Detection of Toxoplasma gondii in wild bivalves from the Kerguelen and Galapagos archipelagos: influence of proximity to cat populations, exposure to marine currents and kelp density.

Oocysts of the protozoan Toxoplasma gondii are found in felid feces and can be washed into coastal waters, where they persist for months, attaching to algae and accumulating in invertebrates. We used wild bivalves to assess contamination of coastal waters of the Kerguelen and Galapagos archipelagos by this zoonotic parasite. Additionally, we leveraged the contrasting situations of these archipelagos to identify some potential drivers of contamination. In the Galapagos, with a cat density reaching 142 per km2, 15.38% of the sampled oysters (Saccostrea palmula) tested positive for T. gondii by quantitative real-time PCR (qPCR) (n = 260), and positive samples were found in all eight sampling sites. In Kerguelen, with 1-3 cats per km2, 40.83% of 120 tested mussels (Mytilus edulis platensis) were positive, and positive samples were found in four out of the five sampling sites. These findings provide evidence of T. gondii contamination in the coastal waters of these archipelagos. Furthermore, T. gondii-positive bivalves were found on islands located 20 km away (Galapagos) and 5 km away (Kerguelen) from the nearest cat population, indicating that T. gondii oocysts can disperse through waterborne mechanisms over several kilometers from their initial deposition site. In the Galapagos, where runoff is infrequent and all sites are exposed to currents, the prevalence of qPCR-positive bivalves did not show significant variations between sites (p = 0.107). In Kerguelen where runoff is frequent and site exposure variable, the prevalence varied significantly (p < 0.001). The detection of T. gondii in Kerguelen mussels was significantly correlated with the site exposure to currents (odds ratio (OR) 60.2, p < 0.001) and the on-site density of giant kelp forests (OR 2.624, p < 0.001). This suggests that bivalves can be contaminated not only by oocysts transported by currents but also by consuming marine aggregates containing oocysts that tend to form in kelp forests.

Identification and expression analysis of two steamer-like retrotransposons in the Chilean blue mussel (Mytilus chilensis).

BACKGROUND: Disseminated neoplasia (DN) is a proliferative cell disorder of the circulatory system of bivalve mollusks. The disease is transmitted between individuals and can also be induced by external chemical agents such as bromodeoxyuridine. In Mya arenaria, we have cloned and characterized an LTR-retrotransposon named Steamer. Steamer mRNA levels and gene copy number correlates with DN and can be used as a marker of the disease. So far, the only mollusk where a retrotransposon expression relates to DN is Mya arenaria. On the other hand, it has been reported that the Chilean blue mussel Mytilus chilensis can also suffers DN. Our aim was to identify retrotransposons in Mytilus chilensis and to study their expression levels in the context of disseminated neoplasia. RESULTS: Here we show that 7.1% of individuals collected in August 2018, from two farming areas, presents morphological characteristics described in DN. Using Steamer sequence to interrogate the transcriptome of M. chilensis we found two putative retrotransposons, named Steamer-like elements (MchSLEs). MchSLEs are present in the genome of M. chilensis and MchSLE1 is indeed an LTR-retrotransposon. Neither expression, nor copy number of the reported MchSLEs correlate with DN status but both are expressed at different levels among individual animals. We also report that in cultured M. chilensis haemocytes MchSLEs1 expression can be induced by bromodeoxyuridine. CONCLUSIONS: We conclude that SLEs present in Mytilus chilensis are differentially expressed among individuals and do not correlate with disseminated neoplasia. Treatment of haemocytes with a stressor like bromodeoxyuridine induces expression of MchSLE1 suggesting that in Mytilus chilensis environmental stressors can induce activation of LTR-retrotransposon.

The Mediterranean Mussel Mytilus galloprovincialis (Mollusca: Bivalvia) in Chile: Distribution and Genetic Structure of a Recently Introduced Invasive Marine Species.

