Semi-quantitative risk assessment of the contamination of Vibrio parahaemolyticus in oysters from Jinshan District, Shanghai / 上海预防医学

Objective To investigate the prevalence of Vibrio parahaemolyticus(VP)in oysters in Jinshan District, Shanghai and make assessment on the risks that may cause, providing the basis for prevention and control of foodborne disease. Methods Raw oyster samples with shells were randomly collected from markets, supermarkets and restaurants in Jinshan District from July to October in 2017. The content of VP in oysters was tested in accordance with the national standard methods. The semi-quantitative risk assessment for VP in oysters was made by Risk Ranger combining with the monitoring results of diet and health status of residents in Jinshan District of Shanghai in 2012-2013. Results The overall positive rate of VP in the 40 oyster samples was 80.0%(32/40). The positive rate of VP in oyster samples from farmer's markets was the highest, 85.7%(12/14), followed by those from restaurants and supermarkets. The relative risk for VP in raw oysters was 63. The probability of illness caused by VP in oysters per day per consumer of interest was 6.85×10-4, and the total predicted patients annual incidence rate in this population was 1 247.8/105. Conclusion The contamination of VP in seafood oysters in Jinshan District is serious. Eating raw oysters is at high risk; consumers are advised to reduce or avoid eating raw oysters, and processing food before eating is an effective method to decrease VP infection. Strengthening surveillance and management is imperative in this regard.

Molecular investigation of Toxoplasma gondii in oysters (Crassostrea spp.) sold on beaches in the State of Pará, Brazil / Investigação molecular de Toxoplasma gondii em ostras (Crassostrea spp.) comercializadas em praias do estado do Pará, Brasil

Abstract The aim of this study was to detect Toxoplasma gondii DNA in oysters (Crassostrea spp.) sold on seven beaches in the State of Pará, Brazil. According to the National Program for Hygiene and Sanitary Control of Bivalve Mollusks, 100 g of the edible part of mollusks is required to analyze contaminating microorganisms. In this study, 12 oysters were assumed to be equivalent to 100 g of edible parts when preparing each pooled sample. In total, 360 oysters were purchased from 30 vendors. From groups of 12 oysters purchased per vendor, 60 pooled samples were obtained, comprising 30 gill tissues and 30 gastrointestinal tracts. For molecular analysis, nested-PCR was conducted to amplify a 155-base-pair product of the B1 gene from T. gondii. All analyzed samples were negative for T. gondii. Our findings indicate that the oyster samples sold on the beaches in the State of Pará were not contaminated by T. gondii.

Resumo O objetivo deste estudo foi detectar DNA de Toxoplasma gondii em ostras (Crassostrea spp.) comercializadas em sete praias do Estado do Pará, Brasil. De acordo com o Programa Nacional de Controle Higiênico Sanitário de Moluscos Bivalves, 100 g da parte comestível dos moluscos são necessários para a análise de microrganismos contaminantes. Neste estudo, 12 ostras foram consideradas equivalentes a 100 g de partes comestíveis na preparação de cada amostra agrupada. No total, 360 ostras foram compradas de 30 vendedores. De grupos de 12 ostras adquiridas por vendedor, foram obtidas 60 amostras agrupadas, compreendendo 30 tecidos branquiais e 30 tratos gastrointestinais. Para a análise molecular, a nested PCR foi realizada para amplificar um produto de 155 pares de bases do gene B1 de T. gondii. Todas as amostras analisadas foram negativas para T. gondii. Os resultados indicam que as amostras de ostras comercializadas em praias do Estado do Pará não foram contaminadas por T. gondii.

Spatio-temporal Distribution Pattern of Oyster Larvae in Guaratuba Bay, Paraná State - Brazil

HIGHLIGHTS Crassostrea oysters larvae patterns in estuarine environments. Larvae distribution patterns in Guaratuba Bay (Paraná State - Brazil).

Abstract Crassostrea oysters present planktonic/planktotrophic larval development. Thus, it is essential knowing larvae distribution patterns in estuarine environments presenting oyster farms and spat collection. The aim of the current research is to investigate the spatio-temporal distribution of oyster larvae in Guaratuba Bay (Paraná State - Brazil) based on monthly samplings conducted with a plankton net (50 cm diameter and 225 µm mesh size) from June 2003 to June 2004 in three different points (Point I - bay´s entrance; Point II - cultivation park; Point III - median sector of the bay). Plankton samples were preserved in 4% buffered formaldehyde. They were concentrated and quantified in the laboratory. The total number of larvae in the sample from each net was counted and corrected to a standard larvae/m3 collection tow. Data about sea water temperature, salinity, transparency and pluviosity were also collected. Based on results about the spatial distribution of larvae, mean density was 33.30 ±42.73 larvae/m³ in Point I; 17.84 ±16.88 larvae/m³ in Point II and 55.53 ±78.31 larvae/m³ in Point III, during the studied period. Spring and Summer were the seasons recording the most expressive mean number of larvae; the largest concentration of them was found in Point III, in the middle section of the bay.

Molecular cloning and characterisation of scavenger receptor class B in pearl oyster Pinctada fuctada martensii

Background: Molluscs can accumulate carotenoids in their body tissues by predominantly feeding on aquatic plant sources. Carotenoid transport and absorption are determined by the regulation of various proteins such as Scavenger receptor class B(SR-BI). We report the identification and characterisation of pearl oyster Pinctada fuctada martensii SR-BI (PmSR-BI). The correlation between total carotenoid content (TCC) and gene expression was also estimated. Results: The full-length cDNA of PmSR-BI was 1828 bp, including an open-reading frame encoding of 1518 bp with a pI value of 5.83. PmSR-BI protein contains a hydrophobic CD36 domain and four centrally clustered cysteine residues for the arrangement of disulphide bridges. The deduced amino acid sequence had an identity of 30% to 60% with the SR-B of other organisms. Reverse transcription polymerase chain reaction analysis showed that mRNA transcripts were expressed in multiple tissues of adult pearl oyster. A higher expression of PmSR-BI gene was observed in the hepatopancreas than in the adductor muscle, gill and mantle. The TCC and gene expression of PmSR-BI were significantly correlated (P b 0.05), with a correlation coefficient of 0.978. Conclusions: The results suggested that PmSR-BI is involved in the absorption of carotenoids in the pearl oyster P. fuctada martensii.

