Evaluation of secretory activity in carp pancreatocytes (Cyprinus carpio L. ) when using lactobacillus-based chelate and probiotic supplemental feed complexes / Avaliação da atividade secretora em pancreatócitos de carpa (Cyprinus carpio L. ) ao usar quelato à base de lactobacilos e complexos suplementares de alimentação probiótica

The experimental research was carried out on the juvenile carp (Cyprinus carpio L.). The impact from supplemental feeds consisting of variable concentrations of chelate compounds, biogenic trace elements and probiotic lactobacillus-based product Bacillus subtilis VKPM B-2335 was evaluated. Optical and qualitative parameters of the lactobacillus base were studied in order to identify the major group of substances potentially able to influence the end result. The purpose of this research was to identify changes in the structure of the zymogen granules and their dimensions at which supplemental feeds produce a stimulating effect on the synthesis of zymogens in exogenous cells of the secretory part of pancreas. At the outcome of the study, for the first time, it was possible to prove that the integrated action of chelates and lactobacillus-based probiotics complemented each other. Metal chelate compounds contributed to enlargement of the zymogen granules, if compared to the control values. The bacterial products accelerated production of the zymogen granules in acinar cells diffusely located in carp hepatopancreas.

A pesquisa experimental foi realizada em carpas juvenis (Cyprinus carpio L.). O impacto de suplementos alimentares consistindo em concentrações variáveis de compostos quelatos, oligoelementos biogênicos e produto probiótico baseado em lactobacilos Bacillus subtilis VKPM B-2335 foi avaliado. Parâmetros ópticos e qualitativos da base de lactobacilos foram estudados a fim de identificar o maior grupo de substâncias potencialmente capazes de influenciar o resultado final. O objetivo dessa pesquisa foi identificar mudanças na estrutura dos grânulos de zimogênio e suas dimensões nas quais alimentos suplementares produzem um efeito estimulante na síntese de zimogênios em células exógenas da parte secretora do pâncreas. No resultado do estudo, pela primeira vez, foi possível comprovar que a ação integrada dos quelatos e dos probióticos à base de lactobacilos se complementava. Compostos quelatos metálicos contribuíram para o aumento dos grânulos de zimogênio, se comparados aos valores de controle. Os produtos bacterianos aceleraram a produção dos grânulos de zimogênio nas células acinares localizadas difusamente no hepatopâncreas da carpa.

Evaluation of secretory activity in carp pancreatocytes (Cyprinus carpio L.) when using lactobacillus-based chelate and probiotic supplemental feed complexes

Abstract The experimental research was carried out on the juvenile carp (Cyprinus carpio L.). The impact from supplemental feeds consisting of variable concentrations of chelate compounds, biogenic trace elements and probiotic lactobacillus-based product Bacillus subtilis VKPM B-2335 was evaluated. Optical and qualitative parameters of the lactobacillus base were studied in order to identify the major group of substances potentially able to influence the end result. The purpose of this research was to identify changes in the structure of the zymogen granules and their dimensions at which supplemental feeds produce a stimulating effect on the synthesis of zymogens in exogenous cells of the secretory part of pancreas. At the outcome of the study, for the first time, it was possible to prove that the integrated action of chelates and lactobacillus-based probiotics complemented each other. Metal chelate compounds contributed to enlargement of the zymogen granules, if compared to the control values. The bacterial products accelerated production of the zymogen granules in acinar cells diffusely located in carp hepatopancreas.

Resumo A pesquisa experimental foi realizada em carpas juvenis (Cyprinus carpio L.). O impacto de suplementos alimentares consistindo em concentrações variáveis de compostos quelatos, oligoelementos biogênicos e produto probiótico baseado em lactobacilos Bacillus subtilis VKPM B-2335 foi avaliado. Parâmetros ópticos e qualitativos da base de lactobacilos foram estudados a fim de identificar o maior grupo de substâncias potencialmente capazes de influenciar o resultado final. O objetivo dessa pesquisa foi identificar mudanças na estrutura dos grânulos de zimogênio e suas dimensões nas quais alimentos suplementares produzem um efeito estimulante na síntese de zimogênios em células exógenas da parte secretora do pâncreas. No resultado do estudo, pela primeira vez, foi possível comprovar que a ação integrada dos quelatos e dos probióticos à base de lactobacilos se complementava. Compostos quelatos metálicos contribuíram para o aumento dos grânulos de zimogênio, se comparados aos valores de controle. Os produtos bacterianos aceleraram a produção dos grânulos de zimogênio nas células acinares localizadas difusamente no hepatopâncreas da carpa.

Masculinization in common carp (Cyprinus carpio) by androgen immersion: The interaction effect of hormone concentration and immersion time / Masculinização na carpa comum (Cyprinus carpio) por imersão em androgênio: Efeito da interação da concentração hormonal e do tempo de imersão

Synthetic androgens (male hormones) administered to fish nursery are being used in aquaculture to avoid sexual differentiation and unwanted spawning at the eggs or the first feeding fry stage of fish. Present trial was conducted with the aim to produce male common carp (Cyprinus carpio) by egg immersion technique. Through this little insight, the effect of different hormone concentrations (17α-methyltestosterone @ HC:150, 300, 450 and 600 µgl-1) with immersion times (IT: 24, 48 and 72 hrs) and their interaction effect (HC x IT) on the hatching percentage of Cyprinus carpio eggs, percent survival and percent of male's production was evaluated specifically. Results showed that egg hatching percentage decreased with increased IT likewise, survival of treated fry was affected by increasing the IT (P<0.001). The main interaction effect of HC x IT showed that the highest percent of male individuals (95%) was obtained at 450-600 µgl-1 HC for 72 hrs IT, followed by 88-92.50% at 150-300 µgl-1 HC for 72-hrsof IT, 87.50% at 48-hrs of IT for rest of the hormone treatments, and lowest 47.50% was recorded in control (P<0.05). Increased percent male of Cyprinus carpio was obtained with increasing HC across all ITs. It was observed that the immersion treatment at 600µgl-1 for 72 hours was more effective to change the sex ratio of pre hatch Cyprinus carpio. A comparative outlook made from this experimental trial that sex induction of Cyprinus carpio by eggs immersion using synthetic male steroid hormone is an alternative safe technique of fish sex reversal in contrast to oral administration of hormone in fish feed.