The genetic characteristics of invasive species have a significant impact on their ability to establish and spread. The blue mussel (Mytilus galloprovincialis), native to the Mediterranean Sea, is a leading invasive species of intertidal coasts throughout much of the world. Here, we used mitochondrial DNA sequence data to investigate the genetic diversity and phylogeographic structure of invasive (M. galloprovincialis) versus native (Mytilus chilensis) populations of blue mussels in Chile. We evaluated whether genetic diversity in invasive populations could be explained by the genetic characteristics of the native sources from which they might be derived. A phylogenetic analysis confirmed two lineages of the invasive M. galloprovincialis, i.e., the NW Atlantic and the Mediterranean lineages. We found no evidence of genetic structure in the invasive range of M. galloprovincialis in Chile, most probably because of its recent arrival. We did, however, detect a spatial mixture of both M. galloprovincialis lineages at sampling locations along the Chilean coast, giving rise to higher levels of genetic diversity in some areas compared to the population of native M. chilensis. The coastal area of the invasion is still small in extent (~100 km on either side of two large ports), which supports the hypothesis of a recent introduction. Further expansion of the distribution range of M. galloprovincialis may be limited to the north by increasing water temperatures and to the south by a natural biogeographic break that may slow or perhaps stop its spread. The use of internal borders as a tool to minimise or prevent M. galloprovincialis spread is therefore a genuine management option in Chile but needs to be implemented rapidly.

Trace Metals in the Mussel Mytilus californianus from the West Coast of Baja California, Mexico.

This study assessed the concentrations of trace metals in tissues of Mytilus californianus from the west coast of Baja California, Mexico, during the cold and warm seasons. The concentrations of silver, cadmium, copper and zinc in mussel tissues were measured using Atomic Absorption Spectrophotometry. Silver concentrations ranged from 0.03 to 0.48 µg/g dry weight (d.w.) with significantly higher values registered at sites close to densely populated areas. Cadmium, copper, and zinc concentrations ranged from 0.57 to 7.83 µg/g d.w., 3.71 to 621 µg/g d.w., and 57.54 to 124.55 µg/g d.w., respectively. According to the WHO, values of copper and zinc were within acceptable limits. The maximum values of cadmium, however, exceeded the recommended limits set by the European Commission (2014). The human health risk of the metals analyzed was low (THQ and HI < 1). The regular monitoring of trace metals in M. californianus is recommended for a more definitive evaluation of contaminants.

Identification and expression analysis of two steamer-like retrotransposons in the Chilean blue mussel (Mytilus chilensis)

Background Disseminated neoplasia (DN) is a proliferative cell disorder of the circulatory system of bivalve mollusks. The disease is transmitted between individuals and can also be induced by external chemical agents such as bromodeoxyuridine. In Mya arenaria, we have cloned and characterized an LTR-retrotransposon named Steamer. Steamer mRNA levels and gene copy number correlates with DN and can be used as a marker of the disease. So far, the only mollusk where a retrotransposon expression relates to DN is Mya arenaria. On the other hand, it has been reported that the Chilean blue mussel Mytilus chilensis can also suffers DN. Our aim was to identify retrotransposons in Mytilus chilensis and to study their expression levels in the context of disseminated neoplasia. Results Here we show that 7.1% of individuals collected in August 2018, from two farming areas, presents morphological characteristics described in DN. Using Steamer sequence to interrogate the transcriptome ofM. chilensis we found two putative retrotransposons, named Steamer-like elements (MchSLEs). MchSLEs are present in the genome of M. chilensis and MchSLE1 is indeed an LTR-retrotransposon. Neither expression, nor copy number of the reported MchSLEs correlate with DN status but both are expressed at different levels among individual animals. We also report that in cultured M. chilensis haemocytes MchSLEs1 expression can be induced by bromodeoxyuridine. Conclusions We conclude that SLEs present in Mytilus chilensis are differentially expressed among individuals and do not correlate with disseminated neoplasia. Treatment of haemocytes with a stressor like bromodeoxyuridine induces expression of MchSLE1 suggesting that in Mytilus chilensis environmental stressors can induce activation of LTR-retrotransposon.

Nocturnal seston: A key to explain the cadmium transfer from seawater to mussels (Mytilus chilensis).