Oyster is an effective transmission vehicle for Cryptosporidium infection in human

OBJECTIVE@#To determine the ability of oysters to trap and maintain viable Cryptosporidium oocysts, and the feasibility of Cryptosporidium multiplication in oysters' organs.@*METHODS@#Seventy oysters were raised in experimentally seeded natural seawater for up to 3 months, with weekly oocysts inoculations. Cryptosporidium oocysts, viable and non-viable, as well as other stages were detected using two immunofluorescence vital staining techniques (Sporo-Glo and Merifluor(®)) with confocal microscopy. Viability rate at various times after inoculations were calculated.@*RESULTS@#Cryptosporidium oocysts were found most concentrated in oysters' digestive organs than in gill and water inside the oysters. Oocysts numbers were 857.33 at 24 h after inoculation and strikingly decreased to 243.00 and 126.67 oocysts at 72 h and 7 days, respectively. The oocysts in oyster were also less viable over time; 70%, 60% and 30% viable at 24 h, 72 h and 7 days after inoculation, respectively. At 77 days, the number of oocysts was very low and none was found at 84 days onwards. Although some oocysts were ruptured with released sporozoites, there was no evidence throughout the study of sporozoites multiplication to indicate that oyster is a biological host. Despite the significant reduction in oocysts number after 7 days of inoculation, the remained viable oocysts can still cause cryptosporidiosis.@*CONCLUSION@#The findings confirm that Cryptosporidium parvum does not multiply in oyster, and is therefore not a biological host. Nevertheless, the results suggest that oyster can be an effective transmission vehicle for Cryptosporidium oocysts, especially within 24-72 h of contamination, with viable oocysts present at up to 7 days post infection. Unless consuming well-cooked oyster dishes, eating raw oyster remains a public health concern and at least 3 days of depuration in clean sea water prior to consumption is recommended.

Oyster is an effective transmission vehicle for Cryptosporidium infection in human

Objective To determine the ability of oysters to trap and maintain viable Cryptosporidium oocysts, and the feasibility of Cryptosporidium multiplication in oysters' organs. Methods Seventy oysters were raised in experimentally seeded natural seawater for up to 3 months, with weekly oocysts inoculations. Cryptosporidium oocysts, viable and non-viable, as well as other stages were detected using two immunofluorescence vital staining techniques (Sporo-Glo and Merifluor

Detection of Perkinsus marinus in the oyster Crassostrea rhizophorae in southern Bahia by proteomic analysis / Detecção de Perkinsus marinus na ostra Crassostrea rhizophorae do sul da Bahia por análise proteômica

This study reports the presence of the pathogen Perkinsus marinus, notifiable to the World Organization for Animal Health (Office International des Èpizooties = OIE) in the oyster Crassostrea rhizophorae in southern Bahia via proteomic analysis. We analyzed Crassostrea brasiliana from a long-line cultivation system and C. rhizophorae from an adjacent mangrove in Porto do Campo, Camamu Bay, Bahia, Brazil. The collections (n = 100) were performed in October 2012. In the laboratory, the oysters were measured and opened to remove the meat, which was steeped in dry ice. For extraction of proteins, adaptation of a protocol used for mussels was used, after which separation in the first dimension was taken by isoelectric focusing (IEF). The peptides were transferred to a Mass Spectrometer. The obtained spectra were analyzed with the ProteinLynx Global Server 4.2 software tool and also by MASCOT (Matrix Science) and compared to the databases of the SWISSPROT and NCBI, respectively. The identification was evidenced by beta-tubulin, Perkinsus marinus ATCC 50983 and protein homology code in the database NCBI = gi | 294889481. This is the first record of P. marinus in Bahia and the fourth in Brazil.(AU)

Este estudo relata a presença do patógeno Perkinsus marinus, de notificação obrigatória à Organização Internacional de Epizootias (OIE) na ostra Crassostrea rhizophorae no sul da Bahia, via análise proteômica. Foram analisadas as ostras Crassostrea brasiliana de um cultivo em espinhel e C. rhizophorae de um manguezal adjacente, na localidade de Porto do Campo, Baía de Camamu, Bahia. As coletas (n = 100) foram efetuadas em outubro de 2012. Em laboratório, as ostras foram medidas e abertas para a retirada da carne, que foi macerada em gelo seco. Para a extração das proteínas, foi adotada a adaptação de um protocolo utilizado para mexilhões, após o que foi realizada a separação na primeira dimensão, por focalização isoelétrica (IEF). Os peptídeos foram transferidos para um Espectrômetro de Massas. Os espectros obtidos foram analisados no software ProteinLynx Global Server 4.2 e também pela ferramenta MASCOT (Matrix Science) e comparados com os bancos de dados do SWISSPROT e do NCBI, respectivamente. A identificação foi evidenciada por meio da beta-tubulina, homologia Perkinsus marinus ATCC 50983 e código da proteína no banco de dados NCBI = gi|294889481. Este é o primeiro registro de P. marinus na Bahia e o quarto no Brasil.(AU)

The reproductive cycle of the oyster Crassostrea gasar / Ciclo reprodutivo da ostra Crassostrea gasar

The purpose of this study was to analyze the reproductive cycle of the oyster Crassostrea gasar (= C. brasiliana) in the field and the laboratory. The reproductive cycle of the animals was evaluated in the field at Sambaqui Beach, Florianópolis, SC (27° 29′18″ S and 48° 32′12″ W) from May 2008 through November 2009. In July, the animals were in the resting stage. The early growth stage began in August and was followed by the late growth stage in October. In November and December, the oysters began to enter the mature stage. Females in spawning condition were predominant during these months. The stages of the reproductive cycle were positively associated with temperature (r=0.77, P<0.01) and negatively associated with salinity (r=−0.56, P=0.042). These findings demonstrated that increased temperature and reduced salinity influence the reproductive development of Crassostrea gasar. The condition index (CI) of the animals was also associated with the seawater temperature. The highest values of the condition index were observed during the months when the temperature of the seawater was gradually increasing. A laboratory experiment was performed to test the effect of salinity on the reproductive cycle of the oysters. The experiment was conducted in standardized tanks. The animals were conditioned using two salinities (24‰ and 34‰). The salinity regime influenced the development of the gonadal tissue of the oysters. A salinity of 24‰ produced greater reproductive development.

