Nannochloropsis spp. as Feed Additive for the Pacific White Shrimp: Effect on Midgut Microbiology, Thermal Shock Resistance and Immunology.

This work aimed to evaluate Nannochloropsis spp. as feed additive in the diet of Pacific white shrimp for their effect on midgut microbiology, thermal shock resistance and immunological parameters. Initially, the digestibility of the microalgae meal was assessed, and the apparent digestibility coefficient (ADC) was determined. The ADC was, in general, high in lipids (78.88%) and eicosapentaenoic fatty acid (73.86%). Then, Nannochloropsis spp. were included in diets at four levels (0, 0.5, 1 and 2% inclusion). The shrimp were reared in 500 L clear water tanks containing 20 shrimp per tank with an initial weight of 6.05 ± 0.06 g and fed four times a day. Shrimp fed with supplemented diets containing Nannochloropsis spp. (0.5 and 2%) presented higher resistance to thermal shock when compared to the non-supplemented group (control). Shrimp fed with 1 and 2% of algae inclusion had a higher production of reactive oxygen species (ROS) when compared to other treatments. No statistical difference was observed in the immunological parameters and microbiology of the intestinal tract. Thus, the inclusion of Nannochloropsis spp. in shrimp diets at 0.5 and 2% levels increases resistance to thermal shock and ROS production in shrimp.

Work Ergonomic Analysis: Application of a Postural Study on the Oysters Cultivation / Análise Ergonômica do Trabalho: Aplicação de um Estudo Postural no Cultivo de Ostras

The cultivation of marine mollusks is an activity of great importance in Brazil, providing employment and income for artisanal fishermen and fishing communities, contributing to local development. However, this type of cultivation is still carried out in a very handmade way with intense use of labor. The objective of this study was to describe the postures and movements of fish farmers during the oyster classification activity. Ten employees of a marine farm participated in the municipality of Florianópolis, SC. For the analysis of the posture and movements, the Rapid Entire Body Assessment (REBA) method and NIOSH method were used along with in situ observation and interviews with the workers. The task of oysters classification presents an average risk of injury or occupational diseases, especially in the cervical region and upper limbs, as it is developed with marked flexion of the head and shoulders, which requires a lot of the muscles involved to maintain the posture and perform repetitive movements. Many workers have muscle pains in the shoulders, lower back, hands, wrists and fingers. Simple and economically viable adaptations should be adopted, because through ergonomic evaluation it is possible to carry out interventions that are within the reach of the workers. It is necessary to transmit knowledge so that the workers are aware of the postures that they must adopt in the accomplishment of this aquaculture activity and thus to promote a better quality of life of this population. (AU).

O cultivo de moluscos marinhos é uma atividade de grande importância no Brasil proporcionando a geração de emprego e renda para pescadores artesanais e comunidades pesqueiras, contribuindo para o desenvolvimento local. Entretanto, este tipo de cultivo ainda é realizado de forma bastante artesanal com intensa utilização de mão de obra. Diante disso, este estudo teve por objetivo realizar uma descrição das posturas e movimentos de maricultores durante a realização da atividade de classificação de ostras. Participaram voluntariamente dez funcionários de uma fazenda marinha no município de Florianópolis, SC. Para análise da postura e movimentos, utilizou-se o método Rapid Entire Body Assessment (REBA) e método NIOSH juntamente com a observação in loco e entrevistas com os maricultores. A tarefa de classificação de ostras apresenta médio risco de lesão ou doenças ocupacionais, principalmente na região cervical e membros superiores, pois é desenvolvida com acentuada flexão de cabeça e ombros, o que exige muito da musculatura envolvida para manter a postura e realizar movimentos repetitivos. Muitos trabalhadores apresentam dores musculares nos ombros, lombar, mãos, punhos e dedos. Adaptações simples e economicamente viáveis devem ser adotadas, pois através da avaliação ergonômica é possível realizar intervenções que estão ao alcance dos trabalhadores. Necessitase transmitir os conhecimentos para que os trabalhadores tenham consciência das posturas que devem adotar na realização desta atividade aquícola e assim promover uma melhor qualidade de vida desta população. (AU).

RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei.

Disease in Pacific white shrimp Litopenaeus vannamei caused by the infectious myonecrosis virus (IMNV) causes significant socioeconomic impacts in infection-prone shrimp aquaculture regions. The use of synthetic dsRNA to activate an RNA interference (RNAi) response is being explored as a means of disease prophylaxis in farmed shrimp. Here, survival was tracked in L. vannamei injected with long synthetic dsRNAs targeted to IMNV open reading frame (ORF) 1a, ORF1b, and ORF2 genome regions prior to injection challenge with IMNV, and real-time RT-PCR was used to track the progress of IMNV infection and mRNA expression levels of the host genes sid1, dicer2, and argonaute2. Injection of dsRNAs targeting the ORF1a and ORF1b genes but not the ORF2 gene strongly inhibited IMNV replication over a 3 wk period following IMNV challenge, and resulted in 90 and 83% shrimp survival, respectively. Host gene mRNA expression data indicated that the Sid1 protein, which forms a transmembrane channel involved in cellular import/export of dsRNA, increased in abundance most significantly in shrimp groups that were most highly protected by virus-specific dsRNA injection. Subclinical IMNV infections present in the experimental L. vannamei used increased markedly in the 2 d between injection of any of the 4 virus-specific or non-specific dsRNAs tested and IMNV challenge. While handling and injection stress are implicated in increasing IMNV replication levels, the underlying molecular factors that may have been involved remain to be elucidated.

