Assessment of total mercury content in tissues of Mullus argentinae in Rio de Janeiro, Brazil / Avaliação de mercúrio total em tecido comestível de Mullus argetinae coletados no Rio de Janeiro, Brasil

This study has as objective to determine total mercury (Total Hg) levels by atomic absorption spectrophotometry in 134 individuals edible part of Mullus argentinae, in two different fishing areas and two seasons in Rio de Janeiro State. Also, proximate composition was performed. Total Hg results in wet weight basis ranged from 0.0867 to 0.7476 µg.g-1 in muscle; 0.0023 to 0,1034 µg.g-1 in flippers; and 0.0177 to 0.1849 µg.g-1 in skin. Mean evaluated moisture was 73.39%; protein was 18.76%; lipid concentration of 5.36%; carbohydrates of 2.35%; and ashes were 0.85%.Results showed that Total Hg contents was lower than accepted limits established by regulatory organization. Higher averages were observed in muscle (0.2441 µg.g-1) when compared with skin (0.2386 µg.g-1) and flippers (0.0195 µg.g-1). In general, samples collected on summer showed higher values of total Hg when comparing to winter. Regarding beach areas there was no significant difference (p>0.05). We can conclude that this specie should be cautious consumed because of total Hg bioaccumulation characteristics, although neither levels were above limits established.

O objetivo deste estudo foi determinar o teor de mercúrio no tecido comestível de Mullus argentinae, conhecido como peixe trilha, espécie amplamente consumida no Rio de Janeiro, Brasil. Foi determinado o teor de mercúrio total (Hg total) por espectrofotometria de absorção atômica em 134 amostras, coletados em duas áreas e estações climáticas diferentes. Além disso, foi avaliada a composição centesimal das amostras. Os resultados de Hg total em peso úmido variaram de 0,0867 a 0,7476 µg.g-1 no músculo; 0,0023 a 0,1034 µg.g-1 nas nadadeiras; e 0,0177 a 0,1849 µg.g-1 na pele. Os valores médios da composição centesimal foram de 73,30% de umidade, 18,76% de proteína, 5,36% de lipídios, 2,35% de carboidratos e 0,85% de matéria mineral. Os resultados das 134 amostras analisadas demostraram que os teores de Hg Total apresentam concentração inferior aos limites aceitos pelos órgãos reguladores. As maiores médias foram observadas no músculo (0,2441 µg.g-1) quando comparadas à pele (0,2386 µg.g-1) e nadadeiras (0,0195 µg.g-1). Em geral, as amostras coletadas no verão apresentaram maiores valores de Hg total em relação ao inverno. Em relação aos locais de coleta não houve diferença significativa (p> 0,05). Podemos concluir que esta espécie deve ser consumida com cautela devido às características de bioacumulação do Hg total, apesar das médias apresentadas estarem abaixo dos limites estabelecidos pela legislação.

Uso de perifiton en un sistema de policultivo en agro acuicultura integrada en la comunidad indígena de Jimaín, Sierra Nevada de Santa Marta (Colombia) / Periphyton-based polyculture in an integrated agri-aquaculture system at the indigenous community of Jimaín, Sierra Nevada de Santa Marta (Colombia)

RESUMEN Los Sistemas de Agro Acuicultura Integrada (SAAI) han sido estudiados como opción para la producción de pescado en comunidades con recursos limitados. Así mismo, el uso de perifiton se ha convertido recientemente en una alternativa viable por el aporte de alimento natural de bajo costo. El objetivo de este estudio fue evaluar el efecto del perifiton sobre el desempeño productivo del policultivo Piaractus sp. (cachama híbrida) y Prochilodus magdalenae (bocachico) en SAAI en la comunidad Indígena de Jimaín, Colombia. Se sembraron 1,5 alevinos de cachama híbrida y 1,5 bocachicos/m2, con peso promedio de 0,49 ± 0,14 g y 1,83 ±1,61 g respectivamente, en seis estaques en tierra de 90 m2 tres de los cuales contaron con varas de madera (3 varas/m2) para la fijación de perifiton. La duración del cultivo fue de 135 días. Los tratamientos (CS: con sustrato y SS: sin sustratos) recibieron una dieta suplementaria dos veces al día, calculada como porcentaje de la biomasa con ajuste quincenal. Se realizaron biometrías quincenales. Los datos productivos por especie y para el policultivo se evaluaron mediante ANOVA con un modelo lineal general (p < 0,05). Los datos de peso (g) y longitud estándar (cm) por especie se evaluaron mediante un modelo mixto de medidas repetidas. No se encontraron efectos del sustrato sobre los parámetros productivos analizados tanto para cada especie, como para el policultivo. Se presentó interacción significativa para el peso del bocachico el día 90 (CS: 74,7 y SS: 47,1). Según los resultados obtenidos, la utilización de perifiton y el manejo en policultivo de especies nativas requiere más estudios.