Andrógenos sintéticos (hormônios masculinos) administrados ao viveiro de peixes estão sendo usados ​​na aquicultura para evitar a diferenciação sexual e a desova indesejada nos ovos ou no primeiro estágio de alimentação dos peixes. O presente estudo foi conduzido com o objetivo de produzir carpa comum masculina (Cyprinuscarpio) pela técnica de imersão em ovos. Com essa pequena percepção, o efeito de diferentes concentrações hormonais (17α-metiltestosterona @ HC: 150, 300, 450 e 600 µgl-1) com tempos de imersão (IT: 24, 48 e 72 horas) e seu efeito de interação (HC x IT) na porcentagem de eclosão dos ovos de Cyprinuscarpio, a porcentagem de sobrevivência e a porcentagem da produção masculina foram avaliadas especificamente. Os resultados mostraram que a porcentagem de incubação de ovos diminuiu com o aumento da TI da mesma forma, a sobrevivência dos alevinos tratados foi afetada pelo aumento da TI (P <0,001). O principal efeito de interação do HC x IT mostrou que o maior percentual de indivíduos do sexo masculino (95%) foi obtido com 450-600 µgl-1 HC por 72 horas de TI, seguido por 88-92,50% com 150-300 µgl-1 HC para 72 horas de TI, 87,50% às 48 horas de TI para o restante dos tratamentos hormonais, e 47,50% mais baixos foram registrados no controle (P <0,05). A porcentagem aumentada de macho de Cyprinuscarpio foi obtida com o aumento do HC em todas as TIs. Observou-se que o tratamento de imersão a 600µgl-1 por 72 horas foi mais efetivo na alteração da razão sexual do Cyprinuscarpio antes da eclosão. Uma perspectiva comparativa feita a partir deste ensaio experimental de que a indução sexual de Cyprinuscarpio por imersão de ovos usando hormônio esteróide masculino sintético é uma técnica alternativa segura de reversão do sexo em peixes, em contraste com a administração oral de hormônio na alimentação de peixes.

Características de replicación y supervivencia del virus de viremia primaveral de la carpa (SVCV) aislado en México / Replication and survival traits of spring viremia of carp virus (SVCV) isolated in Mexico

RESUMEN Objetivo. Realizar el aislamiento del virus de la viremia primaveral de la carpa (SVCV) en ejemplares de carpa común (Cyprinus carpió), evaluar su crecimiento en diferentes tipos de células, así como la supervivencia viral a diferentes temperaturas. Materiales y métodos. Diez carpas de entre 400 500 gramos de una laguna del centro de México fueron procesadas para el diagnóstico de SVCV mediante aislamiento en cultivo de células y RT-PCR semianidado. El virus obtenido se inoculó en células EPC, BF-2, CHSE-214 y RTG-2 para determinar diferencias de crecimiento de SVCV. Además, se evaluó la supervivencia del virus conservado a temperatura ambiente (TA 20-25°C), refrigeración (REF 4°C) y congelación (CONG -80°C) hasta once meses. Los órganos internos se procesaron para análisis histológico. Resultados. Los peces analizados no presentaron signos externos sugestivos de enfermedad, pero interna e histopatológicamente se observaron lesiones sugestivas de infección sistémica. SVCV fue aislado en células EPC y BF-2 y confirmado por RT-PCR semianidado. SVCV únicamente indujo CPE en células EPC y BF-2 y fue negativo en RTG-2 y CHSE-214. El virus conservado a TA perdió viabilidad después de cuatro meses post infección (mpi), siendo total a seis mpi; mientras REF y CONG fueron estables durante los once meses de estudio. Conclusiones. La infección subclínica por SVCV fue confirmada en carpas que presentaron lesiones histológicas asociadas a esta infección. SVCV únicamente causó CPE en células EPC y BF-2 y el virus conservó su viabilidad a 4°C y -80°C hasta once meses; mientras que a TA se perdió en seis meses.

ABSTRACT Objective. To perform the isolation of spring viremia of carp virus (SVCV) in common carp (Cyprinus carpió) and evaluate its growth in different cell types and viral survival at different temperatures. Materials and methods. Ten carps of between 400-500 grams of a lagoon in central Mexico were processed for diagnosis of SVCV by isolation in cell culture and by RT-PCR. The virus obtained was inoculated into EPC, BF-2, CHSE-214 and RTG-2 cells to determine differences in virus growth; the survival of virus stored at room temperature (TA 20-25°C), refrigeration (REF 4°C) and freezing (CONG -80°C) up to eleven months was also evaluated. Internal organ samples were processed for histological analysis. Results. The fish analyzed did not show external signs suggestive of disease but internally and histopathologically lesions suggestive of systemic infection were observed. SVCV was isolated in EPC and BF-2 cells and confirmed by semi-nested RT-PCR. SVCV only induced CPE in EPC and BF-2 cells and was negative in RTG-2 and CHSE-214. The virus conserved at TA lost viability after four months post-infection (mpi), being total at six mpi; while REF and CONG were stable during the eleven months. Conclusions. Subclinical SVCV infection was confirmed in carp that presented histological lesions associated with this infection; SVCV only caused CPE in EPC and BF-2 cells; and the virus kept in refrigeration and at -80°C retained its viability up to eleven months; while TA was lost in six months.

Effect of salinity on lipid profile of Cyprinus carpio reared in inland saline water

Aim: The present study was undertaken to investigate the influence of different salinity levels on gonadal tissue and serum biochemical parameters of Cyprinus carpio reared in inland saline groundwater to standardize the optimal salinity level for better maturation and seed production.Methodology: A 90 days experiment was conducted in non-drainable rectangular earthen ponds (n= 8) to study the effects of different salinity (0, 5, 10 and 15 ppt) levels on distribution of lipid class profile in Cyprinus carpio reared in inland saline groundwater. Gonads and serum were collected on 15 days interval and lipid classes, cholesterol, phospholipid and triacylglycerol were analyzed by using extracted total lipid. Results: The concentration of different forms of lipids were significantly influenced by salinity and their concentration increased with development of maturity stages but lowest level was recorded at ovulation or spermiation stages. It was observed that significantly (p < 0.05) highest level of all lipids was found at 0 and 5 ppt compared to 10 and 15 ppt salinity. Interpretation: The present study revealed that various lipid class fatty acids varied according to salinity and maturity stages of C. carpio in inland saline groundwater. Further, it also indicated that C. carpio has a tendency to adapt, survive, tolerate and reproduce in inland saline environment salinity ranging from 0-15 ppt and was found optimal at 5 ppt based on the reproductive ability