Molecular signatures at imminent death: hemocyte gene expression profiling of shrimp succumbing to viral and fungal infections.

Infectious diseases represent the most serious threat to shrimp farming worldwide. Understanding the molecular mechanisms driving shrimp-pathogen interactions is necessary for developing strategies to control disease outbreaks in shrimp production systems. In the current study, we experimentally reproduced mortality events using standardized infections to characterize the hemocyte transcriptome response of the shrimp Litopenaeus vannamei succumbing to infectious diseases. By using a high-throughput microfluidic RT-qPCR approach, we identified molecular signatures in shrimp during lethal infections caused by the White Spot Syndrome Virus (WSSV) or the filamentous fungus Fusarium solani. We successfully identified gene expression signatures shared by both infections but also pathogen-specific gene responses. Interestingly, whereas lethal WSSV infection induced the expression of antiviral-related genes, the transcript abundance of many antimicrobial effectors was reduced by lethal F. solani infection. To our knowledge, this is the first report of the immune-gene repertoire of infected shrimp at the brink of death.

First report of the protozoan parasite Perkinsus marinus in South America, infecting mangrove oysters Crassostrea rhizophorae from the Paraíba River (NE, Brazil).

The present work aimed to study the infection by Perkinsus sp. in the mangrove oysters Crassostrea rhizophorae from the estuary of the Paraíba River (Paraíba State, Brazil). Perkinsosis was detected by incubation of oyster gill pieces in Ray's fluid thioglycollate medium. The monthly prevalence values were all above 70%, thus infection was not likely to be a transient event. Perkinsus sp. parasites isolated from eight oysters were propagated in vitro. PCR-RFLP analysis of in vitro cultured cells as well as the sequences of the rDNA ITS region allowed the identification of the in vitro propagated parasites as Perkinsus marinus. Phylogenetic analyses using rDNA ITS region sequences strongly supported the Perkinsus sp. from Paraíba in a monophyletic group with P. marinus. Thus, the results confirmed the species affiliation of Paraíba Perkinsus sp. as P. marinus. This is the first report of P. marinus in Brazil and South America and the first report of P. marinus naturally infecting C. rhizophorae.

Parasitological survey of mangrove oyster, Crassostrea rhizophorae, in the Pacoti River Estuary, Ceará State, Brazil.

The mangrove oyster, Crassostrea rhizophorae (Bivalvia, Ostreidae) is commonly collected by fisherwomen in the estuaries of the Ceará State (CE), Northeastern Brazil. Despite the socioeconomic importance of this natural resource, there are few studies on the health of the oysters in this region. This study aimed to survey pathological changes in the mangrove oyster C. rhizophorae in the estuary of the Pacoti River, CE. Adult oysters were collected in August 2008 (N=450) and December 2009 (N=450) at three sites of the Pacoti estuary and in 2010 (N=600) samplings were done quarterly at one site which has showed the higher prevalence de Perkinsus. Macroscopical and histological analyses were used to evaluate pathological changes, Ray's Fluid Thioglycollate Medium (RFTM) to detect Perkinsus spp. and polymerase chain reactions (PCR) and DNA sequencing to identify Perkinsus species. In 2009, RFTM assay detected Perkinsus sp. infecting the tissues of C. rhizophorae with low prevalences of 1.3%, 6.7% e 7.3% in sites 1, 2 and 3, respectively, and in 2010, in site 3, prevalence was 2% (12 of 600 oysters). PCR did not confirm any positive case in 2009 and only 5 in 2010. The phylogenetic analyses strongly indicate that the Perkinsus species infecting oysters C. rhizophorae of this study belongs to Perkinsus beihaiensis. The histology confirmed 11 cases of Perkinsus sp. infecting the C. rhizophorae in 2009, and only two cases in 2010. Nematopsis sp. was the protozoan observed with greater prevalence (up 96.7%). Other found protozoa were: Trichodina, Sphenophrya, Ancistrocoma - like and an unknown ovarian parasite. The metazoa found were the polychaete Polydora with high prevalences, a turbellarian, possibly of the genus Urastoma, an unidentified digenean metacercariae and larvae of cestode Tylocephalum. A continuous monitoring of diseases in bivalves from this natural population is recommended, since the phylogenetic analyses indicate the occurrence of P. beihaiensis infecting oysters C. rhizophorae whose pathogenic potential is unknown.