos , Metalotioneína/metabolismo , Moluscos/metabolismo , Rios , Ucrânia , Unio/metabolismo , Poluentes Químicos da Água/análiseRESUMO
Our understanding of disease processes and host-pathogen interactions in model species has benefited greatly from the application of medium and high-throughput genomic, metagenomic, epigenomic, transcriptomic, and proteomic analyses. The rate at which new, low-cost, high-throughput -omic technologies are being developed has also led to an expansion in the number of studies aimed at gaining a better understanding of disease processes in bivalves. This review provides a catalogue of the genetic and -omic tools available for bivalve species and examples of how -omics has contributed to the advancement of marine bivalve disease research, with a special focus in the areas of immunity, bivalve-pathogen interactions, mechanisms of disease resistance and pathogen virulence, and disease diagnosis. The analysis of bivalve genomes and transcriptomes has revealed that many immune and stress-related gene families are expanded in the bivalve taxa examined thus far. In addition, the analysis of proteomes confirms that responses to infection are influenced by epigenetic, post-transcriptional, and post-translational modifications. The few studies performed in bivalves show that epigenetic modifications are non-random, suggesting a role for epigenetics in regulating the interactions between bivalves and their environments. Despite the progress -omic tools have enabled in the field of marine bivalve disease processes, there is much more work to be done. To date, only three bivalve genomes have been sequenced completely, with assembly status at different levels of completion. Transcriptome datasets are relatively easy and inexpensive to generate, but their interpretation will benefit greatly from high quality genome assemblies and improved data analysis pipelines. Finally, metagenomic, epigenomic, proteomic, and metabolomic studies focused on bivalve disease processes are currently limited but their expansion should be facilitated as more transcriptome datasets and complete genome sequences become available for marine bivalve species.
Assuntos
Bivalves/genética , Genômica , Interações Hospedeiro-Patógeno/fisiologia , Proteômica , AnimaisRESUMO
The antipsychotic chlorpromazine (Cpz) has raised concern as a pharmaceutical effluent due to its wide medical applications. Moreover, its potent pro-oxidant properties and impact on the cell viability of the marine mollusc Mytilus galloprovincialis, even at low concentrations (ng/L), have been noted. Based on this evidence, in this study, we investigated the physiological effects of Cpz on M. galloprovincialis, to elucidate its fate within the organism, in terms of bioaccumulation, biotransformation, byssus changes and stress responses of the cellular thiolome. Histological and indicators of vitality analyses were also performed to better evaluate the influence of the drug on the morphology and cell viability of the digestive gland. To this end, two different concentrations of Cpz (Cpz I (12 ng/L or 37 pM) and Cpz II (12 µg/L or 37 nM)) were administered to mussels over 14 days. Cpz accumulation in the digestive gland significantly increased with water concentration (BCF of Cpz I and Cpz II). Biochemical analyses indicated lysosomal dysfunction, reflected in elevated total Cathepsin D activity and compromised lysosomal membrane stability. Stress-related and metal-buffering proteins (GST and metallothionein) responded to both Cpz concentrations. Cpz I induced phase I biotransformation activity (CYP450-dependent EROD), while Cpz II triggered caspase-3 activation, indicative of detoxification overload. Histological analysis revealed digestive gland atrophy, epithelial thinning, haemocyte infiltration, and brown cell presence. Byssus analysis showed significant alterations. In conclusion, our study underscores Cpz-induced physiological and histological changes in M. galloprovincialis, posing potential implications for mussel health and confirming the utilisation of this mussel as an indication of Cpz ecotoxicity.
Assuntos
Mytilus , Poluentes Químicos da Água , Animais , Mytilus/metabolismo , Clorpromazina/toxicidade , Metais/metabolismo , Biotransformação , Poluentes Químicos da Água/metabolismo , Biomarcadores/metabolismoRESUMO
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.