Characterization of lactic acid bacteria as a fish potential immunostimulant

@#Aims: This study aims to characterize the intestinal carp (Cyprinus carpio L.) bacteria, especially lactic acid bacteria (LAB) and its potential as immune-stimulant to be applied in the prevention of diseases in fish. Methodology and result: The bacteria were isolated from carp intestine and cultured in de Mann Rogosa Sharpe (MRS) and glucose yeast peptone agar + calcium carbonate (GYPA+CaCO3) media. The obtained LABs were identified and characterized by 16S rRNA gene primers. The phylogenetic analysis on DNA sequence was performed by using BioEdit and MEGA 7.0 software. The potential immunostimulant were derived from its ability to resist the growth of Aeromonas sp. and Vibrio sp. as pathogenic bacteria by the paper disc agar diffusion method. The clear zone diameter around the paper disc were measured by using calipers. Forty bacteria that isolated from the carp were selected for lactic acid production and clear zone around the colonies were formed from the GYPA+CaCO3. For further analysis, a total of ten LABs were selected based on different colony forms and the largest clear zone around the colonies. Based on phylogenetic analysis, Enterococcus, Lactobacillus, Streptococcus and Lactococcus were found as the genera of lactic acid bacteria. Conclusion, significance, and impact of study: We discovered that there is a wide diversity among the 40 isolated bacteria. This result indicates that Enterococcus, Lactobacillus, Streptococcus and Lactococcus were the common genera. The most potential LAB as an immuno-stimulant was Lactobacillus sp.

Study on assessment of proximate composition and meat quality of fresh and stored Clarias gariepinus and Cyprinus carpio / Estudo da avaliação da composição centesimal e da qualidade da carne de Clarias gariepinus e Cyprinus carpio

Abstract The organoleptic evaluation and proximate analysis of Clarias gariepinus and Cyprinus carpio were determined in fresh fish and when refrigerated at two different temperatures (-21 °C and 4 °C) for a period of six weeks. A panel of twelve trained judges evaluated the color (live), texture, softness and flavor of fish meat after two minutes' steam cooking. Average score revealed a general decline in organoleptic properties such as color, texture, freshness, and taste of C. gariepinus and C. carpio stored at two temperatures compared to the fresh fish. Proximate analysis revealed a more decrease in crude protein and lipid contents and increase in ash content in C. gariepinus and C. carpio at the two storage temperatures compared to the fresh fish muscle. Moisture content decreased in the fish muscle samples of both the fish species stored at -21 °C but increased in the 4 °C stored samples. pH of fish was found to increase in the two stored temperatures. There were significant differences (P<0.05) in the organoleptic and proximate composition of the ice stored and fresh C. gariepinus and C. carpio, the same temperature and between the two different temperatures. The quality of fish muscle stored at 4 °C deteriorated faster than that of the -21 °C. Thus, storage temperature and duration have adverse effects on the nutritional quality of fish meat.

Resumo A avaliação organoléptica e a análise aproximada de Clarias gariepinus e Cyprinus carpio foram determinadas em peixe fresco e refrigerado a duas temperaturas diferentes (-21 °C e 4 °C) por um período de seis semanas. Um painel de doze juízes treinados avaliou a cor (ao vivo), textura, maciez e sabor da carne de peixe após dois minutos de cozimento a vapor. O escore médio revelou um declínio geral nas propriedades organolépticas, como cor, textura, frescor e sabor de C. gariepinus e C. carpio armazenados a duas temperaturas em comparação com o peixe fresco. A análise imediata revelou uma maior diminuição nos teores de proteína bruta e lipídios e aumento no teor de cinzas em C. gariepinus e C. carpio nas duas temperaturas de armazenamento em comparação com o músculo do peixe fresco. O teor de umidade diminuiu nas amostras de músculo de peixe de ambas as espécies de peixes armazenadas a -21 °C, mas aumentou nas amostras armazenadas a 4 °C. O pH dos peixes aumentou nas duas temperaturas armazenadas. Houve diferenças significativas (P <0,05) na composição organoléptica e próxima do gelo armazenado e fresco C. gariepinus e C. carpio, a mesma temperatura e entre as duas temperaturas diferentes. A qualidade do músculo do peixe armazenado a 4 °C deteriorou-se mais rapidamente do que a temperatura de -21 °C. Assim, a temperatura e duração do armazenamento têm efeitos adversos na qualidade nutricional da carne de peixe.

Impact of Industrial Wastewaters on Fresh Water Fish, Cyprinus carpio (Linnaeus, 1785): A Tool for Ecological Risks Monitoring

Background: In the era of industries, the problem of pollution of aquatic resources has become aggravated due to continuous wastewater disposal. The Aquatic ecosystem health can be considered as an indicator of the environmental state. Furthermore, fishes are an ideal indicators for contaminated environment. Aims: To assess the sensitivity of common carp fishes against industrial wastewaters exposure on the basis of lethal effects and behavioral changes as a tool for ecotoxicology for knowing the possible effects upon environment due to selected industries wastewater. Methodology: In this study, the effects of industrial wastewaters for different ecotoxicological parameters (mortality and behaviour changes) on common carp were studied under static conditions during different time interval (24, 42, 72 and 96 h) followed by sampled industrial wastewater quality analysis for physicochemical parameters. Results: Increased space between the gills and operculum, excessive excretion, as well as increased fish surface activity was found in the wastewater exposed fish compared to control. Conclusion: The overall findings from the present study indicate that the sampled industrial wastewaters are safe for crop irrigation but may have some adverse impact on the biota due to the observed behavioral and histological changes in wastewater exposed fishes.