ABSTRACT Integrated agri-aquaculture systems (IAAS) have been studied as an option for fish production in communities with limited resources. Likewise, the use of periphyton has recently become a viable alternative for its contribution of low-cost natural food. The effect of periphyton use on the productive yield of a hybrid cachama and bocachico polyculture in IAAS, was studied in the Jimaín Indigenous community, Colombia. For this experiment, 1.5 hybrid cachama and 1.5 bocachico fingerlings/m2 were stocked with average weights of 0.49 ± 0.14 g and 1.83 ± 1.61 g, respectively, in six 90 m2 land ponds; three of these ponds had wooden dowels (three dowels/m2) for periphyton fixation. The growth evaluation lasted 135 days. The treatments (WS: with substrate and WOS: without substrates) received a supplementary diet, adjusted as a percentage of the biomass with biweekly adjustment, twice a day. Biometrics were performed every two weeks. The productive data by species and for the polyculture were evaluated using an ANOVA with a general linear model (p <0.05). Weight (g) and standard length (cm) data by species were evaluated using a mixed model of repeated measures. No substrate effects were found on the productive parameters analyzed for each species as well as for the polyculture. A significant interaction was observed for bocachico weight on day 90 (WS: 74.7 and WOS: 47.1). According to the obtained results, optimizing the use of periphyton and the management of native species polyculture requires further studies.

Elaboração de salsicha de peixe pargo Pagrus pagrus de tamanho reduzido proveniente da pesca de arrasto com baixo valor comercial / Development of fish sausage using low commercial value red porgy Pagrus pagrus

O objetivo deste trabalho foi desenvolver salsicha utilizando o pargo Pagrus pagrus de baixo valor comercial capturado na modalidade de pesca de arrasto e classificado como na categoria "mistura" por ter tamanho reduzido para o mercado varejista e realizar análises físico-químicas, microbiológicas, toxicológicas e sensoriais. A salsicha foi elaborada na planta piloto da Embrapa Agroindústria de Alimentos utilizando 50% de surimi e 50% de filé. As análises físico-químicas, microbiológicas e toxicológicas foram realizadas com métodos oficiais. O teste de aceitação foi realizado com 87 provadores que avaliaram o produto utilizando escala hedônica de sete pontos e também arguidos quanto à intenção de compra do produto. O produto foi considerado aceito quando 70% dos provadores atribuíram nota ≥ 4. Os resultados da composição centesimal foram: umidade 71,22%, proteínas 15,34%, lipídios totais 5,55%, carboidratos 6,11%, cinzas 1,78% e o valor energético 135,75 kcal. As avaliações microbiológicas e toxicológicas mostraram que os produtos apresentaram qualidade satisfatória conforme a legislação. A salsicha foi aceita por 82,7% dos provadores e o aspecto global do produto atingiu média de 4,93 (±1,69). Em relação à intenção de compra foi atribuída a maior porcentagem para talvez sim/talvez não (32%), seguido de provavelmente compraria (24%), e 28,1% informaram que provavelmente ou decididamente não comprariam. Com base nos resultados obtidos, conclui-se que o peixe de baixo valor agregado da categoria "mistura" resultou em produto embutido viável para o uso na tecnologia do pescado.

The aim of this work was to develop sausage using the low commercial value red porgy (Pagrus pagrus) caught in the "fish mix" category in the trawl fishery and to perform physicochemical, microbiological, toxicological and sensorial analysis. The sausage was prepared using 50% of surimi and 50% of fillet in Embrapa Food Agroindustry Pilot Plant. The physicochemical, microbiological, toxicological analyses were performed with official methods. The sensorial analyses were performed with 87 testers who evaluated the product using a hedonic scale of seven (7) points and were also accused of buying the product. The product was considered accepted when 70% of the tasters assigned a score ≥ 4. One sausage formulation was tested using 50% surimi. The results of the centesimal composition were: moisture 71.22%, proteins 15.34%, total lipids 5.55%, carbohydrates 6.11%, ash 1.78% and energy value 135.75 kcal. The parameters of microbiological and toxicological are within the current legislation for breaded products. The sausage was accepted obtaining 82.7% and the overall impression of the product reached a mean of 4.93 (± 1.69). Regarding intention to purchase for sausage, the highest percentage was attributed to maybe / perhaps not (32%), followed by likely to buy (24%), and 28.1% reported that they probably or decidedly would not buy. Based on the results obtained, it was concluded that the low commercial value fish of the "fish mix" category resulted in a viable product available for use in fish technology.