O presente estudo teve como objetivo avaliar o ciclo reprodutivo da ostra Crassostrea gasar, em campo e em laboratório. O estádio de desenvolvimento do ciclo reprodutivo dos animais foi acompanhado na Praia do Sambaqui/Florianópolis/SC (27°29′18″S e 48°32′12″W) entre maio de 2008 e novembro de 2009. No mês de julho os animais encontravam-se no estádio de repouso e entre os meses de agosto a outubro, as ostras seguiram nos estádios de pré-maturação e maturação. Entre os meses de novembro e dezembro, as ostras começaram a ser encontradas no estádio maturo, havendo predominância de fêmeas aptas a eliminação de gametas. Houve associação positiva (r=0,77; P<0,01) entre o estádio de desenvolvimento do ciclo reprodutivo das ostras e a temperatura, bem como associação negativa com a salinidade (r=−0,56; P=0,042), demonstrando que tanto o aumento da temperatura como a redução da salinidade são fatores ambientais que influenciam no desenvolvimento reprodutivo das ostras dessa espécie. O índice de condição (IC) dos animais, também teve relação com a temperatura da água do mar, sendo os maiores índices observados nos meses em que a temperatura da água do mar teve aumento gradativo. Também foi realizado um experimento em laboratório visando testar o efeito da salinidade da água sobre o desenvolvimento reprodutivo das ostras. Para tanto, os animais foram condicionados em tanques padronizados e testaram-se duas salinidades (24‰ e 34‰). Observou-se que o regime de salinidade influenciou o desenvolvimento do tecido gonádico das ostras, sendo que a salinidade de 24‰ permitiu aos animais maior desenvolvimento reprodutivo.

Escherichia coli in the surface waters and in oysters of two cultivations of Guaratuba Bay - Paraná - Brazil

The present work aimed to evaluate the contamination of Escherichia coli in the surface waters and oysters from two cultivations of Guaratuba Bay and to analyze the correlation patterns among the concentrations of E. coli in the waters and in the oysters with the local physical-chemical parameters. Samples were collected in the spring of 2007 and summer, autumn and winter of 2008 from two points of the bay (internal point and external point). From each cultivation and sampling period, 18 oysters were collected. The samples of surface water were collected for the measurement of physical-chemical parameters (pH, salinity, temperature, dissolved oxygen, seston, particulate organic matter) and quantification of E. coli. The surface water analyzed in the summer presented the largest most probable number of E. coli, (1,659.22 MPN.100 ml-1 and 958,55 MPN.100 ml-1 at external and internal points, respectively. The oysters from the internal point presented more E. coli, except in the winter sampling. The largest contamination was observed in the spring, at the internal point (979,78 MPN.g-1). The Principal Components Analysis showed direct correlation among the amount of E. coli in the oysters and in the surface water.

Health assessment of the oyster Crassostrea rhizophorae on the southern coast of Bahia, northeastern Brazil / Avaliação da saúde da ostra Crassostrea rhizophorae no litoral sul da Bahia, nordeste do Brasil

This study investigated the health of natural stocks of the oyster Crassostrea rhizophorae on the southern coast of Bahia in northeastern Brazil, during summer and winter 2010, at three localities (sampling points) in the estuaries of the Maraú (Camamu Bay) and Graciosa rivers. A total of 180 oysters (30/sampling point/season) were examined macroscopically for the presence of pathogens and anatomical changes. The specimens were subsequently fixed in Davidson solution, processed for paraffin embedding, sectioned and stained with Harris' hematoxylin and eosin. Histological analysis revealed the presence of Rickettsia-like organisms (RLOs), Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus in the sporocyst phase, a nonspecific metacercaria, and a metacestode of genus Tylocephalum. The prevalence of infection was low except for parasitism by Nematopsis sp. which also caused histopathological changes. The presence of Bucephalus sp. caused parasitic castration. These two pathogens significantly affect the health of C. rhizophorae.

Este estudo investigou a saúde de ostras da espécie Crassostrea rhizophorae de estoques naturais do Litoral Sul do Estado da Bahia, Nordeste do Brasil, durante o verão e o inverno de 2010, em três pontos amostrais distribuídos nos estuários dos rios Maraú (Baía de Camamu) e Graciosa. Um total de 180 ostras (30/ponto amostral/período) foram examinadas macroscopicamente para a presença de patógenos e alterações anatômicas e posteriormente fixadas em solução de Davidson, processadas para inclusão em parafina, seccionadas e coradas com hematoxilina de Harris e eosina. A análise histológica evidenciou a presença de organismos com características similares a Rickettsia (RLOs), Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus em fase esporocística, metacercária inespecífica e metacestóide de Tylocephalum. As prevalências de infecção foram baixas, com exceção do parasitismo por Nematopsis sp., o qual também causou alterações histopatológicas. A presença de Bucephalus sp. causou castração parasitária. Esses dois patógenos têm interferência significativa na saúde de C. rhizophorae.

