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La Escuela Especializada en Ingeniería ITCA FEPADE, MEGATEC La Unión, a través de la carrera Técnico en Manejo Integrado de Recursos Costero Marino con especialidad en Acuicultura y Pesquería, realizó esta investigación en asocio con Camaronera Eben Ezer. El proyecto tuvo como objetivo la caracterización de la calidad física, química y biológica del agua del Golfo de Fonseca y el Estero El Chapernal, previo al desarrollo de dos ciclos de cultivo de camarón marino Litopenaeus vannamei, durante el cultivo y antes de las descargas de agua a los efluentes receptores. La metodología se desarrolló en tres fases, de junio a diciembre de 2021. Fase de campo: toma de parámetros físicos, químicos y biológicos en 8 puntos de muestreos, Estero El Chapernal, Golfo de Fonseca y estanques de producción. Fase de laboratorio: se realizó sembrando muestras de agua en medios de cultivo para aislamiento, crecimiento e identificación de bacterias. Tercera fase: procesamiento de datos y análisis de resultados. Se identificaron tres tipos de bacterias: Psudomonas, Vibrios y Heterótrofas. En el primer ciclo de junio a agosto, éstas no reportaron crecimiento, esto pudo deberse a que la transición de la época seca a época lluviosa influyó en los factores físicos y químicos, como la temperatura y el pH. Durante el primer ciclo, la calidad del agua del Estero El Chapernal mantuvo estándares idóneos para el crecimiento del camarón. El segundo ciclo presentó un crecimiento exponencial de las bacterias Heterótrofas y Pseudomonas sobrepasando los límites de referencia establecidos en la normativa vigente. Como parte de la proyección social se efectuó transferencia de conocimientos y tecnología durante la producción, se establecieron mejoras para la operatividad y se les dotó de un protocolo de producción acuícola.
The Escuela Especializada en Ingeniería ITCA FEPADE, MEGATEC La Unión, through the major of Técnico en Manejo Integrado de Recursos Costero Marino con especialidad en Acuicultura y Pesquería, carried out this research in association with Camaronera Eben Ezer. The objective of the project was to characterize the physical, chemical and biological quality of water in Golfo de Fonseca and Estero El Chapernal, prior to the development of two culture cycles of the marine shrimp Litopenaeus vannamei, during the culture and before discharges of water to receiving effluents. The methodology was developed in three phases, from June to December 2021. Field phase: collection of physical, chemical and biological parameters at 8 demonstration points, Estero El Chapernal, Golfo de Fonseca and production ponds. Laboratory phase: it was carried out by planting water samples in culture media for isolation, growth and identification of bacteria. Third phase: data processing and analysis of results. Three types of bacteria were identified: Psudomonas, Vibrios and Heterotrophs. In the first cycle from June to August, growth was not reported, due to the fact that the transition from the dry season to the rainy season influenced in physical and chemical factors, such as temperature and pH. During the first cycle, the water quality of Estero El Chapernal maintained optimal standards for shrimp growth. The second cycle presented an exponential growth of Heterotrophic and Pseudomonas bacteria, exceeding the reference limits established in current regulations. As part of the social projection, knowledge and technology were transferred during production, they were profoundly improved for operability and they were provided with an aquaculture production protocol.
Subject(s)
Water Quality , Aquaculture/methods , Astacoidea/growth & development , Crop Production , PondsABSTRACT
Abstract Background: The white spot syndrome virus (WSSV) causes high mortalities in aquaculture. The use of immunostimulants increases animal resistance. Objective: To evaluate the WSSV infection percentage and the immunostimulant effect of lactic acid bacteria and yeast (MI= microbial immunostimulants) on WSSV infected Litopenaeus vannamei. Methods: A bioassay was performed for 33 d, with treatments in triplicate. The MI was added to the feed at 8.5 mg/kg feed and offered to shrimp (9.9 ± 3.1 g) daily, every 2 days, or every 3 days. Shrimp were infected with WSSV at 9 and 19 days. The expression of four immune system-related genes was studied using qRT-PCR. Results: No significant differences were observed in growth and survival among treatments. At the end of the bioassay, WSSV infection percentage (low viral load) decreased 8.3 and 25% in treatments III and IV as compared to the control group. Treatments with MI showed significant differences in the relative expression of LvToll, transglutaminase, and prophenoloxidase genes when MI was offered daily as compared to the control group. The MI did not regulate the expression of the superoxide dismutase gene. The WSSV infection percentage decreased when feed with MI was offered every 3 d. Conclusion: The MI decrease WSSV infection percentage in L. vannamei infected with low viral load when it is offered every three days. The MI up-regulates LvToll, Tgase, and proPO genes when it is offered daily. Further research is needed regarding prophylactic treatment with microbial immunostimulants against WSSV in commercial shrimp farms.
