An inhibitory compound produced by Pseudomonas with effectiveness on Vibrio harveyi

Persistence of the antivibrio property of the potential antagonistic probiotics, Pseudomonas MCCB 102 and 103, at di¡erent temperatures, pH and in organic solvents was studied. The antivibrio compound was extracted, puri¢ed and characterized using thin-layer chromatography, high-pressure liquid chromatography, liquid chromatography-mass spectroscopy, UV^ Vis and nuclear magnetic resonance spectroscopy and identi¢ed as N-methyl-1-hydroxyphenazine, a phenazine antibiotic. The toxicity of the compound was tested in Penaeus monodon haemocyte culture and the IC50 valuewas found to be1.4 0.31mg L 1. The compound was found to be bacteriostatic at 0.5mg L 1. Its stability to varying temperature, pH, organic solvents, prolonged shelf-life and vibriostatic nature point to its suitability for prophylatic aquaculture application.

Nitrification in a packed bed bioreactor integrated into amarine recirculating maturation system under different substrate concentrations and flow rates

BACKGROUND: A packed bed bioreactor (PBBR) activated with an indigenous nitrifying bacterial consortia was developed and commercialized for rapid establishment of nitrification in brackish water and marine hatchery systems in the tropics. The present study evaluated nitrification in PBBR integrated into a Penaeus monodon recirculating maturation system under different substrate concentrations and flow rates. RESULTS:Instantnitrificationwasobservedafter integration ofPBBRinto thematuration system.TANandNO2-Nconcentrations were always maintained below0.5 mg L−1 during operation. The TANandNO2-N removalwas significant (P < 0.001) in all the six reactor compartments of the PBBR having the substrates at initial concentrations of 2, 5 and 10 mg L−1. The average volumetric TAN removal rates increased with flow rates from 43.51 (250 L h−1) to 130.44 (2500 L h−1) gTAN m−3 day−1 (P < 0.05). FISH analysis of the biofilms after 70 days of operation gave positive results with probes NSO 190 ((β ammonia oxidizers), NsV 443 (Nitrosospira spp.) NEU (halophilic Nitrosomonas), Ntspa 712 (Phylum Nitrospira) indicating stability of the consortia. CONCLUSION: The PBBR integrated into the P. monodon maturation system exhibited significant nitrification upon operation for 70 days as well as at different substrate concentrations and flow rates. This system can easily be integrated into marine and brackish water aquaculture systems, to establish instantaneous nitrification

Stringed bed suspended bioreactors (SBSBR)for in situ nitrification in penaeid and non-penaeid hatchery systems

For establishing nitrification in prawn (non-penaeid, salinity 10–15 ppt) and shrimp (penaeid, salinity 30–35 ppt) larval production systems, a stringed bed suspended bioreactor (SBSBR) was designed, fabricated, and validated. It was fabricated with 5 mm polystyrene and low density polyethylene beads as the substrata for ammonia and nitrite oxidizing bacterial consortia, respectively, with an overall surface area of 684 cm2. The reactors were activated in a prototype activator and were transported in polythene bags to the site of testing. Performance of the reactors activated with the nitrifying bacterial consortia AMONPCU-1 (ammonia oxidizers for non-penaeid culture) and NIONPCU-1 (nitrite oxidizers for non-penaeid culture) was evaluated in a Macrobrachium rosenbergii larval rearing system and those activated with AMOPCU-1 (ammonia oxidizers for penaeid culture) and NIOPCU-1 (nitrite oxidizers for penaeid culture) in a Penaeus monodon seed production system. Rapid setting up of nitrification could be observed in both the static systems which resulted in a higher relative per cent survival of larvae

Fenneropenaeus indicus is protected from white spot disease by oral administration of inactivated white spot syndrome virus