Ecologia molecular de ostras (Crassostrea spp. )do Atlântico Tropical / Molecular ecology of oysters (Crassostrea spp. ) from Tropical Atlantic

Crassostrea (Sacco, 1897) é o gênero mais importante do mundo de ostras de cultivo e consiste de 34 espécies distribuídas pelas regiões tropicais e temperadas do globo. C. gasar e C. rhizophorae são as duas espécies nativas que estão distribuídas ao longo de toda a costa do Brasil até o Caribe. C. gasar também ocorre na costa da Africa. Ainda que sua distribuição seja extensa e com disponibilidade abundante, o cultivo de ostras nativas no Brasil ainda é incipiente e a delimitação correta dos estoques mantém-se incerta. O sucesso do desenvolvimento da malacocultura, que é recomendada internacionalmente como forma sustentável de aquicultura, depende da resolução desses problemas. Assim, com o objetivo de determinar geneticamente seus estoques no Atlântico como também estimar sua história demográfica, dois diferentes marcadores moleculares foram empregados: sequências de DNA da região controle mitocondrial e loci de microssatélites espécie-especifícos, desenvolvidos no presente estudo. Foram sequenciados fragmentos da região controle de um total de 930 indivíduos de C. gasar e C. rhizophorae coletados em 32 localidades que incluíram o Caribe, a Guiana Francesa, a costa brasileira e a África. Também foram realizadas genotipagens de 1178 indivíduos, e ambas as espécies, com 9 e 11 loci de microssatélites para C. gasar e C. rhizophorae, respectivamente. Os dados genéticos foram analisados através de diferentes abordagens (índices de estruturação (FST) e de (Jost D), análise molecular de variância (AMOVA), análise espacial molecular de variância (SAMOVA), Bayesian Skyline Plots (BSP), análise fatorial de correspondência (AFC) e análise de atribuição Bayesiana (STRUCTURE)). Os resultados indicaram um padrão geral de estruturação, onde dois diferentes estoques foram detectados para ambas as espécies: grupos do norte e do sul, onde o Rio de Janeiro seria a região limitante entre os dois estoques. Os maiores valores dos índices de estruturação foram encontrados ...

Crassostrea (Sacco, 1897) is the most important genus of cultivated oysters in the world and consisting of 34 species distributed by tropical and temperate regions of the globe. C. gasar and C. rhizophorae are the two native species which have wide distribution along the entire Brazilian coast up to the Caribbean. C. gasar also occurs on coast of Africa. Despite its extensive distribution and abundant availability, cultivation of those oysters in Brazil is incipient, and the correct delimitation of the existing stocks is still uncertain. The successful development of malacoculture which is recommended internationally as an environmentally sustainable form of aquaculture depends on the resolution of these issues. Thus, in order to genetically determinate their stocks in the Atlantic and to estimate their demographic history, two different molecular markers were employed: sequences of the mitochondrial DNA control region and species-specific microsatellite loci, developed in the present study. We have sequenced a fragment of the mitochondrial control region from a total of 930 individuals of C. gasar and C. rhizophorae collected in 32 localities including the Caribbean, French Guyana, Brazilian coast and Africa. We have also genotyped 1178 individuals of both species with 9 and 11 loci of microsatellites for C. gasar and C. rhizophorae, respectively. Genetic data were analyzed with different approaches (fixation (FST) and differentiation (Jost D) indices, analysis of molecular variance (AMOVA), spatial analysis of molecular variance (SAMOVA), Bayesian Skyline Plots (BSP), factorial correspondence analysis (AFC) and Bayesian attribution analysis (STRUCTURE)). The results indicated a general structure pattern, where two different stocks were detected for both species: north and south groups, where Rio de Janeiro would be the limited region between them. Higher values of fixation indices were found for C. gasar, indicating that this species would be more ...

Detección de norovirus y rotavirus en ostras comercializadas en Cumaná, Venezuela / Norovirus and rotavirus detection in oysters commercialized in Cumana, Venezuela

Los brotes de gastroenteritis en humanos han sido frecuentemente asociados con el consumo de moluscos bivalvos. En este estudio se analizaron 15 mezclas de muestras de tejido digestivo de ostras (Pinctada imbricata) comercializadas en la avenida Perimetral de Cumaná, estado Sucre, Venezuela, mediante dos métodos de procesamiento y dos métodos de extracción de ácidos nucleicos, con el fin de detectar por Transcripción Reversa de la Reacción en Cadena de la Polimerasa, el ARN de norovirus y rotavirus. Además se utilizaron cinco pares de oligonucleótidos para seleccionar el mejor en la detección de los virus. Norovirus fue detectado en 13 de las 15 mezclas de muestras con la combinación de los métodos de procesamiento A y de extracción de ácidos nucleicos B utilizando el par de oligonucleótidos JV12 y JV13. Esta combinación de métodos resultó ser la más eficaz para la detección de norovirus mediante RT-PCR. No obstante, rotavirus no se detectó en el tejido digestivo de las ostras analizadas con ninguna de las combinaciones de métodos, ni con ninguno de los pares de oligonucleótidos utilizados. La elevada presencia de norovirus en las ostras mostró su potencial de servir como vehículo de infecciones por virus entéricos.

Gastroenteritis outbreaks in humans have been often associated with the ingestion of bivalve mollusks. In this study, 15 mixtures of samples of the digestive tissue of oysters (Pinctada imbricate), commercialized at the Avenida Perimetral in Cumana, Sucre State, Venezuela, were analyzed through two processing methods and two nucleic acid extraction methods, with the purpose of detecting norivirus and rotavirus RNA by the Polymerase Chain Reverse Transcription Method. Five pairs of oligonucleotids were also used, to select the best one for virus detection. Norovirus was detected in 13 of the 15 mixtures of samples using a combination of processing method A and nucleic extraction method B, and the JV12 and JV13 oligonucleotid pair. This combination of methods resulted as the most efficient for norovirus detection through PCR-RT. Nevertheless, rotavirus was not detected in the digestive tissue of any of the oysters analyzed with any of the combination of methods, or any of the oligonucleotid pairs used. The high presence of norovirus in the oysters showed their potential for serving as carriers of enteric virus infections.