Resumen Antecedentes: El virus del síndrome de la mancha blanca (WSSV) ocasiona alta mortalidad en acuacultura. El empleo de inmunoestimulantes incrementa la resistencia de los animales. Objetivo: Evaluar el porcentaje de infección de WSSV y el efecto inmunoestimulante de bacterias ácido lácticas y levadura (MI= inmunoestimulantes microbianos) en Litopenaeus vannamei infectado con WSSV. Métodos: Se realizó un bioensayo durante 33 días con tratamientos por triplicado. Se agregó MI al alimento (8,5 mg/kg de alimento), suministrandolo al camarón (9,9 ± 3,1 g) diariamente, cada 2, o cada 3 días. Los camarones se infectaron con WSSV a los 9 y 19 días. Se estudió la expresión de cuatro genes relacionados con el sistema inmune utilizando qRT-PCR. Resultados: No hubo diferencias significativas en el crecimiento y la sobrevivencia entre los tratamientos. Al final del bioensayo, el porcentaje de infección de WSSV (baja carga viral) disminuyó un 8,3 y un 25% en los tratamientos III y IV en comparación con el grupo control. Los tratamientos con MI mostraron diferencias significativas en la expresión relativa de los genes LvToll, transglutaminasa, y profenoloxidasa cuando se suministró MI diariamente en comparación con el grupo control. Los MI no regularon la expresión del gen de la superóxido dismutasa. El porcentaje de infección de WSSV disminuyó cuando el alimento con MI se aplicó cada 3 d. Conclusiones: Los MI disminuyen el porcentaje de infección de WSSV en L. vannamei infectado con baja carga viral cuando se aplican cada tres días. Además, los MI causan una sobre-expresión de los genes LvToll, Tgase y proPO cuando se administran diariamente. Se necesita más investigación sobre los inmunoestimulantes microbianos como tratamiento profiláctico contra WSSV en granjas comerciales de camarón.
Resumo Antecedentes: O vírus da mancha branca (WSSV) causa alta mortalidade na aqüicultura e o uso de imunoestimulantes aumenta a resistência em animais. Objetivo: Este trabalho avaliou a prevalência do WSSV e o efeito imunoestimulante de bactérias ácido lácticas e levadura (MI= inmunoestimulantes microbianos) em Litopenaeus vannamei infectado con WSSV. Métodos: Se realizou um bioensayo durante 33 dias com tratamientos por triplicado. Se agregou MI al alimento (8,5 mg/kg feed) e se o fornecimento de camarão (9,9 ± 3,1 g) diariamente, cada 2 e cada 3 d. Los camarones foram infectados com WSSV a los 9 y 19 d. Se estudió a expressão de quatro genes relacionados com o sistema imune, use qRT-PCR. Resultados: No hubo diferencias significativas no crescimento e na supervisão entre os tratamientos. Al final del bioensayo, a prevalência do WSSV diminuiu um 8.3 e não 25% nos tratamentos III e IV em comparação com o grupo de controle. Os tratamentos com MI mostraram diferenças significativas na expressão relativa dos genes. LvToll, transglutaminasa e prophenoloxidase quando se administraron em diários de MI em comparação com o controle de grupo. Los MI não regularam a expressão do genótipo da superóxido dismutasa. A prevalência do WSSV diminuiu quando o alimento com MI se aplicó cada 3 d. Conclusões: Los IM diminuir a prevalência de WSSV em L. vannamei infectado com baixa carga viral quando aplicável a cada três dias. Além disso, os IM causaram uma sobre-expressão dos genes LvToll, Tgase e proPO quando se administran diariamente. Há mais informações sobre imunoestimulantes microbianos como tratamento profiláctico contra WSSV en granjas comerciales de camarón.
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ABSTRACT The release of wastewater and the shrimp feed cost are the main challenges faced by the shrimp farming industry. An alternative solution to both problems is biofloc production in a unit external to the farm, in an activated sludge system for effluent treatment. The treatment system's influent was composed of the shrimp farm wastewater supplemented with urea and sugarcane molasses. The results show that the average removal of chemical oxygen demand was 71% and the average biofloc production in the reactor was approximately 1.5g.L-1. Adding molasses to the influent contributed to the increase in the quantity and diversity of existing microorganisms that are beneficial to cultured shrimp. The mass balance of nitrogen compounds confirmed that nitrification occurred in the system. Therefore, the use of the activated sludge system is a viable and environmentally suitable alternative to produce bioflocs and shrimp farming effluent treatment.