Efeito de vitaminas adicionadas ao diluente ACP-104 sobre a qualidade do sêmen criopreservado de carpa comum (Cyprinus carpio) / Effect of vitamins added to the ACP-104 extender on the quality of cryopreserved semen of common carp (Cyprinus carpio)

O objetivo da presente pesquisa foi alcançado com a divisão da pesquisa em dois experimentos: (1) aperfeiçoar o protocolo de congelação utilizando água de coco em pó (ACP-104) como diluente para a criopreservação seminal de carpa comum; (2) avaliar o efeito da suplementação das vitaminas C (ácido ascórbico) ou E (α-tocoferol) sobre os melhores diluidores testados no experimento 1 na qualidade do sêmen pós-descongelado da espécie. Para o experimento 1, foram formados oito pools de sêmen, provenientes de 14 machos selecionados. As amostras seminais coletadas foram avaliadas quanto à motilidade total, à velocidade, ao percentual de espermatozoides normais e à vitalidade espermática antes e depois da criopreservação seminal. Esta foi realizada em meio ACP-104 acrescido de dimetilsulfóxido (DMSO), ou etilenoglicol (EG), ou glicerol, ou metanol, todos à concentração de 10%, diluídos em 1:3 (sêmen:diluidor). As amostras foram, então, congeladas em vapor de nitrogênio líquido em dry shipper e estocadas em nitrogênio líquido (-196°C). Para o experimento 2, foram formados oito pools provenientes da coleta de sêmen de 15 machos. As amostras seminais foram avaliadas seguindo as mesmas análises do experimento 1, acrescentando-se a duração da motilidade total. A criopreservação seminal utilizou-se do meio ACP-104 acrescido de DMSO ou EG, suplementado ou não com vitamina C ou E. Os melhores resultados encontrados no experimento 1 foram obtidos com o DMSO e o EG. Estes não diferiram significativamente entre si para a motilidade total (24% e 28%; P>0,05) e a normalidade espermática (32% e 26%; P>0,05), respectivamente. Para o experimento 2, o EG suplementado com vitamina E produziu significativamente resultados superiores de motilidade total, normalidade espermática e duração da motilidade em relação ao DMSO, concluindo-se que o EG deve ser, portanto, o crioprotetor de escolha a ser utilizado com o ACP-104 suplementado ou não com vitamina E.(AU)

The objective was achieved by dividing the research into two experiments: (1) improving the freezing protocol using powdered coconut water (ACP-104) as a diluent for the cryopreservation seminal of common carp; (2) evaluating the effect of supplementation of vitamins C (ascorbic acid) or vitamin E (α-tocoferol) with the best extenders tested in experiment 1 on the quality of post-thawed. For experiment 1, semen pools from 14 selected males were formed. Seminal samples were evaluated for total motility, velocity, percentage of normal sperm and sperm vitality before and after the seminal cryopreservation. This was done in ACP-104 extender plus dimethyl sulfoxide (DMSO), or ethylene glycol (EG), or glycerol or methanol all at concentration 10% diluted in 1:3 (semen:extender). The samples were frozen in vapors of nitrogen into dry shippers and stored in liquid nitrogen (-196 °C). For experiment 2, eight pools were formed from the 15 males. The semen samples were evaluated following the same analysis of experiment 1 adding duration of total motility. The sperm cryopreservation was performed in extenders ACP-104 plus DMSO or EG supplemented or not with vitamin C or E. The best results found in Experiment 1 were obtained with DMSO and EG. They do not differ significantly for total motility (24% and 28%; P>0.05) and normal sperm (32% and 26%; P>0.05) respectively. For experiment 2, EG supplemented with vitamin E, produced significantly better results overall motility, sperm normality and duration of motility relative to DMSO. In conclusion, EG should be the cryoprotectant of choice for use with the ACP-104 supplemented or not with vitamin E.(AU)

Actividad enzimática de proteasas de Cyprinus carpio (Cypriniformes: Cyprinidae) extraídas de una laguna contaminada en México / Enzymatic activity of proteases from Cyprinus carpio (Cypriniformes: Cyprinidae) captured in a polluted lagoon in Mexico

ResumenLa carpa común (Cyprinus carpio) es un organismo acuático de valor comercial que puede sobrevivir en ambientes contaminados; la carpa contiene enzimas proteolíticas de importancia fisiológica y potencial aplicación industrial. El objetivo de este estudio fue purificar parcialmente y determinar la actividad proteolítica a diferente pH de proteasas de carpas colonizando un ambiente contaminado. Se capturaron tres carpas en diferentes zonas de la laguna contaminada de Zumpango (México) a 1 m de profundidad máxima. El extracto crudo se obtuvo del músculo dorsal mediante extracción acuosa y se fraccionó con (NH4)2SO4 saturado a 20 %, 50 % y 80 %. Posteriormente, se seleccionaron las fracciones obtenidas con (NH4)2SO4 saturado a 50 % y 80 % por su alta actividad proteolítica, se concentraron por ultrafiltración con membranas de corte de peso molecular de 100 kDa, y se analizaron por electroforesis en geles de poliacrilamida con dodecil sulfato de sodio (SDS-PAGE). La actividad proteolítica del extracto crudo fue significativamente mayor (19.7-20.3 U / mg) a pH 2, 5, y 7 (P < 0.001). Las fracciones obtenidas con (NH4)2SO4 saturado a 20 %, 50 % y 80 % presentaron actividades proteolíticas óptimas a pH 5 (2.8 U / mg) y pH 6 (2.2 U / mg); pH 6 (4.3 U / mg) y pH 3-4 (3.6 - 3.7 U / mg); pH 3 (10.8 U / mg) y pH 10 (10.6 U / mg); respectivamente. Las subfracciones con peso molecular < 100 kDa obtenidas con (NH4)2SO4 saturado a 50 % y 80 % tuvieron máxima actividad proteolítica a pH alcalino. La subfracción < 100 kDa, obtenida con (NH4)2SO4 saturado a 80 % tuvo la mayor actividad proteolítica (37.3-43.7 U / mg) a pH 8-10, factor de purificación de 3 y 19.1 % de recuperación. Trece proteínas entre 9.8 a 104.8 kDa se identificaron en el extracto proteolítico crudo. Las proteínas de 31 - 33 y 39 - 41 kDa tuvieron la concentración más alta en las fracciones estudiadas, sugiriendo la posible predominancia de serín y aspartil proteasas, respectivamente. Nosotros sugerimos que la presencia de proteasas con máxima actividad a pH alcalino está relacionada con la adaptación de C. carpio a aguas contaminadas con pH alto. Aunque estos peces son inadecuados para el consumo humano, pueden ser empleados como materia prima para la producción de proteasas destinadas para varias industrias, incluido el tratamiento de aguas residuales.

