Antibacterial and cytotoxicity activity in macroalgae extracts: perspectives for the use against pathogenic bacteria from shrimp farms (Litopenaeus vannamei) / Atividade antibacteriana e citotóxica em extratos de macroalgas: perspectivas de uso contra bactérias patogênicas de ambiente de cultivo de camarões (Litopenaeus vannamei)

Some infections caused by pathogenic microorganisms might shows high prevalence in farmed shrimp environments, compromising production and causing economic losses. Therefore, the search for compounds with antibiotic activity has become intensive, following the record of new antimicrobial-resistant bacteria. The study of those bioactive compounds in marine macroalgae has produced satisfactory results, such as the discovery of antibacterial activity against multiresistant strains. Accordingly, this study aims to research antibiotic activity in macroalgae extracts of Chlorophyta, Phaeophyta and Rhodophyta found in the coast of Ceará and also to evaluate the cytotoxicity activity against bacterial strains (Vibrio sp.) from shrimp farms (Litopenaeus vannamei). The extracts cytotoxicity was also evaluated. The results prove that there was antibacterial activity in ethanolic, acetonic, hexanic and methanolic extracts against bacterial strains of Vibrio with multiple resistance profile as well as displaying low cytotoxicity.

Algumas infecções causadas por micro-organismos patogênicos podem apresentar alta prevalência em ambientes de cultivo de camarões marinhos, comprometendo a produção e causando prejuízos econômicos aos aquicultores. Assim, tem-se tornado intensa a busca por compostos com atividade antibiótica pelo registro cada vez mais frequente de bactérias com perfil de resistência a antimicrobianos. A presença desses compostos com bioatividade em macroalgas marinhas tem revelado resultados satisfatórios, como a descoberta de ação antibacteriana contra cepas multirresistentes. Desta forma, decidiu-se pesquisar as propriedades antibióticas dos extratos de macroalgas das classes Chlorophyta, Phaeophyta e Rhodophyta, coletadas no litoral cearense, bem como avaliar a citotoxicidade destes extratos, frente a cepas bacterianas (Vibrio sp.) isoladas e provenientes de ambientes de cultivo de camarões marinhos (Litopenaeus vannamei). Os resultados comprovaram que houve atividade antibacteriana dos extratos etanólicos, acetônicos, hexânicos e metanólicos contra cepas bacterianas de Vibrio, além de apontar que os extratos de todas as espécies apresentaram baixa citotoxicidade.

Expression studies on NA+/K+-ATPase in gills of Penaeus monodon (Fabricius) acclimated to different salinities.

The decapod crustacean Penaeus monodon survives large fluctuations in salinity through osmoregulation in which Na+/K+-ATPase (NKA) activity in the gills plays a central role. Adult P. monodon specimens were gradually acclimatized to 5, 25 and 35‰ salinities and maintained for 20 days to observe long term alterations in NKA expression. Specific NKA activity assayed in gill tissues was found to be 3 folds higher at 5‰ compared to 25‰ (isosmotic salinity) and 0.48 folds lower at 35‰. The enzyme was immunolocalized in gills using mouse α-5 monoclonal antibody that cross reacts with P. monodon NKA α-subunit. At 5‰ the immunopositive cells were distributed on lamellar tips and basal lamellar epithelium of the secondary gill filaments and their number was visibly higher. At both 25‰ and 35‰ NKA positive cells were observed in the inter-lamellar region but the expression was more pronounced at 25‰. Gill architecture was normal at all salinities. However, the 1.5 fold increase in NKA α-subunit mRNA at 5‰ measured by quantitative RT-PCR (qRT-PCR) using EF1α as reference gene was not statistically significant. The study confirms the osmoregulating ability of P. monodon like other crustaceans at lower salinities. It is likely that significant increase in NKA transcript level happens at an earlier time point. At higher salinities all three methods record only marginal or no change from isosmotic controls confirming the hypothesis that the animal largely osmoconforms in hyperosmotic environment.