Ciclo reproductivo del ostión de manglar Crassostrea rhizophorae (Bivalvia: Ostreidae) en la Bahía de Camamu, Bahia, Brasil

Reproductive cycle of the mangrove oyster Crassostrea rhizophorae (Bivalvia: Ostreidae) in Camamu Bay, Bahia, Brasil. The mangrove oyster Crassostrea rhizophorae is important fishery resource along the entire Brasilian coast with excellent potential for marine culture. The purpose of this paper was to examine the reproductive characteristics of the oyster of the Maraú river estuary in Camamu Bay, Bahia, Brasil. The samples were collected monthly, from September 2006 to August 2007, at two points (I and II) in Camamu Bay. At each site 20 oysters were collected for histological analysis, fixed in Davidson’s solution, embedded in paraffin, dehydrated in an ethanol series, sectioned (7μm thick) and stained with Harris hematoxylin and Eosin (HE). Additionally, 30 oysters were sampled, at each point, for a condition index analysis. The water temperature ranged from 23.5°C to 30°C and the salinity from 15 to 25 ups at Point I (Maraú) and from 25 to 35 at Point II (Tanque Island). The oyster’s height ranged from 30 to 92mm at Point I and from 27 to 102mm at Point II, with an average of 49.0mm±9.1 (n=230) and 49.9mm±9.9 (n=237), respectively. Among the sampled oysters at Point I, 59.1% were females, 31.3% males, 1.3% hermaphrodites and 8.2% of the oysters of undetermined sex. At Point II, 66.2% were females, 30.4% males, 0.8% hermaphrodites and 2.5% (n=237) of undetermined sex. The gonadic stage analysis indicated that the reproduction period of the C. rhizophorae in the Maraú Peninsula was continuous all year, without any regressive phase. The condition index (R) ranged from 8.0% to 17.7%. The peak periods of R coincided with the expressive oyster’s percentage in the maturation and liberation gametic stages. The results of these findings will contribute information for the oyster spat collection and to the process installation of the oyster culture in Camamu Bay. Rev. Biol. Trop. 59 (1): 137-149. Epub 2011 March 01.

El ostión de manglar es un importante recurso pesquero que se distribuye por toda la costa brasileña y una de las especies nativas con mayor potencial para maricultura. Este estudio tuvo como objetivo conocer las características reproductivas del ostión de manglar Crassostrea rhizophorae en el estero del Río Maraú, Bahía de Camamu, Bahia, Brasil. Las muestras fueran recolectadas mensualmente, entre agosto de 2006 y septiembre de 2007, en dos sitios. Las metodologías utilizadas fueron el análisis histológico de las gónadas y el cálculo de rendimiento de carne. La temperatura del agua durante el período de estudio varió de 23.5°C a 30°C y la salinidad osciló entre 15 y 35ups. La altura de los ostiones examinados varió de 27 a 102mm (n=437). Hubo predominancia de hembras en las poblaciones de ambos sitios. Los análisis de estadios gonádicos demostraron que el proceso de reproducción de C. rhizophorae en el estero del Río Maraú es continuo durante el año, sin período de reposo sexual. El rendimiento de carne (R) fue de 8.0% a 17.7% (n=669). Los resultados de este estudio proveen informaciones para la captación de semilla e implantación de la ostricultura en la Bahía de Camamu.

Microrganismos indicadores de qualidade higiênico-sanitária em ostras(Crassostrea gigas) e águas salinas de fazendas marinhas localizadas na Baía Sul da Ilha de Santa Catarina, Brasil / Hygienic sanitary indicator microorganisms in oysters (Crassostrea gigas) and saline waters from sea farms in the Baía Sul of the Santa Catarina’s Island, Brazil

Esta investigação teve como objetivo efetuar o monitoramento da qualidade higiênico-sanitária em ostras (Crassostrea gigas) e águas salinas de fazendas marinhas da Baía Sul da Ilha de Santa Catarina, por meio de microrganismos indicadores, durante o período de um ano. As amostras de ostras e águas salinas foram submetidas a ensaios microbiológicos para enumeração de coliformes a 35°C, coliformes a 45°C e Escherichia coli. Foram avaliados os parâmetros físico químicos de temperatura, salinidade, turbidez e pH nas águas e pH nas ostras. Foram observadas correlações positivas entre as contagens de coliformes a 35°C, coliformes a 45°C e E. coli nas amostras de águas salinas e de carne das ostras; e diferenças estatisticas significativas foram verificadas entre as contagens de coliformes a 45°C nas amostras de águas salina provenientes da região E. Os parâmetros físico-químicos apresentaram pouca ou nenhuma correlação estatística com as contagens bacteriológicas. Considerando-se a Baía Sul em toda sua extensão, observou se a influência do índice pluviométrico sobre as contagens bacteriológicas obtidas nas águas e nas ostras. As contagens de coliformes a 45°C observadas nas águas salinas estavam de acordo com os parâmetros descritos na legislação brasileira, contudo, ressalta-se a importância da implantação de saneamento básico em regiões de cultivo de moluscos bivalves, assim como o constante monitoramento bacteriológico de indicadores de qualidade nas águas e nos moluscos.

This investigation was conducted for one year in order to monitor the hygienic sanitary conditions of oysters(Crassostrea gigas) and saline waters from sea farms at the South Bay of Santa Catarina Island, by means ofmicroorganism indicators. Oysters and saline waters samples were assayed by bacteria counting techniquefor coliforms at 35°C, coliforms at 45°C and Escherichia coli. Physical and chemical parameters were assessedin water samples (pH, salinity, turbidity and temperature) and oyster samples (pH). Positive correlations oncoliform counts at 35°C and at 45°C, and E. coli were observed within saline waters and samples of oyster meat.Statistically significant differences on coliforms counting at 45ºC were observed among saline water samplescollected from the region E. Physical-chemical parameters showed somewhat no statistical correlation with microorganism counting. Nonetheless, the influence of rainfall indices in the Great Florianopolis region over the bacteriological counting in saline waters and oysters could be observed. The coliforms counting at 45°C in saline water samples complied with the parameters established by the Brazilian legislation, although the basicsanitation at bivalve shellfish growing sites and a regular bacteriological quality monitoring by using specificindicators are demanded for the culturing water and for growing oysters.