Bioprospection of proteases from Halobacillus andaensis for bioactive peptide production from fish muscle protein

BACKGROUND: Biologically active peptides produced from fish wastes are gaining attention because their health benefits. Proteases produced by halophilic microorganisms are considered as a source of active enzymes in high salt systems like fish residues. Hence, the aim of this study was the bioprospection of halophilic microorganisms for the production of proteases to prove their application for peptide production. RESULTS: Halophilic microorganisms were isolated from saline soils of Mexico and Bolivia. An enzymatic screening was carried out for the detection of lipases, esterases, pHB depolymerases, chitinases, and proteases. Most of the strains were able to produce lipases, esterases, and proteases, and larger hydrolysis halos were detected for protease activity. Halobacillus andaensis was selected to be studied for proteolytic activity production; the microorganism was able to grow on gelatin, yeast extract, skim milk, casein, peptone, fish muscle (Cyprinus carpio), and soy flour as protein sources, and among these sources, fish muscle protein was the best inducer of proteolytic activity, achieving a protease production of 571 U/mL. The extracellular protease was active at 50°C, pH 8, and 1.4 M NaCl and was inhibited by phenylmethylsulfonyl fluoride. The proteolytic activity of H. andaensis was used to hydrolyze fish muscle protein for peptide production. The peptides obtained showed a MW of 5.3 kDa and a radical scavenging ability of 10 to 30% on 2,2-diphenyl-1-picrylhydrazyl and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) and a ferric reducing ability of plasma. Conclusion: The use of noncommercial extracellular protease produced by H. andaensis for biologically active peptide production using fish muscle as the protein source presents a great opportunity for high-value peptide production.

Aceptabilidad de pre-escolares y escolares a la proteína purificada de pescado en polvo / Acceptability of purified fish-protein powder in pre-school and school age children

RESUMEN Objetivos. Determinar la aceptabilidad de la proteína purificada de pescado (FPi) en bebidas a base de cereales y pseudocereales. Materiales y Métodos. El estudio se realizó en el Centro de Desarrollo Integral Familiar «Año Nuevo¼ del Programa Integral Nacional para el Bienestar Familiar (INABIF), del distrito de Comas en Lima. La FPi (0-4 g por ración) se administró diariamente durante tres meses mezclada en las bebidas a base de cereales y pseudocereales. Se midió la aceptabilidad de la FPi mediante pesado directo en niños menores de 6 años y por escala hedónica facial en escolares (6 a 16 años). Resultados. Se enroló a 158 participantes y se administró de manera supervisada 4211 desayunos. El aporte calórico de las bebidas a base de cereales y pseudocereales luego de adicionar la FPi estuvo entre 117 y 140 Kcal y el aporte proteico entre 3,4 y 5,4 g por ración. La aceptabilidad en los niños mayores de 6 años de las bebidas suplementadas con la FPi (juntando los dos mayores puntajes) estuvo entre 55-74 % para la avena, alrededor del 47 % para la quinua y entre 40-58 % para la kiwicha. Mientras que el 90 % de los niños menores de 6 años consumieron más del 70 % de lo ofrecido. Conclusiones. Se encontró una buena aceptabilidad de la FPi en las bebidas a base de cereal y pseudocereales en menores de 6 años. La FPi es una alternativa para mejorar el aporte de proteínas de origen animal en los niños, sobre todo en los preescolares.

ABSTRACT Objective. To determine the acceptability of purified fish protein (FPi) in cereal and pseudocereal-based drinks. Materials and Methods. The study was carried out at Centro de Desarrollo Integral Familiar "Año Nuevo" belonging to the INABIF Program in the district of Comas in Lima. FPi (0-4 g per serving) was administered daily for 3 months mixed with cereal and pseudocereal-based drinks. The acceptability of FPi was measured by direct weighing in children under 6 years of age and by the hedonic facial scale in school-age children. Results. One hundred and fifty-eight (158) participants were enrolled and 4,211 breakfasts were served. The caloric intake of the cereal and pseudocereal drinks, after adding the FPi, ranged between 117 and 140 Kcal, and protein intake ranged between 3.4 g and 5.4 g per serving. The acceptability of drinks supplemented with FPi in children older than 6 years of age (the 2 highest scores together) ranged from 55% to 74% for oats, around 47% for quinoa and 40% to 58% for "kiwicha" (amaranth). Whereas 90% of children under 6 consumed more than 70% of the drinks offered. Conclusions. We found a good acceptability of FPi in cereal and pseudocereal beverages in children under 6 years of age. FPi is an alternative to improve the consumption of animal protein in the nutrition of children, especially in pre-schoolers.

