Elevated seasonal temperature disrupts prooxidant-antioxidant homeostasis and promotes cellular apoptosis in the American oyster, Crassostrea virginica, in the Gulf of Mexico: a field study.

One of the major impacts of climate change has been the marked rise in global temperature. Recently, we demonstrated that high temperatures (1-week exposure) disrupt prooxidant-antioxidant homeostasis and promote cellular apoptosis in the American oyster. In this study, we evaluated the effects of seasonal sea surface temperature (SST) on tissue morphology, extrapallial fluid (EPF) conditions, heat shock protein-70 (HSP70), dinitrophenyl protein (DNP, an indicator of reactive oxygen species, ROS), 3-nitrotyrosine protein (NTP, an indicator of RNS), catalase (CAT), superoxide dismutase (SOD) protein expressions, and cellular apoptosis in gills and digestive glands of oysters collected on the southern Texas coast during the winter (15 °C), spring (24 °C), summer (30 °C), and fall (27 °C). Histological observations of both tissues showed a notable increase in mucus production and an enlargement of the digestive gland lumen with seasonal temperature rise, whereas biochemical analyses exhibited a significant decrease in EPF pH and protein concentration. Immunohistochemical analyses showed higher expression of HSP70 along with the expression of DNP and NTP in oyster tissues during summer. Intriguingly, CAT and SOD protein expressions exhibited significant upregulation with rising seasonal temperatures (15 to 27 °C), which decreased significantly in summer (30 °C), leaving oysters vulnerable to oxidative and nitrative damage. qRT-PCR analysis revealed a significant increase in HSP70 mRNA levels in oyster tissues during the warmer seasons. In situ TUNNEL assay showed a significant increase in apoptotic cells in seasons with high temperature. These results suggest that elevated SST induces oxidative/nitrative stress through the overproduction of ROS/RNS and disrupts the antioxidant system which promotes cellular apoptosis in oysters.

First report of detection of Toxoplasma gondii DNA in oysters (Crassostrea sp) in the state of Maranhão / Primeiro relato da detecção de DNA de Toxoplasma gondii em ostras (Crassostrea sp) no Estado do Maranhão

The aim of this study was to report on detection of Toxoplasma gondii DNA in oysters (Crassostrea sp.) in the state of Maranhão. To conduct this study, 200 farmed oysters were acquired in the municipality of Raposa and 100 in Paço do Lumiar; and a further 100 oysters were taken from the natural stock in the municipality of Primeira Cruz. This total of 400 specimens sampled was divided into 80 pools composed of five animals each. The gills and visceral mass of each oyster were removed for DNA extraction (per pool of oysters), using a commercial kit. The nested PCR technique (with the primer SAG-1) was then used to investigate any presence of protozoa. This molecular technique demonstrated the presence of DNA of T. gondii in 2.5% of the pools of oysters (n = 2/80): these oysters were exclusively from farms. The results from this study allow the conclusion that oysters of the genus Crassostrea that are farmed in the state of Maranhão are capable of filtering oocysts of T. gondii and maintaining them in their tissues. They are therefore potential sources of contamination for humans and other animals.(AU)

Objetivou-se com este estudo relatar a detecção do DNA de Toxoplasma gondii em ostras (Crassostrea sp.) no estado do Maranhão. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa, e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz. Do total de 400 exemplares amostrados, formaram-se 80 pools em que cada pool foi constituído por cinco animais. De cada ostra foi procedida à retirada das brânquias e massa visceral, seguido da extração de DNA de cada pool de ostras, com a utilização de kit comercial. Posteriormente, realizou-se a pesquisa do protozoário por meio da técnica de nested PCR (primer SAG-1). Com a técnica molecular utilizada, foi diagnosticado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools de ostras oriundas exclusivamente de cultivo. Com os resultados obtidos neste estudo, conclui-se que ostras do gênero Crassostrea sp., cultivadas no estado do Maranhão, são capazes de filtrar e manter nos seus tecidos oocistos de T. gondii, sendo, portanto, fontes potenciais de contaminação para seres humanos e outros animais.(AU)

Recruitment of oyster in artificial collectors on the Amazon macrotidal mangrove coast / Recrutamento de ostra em coletores artificiais na costa de manguezais de macromaré amazônico

The purpose of this study was to determine the moment of the year for the oyster recruitment and define the type of collector and environmental conditions that maximize recruitment. Collections were conducted, during 12 months, on Amazon Macrotidal Mangrove at two different sites: raft (point I) and mangrove (point II). In each location three types of collectors were used (1) transparent PET bottles, (2) green PET bottles, and (3) PVC sheets, each with three replicates. Spats were counted and measured at 45-day intervals, while the environmental data were measured every two weeks. Identification of oyster species occurred by genetic testing (multiplex PCR) by randomly selecting individuals by sampling. Results indicated spat capturing was significantly influenced by the collector type, location and period of collection (P<0.05, MANOVA) with significantly higher recruitment in the PVC collector (P<0.05, Tukey test). Oyster recruitment occurred throughout the year, suggesting that these individuals reproduce during all months; however, months with less rain and greater salinity were the best for spat collection, while the rainy period with lower salinity proved to be the best for individuals growth. The location in interaction with the environmental variables, mainly salinity, has a significant effect on the recruitment rate of spat and on their size, so that point II (mangrove) had the best results for recruitment and point I (raft) provided the spats of the largest size. Genetic identification verified two native oysters species (Crassostrea gasar and Crassostrea rhizophorae) in both points (I and II).(AU)