The objective of this work was to discover a biochemical pathway to explain the transfer of cadmium, a toxic element, from seawater to cultured mussels. Understanding the intricacies of this transfer is crucial for global mussel crops, as it has the potential to mitigate risks to human health and prevent economic losses in the industry. We focused our investigation on Yal Bay, a typical area with intense mussel aquaculture activity (16,000 t y-1) in the inland sea of southern Chile. Seasonal samples of blue mussels (Mytilus chilensis) were collected and analyzed from September 2014 to December 2015 at two integrated depths (0-5 m and 5-10 m). Diurnal and nocturnal seston, seawater, benthic sediments and decanted suspensions from the water column were recorded. Our findings indicate that nocturnal seston satisfactorily explains the presence of cadmium in Mytilus chilensis aquaculture throughout its annual temporal distribution (Spearman rs = 0.63, p = 0.002).

El mejillón Semimytilus patagonicus (Mytilida: Mytilidae) y el gusano Pseudonereis gallapagensis (Phyllodocida: Nereididae) como herramientas ecotoxicológica para la contaminación por detergentes en ambientes marinos / The mussel Semimytilus patagonicus (Mytilida: Mytilidae) and the worm Pseudonereis gallapagensis (Phyllodocida: Nereididae) as ecotoxicological tools for detergent pollution in marine environments

Introduction: The bivalve Semimytilus patagonicus is a potentially useful bioindicator because of its feeding mechanism, and the worm Pseudonereis gallapagensis is also interesting as a bioindicator because it is benthonic, abundant, and a food source for the squid Doryteuthis gahi. However, their sensitivity to contaminants has not been sufficiently studied. Objective: To test the usefulness of the mussel Semimytilus patagonicus and the polychaete Pseudonereis gallapagensis as ecotoxicological tools for detergents in the marine environment. Methods: We used 120 individuals of S. patagonicus from Miraflores and 120 of P. gallapagensis from Barranco (both near the city of Lima, Peru). For the bioassays, we used two anionic detergents (active ingredient, ai, Sodium Dodecylbenzene Sulfonate). For S. patagonicus, with an average valve length of 32.3 ± 6.4 mm, we tested "Double power Ariel®" (90 %) at concentrations of 17.5, 35, 70 and 140 mg ai l-1, evaluated after 48 and 72 h of exposure; and for P. gallapagensis, with a total body length of 20.4 ± 8.8 mm, we tested "Caricia®" at 62.5, 125, 250, 500 and 1 000 mg of ai l-1 at 24, 48 and 72 h of exposure. Results: The LC50 values (Mean Lethal Concentration) were 34.95 mg ia l-1 for S. patagonicus and 102.48 mg ia l-1 for P. gallapagensis at 72 h of exposure. The detergents were toxic for S. patagonicus and slightly toxic for P. gallapagensis. The risk classification for S. patagonicus is "harmful" and for P. gallapagensis "not classifiable". Conclusions: These two bioindicators allow evaluating the acute toxicity of SDBS-based commercial detergents in the marine aquatic environment.

Introducción: El bivalvo Semimytilus patagonicus es un bioindicador potencialmente útil por su mecanismo de alimentación, y el gusano Pseudonereis gallapagensis también es interesante como bioindicador por ser bentónico, abundante y fuente de alimento para el calamar Doryteuthis gahi. Sin embargo, su sensibilidad a los contaminantes no ha sido suficientemente estudiada. Objetivo: Probar la utilidad del mejillón S. patagonicus y el poliqueto P. gallapagensis como herramientas ecotoxicológicas para detergentes en el medio marino. Métodos: Se utilizaron 120 individuos de S. patagonicus de Miraflores y 120 de P. gallapagensis de Barranco (ambos cerca de la ciudad de Lima, Perú). Para los bioensayos se utilizaron dos detergentes aniónicos (ingrediente activo, ia, dodecilbenceno sulfonato de sodio). Para S. patagonicus, con una longitud valver promedio de 32.3 ± 6.4 mm, probamos Ariel Doble Poder® (90 %) a concentraciones de 17.5, 35, 70 y 140 mg·ia·l-1, evaluadas a las 48 y 72 h de exposición; y para P. gallapagensis, con una longitud corporal total de 20.4 ± 8.8 mm, probamos Caricia® a 62.5, 125, 250, 500 y 1 000 mg·ia·l-1 a las 24, 48 y 72 h de exposición. Resultados: Los valores de CL50 (Concentración Letal Media) fueron de 34.95 mg·ia·l-1 para S. patagonicus y 102.48 mg·ia·l-1 para P. gallapagensis a las 72 h de exposición. Los detergentes fueron tóxicos para S. patagonicus y levemente tóxicos para P. gallapagensis. La clasificación de riesgo para S. patagonicus es "nocivo" y para P. gallapagensis "no clasificable". Conclusiones: Estos dos bioindicadores permiten evaluar la toxicidad aguda del detergente comercial a base de SDBS en el ambiente acuático marino.