AbstractCommon carp (Cyprinus carpio) is an aquatic organism of commercial value able to survive in polluted environments; carps contain proteolytic enzymes of physiological importance and potential industrial application. The objective of this work was partially purify and study the proteolytic activity at different pH of carp proteases living in a polluted environment. Three carps were captured in different zones of Zumpango polluted lagoon (Mexico) at 1 m of maximum deep. Protease crude extracts were obtained from dorsal muscle by aqueous extraction and fractionated by 20 %, 50 %, 80 %-saturated (NH4)2SO4. Fractions extracted with 50 % and 80 %-saturated (NH4)2SO4 were selected for their high proteolytic activity and concentrated by ultrafiltration through 100 kDa molecular weight cutoff membranes and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The crude proteolytic extract had significantly higher activity (19.7 - 20.3 U / mg) at pH 2, 5, and 7 (P < 0.001). Fractions obtained with 20 %, 50 % and 80 % - saturated (NH4)2SO4 showed peak activity at pH 5 (2.8 U / mg) and pH 6 (2.2 U / mg); pH 6 (4.3 U / mg) and pH 3 - 4 (3.6 - 3.7 U / mg); pH 3 (10.8 U / mg) and pH 10 (10.6 U / mg); respectively. Subfractions of < 100 kDa, obtained with 50 % and 80 %-saturated (NH4)2SO4, had peak proteolytic activity at alkaline pH. A < 100 kDa fraction, obtained with 80 %-saturated (NH4)2SO4, had the highest proteolytic activity (37.3 - 43.7 U / mg) at pH 8 - 10, purification factor of 3 and 19.1 % recovery. Thirteen proteins between 9.8 to 104.8 kDa were identified in the crude extract. Peak protein concentration was observed for 31 - 33 and 39 - 41 kDa, suggesting the possibility predominance of serine- and aspartyl- proteases, respectively. We suggest this protease with maximum activity at alkaline pH is related to the adaptation of C. carpio to polluted waters with high pH. Although unsuitable for human consumption, these organisms can be a source of protease production aimed to several uses as in the industry and waste water treatment among others.

Study on assessment of proximate composition and meat quality of fresh and stored Clarias gariepinus and Cyprinus carpio

Abstract The organoleptic evaluation and proximate analysis of Clarias gariepinus and Cyprinus carpio were determined in fresh fish and when refrigerated at two different temperatures (-21 °C and 4 °C) for a period of six weeks. A panel of twelve trained judges evaluated the color (live), texture, softness and flavor of fish meat after two minutes steam cooking. Average score revealed a general decline in organoleptic properties such as color, texture, freshness, and taste of C. gariepinus and C. carpio stored at two temperatures compared to the fresh fish. Proximate analysis revealed a more decrease in crude protein and lipid contents and increase in ash content in C. gariepinus and C. carpio at the two storage temperatures compared to the fresh fish muscle. Moisture content decreased in the fish muscle samples of both the fish species stored at -21 °C but increased in the 4 °C stored samples. pH of fish was found to increase in the two stored temperatures. There were significant differences (P 0.05) in the organoleptic and proximate composition of the ice stored and fresh C. gariepinus and C. carpio, the same temperature and between the two different temperatures. The quality of fish muscle stored at 4 °C deteriorated faster than that of the -21 °C. Thus, storage temperature and duration have adverse effects on the nutritional quality of fish meat.

Resumo A avaliação organoléptica e a análise aproximada de Clarias gariepinus e Cyprinus carpio foram determinadas em peixe fresco e refrigerado a duas temperaturas diferentes (-21 °C e 4 °C) por um período de seis semanas. Um painel de doze juízes treinados avaliou a cor (ao vivo), textura, maciez e sabor da carne de peixe após dois minutos de cozimento a vapor. O escore médio revelou um declínio geral nas propriedades organolépticas, como cor, textura, frescor e sabor de C. gariepinus e C. carpio armazenados a duas temperaturas em comparação com o peixe fresco. A análise imediata revelou uma maior diminuição nos teores de proteína bruta e lipídios e aumento no teor de cinzas em C. gariepinus e C. carpio nas duas temperaturas de armazenamento em comparação com o músculo do peixe fresco. O teor de umidade diminuiu nas amostras de músculo de peixe de ambas as espécies de peixes armazenadas a -21 °C, mas aumentou nas amostras armazenadas a 4 °C. O pH dos peixes aumentou nas duas temperaturas armazenadas. Houve diferenças significativas (P 0,05) na composição organoléptica e próxima do gelo armazenado e fresco C. gariepinus e C. carpio, a mesma temperatura e entre as duas temperaturas diferentes. A qualidade do músculo do peixe armazenado a 4 °C deteriorou-se mais rapidamente do que a temperatura de -21 °C. Assim, a temperatura e duração do armazenamento têm efeitos adversos na qualidade nutricional da carne de peixe.

Masculinization in common carp (Cyprinus carpio) by androgen immersion: The interaction effect of hormone concentration and immersion time

Abstract Synthetic androgens (male hormones) administered to fish nursery are being used in aquaculture to avoid sexual differentiation and unwanted spawning at the eggs or the first feeding fry stage of fish. Present trial was conducted with the aim to produce male common carp (Cyprinus carpio) by egg immersion technique. Through this little insight, the effect of different hormone concentrations (17-methyltestosterone @ HC:150, 300, 450 and 600 µgl-1) with immersion times (IT: 24, 48 and 72 hrs) and their interaction effect (HC x IT) on the hatching percentage of Cyprinus carpio eggs, percent survival and percent of males production was evaluated specifically. Results showed that egg hatching percentage decreased with increased IT likewise, survival of treated fry was affected by increasing the IT (P 0.001). The main interaction effect of HC x IT showed that the highest percent of male individuals (95%) was obtained at 450-600 µgl-1 HC for 72 hrs IT, followed by 88-92.50% at 150-300 µgl-1 HC for 72-hrsof IT, 87.50% at 48-hrs of IT for rest of the hormone treatments, and lowest 47.50% was recorded in control (P 0.05). Increased percent male of Cyprinus carpio was obtained with increasing HC across all ITs. It was observed that the immersion treatment at 600µgl-1 for 72 hours was more effective to change the sex ratio of pre hatch Cyprinus carpio. A comparative outlook made from this experimental trial that sex induction of Cyprinus carpio by eggs immersion using synthetic male steroid hormone is an alternative safe technique of fish sex reversal in contrast to oral administration of hormone in fish feed.