Fenneropenaeus indicus could be protected from white spot disease (WSD) caused by white spot syndrome virus (WSSV) using a formalin-inactivated viral preparation (IVP) derived from WSSV-infected shrimp tissue. The lowest test quantity of lyophilized IVP coated onto feed at 0.025 g–1 (dry weight) and administered at a rate of 0.035 g feed g–1 body weight d–1 for 7 consecutive days was sufficient to provide protection from WSD for a short period (10 d after cessation of IVP administration). Shrimp that survived challenges on the 5th and 10th days after cessation of IVP administration survived repeated challenges although they were sometimes positive for the presence of WSSV by a polymerase chain reaction (PCR) assay specific for WSSV. These results suggest that F. indicus can be protected from WSD by simple oral administration of IVP

The importance of dietary calcium consumption in two species of semi-terrestrial grapsoid crabs

Calcium (Ca) is essential for crustaceans, due to calcium carbonate (CaCO(3)) deposition in the new exoskeleton to harden it. The purpose of this work was to study short term Ca balance in terms of dietary Ca ingestion in two phylogenetically related crabs (Superfamily Grapsoidea) showing different degrees of terrestrial adaptations: Sesarma rectum Randall, 1840 and Neohelice granulata (Dana, 1851). Dietary Ca ingestion was studied using purified diets with different Ca concentrations (0, 2.2 and 6.66% Ca), together with measurements of Ca excretion and Ca hemolymph levels. The results showed that both crabs had the same response to foods containing different levels of Ca, with both species eating more of the high Ca diet. However, S. rectum consumed more per mg body mass at all Ca concentrations (6 mg.g(-1) for S. rectum against 3 mg.g(-1) for N. granulata). Both species excreted/egested Ca differently: S. rectum excreted Ca proportionally to ingestion, whereas N. granulata maintained constant faecal Ca output at all dietary Ca levels. Moreover, Ca hemolymph levels for crabs fed the different diets were independent of dietary Ca. In conclusion, both S. rectum and N. granulata seem to regulate the consumption of diets containing more Ca, which suggests a fine balance for Ca intake.

Caracterização de microorganismos aquáticos por processamento digital de imagens e redes neurais artificiais

A identificação e o monitoramento de microorganismos aquáticos, como bactérias e microalgas, tem sido uma tarefa árdua e morosa. Técnicas convencionais, com uso de microscópios e corantes, são complexas, exigindo um grande esforço por parte dos técnicos e pesquisadores. Uma das maiores dificuldades nos processos convencionais de identificação via microscopia é o elevado número de diferentes espécies e variantes existentes nos ambientes aquáticos, muitas com semelhança de forma e textura. O presente trabalho tem por objetivo o desenvolvimento de uma metodologia para a caracterização e classificação de microorganismos aquáticos (bactérias e microalgas), bem como a determinação de características cinemáticas, através do estudo da mobilidade de microalgas que possuem estruturas que permitem a natação (flagelos). Para caracterização e reconhecimento de padrões as metodologias empregadas foram: o processamento digital de imagens e redes neurais artificiais (RNA). Para a determinação da mobilidade dos microorganismos foram empregadas técnicas de velocimetria por processamento de imagens de partículas em movimento (Particle Tracking Velocimetry - PTV). O trabalho está dividido em duas partes: 1) caracterização e contagem de microalgas e bactérias aquáticas em amostras e 2) medição da velocidade de movimentação das microalgas em lâminas de microscópio. A primeira parte envolve a aquisição e processamento digital de imagens de microalgas, a partir de um microscópio ótico, sua caracterização e determinação da densidade de cada espécie contida em amostras. Por meio de um microscópio epifluorescente, foi possível, ainda, acompanhar o crescimento de bactérias aquáticas e efetuar a sua medição por operadores morfológicos. A segunda parte constitui-se na medição da velocidade de movimentação de microalgas, cujo parâmetro pode ser utilizado como um indicador para se avaliar o efeito de substâncias tóxicas ou fatores de estresse sobre as microalgas. O trabalho em desenvolvimento contribuirá para o projeto "Produção do Camarão Marinho Penaeus Paulensis no Sul do Brasil: Cultivo em estruturas Alternativas" em andamento na Estação Marinha de Aquacultura - EMA e para pesquisas no Laboratório de Ecologia do Fitoplâncton e de Microorganismos Marinhos do Departamento de Oceanografia da FURG. O trabalho propõe a utilização dos níveis de intensidade da imagem em padrão RGB e oito grandezas geométricas como características para reconhecimento de padrões das microalgas O conjunto proposto de características das microalgas, do ponto de vista de grandezas geométricas e da cor (nível de intensidade da imagem e transformadas Fourier e Radon), levou à geração de indicadores que permitiram o reconhecimento de padrões. As redes neurais artificiais desenvolvidas com topologia de rede multinível totalmente conectada, supervisionada, e com algoritmo de retropropagação, atingiram as metas de erro máximo estipuladas entre os neurônios de saída desejados e os obtidos, permitindo a caracterização das microalgas.