Morphological, histopathological and molecular assessments of Prosorhynchoides sp. (Digenea: Bucephalidae) in Perna perna (Bivalvia: Mytilidae) mussels sampled off the coast of Rio de Janeiro, southeastern Brazil.

Mussel production is expanding worldwide, and in Brazil the main species currently produced is the mussel Perna perna. Bucephalid trematodes have been recorded in P. perna but their larval identification is problematic. In this context, the aims of this paper were to evaluate the prevalence of bucephalids in P. perna, perform taxonomic and phylogenetic trematode studies, and analyze potential histopathological alterations in the infected host. Mussels obtained by fishers from Guanabara Bay, Rio de Janeiro, Brazil were weighed and measured, and internal organ tissues and parasites were collected. Of the 69 analyzed mussels, 24.6 % (17/69) were parasitized by bucephalid larvae. Sporocysts were located mainly in host mantle. Mussels presented sporocysts and cercaria within the connective tissue of mantle, all without associated inflammatory reactions. Parasite loads varied from less than 5 % to > 50 % of parasitized tissue. Histopathological examinations indicated that male or female gonads were not observed in 77 % (10/13) of parasitized mussels and in 4 % (2/56) identified as non-parasitized in the histology but previously classified as parasitized in the stereomicroscopic analysis. Thus, the absence of gonads may be associated with parasitism. Prosorhynchoides sp. is reported herein for the first time in mussels sampled on the coast of Rio de Janeiro, with genetic and histological data reported for the intermediate host, sporocysts and cercariae. New 28S rDNA, 18S rDNA and ITS1, 5.8S and ITS2 sequences are provided.

Abundance of coral-associated fauna in relation to depth and eutrophication along the leeward side of Curaçao, southern Caribbean.

Coral-associated invertebrates contribute much to the biodiversity of Caribbean coral reefs. Although the nature of their symbiotic relation is usually not fully understood, they can cause damage to their hosts, especially when they occur in high densities. The abundance of seven groups of coral-associated invertebrates was investigated on reefs along the leeward side of Curaçao, southern Caribbean. In particular, coral barnacles (Pyrgomatidae), boring mussels (Mytilidae: Leiosolenus spp.), gall crabs (Cryptochiridae), and Christmas tree worms (Serpulidae: Spirobranchus spp.) were recorded together with their host corals by means of a photo survey at four depths (5, 10, 15, 20 m) and across seven sites with high and five sites with low eutrophication values (based on δ15N isotope data). Feather duster worms (Sabellidae: Anamobaea), coral blennies (Chaenopsidae: Acanthemblemaria), and worm snails (Vermetidae: Petaloconchus) were insufficiently abundant for thorough quantitative analyses. The results show a decrease in the number of barnacles and Christmas tree worms per host over depth, which could be related to the availability of their host corals. Sites with high δ15N values show a higher abundance of barnacles and Christmas tree worms per host than sites with low values. This indicates that eutrophication could be favourable for these filter feeding organisms but when their densities become too high, they tend to overgrow their hosts and may become a threat to them.

Prevalence, distribution and environmental effects on faecal indicator bacteria and pathogens of concern in commercial shellfish production areas in a subtropical region of a developing country (Santa Catarina, Brazil).