Characterization of digestive enzymes from captive Brazilian flounder Paralichthys orbignyanus / Caracterização de enzimas digestivas do linguado brasileiro de cativeiro Paralichthys orbignyanus

Abstract Knowledge of specific enzyme activity, along with animal habits and digestive capacity is essential in formulating an appropriate diet for any species. In this study, we evaluated and characterized the activity of digestive enzymes present in the liver, intestine, and stomach of Paralichthys orbignyanus. The effects of pH and temperature on enzyme activity were also evaluated via the use of specific substrates. The use of specific substrates and inhibitors showed strong evidence of the presence of trypsin (BApNA= 0.51 ± 0.2 mU mg-1), chimotrypsin (SApNA= 2.62 ± 1.8 mU mg-1), and aminopeptidases (Leu-p-Nan =0.9709 ± 0.83 mU mg-1) in the intestine. Optimum pH for the activity of trypsin, chemotrypsin, leucino aminopeptidase, amilase, and pepsin were 9.5, 9.0, 8.0, 7.5, and 3.5, respectively, while optimum temperatures were 50, 50, 50, 40, and 45 °C, respectively. These results provide additional information regarding the biology of Brazilian flounder and can be used as a basis for further studies regarding fish feeding physiology.

Resumo O conhecimento da atividade enzimática é essencial para formular uma correta dieta específica para espécie, além de estarem correlacionadas com o hábito da alimentação e capacidade digestive. Neste estudo determinamos e caracterizamos a atividade enzimática presente no intestino, estômago e fígado do linguado Paralichthys orbignyanus. Os efeitos da temperatura e pH sobre a atividade enzimática também foram avaliados utilizando substratos específicos. O uso de substratos e inibidores específicos mostrou uma forte evidência da presença da tripsina (BApNA = 0,51 ± 0,2 mU mg-1), quimotripsina (SAPNA = 2,62 ± 1,8 mU mg-1), e as aminopeptidases (Leu-p-Nan = 0,97 ± 0,83 mU mg-1) no intestino. O pH ótimo observado para a atividade de tripsina, quimotripsina, leucino aminopeptidase, amilase e pepsina foi 9,5, 9,0, 8,0, 7,5 e 3,5, respectivamente. A temperatura ótima observada foi 50, 50, 50, 40 e 45 °C, respectivamente. Estes resultados fornecem informações adicionais sobre a biologia do linguado brasileiro e pode ser usado como base para novos estudos sobre fisiologia alimentar.

Hidrolisado proteico de resíduo de sardinha como atrativo alimentar para juvenis de jundiá / Sardine waste protein hydrolisate as feeding stimulant for silver catfish juveniles

O objetivo deste estudo foi avaliar a utilização do hidrolisado proteico de resíduo de sardinha como atrativo na alimentação do Rhamdia quelen. No experimento 1, foram utilizados os seguintes atrativos alimentares: 1. extrato aquoso de músculo de tilápia-do-Nilo (controle positivo); 2. hidrolisado proteico de resíduo de sardinha com baixo grau de hidrólise (GH); 3. hidrolisado proteico de resíduo de sardinha com alto GH; 4. hidrolisado proteico de resíduo de sardinha com alto GH diluído (10% da concentração) e 5. controle usando somente água destilada. Após jejum de 48 horas, o comportamento foi registrado em vídeo por um período basal de dois minutos e por mais 18 minutos após a inoculação do atrativo. O delineamento foi inteiramente ao acaso, com três tratamentos e 20 repetições. O experimento 2 foi realizado para avaliar a capacidade do hidrolisado proteico de estimular a ingestão de alimento em juvenis de jundiá. Para isso, foram confeccionados pellets de ágar contendo ou não hidrolisado proteico de resíduo de sardinha. Os peixes foram avaliados individualmente e tiveram um período de adaptação de sete dias. Os resultados foram analisados por meio do teste de proporção de Goodman (1964). A inoculação dos hidrolisados com alto e baixo GH aumentou o tempo de movimentação dos barbilhões. O hidrolisado com alto GH diluído proporcionou os mesmos resultados que o hidrolisado com baixo GH , mas as médias não diferiram das obtidas para a água destilada (controle negativo) e do extrato de músculo. O incremento na movimentação de um lado para outro do aquário foi maior (P<0,05) para os hidrolisados com alto e baixo GH. No experimento 2, a proporção de peixes que ingeriu os pellets contendo hidrolisado proteico de resíduo de sardinha com alto GH foi maior (P<0,05) em relação aos que ingeriram os pellets contendo água destilada. O hidrolisado proteico foi eficiente para estimular o comportamento associado à alimentação em juvenis de Rhamdia quelen.(AU)