Diel variation in abundance and size of the seabob shrimp Xiphopenaeus kroyeri (Crustacea, Penaeoidea) in the Ubatuba region, Southeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aplicação dos isótopos estáveis em aqüicultura

O produto mais importante do conhecimento é a ignorância. A situação sugere a imagem de alguém que vive numa ilha de conhecimento cercado por um mar de ignorância. Quando a área dessa ilha aumenta pelo progresso da ciência, também aumenta o litoral, - a fronteira com o desconhecido. Esta expansão é o fruto mais valioso do conhecimento (David Gross, Nobel em Física, 2004). O uso dos isótopos estáveis dos bioelementos (CHON'S) presentes na Atmosfera, Hidrosfera, Litosfera e Biosfera enquadra-se neste contexto, surgindo como ferramenta de alto potencial em pesquisas nas diversas áreas das Ciências da Vida. Com organismos aquáticos, a aplicação dos isótopos estáveis do carbono (δ 13C) e do nitrogênio (δ 15N) tem ocorrido em diversas áreas. em larvas de pacu Piaractus mesopotamicus, a técnica evidenciou a escolha voluntária entre organismos vivos e dieta seca pelos animais, desde as primeiras alimentações até o final da larvicultura. Na carcinicultura, o crescimento do Penaeus vannamei resultou do aproveitamento dos alimentos oferecidos e da produtividade primária do lago (biota). Por meio do balanço de massa e isotópico foi possível determinar a contribuição da biota, da ordem de 44 a 86% do carbono assimilado pelos animais durante o crescimento. Outro exemplo é com o caranguejo de mangue Sesarma rectum, em que sua preferência alimentar por Spartina alterniflora (C4) versus Rhizophora mangle (C3) ou Hibiscus tiliaceus (C3) foi evidenciada pelo modelo isotópico de duas fontes alimentares e os isótopos estáveis do carbono (δ13C). A técnica dos isótopos estáveis associada ao conhecimento dos conceitos básicos da variabilidade isotópica natural e ao uso criterioso das razões 13C/12C; ²H/¹H; 18O/16O; 15N/14N e 36S/34S revela-se como um avanço na fronteira dos conhecimentos, nas diversas reentrâncias do litoral da aqüicultura.

Composition and abundance of marine shrimps (Crustacea, Decapoda, Penaeidea) in Ubatuba bay, Ubatuba (SP), Brazil

This work aims to analyse the composition, abundance and distribution of the Penaeidea species which occur in the Ubatuba Bay (23 degrees 26'S and 45 degrees 02'W). The samples were monthly collected from October/1992 to September/1993. Each collect was composed of two parallel radials: the first (radial ''A'') was carried out in the mid region of the bay and the second one (radial ''B'') in the bay mouth. The trawls took one hour in a boat equipped with one otter-trawl (10 mm of mesh). The registered environmental factors were depth, bottom water temperature, granulometric composition and organic content of the sediment. After the trawls, the shrimp were separated from other marine organisms and counted. Eight species of shrimp were obtained: Xiphopenaeus kroyeri; Artemesia longinaris; Penaeus (L.) schmitti; P. (F.) brasiliensis; Trachypenaeus; constrictus; Sicyonia typica; S. dorsalis and Pleoticus muelleri. The most abundant species were X. kroyeri, A. longinaris, P. muelleri and T. constrictus. It was verified a very strong seasonality among the species. The X. kroyeri species occurred in both radials along the months but its abundance decreased from a November to March. Such fact is attributed to the temperature which reached a minimum value of 20 degrees C during this period. The species A. longinaris and P. muelleri were more frequent in the radial B which was caracterized by 14 +/- 1.3 m of depth, low organic content in the sediment (2.97%) and granulometric composition of medium sand. The distribution and abundance of these shrimps in the bay, beyond the hydrological features can be related to biotic factors as food availability, migration, inter and intra specific relations (competition, predation, etc.).