This paper reviews recent literature on the abundance and distribution of faecal indicator bacteria and pathogens in shellfish production areas in the state of Santa Catarina, on the subtropical coast of Brazil. This state supplies > 95% of the national production of shellfish. Microbiological monitoring data were mapped using GIS and the results compared with those from other countries. Coastal human population is the main predictive parameter for faecal bacteria in the production areas. Temporal variations of the bacteria can also be predicted by solar radiation and rainfall. The prevalence of pathogens such as hepatitis A virus, human norovirus, Salmonella spp. and Vibrio spp. does not differ substantially from that in developed countries. The information reported here can be used to inform development of microbiological risk profiles for shellfish production areas.

The Role of the Ecotoxicology Applied to Seafood as a Tool for Human Health Risk Assessments Concerning Polycyclic Aromatic Hydrocarbons.

BACKGROUND: Polycyclic aromatic hydrocarbons (PAHs) are persistent pollutants routinely detected in aquatic ecosystems. It is, therefore, necessary to assess the link between deleterious marine biota PAH effects, especially in commercialized and consumed animals, environmental health status, and potential human health risks originating from the consumption of contaminated seafood products. Thus, this review seeks to verify the relationships of ecotoxicological studies in determining effect and safety concentrations on animals routinely consumed by humans. METHODS: A total of 52 published studies between 2011 and 2021, indexed in three databases, were selected following the PICO methodology, and information on test animals, evaluated PAH, and endpoints were extracted. RESULTS: Benzo(a)pyrene and phenanthrene were the most investigated PAHs in terms of biomarkers and test organisms, and mussels were the most evaluated bioindicator species, with an emphasis on reproductive responses. Furthermore, despite the apparent correlation between environmental PAH dynamics and effects on aquatic biota and human health, few assessments have been performed in a multidisciplinary manner to evaluate these three variables together. CONCLUSIONS: The links between human and environmental sciences must be strengthened to enable complete and realistic toxicity assessments as despite the application of seafood assessments, especially to mussels, in bioassays, the connection between toxicological animal responses and risks associated with their consumption is still understudied.

Norovirus GII and astrovirus in shellfish from a mangrove region in Cananéia, Brazil: molecular detection and characterization.

In recent years, annual cases of gastroenteritis have been reported in the world at high rates, suggesting an association with the consumption of shellfish with enteric viruses in their tissues. Anthropic activities are considered a source of environmental pollution and the main responsible for contamination by pathogenic microorganisms in aquatic environments. The objective of this study was to evaluate, by RT-semi-nested PCR, the presence of astrovirus (AstV) and norovirus genogroup II (NoV GII) in mussels (Mytella falcata) and oysters (Crassostrea brasiliana) collected in two sites of the Lagunar Complex of Cananéia, State of São Paulo, Brazil. A total of 150 samples of mussels and oysters (75 samples each) were analyzed. AstV was not identified in any shellfish sample. NoV GII was detected in 21 samples (14%), 8 mussel samples (38%), and 13 oyster samples (62%). From the 21 positive samples, 16 were analyzed by nucleotide sequencing. The molecular characterization revealed that Brazilian samples were grouped into clades along with other sequences from Brazil, Japan, and Mexico. There was 93.8-100% amino acid sequence similarity among the samples in this study and > 94.9% when compared with the strains isolated from clinical cases in Brazil. The screening of shellfish for the presence of health-significant enteric viruses can help prevent outbreaks among consumers and contribute to the improvement of the estuarine environment.

Occurrence of pharmaceuticals in seafood from two Brazilian coastal areas: Implication for human risk assessment.