The aim of this study was to evaluate the use of the sardine waste hydrolysate as a feeding stimulant for Rhamdia quelen juveniles. In experiment 1 the following feeding stimulants were evaluated: 1. Aqueous extract of Nile tilapia muscle; 2. sardine waste protein hydrolysate with a low degree of hydrolysis (DH); 3. Sardine waste protein hydrolysate with high GH; 4. sardine waste protein hydrolysate with high GH diluted (10% concentration) and 5. control using only distilled water. The fish were evaluated individually. After 48 hours fasting, the behavior was videotaped for a baseline period of 2 minutes, and for another 18 minutes after attractive inoculation. The design was completely randomized with three treatments and twenty repetitions. Experiment 2 was conducted to evaluate the effect of the sardine waste protein hydrolysate on the food intake of silver catfish. For this purpose agar pellets were produced containing or not sardine waste protein hydrolysate. The fish were evaluated individually and had an adjustment period of 7 days. The results were analyzed using the Goodman test (1964). Inoculation of the sardine waste protein hydrolysate with high and low GH increased the barbel movement time. The sardine waste protein hydrolyzate diluted with high GH yielded the same results as the hydrolysate with low GH, but did not differ from the average obtained for distilled water (negative control) and muscle extract. The increase in moving side to side in the aquarium was higher (P<0.05) for sardine waste protein hydrolysate with high and low GH. In experiment 2 the proportion of fish that ingested the pellets containing sardine waste protein hydrolysate was higher (P<0.05) than the proportion of fish that ingested the pellets containing distilled water. The sardine waste protein hydrolysate was efficient to stimulate the feeding associated behavior in Rhamdia quelen juveniles.(AU)

Nano-LC-ESI MS/MS analysis of proteins in dried sea dragon Solenognathus hardwickii and bioinformatic analysis of its protein expression profiling / 中国天然药物

The sea dragon Solenognathus hardwickii has long been used as a traditional Chinese medicine for the treatment of various diseases, such as male impotency. To gain a comprehensive insight into the protein components of the sea dragon, shotgun proteomic analysis of its protein expression profiling was conducted in the present study. Proteins were extracted from dried sea dragon using a trichloroacetic acid/acetone precipitation method and then separated by SDS-PAGE. The protein bands were cut from the gel and digested by trypsin to generate peptide mixture. The peptide fragments were then analyzed using nano liquid chromatography tandem mass spectrometry (nano-LC-ESI MS/MS). 810 proteins and 1 577 peptides were identified in the dried sea dragon. The identified proteins exhibited molecular weight values ranging from 1 900 to 3 516 900 Da and pI values from 3.8 to 12.18. Bioinformatic analysis was conducted using the DAVID Bioinformatics Resources 6.7 Gene Ontology (GO) analysis tool to explore possible functions of the identified proteins. Ascribed functions of the proteins mainly included intracellular non-membrane-bound organelle, non-membrane-bounded organelle, cytoskeleton, structural molecule activity, calcium ion binding and etc. Furthermore, possible signal networks of the identified proteins were predicted using STRING (Search Tool for the Retrieval of Interacting Genes) database. Ribosomal protein synthesis was found to play an important role in the signal network. The results of this study, to best of our knowledge, were the first to provide a reference proteome profile for the sea dragon, and would aid in the understanding of the expression and functions of the identified proteins.

Expression of Pleurocidin from winter flounder in Escherichia coli and optimization of culture conditions / 生物工程学报

To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.

Connection of hepcidin genes from two fish species and their expression in Pichia pastoris / 生物工程学报

Hepcidin are small cationic peptides with antibacterial activity expressed mainly in the liver of living organisms, and they play important roles in the host's immune response against microbial invasion and regulation of iron metabolism. Thus, they are considered to be good substitutes for traditional antibiotics. It is a good choice that the antimicrobial peptides are prepared by recombinant DNA expression. In the present study, two hepcidin mature peptide cDNAs from channel catfish (Ictalurus punctatus) (mCH) and tilapia (Oreochromis niloticus) (mTH) were connected by SOE-PCR in order to obtain more recombinant hepcidin with broad antimicrobial spectrum, and EcoR I and Not I sites were added to 5'- and 3'- ends of the fragment, respectively. The recombinant eukaryotic expression vector "pPIC9K-mCH-mTH" was successfully constructed, and transformed into Pichia pastoris GS115. The transformants containing multicopy gene insertion were selected by using different concentrations of G418 and other specific mediums, and identified by PCR for yeast genomic DNA. Expression was induced by adding 1% methanol at 30 degrees C for different times. Tricine-SDS-PAGE analysis demonstrated that the most appropriate expression time was 72 h, at which a high expression yield (77 mg/L) for the target protein was exhibited. The highly purified target protein was obtained from the fermentation supernatant by SP-Sepharose cation exchange chromatography. Bacteriostatic activity assay demonstrated that the fermentation supernatant containing the target protein and purified recombinant target protein had bacteriostatic activities against gram-positive and gram-negative bacterium. The present result provides the important initial value for industrial production of hepcidin antimicrobial peptide.