Resumo Andrógenos sintéticos (hormônios masculinos) administrados ao viveiro de peixes estão sendo usados na aquicultura para evitar a diferenciação sexual e a desova indesejada nos ovos ou no primeiro estágio de alimentação dos peixes. O presente estudo foi conduzido com o objetivo de produzir carpa comum masculina (Cyprinuscarpio) pela técnica de imersão em ovos. Com essa pequena percepção, o efeito de diferentes concentrações hormonais (17-metiltestosterona @ HC: 150, 300, 450 e 600 µgl-1) com tempos de imersão (IT: 24, 48 e 72 horas) e seu efeito de interação (HC x IT) na porcentagem de eclosão dos ovos de Cyprinuscarpio, a porcentagem de sobrevivência e a porcentagem da produção masculina foram avaliadas especificamente. Os resultados mostraram que a porcentagem de incubação de ovos diminuiu com o aumento da TI da mesma forma, a sobrevivência dos alevinos tratados foi afetada pelo aumento da TI (P 0,001). O principal efeito de interação do HC x IT mostrou que o maior percentual de indivíduos do sexo masculino (95%) foi obtido com 450-600 µgl-1 HC por 72 horas de TI, seguido por 88-92,50% com 150-300 µgl-1 HC para 72 horas de TI, 87,50% às 48 horas de TI para o restante dos tratamentos hormonais, e 47,50% mais baixos foram registrados no controle (P 0,05). A porcentagem aumentada de macho de Cyprinuscarpio foi obtida com o aumento do HC em todas as TIs. Observou-se que o tratamento de imersão a 600µgl-1 por 72 horas foi mais efetivo na alteração da razão sexual do Cyprinuscarpio antes da eclosão. Uma perspectiva comparativa feita a partir deste ensaio experimental de que a indução sexual de Cyprinuscarpio por imersão de ovos usando hormônio esteróide masculino sintético é uma técnica alternativa segura de reversão do sexo em peixes, em contraste com a administração oral de hormônio na alimentação de peixes.

New Record of Aspidogaster ijimai Kawamura, 1913 (Trematoda: Aspidogastridae) from Cyprinus carpio in Korea

Aspidogastrid trematodes (Subclass Aspidogastrea) are a relatively small group with a characteristic adhesive disc and parasitize in a variety of cold-blooded hosts. Until now, only 2 species in the genus Cotylaspis, i.e., C. coreensis and C. sinensis, have been reported as the aspidogastrid trematode in the Republic of Korea (=Korea). In the present study, we intended to describe a species of aspidogastrids collected from the small intestine of the common carp, Cypri-nus carpio, in a faunistic point of view. Total 51 specimens were collected from a carp caught in Nakdong-gang (River) on May 2015. Some of them were prepared as the specimens for light microscopic observations, and some others were prepared for SEM. They were slightly elongated without head lobes, 2,432×840 μm in average size, and had characteristic adhesive discs with 4 rows and 46 alveoli in average. The ovary was reniform and was located in the posterior-upper part of the body. The single testis was larger than the ovary and was located below the ovary. The uterus was coiled containing numerous eggs and distributed in the posterior 2/3 of the body. The vitellaria were follicular, and distributed from the mid-level of testis to near the posterior end. The morphological characters with dimensions of our specimens were closely identical with those of Aspidogaster ijimai previously described. A new aspidogastrid is added among the Korean trematode fauna by the present study.

Avaliação da qualidade da água do Rio dos Sinos (Brasil) por meio do teste de micronúcleos em Cyprinus carpio e de análises físico-químicas e microbiológicas / Evaluation of water quality of the Sinos River (Brazil) by micronucleus test in Cyprinus carpio and physicochemical and microbiological analysis

O Rio dos Sinos está localizado na região sul do Brasil, possui aproximadamente 190 km de extensão e fornece água para atividades agrícolas, industriais e para o consumo de mais de um milhão de habitantes. Este rio é considerado um dos mais poluídos do Brasil e, portanto, estudos para avaliar a qualidade da água ao longo de sua extensão são necessários. O teste de micronúcleos em eritrócitos de peixes tem sido utilizado com sucesso para detectar a presença de poluentes mutagênicos nos ambientes aquáticos. O objetivo do presente estudo foi avaliar a qualidade da água do Rio dos Sinos por meio do teste de micronúcleos em Cyprinus carpio bem como de análises físico-químicas e microbiológicas. Foram coletadas amostras de água nos trechos superior (Caraá), médio (Parobé) e inferior (Novo Hamburgo) do Rio dos Sinos em dezembro de 2013. As amostras foram transportadas para o laboratório para análise de 14 parâmetros de qualidade da água bem como para a exposição de espécimes de C. carpio por 72h em aquários. Não foram verificadas diferenças significativas na frequência de micronúcleos e anormalidades nucleares nos grupos expostos a água do rio em comparação ao grupo controle. Apenas um parâmetro de qualidade da água foi observado em desacordo com a legislação no trecho superior do rio, enquanto que nos trechos médio e inferior foram seis e cinco parâmetros, respectivamente. Os resultados demonstram que o Rio dos Sinos apresenta redução da qualidade da água ao longo de sua extensão e indicam ausência de potencial genotóxico no período amostrado.

The Sinos River is located in South of Brazil, it has about 190 km of extension and provides water for agricultural and industrial activities, and for consumption of more than one million inhabitants. This river is considered one of the most polluted rivers in Brazil; therefore, studies aiming to evaluate the water quality along its course are necessary. The micronucleus test in fish erythrocytes has been successful used to detect mutagenic pollutants in the aquatic environment. The objective of this study was to evaluate the water quality of the Sinos River by the micronucleus test in Cyprinus carpio, as well as physicochemical and microbiological analyses. Water samples were collected in the upper (Caraá municipality), middle (Parobé municipality) and lower (Novo Hamburgo municipality) sections of the Sinos River, in December 2013. The samples were transported to the laboratory for analysis of 14 water quality parameters and for exposure of C. carpio for 72 hours in aquaria. Significant differences in micronucleus and nuclear abnormalities frequencies were not found between the control and the exposed groups. Only one parameter of water quality exceeded the limit of the legislation in the upper section of the river, while in the middle and lower sections, six and five parameters, respectively. These results show that the Sinos River presents a reduction in water quality along its extension and indicates the absence of genotoxic potential in the sampled period.