Assuntos
Bivalves , Infecções por Caliciviridae , Mastadenovirus , Norovirus , Ostreidae , Rotavirus , Animais , Humanos , Brasil/epidemiologia , Cidades , Rotavirus/genética , Norovirus/genética , Genótipo , Filogenia , FezesRESUMO
SARS-CoV-2 can be detected in the feces of infected people, consequently in wastewater, and in bivalve mollusks, that are able to accumulate viruses due to their ability to filter large amounts of water. This study aimed to monitor SARS-CoV-2 RNA presence in 168 raw wastewater samples collected from six wastewater treatment plants (WWTPs) and 57 mollusk samples obtained from eight harvesting sites in Campania, Italy. The monitoring period spanned from October 2021 to April 2022, and the results were compared and correlated with the epidemiological situation. In sewage, the ORF1b region of SARS-CoV-2 was detected using RT-qPCR, while in mollusks, three targets-RdRp, ORF1b, and E-were identified via RT-dPCR. Results showed a 92.3% rate of positive wastewater samples with increased genomic copies (g.c.)/(day*inhabitant) in December-January and March-April 2022. In the entire observation period, 54.4% of mollusks tested positive for at least one SARS-CoV-2 target, and the rate of positive samples showed a trend similar to that of the wastewater samples. The lower SARS-CoV-2 positivity rate in bivalve mollusks compared to sewages is a direct consequence of the seawater dilution effect. Our data confirm that both sample types can be used as sentinels to detect SARS-CoV-2 in the environment and suggest their potential use in obtaining complementary information on SARS-CoV-2.
Assuntos
Bivalves , COVID-19 , Humanos , Animais , Águas Residuárias , RNA Viral/genética , SARS-CoV-2/genética , COVID-19/epidemiologia , Itália/epidemiologiaRESUMO
The common cockle is a valuable bivalve species inhabiting the Atlantic European coasts. The parasite Marteilia cochillia has devastated cockle beds in the southern Galician (NW Spain) rias since 2012. Previous data suggested that cockles from Ría de Arousa acquired some resilience to this parasite through natural selection after consecutive annual marteiliosis outbreaks and candidate markers associated with marteiliosis resilience were identified using population genomics and transcriptomics approaches. Here, a common garden experiment was performed using a naïve stock (from Ría de Muros-Noia) and an affected stock (from Ría de Arousa) to test this hypothesis. Breeders from both stocks were used to produce seed cohorts at hatchery, which were pre-grown in a raft (outdoor nursery stage) and deployed in two shellfish beds affected by marteiliosis in Ría de Arousa (growing-out stage). In both beds, the naïve stock showed high marteiliosis prevalence and was fully depleted in a short period, while the affected stock barely showed evidence of marteiliosis. A set of 45 SNPs putatively associated with marteiliosis resilience were fitted for MassARRAY genotyping to check their role in the differential resilience detected between both stocks. Though no significant differentiation was found between the naïve and the affected stocks with neutral markers, 28 SNPs showed significant divergence between them, suggesting that these SNPs were involved in directional selection during eight generations (to the most) of marteiliosis pressure (long-term selection). Furthermore, signals of selection were also detected in the naïve stock along the marteiliosis outbreak in the growing-out stage (short-term selection) and six SNPs, all shared with the long-term evaluation, showed consistent signals of differentiation according to the infection severity. Some of these SNPs were located within immune genes pertaining to families such as proteasome, ubiquitin, tumor necrosis factor, and glutathione S-transferase. These resilience-associated markers will be useful to recover cockle production in Galicia.
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A growing body of evidence shows that microplastic pollution is ubiquitous in bivalve mollusks globally and is of particular concern due to its potential impact on human health. However, non-standardized sampling, processing, and analytical techniques increased the difficulty of direct comparisons among existing studies. Based on 61 peer-reviewed papers, we summarized the current knowledge of microplastics in bivalve mollusks globally and provided an in-depth analysis of factors affecting the outcome of microplastic data, with the main focus on the effects of different species and methodologies. We found no significant differences in microplastic abundance among genera from the same family but significant differences among bivalve families, indicating habitats play an important role in microplastic ingestion by bivalve mollusks. This also provided foundational knowledge for using epifaunal and infaunal bivalves to monitor microplastic pollution in water and sediment, respectively. Recommendations for microplastic monitoring protocol in bivalve mollusks were proposed according to the results of this review, covering (i) a sample size of at least 50 bivalves in the study area, (ii) the use of 10 % KOH as the digestion solution, and (iii) the pore size of a filter membrane of < 5 µm. Acknowledging the need for a standard procedure, more efforts towards protocol standardization used in long-term and large-scale microplastic monitoring programs in bivalve mollusks are needed.