DNA barcoding application of mitochondrial COI gene sequence in medicinal fish of Culter (Pisces: Cyprinidae) / 中国中药杂志

The sequence variation of medicinal fish of Culter (Pisces: Cyprinidae) was analyzed by using cytochrome c oxidase subunit I (COI) sequencing collected from different regions of the Yangtze River basin, and we examine whether barcoding of COI can be used to discriminate medicinal fish of Culter. The AT content in the COI region of medicinal fish of Culter was higher than that of GC, which was similar with other species of Cypriniformes. Ninty-six percent of nucleotide changes were observed at the 3rd codon position of COI sequence, but the amino acid compositions translated by COI sequences of all Culter fish stayed the same. It is suggested that most synonymous mutations might occur at the 3rd position. The average Kimura-2-parameter (K2P) distance within-species was lower than 1%, and the K2P distance of pairwise-species was 10 times as much as that of within-species. The phylogenetic tree estimated by Neighbour-joining method indicated that species within genera invariably clustered, and generally so did individuals within species. Individuals from operational taxonomic units designated as different Culter species, supporting morphological evidence for each of these being separate species. It is suggested that the COI barcoding can be used to identify medicinal fish species of Culter.

Optimization of expression conditions of recombinant Fuantai-03 and detection of its biological activities / 生物医学工程学杂志

Fuantai-03(FAT-03), isolated from the Dasyatis akajei, has a strong antiangiogenic activity. The recombinant Fuantai-03 (GST/rFAT-03) fusion protein can be obtained with the DNA recombination technology. In this study, expression conditions of GST/rFAT-03 were optimized by response surface experimental design method. The constructed engineering bacteria containing GST/rFAT-03 plasmid was induced by isopropy-beta-D-thiogalactosid (IPTG), the GST affinity column was used for isolation and purification, and then the effects of different culture time, IPTG concentration, induction temperature and induction time on the amount of soluble GST/rFAT-03 fusion protein were compared. The culture time for optimal expression was 6.13 h, IPTG concentration was 0.36 mmol/L, induction temperature was 19.71 degrees C, and induction time was 13.60 h. The amount of soluble GST/rFAT-03 fusion protein was 7.57 mg/L under above mentioned expression conditions. The results also showed that rFAT-03 significantly inhibited angiogenesis in chicken chorioallantoic membrane in a dose-dependent manner. Moreover, the soluble form of the target protein is useful for further work on purification and on studying its biological function.

Caracterización bioquímica, fisicoquímica y funcional de turbot (Scophthalmus maximus) y estudio de sus modificaciones durante el almacenamiento a 4ºC / Biochemical, physicochemical and functional characterization of turbot (Scophthalmus maximus). Study of the changes occurring during the 4ºC storage

El turbot es un pez plano (Scophthalmus maximus), es uno de los peces que se adapta muy bien al cultivo intensivo en granja. Es necesario conocer sus propiedades fisicoquímicas-bioquímicas-tecnológicas y cómo éstas cambian durante su almacenamiento una vez que han sido cosechados. El objetivo de este estudio fue evaluar la composición centesimal, propiedades bioquímicas, fisicoquímicas y funcionales, y cómo se ven influidas por el almacenamiento a 4ºC durante 16 días. Se determinó su composición proximal, electroforesis en geles de poliacrilamida desnaturantes (PAGE-SDS), estabilidad térmica proteica a través de calorimetría diferencial de barrido (DSC). Se evaluó pH, nitrógeno básico volátil total (NBV-T), pérdida por goteo, textura, capacidad de retención de agua (CRA), emulsificación (CE) y gelificación (CG). El análisis proximal fue: humedad 76,3%; proteínas 19,6%; lípidos 2,71%; cenizas 1,3%. Se encontraron dos transiciones térmicas (DSC) a 47,5°C (miosina) y 76,9ºC (actina). PAGE-SDS mostraron perfiles correspondientes a proteínas miofibrilares y sarcoplasmáticas y durante el almacenamiento no se vieron alterados El pH cambió desde 5,9 a 6,6; las NBV-T variaron desde 20,0 a 39,5 mg NBVT/100 g músculo al día 16. CRA presentó valores de 10,5% y llegando al final a 58,1%, sin embargo no presentó CG. La CE y CRA aumentaron por el almacenamiento debido a cambios conformacionales de las proteínas. El esfuerzo de compresión fluctuó entre 111,2 a 106,0 N; el esfuerzo de cizalla fue disminuyendo en el tiempo de almacenamiento desde 148,6 a 95,2 N. La pérdida por goteo varió entre 1,7% a 2,5%. El estudio de almacenamiento refrigerado por 16 días, permitió concluir que el parámetro de frescura de NBV-T, sobrepasa el límite permitido de comercialización a los 14 días de almacenamiento, determinando de esa forma su vida útil.