Metabolismo energético do camarão da amazonia, Macrobrachium amazonicum

Pós-graduação em Biologia Geral e Aplicada - IBB

Variação espaço temporal da comunidade zooplanctônica em viveiros de cultivo de camarão branco, Litopenaeus vannamei (Boone, 1931), no município Curuçá, Pará-Brasil

Ao longo das últimas décadas, a carcinicultura vem apresentando um grande crescimento em diversas partes do mundo, com o Brasil seguindo esta tendência mundial (FAO, 2004). Nesta atividade três espécies de camarão têm se destacado como as mais cultivadas, sendo elas Penaeus monodon (Fabricius, 1798), Fenneropenaeus chinensis (Osbeck, 1765) e Litopenaeus vannamei (Boone, 1931), responsáveis por cerca de 80% da produção mundial (FAO, 2004). No Brasil L. vannamei é a espécie mais cultivada, com a produção brasileira correspondendo a 5% da produção mundial (FAO, 2004). L. vannamei é uma espécie marinha originária do Oceano Pacífico, distribuída do México ao Peru. Por ser eurihalino, este camarão pode se adaptar às mais diversas condições de cultivo, desde águas salgadas até de menores salinidades (BRAY et al., 1993; PONCE-PALAFOX et al., 1997), característica que tem aumentado o interesse dos produtores. Embora seja exótica no Brasil, L. vannamei, mostra maior resistência à variação de temperatura e salinidade do que outros camarões peneídeos nativos (BRITO et al., 2000). O alimento do camarão e as estratégias de seu fornecimento têm merecido uma atenção especial do setor, gerando novas técnicas ou seu aperfeiçoamento. A ração nos sistemas de cultivo intensivo e semi-intensivo, por exemplo, é responsável por 50-60% dos custos totais de produção, demonstrando a importância de novas estratégias para minimizar sue uso. O aumento da biomassa do plâncton (alimento natural), e conseqüentemente, da cadeia alimentar, reduz os custos com a alimentação suplementar, influenciando diretamente os custos finais de produção (AVAULT, 2003). Segundo Nunes (1995), o incremento da produtividade natural é tão importante quanto o uso de uma ração nutricionalmente completa e bem balanceada. Logo após a introdução nos viveiros de cultivo, a base da alimentação de L. vannamei é composta, em parte, pelo alimento natural disponível (NUNES et al. 1997; MARTINEZ-CORDOVA et al. 1997; ROTHLISBERG, 1998) complementada com ração comercial. Martinez-Cordova et al. (2002) mostraram que as concentrações de clorofila ‘a’ diminuem cerca de 50% do início ao fim do cultivo, provavelmente devido a pastagem pelo zooplâncton e por alguns invertebrados bentônicos. Além da importância do zooplâncton como alimento para as pós-larvas de camarão nos viveiros de engorda, o uso destes organismos (principalmente copépodes) como alimento vivo na aqüicultura marinha vem recebendo grande atenção nos últimos anos (DELBARE et al. 1996). Tal fato ocorre por serem ricos em fosfolipídios, ácidos graxos essenciais altamente insaturados e antioxidantes naturais, sendo nutricionalmente superiores aos rotíferos e aos náuplios de artemia, comumente usados na larvicultura marinha (SARGENT et al. 1997, STOTTRUP e NOSKER, 1997) promovendo o sucesso as larviculturas de camarão (PAYNE et al. 1998; SCHIPP et al. 1999; PAYNE e RIPPINGALE, 2000). Desta forma, estudos sobre o cultivo intensivo de camarões marinhos que enfoquem a composição da comunidade planctônica, as variáveis bióticas e abióticas no sistema, e a característica dos efluentes gerados, são de grande importância. Assim, os resultados obtidos podem incrementar a produtividade aquática no cultivo, alem de fornecer subsídios para pesquisas posteriores de avaliação e mitigação dos impactos ambientais causados por esta atividade.