Assuntos
Bivalves , Poluentes Químicos da Água , Animais , Monitoramento Ambiental/métodos , Humanos , Microplásticos , Plásticos/análise , Padrões de Referência , Poluentes Químicos da Água/análiseRESUMO
The vulnerability of bivalve mollusks to micropollutants is estimated mainly in single model exposures. However, chronic environmental stress and complex exposures can modulate their responses. To evaluate the impact of population-dependent adaptations on the ability to react to common micropollutants, we compared freshwater bivalves Unio tumidus from two distinct populations, pure (Pr) and contaminated (Ct), in their exposures to microplastics (MP, 1 mg L-1, size 0.1-0.5 mm), pharmaceutical ibuprofen (IBU, 0.8 µg L-1), or their combination (Mix) for 14 days. Control groups from both sites showed remarkable differences, with lower levels of total antioxidant capacity (TAC), metallothionein protein (MTSH), NADH and NAD+, cytochrome P450-related EROD, glutathione-S transferase (GST), and citrate synthase (CS) but higher levels of GSH, GSSG, caspase-3 and cathepsin D (CTD) in the Ct-control group. These data indicate a chronic stress impact in the Ct population. Under exposures, we found an almost common strategy in both populations for NAD+/NADH and MTSH suppression and CTD induction. Additionally, Mix exposure caused an increase in CS, and IBU did not change GSH in both populations. However, the expected response to IBU - the suppression of caspase-3 - was indicated only in PrIBU- and PrMix-mollusks. CTD efflux increased dramatically only in PrMP- and PrMix- groups, and suppression of EROD and GST was detected in the PrMix-group. According to discriminant analysis, exposed Pr-groups were highly differentiated from control, whereas Ct-control and exposed groups had common localization demonstrating high resistance to environmental stress. Thus, the same exposures resulted in different adverse outcome pathways depending on the population.
Assuntos
Bivalves , Unio , Poluentes Químicos da Água , Animais , Apoptose , Biomarcadores/metabolismo , Bivalves/metabolismo , Caspase 3/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Glutationa Transferase/metabolismo , Ibuprofeno/toxicidade , Microplásticos , NAD/metabolismo , Plásticos , Unio/metabolismo , Poluentes Químicos da Água/metabolismo , Poluentes Químicos da Água/toxicidadeRESUMO
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.
Assuntos
Bivalves , Terapia a Laser , Animais , Chumbo , Espectrometria de Massas/métodos , Isótopos/análise , Bivalves/químicaRESUMO
Toxoplasma gondii is a major foodborne pathogen capable of infecting all warm-blooded animals, including humans. Although oocyst-associated toxoplasmosis outbreaks have been documented, the relevance of the environmental transmission route remains poorly investigated. Thus, we carried out an extensive systematic review on T. gondii oocyst contamination of soil, water, fresh produce, and mollusk bivalves, following the PRISMA guidelines. Studies published up to the end of 2020 were searched for in public databases and screened. The reference sections of the selected articles were examined to identify additional studies. A total of 102 out of 3201 articles were selected: 34 articles focused on soil, 40 focused on water, 23 focused on fresh produce (vegetables/fruits), and 21 focused on bivalve mollusks. Toxoplasma gondii oocysts were found in all matrices worldwide, with detection rates ranging from 0.09% (1/1109) to 100% (8/8) using bioassay or PCR-based detection methods. There was a high heterogeneity (I2 = 98.9%), which was influenced by both the sampling strategy (e.g., sampling site and sample type, sample composition, sample origin, season, number of samples, cat presence) and methodology (recovery and detection methods). Harmonized approaches are needed for the detection of T. gondii in different environmental matrices in order to obtain robust and comparable results.
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The crystallographic texture of the whole valves of bivalve mollusks from the family Unionidae Unio pictorum Linnaeus, 1758 and Anodonta cygnea Linnaeus, 1758 is studied using pole figures measured using neutron diffraction. The use of neutron diffraction, in contrast to X-rays, makes it possible to study the valves without destroying them. Thus, we can discuss the study of the global texture of the entire valve. It was revealed that the pole figures of aragonite in the valves repeat their shape. The pole density maxima for U. pictorum from the Danube Delta and the Gulf of Finland in the Baltic Sea, living at different salinities and temperatures, differs by 0.41 mrd. The maximum value of the crystallographic texture for A. cygnea from the Danube Delta was also measured (5.07 mrd). In terms of texture sharpness, it surpasses the shell of marine bivalve mollusks, which are partially or completely composed of aragonite. Although U. pictorum and Mya arenaria Linnaeus, 1758 have different microstructures, their pole figures are very similar in isolines pattern, but differ in pole density maxima. No relationship was found between the crystallographic texture and the microstructure in U. pictorum. In addition, we report good qualitative agreement between aragonite X-ray pole figures of Sinanodonta woodiana Lea, 1834 from the Czech river Luznice, and neutron pole figures of U. pictorum from the Danube Delta.