The turbot (Scophthalmus maximus) is a kind of flatfish that is well adapted to intensive farm culture. After the harvest it is necessary to know the physicochemical, biochemical and technological properties and if during the refrigerated storage, changes of these properties are developed. The main objective of the study was the assessment of the proximal composition, the biochemical, physicochemical and functional properties, and how they are influenced during the 16 days storage at 4º C. Parameters such as centesimal composition, PAGE-SDS, and protein thermal stability through a DSC were carried out. pH, total volatile base-nitrogen (TVBN) dripping, texture, holding water capacity (WHC), emulsification (EC) and gelification (GC) were also determined. Results for the proximal composition were: humidity 76,3%; fat content 2,71%; proteins 19,6%; and ashes 1,3%. Two different thermal transitions at 47,5°C (myosin) and 76,9°C (actin) were observed. The PAGE-SDS showed profiles corresponding to myofibrilars and sarcoplasmatic proteins, which presented no changes during the storage. pH experimented an increase from 5,9 to 6,6; TVB-N showed a variation from 20,0 to 39,5 mg TVB-N/100 g of muscle at the day 16. The WHC started with a 10,5% and ended in 58,1%; no GC was observed. The increase of the EC and WHC during the storage was due to conformational changes of proteins. The compression force presented a fluctuation between 111,2 and 106,0 N and the shear strain decreased during the storage from 148,6 to 95,2 N. The dripping showed a variation between 1,7% and 2,5%. According to the results of the storage during 16 days at 4ºC, the NBV-T content determined a shelf life of 14 days.

Germ cell sex prior to meiosis in the rainbow trout

Germ cells make two major decisions when they move from an indeterminate state to their final stage of gamete production. One decision is sexual commitment for sperm or egg production, and the other is to maintain mitotic division or entry into meiosis. It is unclear whether the two decisions are made as a single event or separate events, because there has been no evidence for the presence of germ cell sex prior to meiosis. Here we report direct evidence in the fish rainbow trout that gonia have distinct sexuality. We show that dazl expression occurs in both male and female gonia but exhibits differential intracellular distribution. More strikingly, we show that boule is highly expressed in male gonia but absent in female gonia. Therefore, mitotic gonia possess sex, sperm/egg decision and mitosis/meiosis decision are two independent events, and sperm/egg decision precedes mitosis/meiosis decision in rainbow trout, making this organism a unique vertebrate model for mechanistic understanding of germ cell sex differentiation and relationship between the two decisions.

Fish interferon response and its molecular regulation: a review / 生物工程学报

Interferon response is the first line of host defense against virus infection. Recent years have witnessed tremendous progress in understanding of fish innate response to virus infection, especially in fish interferon antiviral response. A line of fish genes involved in interferon antiviral response have been identified and functional studies further reveal that fish possess an IFN antiviral system similar to mammals. However, fish virus-induced interferon genes contain introns similar to mammalian type III interferon genes although they encode proteins similar to type I interferons, which makes it hard to understand the evolution of vertebrate interferon genes directly resulting in a debate on nomenclature of fish interferon genes. Actually, fish display some unique mechanisms underlying interferon antiviral response. This review documents the recent progress on fish interferon response and its molecular mechanism.

Effect of stress during handling, seawater acclimation, confinement, and induced spawning on plasma ion levels and somatolactin immunoreactivity in mature female thin -lipped gray mullet, Liza ramada

Understanding the physiological picture of fish during the reproductive cycle, seawater acclimation, and induced spawning is of essential value to know the possible reasons of preoviposition mortality and to develop successful hatchery technology. Determination of the effect of different stress factors on hydro-mineral balance as well as changes in somatolactin [SL] immunoreactive cells in mature Liza ramada females. Water chemistry and the different plasma ion levels were measured. Immunocytochemical staining for the sections of the pituitary gland was performed to describe the activity of SL immunoreactive cells. The plasma levels of PO[3-4], Na[+], K[+], Ca[2+], and Mg[2+] showed a slight increase during transportation without anesthesia. The concentrations of these minerals returned to the initial levels by using clove oil [5 mg/I] as anesthetic. However, their levels decreased during seawater acclimation and gradually increased with confinement to reach the initial values. Furthermore, the levels of PO[3-4], Na[+], K[+], Ca[2+], and Mg[2+] were significantly [P