Fatty acids in an estuarine mangrove ecosystem

Fatty acids have been successfully used to trace the transfer of organic matter in coastal and estuarine food webs. To delineate these web connections, fatty acid profiles were analyzed in species of microbes (Azotobacter vinelandii, and Lactobacillus xylosus), prawns (Metapenaeus monoceros and Macrobrachium rosenbergii) and finfish (Mugil cephalus), that are associated with decomposing leaves of two mangrove species, Rhizophora apiculata and Avicennia marina. The fatty acids, except long chain fatty acids, exhibit changes during decomposition of mangrove leaves with a reduction of saturated fatty acids and an increase of monounsaturated fatty acids. The branched fatty acids are absent in undecomposed mangrove leaves, but present significantly in the decomposed leaves and in prawns and finfish, representing an important source for them. This revealed that the microbes are dominant producers that contribute significantly to the fishes and prawns in the mangrove ecosystem. This work has proved the fatty acid biomarkers as an effective tool for identifying the trophic interactions among dominant producers and consumers in this mangrove. Rev. Biol. Trop. 58 (2): 577-587. Epub 2010 June 02.

Los ácidos grasos se han utilizado con éxito para estudiar la transferencia de materia orgánica en las redes alimentarias costeras y estuarinas. Para delinear las interacciones tróficas en las redes, se analizaron perfiles de ácidos grasos en las especies de microbios (Azotobacter vinelandii y Lactobacillus xylosus), camarones (Metapenaeus monoceros y Macrobrachium rosenbergii) y peces (Mugil cephalus), que están asociadas con la descomposición de las hojas de dos especies de mangle, Rhizophora apiculata y Avicennia marina. Los ácidos grasos, con excepción de los de cadena larga, exhiben cambios durante la descomposición de las hojas de mangle, con una reducción de los ácidos grasos saturados y un aumento de los monoinsaturados. Los ácidos grasos ramificados están ausentes en las hojas de mangle sin descomponer, pero presentes de manera significativa en las hojas descompuestas, en camarones y peces, representando una fuente importante para ellos. Esto revela que los microbios son productores dominantes que contribuyen significativamente con los peces y camarones en el ecosistema de manglar. Este trabajo demuestra que los marcadores biológicos de los ácidos grasos son una herramienta eficaz para la identificación de las interacciones tróficas entre los productores dominantes y consumidores en este manglar.

Propriedades físico-químicas, sensoriais e bacteriológicas de camarões (Litopenaeus brasiliensis) irradiados e armazenados sob refrigeração / Physical, chemical, sensorial and bacterial properties of irradiated shrimp (Litopenaeus brasiliensis) stored under refrigeration

A irradiação de alimentos é uma opção tecnológica para minimizar problemas de perdas de alimentos, pois reduz asalterações provenientes da atividade bacteriana, estendendo a validade do produto. Neste estudo foram irradiados camarões(Litopenaeus brasiliensis) com doses de 1,5 e 2,5 kGy, após 30 dias de armazenamento sob refrigeração (2°C ± 2°C), como objetivo de avaliar os efeitos da irradiação sobre a validade comercial de camarões através de parâmetros físico-químicos,microbiológicos e sensoriais. Foram avaliados pH, bases voláteis totais (BVT), contagem de bactérias heterotróficas aeróbiasmesófilas (BHAM) e heterotróficas aeróbias psicrófilas (BHAP) e teste de aceitação sensorial. Os valores de pH não variaramentre as amostras controle e as amostras irradiadas apresentando média de 7,08 ± 0,21 durante os 30 dias de armazenamento.A produção de BVT nas amostras controle extrapolou o limite aceitável para o consumo no 15o dia de armazenamentoenquanto nas amostras irradiadas com 2,5 kGy, este limite não foi alcançado em 28 dias de armazenamento, demonstrandoo efeito positivo da utilização da irradiação. Na análise sensorial, não ocorreu diferença significativa (p> 0,01) na aceitação,quanto aos atributos de sabor, aparência, aroma e impressão global entre as amostras.

Food irradiation is currently regarded as a technological option to mitigate food losses due to its potential in reducing microbiological load and its associated spoilage effects, consequently extending the shelf life of many products. Samples of shrimp (Litopenaeus brasiliensis) have been exposed to gamma doses of 0, 1.5 and 2.5 kGy and monitored changes in sensorial characteristics (30 judges), pH and total volatile bases (TVB) levels. All samples were kept at 2°C ± 2°C through the experiments. No significant change in pH has been observed during the experiments regardless of the gamma dose applied. There was statistical difference between the non-irradiated samples and those irradiated with 2.5 kGy (p 0.01), though no significant difference has been found between the non-irradiated samples and those irradiated with 1.5 kGy nor between the samples irradiated with 1.5 kGy and those irradiated with 2.5 kGy. Based on the upper commercial limit for NB, non-irradiated samples achieved the limit in 15 days while the samples irradiated with 2.5 kGy didnt achieve the limit in 28 days. In sensorial analyses, there was no significant difference (p> 0.01) in the acceptance, according to the attributes flavor, appearance, aroma and global impression among the samples.