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The structure and composition of the mitogenome of a bivalve mollusk, denoted as Mactra sp. (MT780813), has been obtained. The genome has a variable organization: it includes 12 protein-coding genes, 28 tRNAs, and two rRNA genes. Its content is sufficiently different from that of nearest specimen, accessed from GenBank, supposedly belonging to the other gender. All genes are encoded on the "+"-strand. All protein-coding genes are initiated with ATG codon. Analysis confirms the close topological position of the GenBank Mactra chinensis (KJ754823) and our M. sp. specimen on gene tree. Above data suggesting female- vs. male-type mitogenomes or cryptic species presence.
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Vibrio neptunius is an important pathogen of bivalve mollusks worldwide. Several metalloproteases have been described as virulence factors in species of Vibrio that are pathogenic to bivalves, but little is known about the contribution of these potential virulence factors to Vibrio neptunius pathogenesis. In silico analysis of the genome of V. neptunius strain PP-145.98 led to the identification of two hitherto uncharacterized chromosomal loci encoding a probable vibriolysin-like metalloprotease and a putative collagenase, which were designated VnpA and ColA, respectively. Single defective mutants of each gene were obtained in V. neptunius PP-145.98, and the phospholipase, esterase and collagenase activities were studied and compared with those of the wild-type strain. The results showed that the single inactivation of vnpA resulted in a 3-fold reduction in phospholipase/esterase activity. Inactivation of colA reduced the collagenase activity by 50%. Finally, infection challenges performed in oyster larvae showed that ΔvnpA and ΔcolA-single mutant strains of V. neptunius-are between 2-3-fold less virulent than the wild-type strain. Thus, the present work demonstrates that the production of both VnpA and ColA is required for the full virulence of the bivalve pathogen V. neptunius.
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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.
Assuntos
Apoptose/efeitos dos fármacos , Toxinas Bacterianas/toxicidade , Crassostrea/efeitos dos fármacos , Hemócitos/efeitos dos fármacos , Toxinas Marinhas/toxicidade , Animais , Toxinas Bacterianas/isolamento & purificação , Caspases/metabolismo , Montagem e Desmontagem da Cromatina/efeitos dos fármacos , Crassostrea/imunologia , Crassostrea/metabolismo , Quebras de DNA de Cadeia Dupla , Hemócitos/imunologia , Hemócitos/metabolismo , Hemócitos/patologia , Fosfatidilserinas/metabolismo , Vibrio/metabolismo , Vibrio parahaemolyticus/metabolismoRESUMO
Metal release into the environment from anthropogenic activities may endanger ecosystems and human health. However, identifying and quantifying anthropogenic metal bioaccumulation in organisms remain a challenging task. In this work, we assess Cu isotopes in Pacific oysters (C. gigas) as a new tool for monitoring anthropogenic Cu bioaccumulation into marine environments. Arcachon Bay was taken as a natural laboratory due to its increasing contamination by Cu, and its relevance as a prominent shellfish production area. Here, we transplanted 18-month old oysters reared in an oceanic neighbor area into two Arcachon Bay mariculture sites under different exposure levels to continental Cu inputs. At the end of their 12-month long transplantation period, the oysters' Cu body burdens had increased, and was shifted toward more positive δ65Cu values. The gradient of Cu isotope compositions observed for oysters sampling stations was consistent with relative geographic distance and exposure intensities to unknown continental Cu sources. A binary isotope mixing model based on experimental data allowed to estimate the Cu continental fraction bioaccumulated in the transplanted oysters. The positive δ65Cu values and high bioaccumulated levels of Cu in transplanted oysters support that continental emissions are dominantly anthropogenic. However, identifying specific pollutant coastal source remained unelucidated mostly due to their broader and overlapping isotope signatures and potential post-depositional Cu isotope fractionation processes. Further investigations on isotope fractionation of Cu-based compounds in an aqueous medium may improve Cu source discrimination. Thus, using Cu as an example, this work combines for the first time a well-known caged bivalve approach with metal stable isotope techniques for monitoring and quantifying the bioaccumulation of anthropogenic metal into marine environments. Also, it states the main challenges to pinpoint specific coastal anthropogenic sources utilizing this approach and provides the perspectives for further studies to overcome them.