Cloning and functional analysis of the collectin gene from the grass carp Ctenopharyngodon idella / 生物工程学报

The grass carp (Ctenopharyngodon idella) collectin gene was cloned from mixed liver and kidney cDNA library. The full length sequence of grass carp collectin was 1128 bp, contained a 5' untranslated region of 229 bp and a 3' untranslated region of 104 bp. The open reading frame of grass carp collectin was 795 bp which could code a 264 amino acids polypeptide, including a terminal codon. Phylogenetic analyses showed that grass carp collectin shared the highest homology with that of zebrafish (Danio rerio). To understand the function of grass carp collectin, we expressed and purified the recombinant protein (P(CRD)) that comprised carbohydrate recognition domain (CRD). Agglutination of Aeromonas hydrophila and Staphylococcus aureus etc. and sugars inhibition experiments showed that: galactose, glucose, mannose and maltose could inhibit the agglutination of Aeromonas hydrophila. Maltose could lower the agglutination of Staphylococcus aureus, whereas peptidoglycan and glucose inhibited it well. In addition, the activity of grass carp collectin could not dependent on Ca(2+).

[ role of coelomic fluid in fertilization of rainbow trout [oncorhynchus mykiss] oocytes]

Iran is one of the most important Rainbow trout [Oncorhynchus mykiss] producing countries in the world and fingering production is the key factor for this industry. The present experiment was conducted to determine the role of the coelomic fluid in artificial fertilization and to study the interactive effect of different brood fish coelomic fluid on fertilization process during 60 min storage at 4°C.The egg quality was evaluated through the rate of eyed and hatched eggs where these indices applied with and without coelomic fluid. In the first two trials, the rat of egg fertilization with and without coelomic fluid did not exhibit significant difference [p>0.05]. Eyeing rates were 97.1% and 97.6% for trial 1 and 99.1% and 97.7% for trial II. respectively. In next trial the eyeing and hatching rate were 88.7% and 86.8% for the eggs fertilized immediately alter stripping wheas for those fertilized 60 min. after integrated storage these values were 86.3% and 84.6% respectively [p>0.05]. we concluded that the coelomic fluid has no interative effect on artificial fertilization and the quality of egg fecundity is not influenced by the fertilization process during 60 min storage at 4°C

Histological and enzyme histochemical studies on the transitional gonads of giant grouper Epinephelm tauvina [perciforms: serranidae] from the Arabian Gulf coast of Saudi Arabia

The present work was carried out using twenty mature Epinephelus tauvina [Perciforms: Serranidae] collected from Arabian Gulf coast at Dammam City. Fishes of this species are known to undergo sex change during certain stage of their life cycle. Histological and enzyme histochemical studies were performed on gonads of the collected fishes. The examination of the gonads of E. tauvina revealed the presence of three developmental phases during the sex change process. These were: Female, early transition phase and late transition phase each one may subdivide into two stages. In female phase several developmental stages of the oocytes were recognized. At the beginning of early transition phase perinucleolar mature oocytes began degeneration while late transition phase characterized by rapid proliferation of spermatogenic cells. The histochemical studies demonstrated that alkaline phosphatase enzyme gave intense reaction in granulosa cells of mature oocytes while acid phosphatase gave an intense reaction in the interstitial cells and atretic follicles but delta S -3 beta hydroxysteroid dehydrogenase enzyme gave intense reaction to special cells in capsule of gonads

Structure-antimicrobial activity relationship between pleurocidin and its enantiomer

To develop novel antibiotic peptides useful as therapeutic drugs, the enantiomeric analogue of pleurocidin (Ple), which is a well known 25-mer antimicrobial peptide, was designed for proteolytic resistance by D-amino acids substitution. The proteolytic resistance was confirmed by using HPLC after the digestion with various proteases. To investigate the antibiotic effect of L- and D-Ple, the antibacterial activity and hemolytic effect were tested against human erythrocytes. The D-Ple showed a decreased antibacterial activity and a dramatically decreased hemolytic activity compared with L-Ple. The hemolytic effect of analogue was further confirmed by using calcein leakage measurement with liposome. To elucidate these results, the secondary structure of the peptides was investigated by using circular dichroism spectroscopy. The results revealed that D-Ple, as well as L-Ple, had typical ?-helical structures which were mirror images, with a different helicity. These results suggested that the discrepancy of the structure between the two peptides made their antibacterial activity distinct.