Polidiariose em ostras Crassostrea gigas cultivadas na praia da Ponta do Sambaqui, Florianópolis, Santa Catarina, Brasil / Polydiariosis in cultured oyster Crassostrea gigas from Ponta do Sambaqui, Florianópolis, Brasil

A infestação por poliquetas do gênero Polydora em ostras C. gigas de cultivo foi acompanhada por meio de coletas mensais enquanto os parâmetros físico-químicos da água do mar foram monitorados semanalmente. Foi determinada a área total ocupada pelas bolhas e tubos na superfície interna das valvas, observando os possíveis danos às principais características fisiológicas da ostra. Houve prevalência do anelídeo Polydora sp. (Polychaeta: Spionidae) em todos os meses amostrados, chegando a 100% no período de águas mais frias. As análises histológicas mostraram que os tecidos e órgãos estavam intactos, ocorrendo a formação de gametas. Nas ostras severamente infestadas, foi possível observar intensa fibrose na região do manto, mostrando reparo do tecido.

The infestation of polychaetes Polydora in oyster C. gigas within cultivation was accompanied by monthly samples whereas the physical and chemical parameters of seawater were monitored weekly. The total area occupied by bubbles and tubes in the inner surface of the valve was determined, indicating the possible damage to the main physiological characteristics of oysters. It was observed prevalence of annelid Polydora sp. (Polychaeta: Spionidae) in all samples each month, reaching 100% in the period of cooler waters. The histological analysis showed that the tissues and organs were intact, occurring the formation of gametes. In oysters severely infested, it was possible to observe intense fibrosis in the mantle, showing repair of the tissue.

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.

Treatment of shrimp effluent by sedimentation and oyster filtration using Crassostrea gigas and C. rhizophorae

Efficiency in removing particulate matter from Litopenaeus vannamei shrimp culture effluent was assessed in laboratory scale employing sedimentation and oysters Crassostrea gigas and C. rhizophorae filtration processes. Cylindroconical tanks (100 L) were used in duplicate for sedimentation and 50-L in triplicate for oyster filtration. Fifteen oysters of each species weighing 76-80 g were stocked in each of the filtration treatment experimental units (biomass of 1065 - 1174 g oyster per unit). The control treatment was a tank similar to those used in the filtration treatment but with empty oyster shells. Hydraulic retention time of the effluent was of 6 hours in each treatment. First, effluent went through sedimentation, and then the supernatant went through the filtration tanks. Temperature, pH, dissolved oxygen, salinity, turbidity, total suspended solids, total volatile solids, chlorophyll a and BOD5 were evaluated. During sedimentation and filtration, temperature, pH, salinity and dissolved oxygen concentration remained stable. Sedimentation removed 18, 5.6, 27.5, 45.40 and 23.2 percent of turbidity, total suspended solids, total volatile solids, chlorophyll a and BOD5, respectively. Chlorophyll a and BOD5 after sedimentation presented significant difference (P<0.05) from the farm crude effluent. For the filtration treatment, C. rhizophorae was more efficient removing 62.1, 70.6, 36.1, 100 and 17.2 percent of turbidity, total suspended solids, total volatile solids, chlorophyll a and BOD5, respectively, whereas C. gigas removed 56.3, 41.2, 27.8, 51.4 and 8.0 percent of the same parameters. Statistically comparing C. rhizophorae and C. gigas performances, there were differences (P<0.05) in removing total suspended solids, total volatile solids and chlorophyll a.

Em escala laboratorial, foi comparada a eficiência de remoção de material particulado presente no efluente do cultivo de camarão branco Litopenaeus vannamei, mediante o processo de sedimentação e filtração com ostra nativa Crassostrea rhizophorae e com ostra do pacifico Crassostrea gigas. No processo de sedimentação foram empregados tanques cilindro cônico, em duplicata, de cor preta com 100 L de capacidade total. Para o processo de filtração foram empregados tanques cilindro cônicos, em triplicata, de cor preta de 50 L de volume total. No tratamento de filtração cada unidade experimental foi estocada com 15 indivíduos de ostras de ambas as espécies, com peso médio entre 76 - 80 g, mantendo uma biomassa entre 1.065 e 1.174 g ostra por unidade. Também foi empregado um tanque com as mesmas características ao de filtração, como controle, contendo apenas conchas de moluscos sem animal. O tempo de retenção hidráulica do efluente, em cada tratamento, foi de 6 horas, passando primeiro pelo processo de sedimentação e posteriormente o sobrenadante foi transferido para a filtração. As variáveis avaliadas no estudo foram pH, temperatura, oxigênio dissolvido, salinidade, turbidez, sólidos suspensos totais, sólidos voláteis totais, clorofila a e DBO5. No processo de sedimentação e de filtração, as variáveis temperatura, pH, salinidades e oxigênio dissolvido se mantiveram estáveis. O tratamento de sedimentação conseguiu uma remoção de 18,7 por cento; 5,6 por cento; 27,5 por cento; 45,4 por cento e 23,2 por cento, para a turbidez, sólidos suspensos totais, sólidos voláteis totais, clorofila a e DBO5, respectivamente, sendo que a clorofila a e a DBO5 foram as variáveis que no processo de sedimentação apresentaram diferenças estatísticas (P<0,05) em relação ao efluente bruto da fazenda. No processo de filtração, a ostra C. rizophorae resultou ser mais eficiente na remoção do material particulado do que a ostra C. gigas, com valores de 62,1 por cento; ...