Descrição da fauna parasitária de raias e tubarões marinhos (Elasmobranchii), fauna acompanhante da pesca comercial no litoral Sul e Sudeste do Brasil

Pós-graduação em Medicina Veterinária - FCAV

Food Content of Six Commercial Fishes from Mississippi Sound

Specific dietary contents from six fishes collected in Mississippi Sound are recorded. In order of their importance, primary components grouped in major taxonomic categories were fishes, penaeid shrimps, and other crustaceans for Cynoscion nebulosus; Crustaceans and fishes for C. arenarius; fishes and crustaceans for C. nothus; crustaceans, pelecypods, and polychaetes for Pogonias cromis; crustaceans, molluscs, polychaetes, and fishes for Archosargus probatocephalus; and fishes and penaeid shrimps for Paralichthys lethostigma. Principal items in the diets of most of the fishes included Anchoa mitchilli, Penaeus aztecus, P. setiferus, and Callinectes sapidus. Those crustaceans show that competition exists for commercial shellfishes in Mississippi Sound. Ratios among the different dietary items vary, according at least to species of fish, length of fish, season, specific location, and abundance of available prey. Some of these variations are documented and are additionally related to selected findings by other authors sampling different localities. We suggest that examination of food items in Archosargus probatocephalus can serve as a practical means to sample and assess seasonal prevalence and abundance of a wide range of invertebrates throughout different habitats in Mississippi Sound and elsewhere.

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller`s dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 +/- 0.5 degrees C, 32.2 +/- 0.4%.) with juvenile L vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg(-1) CP: 8.03 g kg(-1) lipid; 2.01 kcal g(-1)) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 degrees C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx2), n = 26. Best in vitro APD predictions were obtained at 25 degrees C, 4 U: 80 mg CP both for ingredients (R(2) = 0.86: P = 0.001) and test diets (R(2) = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD - determined APD) for corn gluten meal, feather meal. poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD>80% showed higher prediction precision (R(2) = 0.98: P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R(2) = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information. (C) 2009 Elsevier B.V. All rights reserved.

Digestive peptidases and proteinases in the midgut gland of the pink shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae)

Proteases from the midgut gland of the Farfantepenaeus paulensis juveniles were assessed. Enzyme activity was determined using protease substrates and inhibitors. The effect of pH, temperature and calcium on proteolytic activity was assayed. Caseinolytic activity was analysed in substrate-sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Trypsin, chymotrypsin and leucine aminopeptidase activity was detected. Proteolytic activity was strongly inhibited by the specific trypsin inhibitors. Tosyl-phenylalanine chloromethyl ketone inhibited 59.3% of chymotrypsin activity. The greatest trypsin-like activity occurred at pH 8.0 and 45 degrees C. Chymotrypsin-like activity reached maximal values at alkaline pH (7.2-9.0) and 55 degrees C. CaCl(2) did not increase trypsin-like activity, but rather inhibited it at concentrations of 30 (20%), 50 (30%) and 100 mM (50%). The substrate-SDS-PAGE zymogram revealed eight proteinase bands. Two possibly thermal-resistant (85 degrees C, 30 min) chymotrypsin isoforms were found, which were inhibited by phenyl-methyl-sulphonyl-fluoride. Aminopeptidase activity of enzyme extracts (Arg, Leu, Lys, Phe and Val) and the recommended concentrations of these essential amino acids in penaeid shrimp diets were positively correlated (P < 0.05). Beause protein digestion involves the combined action of different enzymes, adequate knowledge of shrimp digestion and enzyme characteristics is required for the assessment of the digestive potential of different feed sources and development of in vitro digestibility protocols.