Immunostimulatory and growth promoting potential of Tinospora cordifolia (Thunb.) Miers on fingerlings of Amur carp.

The enhancement of immunity of fish through application of natural immunostimulants would help in minimizing the risk of occurrence of diseases in aquaculture. The present study was designed to evaluate the growth promoting and immunostimulatory effect of Guduchi, Tinospora cordifolia (Thunb.) Miers ex Hook F. leaf powder in fingerlings of Amur carp (Cyprinus carpio haematopterus Martens). The fingerlings were divided into four treatment groups, T1, T2, T3 and T4. Experimental diets were prepared by mixing rice bran, deoiled mustard cake, soybean meal and vitamin mineral mixture. The leaf powder of T. cordifolia was incorporated into diets D2, D3 and D4 @ 0.25%, 0.5% and 0.75%, respectively. In control diet D1, leaf powder of T. cordifolia was not incorporated. T1 group fishes were fed with D1 diet, T2 with D2, T3 with D3 and T4 with D4 @ 5% body weight per day for 90 days. Fingerlings fed with diet D4 achieved significantly improved Specific Growth Rate (1), Feed Conversion Ratio (2.76), and Gross Conversion Efficiency (0.361) as compared to the control and all other treatments (P <0.05). Hematological and biochemical parameters, total leuckocyte count (53.849×103/µL), total erythrocyte count (3.50×106/µL), hemoglobin concentration (17.17%), total serum protein, albumin and globulin (12.35, 1.56 and 10.78 g/dL) in D4 diet fed fishes showed increase as compared to the control and all other treatments (P <0.05). The above research revealed that Tinospora cordifolia leaf powder has significant growth promoting and immunostimulatory potential in Amur carp raising.

Cinética e morfologia de espermatozoides de carpa comum criopreservados em água de coco em pó acp-104 / Kinetics and morphology of common carp sperm cryopreserved in powdered coconut water acp-104

O objetivo deste estudo foi avaliar o efeito de diferentes taxas de diluição seminal sobre a motilidade, velocidade e presença de anormalidades espermáticas no sêmen descongelado de carpa comum (Cyprinus carpio). Utilizaram-se 20 machos sexualmente maduros, pertencentes ao Departamento Nacional de Obras Contra as Secas (DNOCS). A partir do sêmen coletado, foi formado um total de nove pools. O sêmen de cada pool foi avaliado quanto à motilidade, velocidade e morfologia espermática antes e depois da criopreservação. A criopreservação seminal foi realizada em meio ACP-104 + DMSO 10% diluído em 1:1 ou 1:3 (sêmen:diluidor). As amostras foram envasadas em palhetas de 0,25mL, congeladas em vapor de nitrogênio líquido (caixa de poliestireno) e estocadas em nitrogênio líquido (-196 °C). Não houve diferença significativa (P>0,05) entre as taxas de diluições sobre os parâmetros cinéticos e morfológicos de sêmen descongelado de carpa comum. Entretanto, esses parâmetros apresentaram diferença significativa (P<0,05) em relação ao sêmen fresco (controle). Baixos valores de velocidade foram registrados no sêmen descongelado (VCL 42,6-46,5μm/s; VSL 33,2-37,1μm/s; VAP 38,9-43,2μm/s); contudo, observaram-se motilidades acima de 59% (59,8-67,8%) e baixos índices de anormalidades espermáticas (19,3-19,5%), sugerindo que o ACP pode ser usado como um meio favorável à criopreservação do sêmen de carpa comum em qualquer uma das taxas de diluição testadas.

The aim of this study was to determine the effects of various dilution ratios on thaw sperm motility, velocity and presence of spermatic abnormalities in the common carp. 20 males with three years of age, raised at the Departamento Nacional de Obras Contra as Secas (DNOCS) were used. From the collected semen a total of nine pools were formed. The semen from each pool was evaluated for motility, velocity and spermatic morphology before and after cryopreservation. The sperm cryopreservation was performed in medium ACP-104 + DMSO 10% diluted in 1:1 or 1:3 (sperm:extender). The samples were loaded into 0.25mL straws, frozen in liquid nitrogen vapor (Styrofoam) and stored in liquid nitrogen (-196 °C). There was no significant difference (P>0.05) between the ratios of dilution on the kinetic and morphological parameters of thawed semen from common carp. However, these parameters were significantly different (P<0.05) compared to fresh semen (control). Low velocity values were recorded in thawed semen (VCL 42.6-46.5μm/s; VSL 33.2-37.1μm/s; VAP 38.9-43.2μm/s), however, observed motility above 59% (59.8-67.8%), and low levels of spermatic abnormalities (19.3-19.5%) suggested that the ACP-104 can be used as a suitable medium for cryopreservation of common carp semen in any of the tested dilution ratios.

Estabilidade lipídica de filés de carpa húngara congelados tratados com extratos de Lippia Alba / Lipid stability of frozen common carp fillets treated with Lippia alba extract

O objetivo do presente estudo foi avaliar o efeito da aspersão de extratos de Lippia alba na estabilidade lipídica de filés de carpa húngara armazenados a -18±2°C. Filés não tratados (controle sem aspersão) ou aspergidos (1mL 10g-1 de filé) com água destilada (controle água destilada) ou com extratos de L. alba (0,10g mL-1) hidrometanólico ou aquoso foram analisados durante o armazenamento nos dias zero, 90 e 180. Independente do tempo de congelamento, o extrato hidrometanólico reduziu os valores de dienos conjugados (DC) dos filés em relação aos demais tratamentos, além de reduzir os valores de ácidos graxos livres aos 90 dias (P<0,05). O extrato aquoso resultou em maior teor de peróxidos após 180 dias de congelamento comparado aos demais tratamentos (P<0,05). Os extratos hidrometanólico e aquoso reduziram os valores de substâncias reativas ao ácido tiobarbitúrico (TBARS) após 180 dias, comparados aos filés tratados com água destilada e sem aspersão (P<0,05). Ambos os extratos de L. alba retardaram a oxidação lipídica, sendo que o extrato aquoso retardou a degradação de produtos primários da oxidação lipídica (peróxidos) em produtos secundários (TBARS), enquanto o extrato hidrometanólico parece ser mais eficiente, pois inibiu de forma similar tanto a formação de DC e peróxidos, quanto a sua degradação em produtos secundários (TBARS).