O estudo teve como objetivo determinar o período do ano para coletar sementes de ostras, definir o tipo de coletor e as condições ambientais que maximizam o recrutamento das sementes. As coletas foram conduzidas, durante 12 meses, no manguezal amazônico de macromaré em dois locais distintos: balsa (ponto I) e manguezal (ponto II). Em cada ponto de coleta foram utilizados três tipos de coletores (1) garrafas PET transparentes, (2) garrafas PET verdes e (3) forro de PVC, com três réplicas cada. As sementes foram contadas e medidas a cada 45 dias e os dados ambientais mensurados a cada duas semanas. As espécies de ostras foram identificadas por teste genético (PCR multiplex) com indivíduos aleatoriamente selecionados por coleta. Os resultados indicaram que a captura de sementes foi influenciada significativamente pelo tipo de coletor, localização e período de coleta (P<0,05, MANOVA) com recrutamento significativamente maior no coletor de PVC (P<0,05, teste de Tukey). O recrutamento ocorreu durante todo ano, sugerindo reprodução mensal, contudo, os meses de menor pluviosidade e maior salinidade foram melhores para coletar sementes, enquanto o período chuvoso, com menor salinidade, favoreceu um crescimento superior. O local de coleta em interação com variáveis ambientais, principalmente salinidade, apresentou efeito significativo sobre a fixação e tamanho das sementes de maneira que o ponto II obteve melhores resultados de fixação e o ponto I sementes de maior tamanho. A análise genética identificou duas espécies nativas (Crassostrea gasar e Crassostrea rhizophorae) em ambos os pontos (I e II).(AU)

Molecular identification, phylogeny and geographic distribution of Brazilian mangrove oysters (Crassostrea)

Oysters (Ostreidae) manifest a high degree of phenotypic plasticity, whereby morphology is of limited value for species identification and taxonomy. By using molecular data, the aim was to genetically characterize the species of Crassostrea occurring along the Brazilian coast, and phylogenetically relate these to other Crassostrea from different parts of the world. Sequencing of the partial cytochrome oxidase c subunit I gene (COI), revealed a total of three species of Crassostrea at 16 locations along the Brazilian coast. C. gasar was found from Curuçá (Pará state) to Santos (São Paulo state), and C. rhizophorae from Fortim (Ceará state) to Florianópolis (Santa Catarina state), although small individuals of the latter species were also found at Ajuruteua beach (municipality of Bragança, Pará state). An unidentified Crassostrea species was found only on Canela Island, Bragança. Crassostrea gasar and C. rhizophorae grouped with C. virginica, thereby forming a monophyletic Atlantic group, whereas Crassostrea sp. from Canela Island was shown to be more similar to Indo-Pacific oysters, and either arrived in the Atlantic Ocean before the convergence of the Isthmus of Panama or was accidentally brought to Brazil by ship.

Trans-atlantic distribution of a mangrove oyster species revealed by 16S mtDNA and karyological analyses.

Three species of mangrove oysters, Crassostrea rhizophorae, C. brasiliana, and C. gasar, have been described along the Atlantic shores of South America and Africa. Because the distribution of these molluscs is of great biological and commercial interest, their taxonomy and distribution deserve further clarification. Therefore, 15 populations were sampled from both continents. Their 16S mitochondrial polymorphism was studied by sequencing and PCR-RFLP analysis. Two haplotypes were identified. Haplotype a was the only one observed in Africa, but it was also observed in South America together with haplotype b. Because C. gasar is the only mangrove oyster identified on the west coast of Africa, haplotype a was attributed to this species, which has thus been shown to occur in South America. Haplotype b is attributed to C. rhizophorae. The karyotypes of specimens of C. gasar, from Africa and from South America, were very similar, and both species were observed at the same location in Brazil. The occurrence of C. gasar in South America adds a third species-in addition to C. rhizophorae and C. brasiliana-to the list of species present along these coasts. The predominant surface circulation patterns in this part of the Atlantic Ocean favor the hypothesis that C. gasar was transported from Africa to America. Finally, a phylogenetic tree built with seven 16S sequences from Crassostrea and Saccostrea species showed that C. gasar is intermediate between the American Crassostrea species (C. virginica and C. rhizophorae) and the Asian species (C. gigas and C. ariakensis).