Pharmaceuticals (PhACs) are considered emerging contaminants with potential accumulation in aquatic organisms. Thus, seafood consumption may cause long-term effects and health risk for consumers. In the present study, the occurrence of PhACs in seafood from two Brazilian coastal areas, Sepetiba Bay (n = 43) and Parnaiba Delta River (n = 48), was determined for the first time, and their potential risk for human health was assessed. An eco-friendly multi-analytes method was used, after being validated for the different types of matrices (mussels, fatty and lean fish). All compounds under study were detected at least in four seafood species, including chloramphenicol, an antibiotic prohibited in animal foods. Most PhACs had mean concentrations below limit of quantification. Ibuprofen and other nonsteroidal anti-inflammatory drugs (NSAIDs), as well as simvastatin and carbamazepine were the main PhACs bioaccumulated in edible parts of seafood species from Brazil. The high trophic level carnivorous species, snook, was the most contaminated by NSAIDs, while bivalves were the seafood more contaminated by lipid regulators. The profile of contamination did not vary among different types of matrix, except in relation to carbamazepine and ketoprofen. These PhACs were more abundant in species from Sepetiba Bay, an area highly impacted by human influence. The estimated daily exposure for Brazilian population that consumes the studied species was up to 20.3 ng/kg bw/day via carib pointed-venus and 25.7 ng/kg bw/day via snooks, lower than acceptable daily intake. Thus, consumption of seafood species from Sepetiba Bay and Parnaiba Delta River seems to be safe to the population in what concerns the PhACs studied.

Concentration of microplastics in bivalves of the environment: a systematic review.

The aim of this review was to identify the current knowledge regarding the concentration of microplastics in bivalves in the marine, estuarine, and freshwater environments. For this purpose, researches were conducted from September 2020 to February 2021 in the Scopus, Web of Science, and Google scholar databases, following a meticulous selection of articles. To comprehensively understand the selected articles, an extensive review was carried out in order to identify the methodologies employed, sampling sites, species evaluated, characteristics of the microplastics (concentrations, shapes, sizes, and polymers) and their relationship with the concentration of this particles in the environment. A total of 93 articles were selected, with an exponential growth in the number of articles from April 2014 to February 2021. Worldwide, 80 articles were realized in the Northern Hemisphere and thirteen in the Southern Hemisphere. The samplings of organisms were carried out in 36 countries, besides one in Antarctica. The concentration of microplastics were studied in 70 species, with mussels Mytilus spp. and the oysters Crassostrea spp. being the main genus studied. Due to the different methodologies used to digest the tissues of organisms and identify microplastics and species, it is difficult to make comparisons between the results of different studies. In addition, data on the concentrations of microplastics in the environment, as well as their composition and characteristics, are needed, enabling the verification of relationships with the concentrations identified in organisms, which does not occur in most studies. Thus, we suggest an increase in the number of studies to be realized in the southern hemisphere, future studies use the same methodology of digestion, the polymer identification of microplastics and samplings of the surrounding environment, enabling a greater comparison between studies.

Trace Metal Residues in Marine Mussels: A Global Survey.

Pressures from anthropogenic activities are causing degradation of estuarine and coastal ecosystems around the world. Trace metals are key pollutants that are released and can partition in a range of environmental compartments, to be ultimately accumulated in exposed biota. The level of pressure varies with locations and the range and intensity of anthropogenic activities. The present study measured residues of trace metals in Mytilus mussel species collected from a range of locations around the world in areas experiencing a gradient of anthropogenic pressures that we classified as low, moderate, or high impact. The data showed no grouping/impact level when sampling sites in all countries were incorporated in the analysis, but there was significant clustering/impact level for most countries. Overall, high-impact areas were characterized by elevated concentrations of zinc, lead, nickel, and arsenic, whereas copper and silver were detected at higher concentrations in medium-impact areas. Finally, whereas most metals were found at lower concentrations in areas classified as low impact, cadmium was typically elevated in these areas. The present study provides a unique snapshot of worldwide levels of coastal metal contamination through the use of Mytilus species, a well-established marine biomonitoring tool. Environ Toxicol Chem 2021;40:3434-3440. © 2021 SETAC.

Comparing the concentrations of heavy metals on two bivalve species in Santos Bay, Brazil: Subsidies to understanding the assimilation dynamic of bivalve contaminants.