The effect of sprinkling with Lippia alba extracts was evaluated on the lipid stability of common carp fillets stored at -18±2°C. Fillets that received no treatment (no spray control) or that were sprayed (1mL 10g-1 fillet) with distilled water (water control) or with hydro-methanolic or aqueous extract of L. alba (0.10g mL-1) were evaluated immediately (time zero) and after 90 and 180 days. Regardless of the storage time, the hydro-methanolic extract reduced the conjugated dienes (CD) values of fillets compared to the other treatments, and reduced the free fatty acid levels at 90 days (P<0.05). The aqueous extract caused higher peroxide value after 180 days of frozen storage compared to the other treatments (P<0.05). The hydro-methanolic and aqueous extracts reduced thiobarbituric acid reactive substances (TBARS) values after 180 days compared to the non-treated fillets or to the water-sprayed fillets (P<0.05). Both extracts have delayed lipid oxidation. While the aqueous L. alba extract delayed the degradation of primary oxidation products (peroxides) into secondary products (TBARS), the hydro-methanolic extract was more efficient as it inhibited both the CD and peroxide formation and its degradation into secondary products (TBARS).

Fontes energéticas vegetais para juvenis de jundiá e carpa / Plant-energy sources for jundiá and carp juvenile

Para avaliar o desempenho, o metabolismo e a composição corporal de juvenis de jundiá (Rhamdia quelen) e de carpa húngara (Cyprinus carpio) alimentados com fontes energéticas vegetais, foi conduzido experimento de 60 dias. Três dietas compostas por aveia descascada, farelo de arroz desengordurado estabilizado (FADE) ou por farelo de trigo e milho (dieta-controle) foram ofertadas três vezes ao dia a 360 juvenis de jundiá (5,59±0,06g) ou de carpa húngara (5,82±0,14g) distribuídos em 18 tanques (280L), totalizando nove tanques por espécie. Não foram observadas diferenças significativas no desempenho das espécies estudadas. Em relação aos parâmetros metabólicos, as menores concentrações séricas de colesterol total no jundiá foram verificadas nas dietas com aveia (214,49mg/dL) e FADE (216,3mg/dL), enquanto para a carpa húngara as menores concentrações observadas foram nos peixes alimentados com FADE (222,54mg/dL). Quanto à composição centesimal, os valores de gordura no filé foram menores nos jundiás dos tratamentos controle (3,76%) e aveia (3,95%). A inclusão de aveia proporcionou menor deposição de gordura corporal em ambas as espécies. Conclui-se que 20% de aveia descascada ou de farelo de arroz desengordurado podem ser incluídos na dieta de juvenis de jundiá e de carpa húngara sem prejuízos para o crescimento.

In order to evaluate the performance, metabolism and body composition of jundiá (Rhamdia quelen) and common carp (Cyprinus carpio) juvenile fed with plant-energy sources, the experiment was conducted during 60 days. Three diets composed of husked oats, defatted rice bran stabilized (FADE) or wheat bran and corn (control diet) were provided three times daily to 360 jundiá juveniles (5.59±0.06g) or common carp (5.82±0.14g) distributed into 18 tanks (280L), totaling nine tanks per species. There were no significant differences in performance parameters in both species. For metabolic parameters, the lowest serum levels of total cholesterol in jundiá were observed in diets containing oats (214,49mg/dL) and FADE (216,3mg/dL), while for the common carp the lowest levels were observed in fish fed FADE (222,54mg/dL). As for the composition, the fat in the fillet was lower in the jundiá from control (3,76%) and oats (3,95%) treatments, providing the lowest body fat deposition in both species. The inclusion of oats showed lower body fat deposition in both species. It is concluded that the inclusion of 20% stabilized husked oats or defatted rice bran in the diet of jundiá and common carp juvenile does no harm to growth.

Assessment of the toxicity of waste water from a textile industry to Cyprinus carpio.

Static, short-term, acute toxicity tests were performed over a period of 96 hrs using different concentrations of influent and effluent of textile industry waste water with the objective of evaluating their acute toxicity on fresh water fish, Cyprinus carpio (common carp). The LC50 24, 48, 72 and 96 hr of influent and effluent were 25.9, 21.10, 15.66, 11.11% (v/v) and 63.18, 54.89, 48.62, 36.04% (v/v), respectively. The acute toxic unit TUa values for 24, 48, 72, 96 hr for influent and effluent are 3.85, 4.73, 6.38, 8.99 and 1.58, 1.82, 2.05, 2.77, respectively. Correspondingly, the TF was found to be 1, 1.22, 1.65 and 2.33 for influent, and for effluent 1, 1.15, 1.29 and 1.75. Total efficiency of the treatment was 69.16% and the safe concentration of effluent is set to be 3.60%. These data are highly useful in establishing limits of acceptability by the aquatic animals. The need to introduce toxicity evaluation assay for confirming the quality of effluent from the point view of effective environmental safe limits and to ensure integrity of aquatic environment, is stressed.

Structurally conserved C-RFa revealed prolactin releasing activity in vitro and gene expression changes in pituitary of seasonally acclimatized carp

Here we show the cloning and characterization of a novel homolog of prepro C-RFa cDNA from Cyprinus carpió. The deduced preprohormone precursor of 115 amino acids leads to a mature bioactive peptide of 20 amino acids with identical sequence to other teleost C-RFa. Modeling of the mature C-RFa peptide highlighted significant similarity to homologous human PrRP20, specifically the conserved amphipathic system defined by the C-terminal alpha-helix. Clearly, the synthetic C-RFa peptide stimulated prolactin release from primary cultured fish pituitary cells. For the first time, significant variation was shown in C-RFa mRNA and protein levels in the hypothalamus and pituitary between summer- and winter-acclimatized carp. Furthermore, C-RFa protein distribution in carp central nervous tissue was visualized by immunodetection in fibers and cells in hypothalamus, olfactory tract, cerebellum and pituitary stalk. In conclusion, we demonstrated the structure conservation of C-RFa in teleosts and mammals and immunopositive cells and fibers for C-RFa in brain areas. Finally, the increase of C-RFa expression suggests the participation of this hypothalamic factor in the mechanism of modulation in PRL expression in carp.