Fate of copper in intensive shrimp farms: bioaccumulation and deposition in pond sediments / Dinâmica do cobre na carcinicultura intensiva: bioacumulação e deposição em sedimentos de tanques de criação

We present the distribution of Cu in water, sediments and biomass from intensive shrimp (Litopenaeus vannamei Boone, 1931)) farming in northeastern Brazil. The results show no difference in dissolved Cu concentrations between waters entering and leaving the ponds. However, there was a large export of particulate Cu to adjacent environments, showed by extremely high particulate (112 µg.L-1) Cu concentrations in draining waters. Copper concentrations in the bottom sediments of the pond varied with depth from 10 to 20 µg.g-1, being 5 to 7 times higher than the local background. Pond management procedures result in a peculiar vertical distribution of Cu with peak values occurring at sub-surface depths in pond bottom sediments. Bioavailable Cu reaches about 20 percent of the total Cu content in sediments. Notwithstanding the relatively high Cu bioavailability, concentrations in shrimp muscle (23.2 to 63.4 µg.g-1 d.w.) were similar to reported values for reared and natural L. vannamei populations, and much lower than acceptable maximum limits for human consumption. Concentrations in the exoskeleton were always higher than in muscle. Increasing Cu mass, and to a lesser extent Cu concentrations in muscle tissues, occurred simultaneously to a decrease in Cu mass and concentrations in the exoskeleton and vice versa, suggesting a dynamical exchange of Cu between the two compartments. Although Cu fate in intensive shrimp culture does not result in increasing Cu exposure to consumers, the activity is an important source of this metal to adjacent ecosystems, in particular in the pristine environments where most aquaculture activities in NE Brazil takes place.

Este trabalho apresenta a distribuição de Cu em água, sedimentos e biomassa em uma fazenda de cultivo intensivo do camarão Litopenaeus vannamei Boone, 1931, no Nordeste do Brasil. Os resultados mostraram não haver diferenças entre as concentrações de Cu nas águas afluentes e efluentes da fazenda. Entretanto, foi verificada uma grande exportação de Cu para áreas adjacentes associada ao material em suspensão. Nas águas efluentes, a concentração de Cu particulado atingiu até 112 µg.L-1. Em sedimentos, as concentrações de Cu variaram com a profundidade da coluna sedimentar de 10 a 20 µg.g-1, sendo estes valores de 5 a 7 vezes mais altos que as concentrações naturais dos solos e sedimentos locais. O manejo dos tanques de criação resulta em perfis de distribuição de Cu em sedimentos com maiores concentrações em subsuperfície. A biodisponibilidade do Cu nestes sedimentos é elevada e atinge cerca de 20 por cento da concentração total. Apesar da biodisponibilidade elevada, as concentrações de Cu em músculo do L. vannamei (23.2 a 63.4 µg.g-1 peso seco) são similares aos valores reportados para populações naturais da espécie e em diversos de cultivo em outros países, e estão bem abaixo dos limites máximos permitidos para consumo humano. As concentrações de Cu no exoesqueleto foram maiores que no músculo. Um aumento da massa de Cu em músculo ocorreu simultaneamente à diminuição do Cu no exoesqueleto e vice-versa sugerindo uma troca dinâmica do Cu entre os dois compartimentos. Embora a presença do Cu no cultivo intensivo do L. vannamei não represente exposição significativa para os consumidores, a atividade é fonte importante de Cu para os sistemas aquáticos adjacentes.

Influence of Vibrio parahaemolyticus MTCC 451 on the levels of ascorbic acid and histamine in Penaeus monodon (Fabricius).

Juvenile shrimps (Penaeus monodon) weighing about 15-20 gms were procured from the grow-out ponds and reared under laboratory conditions. They were challenged with Vibrio parahaemolyticus MTCC 451 and their LD50 value was evaluated. The ascorbic acid and the histamine activity of both the control and experimental groups were assessed. The results showed a marked decline in both the parameters at all intervals. This reveals the pathogenesis of the inoculated bacterial strain, which affects the defense mechanism by reducing the glutathione content by the decreased level of ascorbic acid and letharginess by the decreased histamine content of the experimental prawns.