Contaminant substances consist of chemical elements that present the potential to adversely impact the ecology of the environment, thus representing a threat to local fauna and flora. In this context, heavy metals are critical agents that, depending on the nature and level, are potentially toxic to living organisms. In order to evaluate the bioaccumulation of heavy metals in the Santos estuary and to determine the potential influence of salinity gradient on the heavy metal bioconcentration, the present study measured the concentrations of As, Cd, Pb, Cu, Cr, Fe, Ni, and Zn in two bivalve species (Crassostrea rhizophorae and Perna Perna) sampled at different sites of Santos Bay, in the southeastern region of Brazil. Throughout the study, the "sentinel species" used were effective in bioaccumulating contaminants. In oysters, based on the Brazilian legislation, critical limits were exceeded for As, Zn, Cu, and Cr. In the case of mussels, on the other hand, only for As, Zn, and Cr, the critical limits were overcome. In the present study, obtained data suggested salinity as a determinant parameter in As incorporation processes of bivalve mollusks. PRACTITIONER POINTS: The present study presents important results for the development of environmental management policies in estuarine environments. The present study points out differences between different organisms as biomonitors, providing subsidies for the decision of an effective biomonitoring program. The present study discusses values of contaminants as a danger to public health in Santos Bay, which can be extrapolated to other similar environments around the world.

Longitudinal Study of Total and Pathogenic Vibrio parahaemolyticus (tdh+ and/or trh+) in Two Natural Extraction Areas of Mytilus chilensis in Southern Chile.

Vibrio parahaemolyticus is the leading cause of seafood-associated bacterial gastroenteritis worldwide. Although different studies have focused on its pattern of variation over time, knowledge about the environmental factors driving the dynamics of this pathogen, within the Chilean territory, is still lacking. This study determined the prevalence of total and pathogenic V. parahaemolyticus strains (tdh and/or trh genes) in mussels (Mytilus chilensis) collected from two natural growing areas between 2017 and 2018, using selective agar and PCR analysis. V. parahaemolyticus was detected in 45.6% (93/204) of pooled samples from the Valdivia River Estuary. The pathogenic strains carrying the tdh and/or trh gene were detected in 11.8% (24/204): tdh in 9.8% (20/204), trh in 0.5% (1/204), and 1.5% (3/204) presented both genes. In Reloncaví Fjord, V. parahaemolyticus was detected in 14.4% (30/209) of the samples, pathogenic V. parahaemolyticus carrying the trh gene was detected in 0.5% (1/209) of the samples, while the tdh gene was not detected in the samples from this area. The total count of mauve-purple colonies typical of V. parahaemolyticus on CHROMagar was positively associated by multivariate analysis with area, water temperature, and salinity. Similarly, V. parahaemolyticus detection rates by PCR had a positive correlation with the area and water temperature. The chances of detecting total V. parahaemolyticus in the Valdivia River Estuary are significantly higher than in the Reloncaví Fjord, but inversely, during spring-summer months, the interaction factor between the area and temperature indicated that the chances of detecting V. parahaemolyticus are higher in the Reloncaví Fjord. Interestingly, this period coincides with the season when commercial and natural-growing shellfish are harvested. On the other hand, pathogenic V. parahaemolyticus tdh+ was significantly correlated with an increase of water temperature. These environmental parameters could be used to trigger a warning on potential hazard, which would influence human health and economic losses in aquaculture systems.

Integrated biomarker response in Mytilus chilensis exposed to untreated urban discharges along the coast of Ushuaia Bay (Beagle Channel, Argentina).

The short-term effects of coastal untreated effluents from Ushuaia Bay, Beagle Channel, on the biochemical and physiological biomarkers of Mytilus chilensis were assessed. An integrated biomarker response (IBR) index was calculated as a helpful tool to represent the general stress of the experimental organisms. Cultured mussels were exposed during 96 h to three coastal sites impacted by sewage effluents. At the beginning (T0) and every 24 h, mussels were subsampled from each site and different biochemical and physiological biomarkers were determined. There was no mortality registered in the experiments during the 96 h. However, biochemical and physiological biomarkers presented significant variations. Lipid peroxidation mean levels in mussels decreased in mantle and increased in digestive gland with respect to T0 in almost all cases. Acetylcholinesterase activity was inhibited in all sites, reaching a maximal decrease of 35% with respect to T0. Catalase remained stable and glutathione-S-transferase was activated. Oxygen consumption and ammonia excretion rates increased in organisms from two sites and, consequently, O:N ratio decreased, denoting a symptom of stress. IBR values showed the existence of different stress levels between exposed and unexposed mussels. These results exhibited an alteration of the general metabolism of mussels exposed for a short period of time to untreated coastal wastewater, suggesting the use of these organisms as early indicators of changes in the environmental quality of coastal waters of Ushuaia Bay.