Avaliação quantitativa do risco de doença, causada por Vibrio arahaemolyticus, associado ao consumo de ostras (Crassostrea brasiliana) cruas cultivadas e comercializadas no Estado de São Paulo / Quantitative risk assessment of illness, caused by Vibrio parahaemolyticus, associated with the consumption of raw oysters (Crassostrea brasiliana) farmed and commercialized in the State of São Paulo

Vibrio parahaemolyticus (Vp) é uma bactéria naturalmente presente em regiões estuarinas, sendo a principal causa de gastrenterite de origem bacteriana associada a pescados, principalmente ostras cruas. Nesta pesquisa, foi desenvolvida uma avaliação quantitativa de risco para avaliar a probabilidade de Vp causar doença após o consumo de ostra crua, produzida e comercializada no Estado de São Paulo. O estudo incluiu a identificação e caracterização do perigo, a avaliação da exposição e a caracterização do risco. Um modelo matemático foi desenvolvido. Este modelo leva em consideração o comportamento de Vp em ostras na cadeia produtiva, em cada estação do ano, além da relação entre a dose de Vp ingerida e a probabilidade de desenvolver a doença. A avaliação da exposição foi desenvolvida em três etapas: cultivo, pós-coleta e consumo. Na etapa de cultivo foram considerados os fatores que influenciam a prevalência e o número de Vp em ostras no momento da coleta. Na etapa pós-coleta, foram descritas as práticas da indústria e foram considerados os fatores associados ao processamento, transporte e manipulação. Já na etapa de consumo foram considerados os fatores como a quantidade de ostras consumidas por porção, o peso médio por ostra consumida e a população de Vp patogênico no momento do consumo. O resultado do modelo quantitativo da avaliação da exposição foi, então, integrado ao modelo dose-resposta, Beta-Poisson, para se obter uma estimativa do risco. Esta estimativa expressa o impacto da exposição humana a Vp, sobre a saúde pública, associada ao consumo de ostras. A simulação de Monte Carlo foi utilizada para avaliar o efeito da variabilidade e incerteza das variáveis do modelo sobre a estimativa do risco. O modelo prediz uma probabilidade de ocorrência de doença de 4,6x10-4, por porção de ostra, consumida ao longo do ano. As variáveis que possuem maior influência sobre o risco de ocorrência de doença são a população de Vp em ostras no cultivo, a temperatura de transporte das ostras até o varejo e a porcentagem de Vp patogênico em ostra, no momento do seu consumo. O modelo evidencia que uma das maneiras de reduzir o risco de ocorrência de doença seria intervir nas condições de transporte de ostras até o varejo por meio da sua refrigeração. Com o modelo é possível identificar fatores e simular cenários para avaliar o comportamento de V. parahaemolytícus como um perigo microbiológico, ao longo da cadeia produtiva de ostra até o momento do seu consumo. Também é possível avaliar o impacto de medidas de intervenção na cadeia produtiva. As suposições adotadas limitam a aplicabilidade do modelo. Portanto, é necessário que o modelo seja validado, particularmente com relação ao número de casos de doença causados por Vp, cujos dados de vigilância epidemiológica inexistem no Brasil

Vibrio parahaemolyticus (Vp) is naturally present in estuarine regions and is the main cause of gastroenteritis associated with the consumption of bivalve molluscan shellfish, specially raw oysters. In this research, a quantitative risk assessment was developed to evaluate the probability of Vp causing disease after consumption of raw oyster, produced and commercialized in the state of Sao Paulo. The study included the identification and characterization of the hazard, exposure assessment and risk characterization. A mathematical model was developed. This model takes into account the behavior of Vp in oysters in the productive chain, for each season of the year, besides the relationship between the number of cells of Vp ingested and the probability of developing the disease. The exposure assessment was done in three steps: farming, after harvesting and consumption. At the farming step, the factors that influence the prevalence and the population of Vp at the time of harvesting were considered. At the after harvesting step, the factors associated with transportation, handling and processing were considered. At the consumption step, factors related to the amount of oysters and the average weight per oyster consumed and the density of pathogenic Vp at the time of consumption were considered. Then, the quantitative model of exposure assessment was integrated to the dose-response model, BetaPoisson, in order to obtain a risk estimate. This calculation expresses the impact of the human exposure to Vp associated with the consumption of oysters on public health. The Monte Carlo simulation was used to evaluate the effect of variability and uncertainty of variables of the model in the risk estimation. The model predicts a probability of occurrence of the disease of 4,6x10-4 per serving of oyster consumed during one year. The variables showing the greatest influence on the risk of occurrence of disease are the density of Vp in oyster in the farming step, the temperature during transportation of oysters to the retail market and the percentage of pathogenic Vp strains in oysters,' at the moment of consumption. The model indicates that the use of refrigeration during transportation of oysters to retail could reduce the risk of disease. The model allows the identification of factors and the simulation of scenarios in order to evaluate the behavior of V. parahaemolyticus, as a microbiologícal hazard, in the productive chain of oyster to the consumption. It is also possible to evaluate the impact of intervention measures in the productive chain. The assumptions Iimit the application of the model. Therefore, it is necessary to validate the model, particularly in relation to the number of cases of dísease caused by V. parahaemolyticus of which the data on epidemiologic surveillance do not exist in Brazil

Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR

Shellfish are readily contaminated with viruses present in water containing sewage due to the concentrating effect of filter feeding. Enteroviruses are generally used as a model for the detection of viruses from shellfish due to their public health significance. In the present work, oysters were placed in glass aquaria containing seawater plus unicellular algae. Two experiments were performed: 1) oysters bioaccumulating four different poliovirus type 2 concentrations: 5 x 10(4), 2.5 x 10(4), 5 x 10³ and 5 x 10² PFU/mL during 20h; 2) oyster tissues directly inoculated with 6.0 x 10(5) and 1.0 x 10(5) PFU/mL. After viruses seeding, tissue samples were processed by an adsorption-elution-precipitation method. Positive controls were performed by seeding 6.0 x 10(5) PFU/mL of poliovirus type 2 directly on the final oyster tissue extracts. Oyster extracts were assayed for viruses recovery by plaque assay, RT-PCR and integrated cell culture-PCR methodologies (ICC/PCR). The last one was based on the inoculation of the samples onto VERO cell monolayer followed by RT-PCR analysis of the infected cell fluid. In the first experiment (20h bioaccumulation) until 5 x 10³ PFU were detected after 24 and 48h growth on VERO cells. Direct RT-PCR and ICC/PCR were able to detect 3 and 0.04 PFU of poliovirus, respectively, when bioaccumulation assay was used. When direct tissue virus seeding was performed, the plaque assays showed that polioviruses were recovered in all tested concentrations. Based on these results, it is possible to conclude that viable polioviruses can be detected in oysters after bioaccumulation and these techniques can be directly applied for monitoring virus contamination in environmental samples.

Devido ao hábito alimentar filtrante, os moluscos bivalves são contaminados por vírus presentes em águas contaminadas por esgoto. Os enterovírus são geralmente usados como modelos para a detecção de vírus em moluscos bivalves devido a sua importância em saúde pública. No presente estudo, ostras foram colocadas em aquários de vidro contendo água do mar adicionada de algas unicelulares. Dois tipos de experimentos foram realizados: a) ostras bioacumulando quatro diferentes concentrações de poliovírus: 5 x 10(4), 2,5 x 10(4), 5 x 10³, 5 x 10² PFU/mL durante 20h; b) tecidos de ostras inoculados diretamente com 6,0 x 10(5) e 1,0 x 10(5) PFU/mL. Após a semeadura, os tecidos foram processados por um método de adsorção-eluição-precipitação. Controles positivos foram realizados por inoculação de 6,0 x 10(5) PFU/mL de poliovírus diretamente nos tecidos processados das ostras. Os extratos teciduais foram testados para presença do vírus por ensaios de placa de lise (PFU), RT-PCR e cultura celular integrada ao PCR (ICC/PCR). Este último consistiu na inoculação das amostras sobre monocamadas de células VERO seguida de RT-PCR do fluido celular infeccioso. No primeiro experimento (ensaio de bioacumulação por 20h), foram detectados até 5 x 10³ PFU de poliovírus, após 24 e 48h de replicação nas células. Os ensaios de RT-PCR e ICC/PCR foram capazes de detectar 3 e 0,04 PFU de poliovírus, respectivamente nos ensaios de bioacumulação. Quando os extratos teciduais processados foram semeados, os ensaios de placa de lise demonstraram recuperação de vírus infecciosos em todas as concentrações testadas. Pudemos concluir que partículas viáveis de poliovírus podem ser detectadas em ostras após bioacumulação e que estas técnicas podem ser diretamente aplicadas na detecção de vírus em amostras ambientais.

Incorporação experimental de Vibrio cholerae O1 não toxigênico em ostras (Crassostrea brasiliana), vivas1 / Experimental incorporation of vibrio cholerae 01 non toxigenic in live oysters (Crassostrea brasiliana)

As ostras são moluscos bivalves que, com freqüência, são consumidas cruas. Podem ser objeto de processos de depuração (descontaminações físicas e químicas), pois este hábito de consumo está associado a eventos de doenças transmitidas por alimentos em vários países. No presente trabalho, foram realizados testes para verificar a possibilidade de contaminação de ostras em laboratório, com o objetivo de estabelecer modelo para avaliações de processos de depuração. As ostras foram mantidas vivas em água do mar previamente ozonizada. Foi procedida a contaminação da água com Vibrio cholerae EI Tor 01 não toxigênico. Foram realizadas determinações analíticas da água e das ostras imediatamente após e depois de 2, 4, 12, 18 e 24 horas da contaminação. Após 2 horas, as ostras apresentaram positividade para o V. cholerae usado no experimento, em até a diluição 107, igual ao número inicial de células viáveis imediatamente após a contaminação. Os níveis de presença da cepa testada manteve-se elevada, tanto na água como nas ostras, durante todo o período de observação (até a diluição 109 na água e 108 nas ostras). Os resultados obtidos indicam que este método é útil, pois permite a manutenção da viabilidade das ostras. A avaliação da eficácia e eficiência de processos tecnológicos de depuração (descontaminação), como o uso de ozônio e de radiação ionizante, pode ser conduzi da, mantendo os parâmetros reais, inclusive com a presença da microbiota autóctone destes moluscos. A comparação das diluições positivas no decorrer do experimento, demonstram que a cepa adaptou-se às condições do experimento, apresentando inclusive multiplicação. Foram realizados três testes independentes (triplicata), para observar possíveis variações na incorporação da cepa. (AU)

Oyster are frequently eaten raw and alive. Because of this, are often Object of depuration process (physical an chemica1 decontamination), since its consumption habit has importante epidemiological implications. It was perfomed essas to verify the possibility of live oyster contamination at aboratoria 1 level: oysters was maintened alive in sea water previously ozonized and then contamined with Vibrio cholerae EI Tor, 01, Ogawa, non toxigenic. Quantitative Analytical determination was realized in sea water and oysters, immediately and 2,4,12,18 and 24h after the contamination. After 2h, oysters presented high numbers of V cholerae, namely positivity in 10. 7 dilution, compatib1e with the initial sea water contamination.The leve1s of the strain used in this experimente was constantly high during the period of observation (positivitytill 10 dilution in water and 10 in oysters).The results obtained indicate that this contamination may be useful for laboratorial observation of etficacy and efficiencyof tecnhological depuration (decontamination) process of oysters, sinceallow measurements at real condictions. Number of viables cells obtained at ditferents periods of time shows that the strain was capable of adaptation, including multiplication, in this experimente. It was realized three independents experiments. (AU)