First Report of Zoonotic Genotype of Giardia duodenalis in Mussels (Mytilus edulis) from Patagonia Argentina.

Blue mussels (Mytilus edulis) are among the most consumed fishery products globally. Foodborne outbreaks of mussel-associated infections by viral, bacterial, and parasitic pathogens have been reported in the last years. In this study, we investigated the occurrence, genetic diversity, and zoonotic potential of the diarrhea-causing enteric protozoan Giardia duodenalis in blue mussels from Caleta Córdova in Chubut Province, southeast Patagonia, Argentina. A total of 344 free-living blue mussels were collected and distributed in 53 aliquots of pooled mussel tissue (each containing 5‒7 specimens) during the period 2015‒2018. Conventional optical microscopy was used as screening method for the detection of G. duodenalis cysts in pooled, homogenized tissues. Samples with a positive result were assessed by a multilocus sequence genotyping scheme based on the amplification of partial fragments of the glutamate dehydrogenase and ß-giardin genes of the parasite. G. duodenalis cysts were found in 30.2% (16/53) of the aliquots of pooled mussel tissue tested. PCR and sequencing analyses revealed the presence of G. duodenalis subassemblage BIV in selected aliquots. To the best of our knowledge, this is the first description of zoonotic subassemblage BIV from blue mussels in Argentina.

Stress responses to warming in the mussel Brachidontes rodriguezii (d'Orbigny, 1842) from different environmental scenarios.

The intertidal mussel B. rodriguezii is a representative species from hard bottom substrates where both anthropogenic and natural stressors are present. Pre-exposure to these different stressors can modify the tolerance to additional stressors such as warming. Moreover, this tolerance can vary depending on intraspecific variables such as the organism's sex. The effects of warming and its intraspecific variability in representative coastal species are crucial to understanding the tolerance to future environmental scenarios. The mussels were collected in different environmental scenarios, including low (Control), chemical (Harbour) and natural stressed (Estuary) sites, and then exposed to different water temperatures (10-30 °C) for 14 days. Lethal and sublethal responses were evaluated in different mussel populations. Thus, cumulative death rate, air survival time, heat shock proteins (HSC70/HSP70), total ubiquitin, catalase (CAT), glutathione-s-transferase (GST) and lipid peroxidation (TBARS) were assessed in mussels from different areas and different sexes. The results revealed diminished air survival time and high cumulative mortality rate in mussels collected at the harbour and those exposed to higher temperatures, respectively. The sublethal responses of the field animals showed different patterns according to the different areas investigated. Besides, the results revealed that these differences were also observed between sexes. Regarding the sublethal responses in mussels exposed to warming, the interactive effects of temperature and sites showed a strong influence on all biochemical parameters analyzed (p < 0.001). Therefore, harbour mussels showed a distinct pattern compared to other locations and reflecting the most damaging effects of warming. The influence of sex and its interactions with warming were also crucial in most of the sublethal responses (p < 0.05). Multivariate analysis was performed with all sublethal responses, and the different warming scenarios showed different groups according to the sites. In the predicted warming scenarios, males showed no differences between sites. In contrast to males, females showed differences between sites in the predicted and the worse-case warming scenarios. Our results highlight the importance of compensatory mechanisms in the mussel warming tolerance like HSP70. The influence of sex is also crucial in understanding warming tolerance in mussels chronically exposed to pollutants in their natural environment. Also, lethal endpoints are essential for understanding the non-reversibility signature of the observed biochemical responses.