RESUMO A geração de efluentes e o custo com a alimentação do camarão são os principais desafios enfrentados pela carcinicultura. Uma solução conjunta para ambos os problemas é a produção de bioflocos em um sistema de lodo ativado para tratamento de efluentes. Neste trabalho, o afluente ao sistema estudado era composto pelas águas residuais do cultivo de camarão suplementadas com ureia e melaço de cana-de-açúcar. Os resultados mostraram que a remoção média de demanda química de oxigênio foi de 71% e a produção média de bioflocos no reator foi de aproximadamente 1,5 g.L-1. A adição de melaço ao afluente contribuiu para o aumento da quantidade e da diversidade de microrganismos benéficos para a produção de camarão. Houve remoção de amônia e nitrificação confirmada pelo balanço de massa. Portanto, a utilização do sistema de lodos ativados é uma alternativa viável e ambientalmente adequada para produzir bioflocos e tratar efluentes de cultivo de camarão.
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Abstract Release or escapes of aquaculture organisms may impact the genetic composition and variability of wild populations, leading to diverse issues that may compromise long-term wild stock fitness. Therefore, it is relevant to determine if farmed stocks are currently interacting with wild populations. Shrimp farming is an aquaculture activity taking place along the tropical Pacific coast of the Americas, and represents the most important culture business of Northwestern Mexico. In this study, wild and farmed whiteleg shrimp Litopenaeus vannamei from the State of Sinaloa were genetically evaluated to determine admixture levels. A newly developed set of 14 microsatellite markers (mean number of alleles per locus 11.8, and 0.836 expected heterozygosity) was obtained by Next Generation Sequencing to characterize samples. Sampling consisted of 32 wild shrimps collected during three years (2002, 2012, and 2013) and three different sites, and two hatchery stocks from 2007. No significant differences were observed among years in the wild samples, but cluster analyses showed that hatchery-produced individuals were different from wild specimens. Deviations from Hardy-Weinberg Equilibrium and genotype assignment tests indicated that a fraction from each sample could contain individuals from hatchery origin. Even though the estimated fraction of escaped farmed individuals in the most recent samples (2012-2013; mean = 7.1 %) is considered of low genetic risk, management recommendations for hatcheries and farms were provided. Besides, the reasons that explain the intended and unintended farmed shrimp release into the wild were discussed. Rev. Biol. Trop. 66(1): 381-393. Epub 2018 March 01.
Resumen La liberación o escape de lotes de cultivo pueden impactar la composición y variabilidad genética de las poblaciones silvestres, dando lugar a diversos problemas que pueden comprometer la eficacia biológica a largo plazo. Por lo tanto, es relevante determinar si las poblaciones de cultivo se encuentran actualmente interactuando con las poblaciones silvestres. El cultivo de camarón es una actividad de acuicultura que tiene lugar a lo largo de la costa del Pacífico tropical de América, y es la más importante en el noroeste de México. En este estudio, el camarón blanco Litopenaeus vannameisilvestre y de cultivo proveniente del Estado de Sinaloa, México, fueron evaluados genéticamente para determinar los niveles de mezcla. Se desarrolló un lote de 14 marcadores microsatélites nuevos (número de alelos promedio por locus de 11.8 y heterocigosidad esperada promedio de 0.836), mediante secuenciación de nueva generación, para la caracterización de las muestras. El muestreo consistió en camarón silvestre recolectado durante tres años (2002, 2012 y 2013) y dos lotes de unidades productoras de larva del 2007. No se observaron diferencias significativas entre años en las muestras silvestres, pero el análisis de agrupamiento indicó que los lotes de las unidades productoras de larva fueron distintos a los ejemplares silvestres. Desviaciones del equilibrio de Hardy-Weinberg y los análisis de asignación de genotipos indicaron que una fracción de cada una de las muestras silvestres podría contener individuos originados del larvicultivo. Se discuten las razones que explican la liberación de camarón de cultivo intencional y no intencional al medio silvestre. Aun cuando la fracción estimada de individuos de origen de cultivo en las muestras silvestres más recientes (2012-2013; promedio = 7.1 %) se considera de bajo riesgo, se dan recomendaciones de manejo para unidades de larvicultura y granjas de cultivo.
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En El Salvador el 59.3% del camarón marino de la especie Litopenaeus vannamei se produce en la Bahía de Jiquilisco, departamento de Usulután, el cultivo tiene una duración de 60 a 90 días y el peso de cosecha oscila entre 7 y 12 gramos. El bajo crecimiento y las altas mortalidades son problemas que preocupan a este sector productivo. Por esta razón la Escuela Especializada en Ingeniería ITCA-FEPADE a través de la carrera de Gestión Integral de Recursos Marino Costeros, desarrolló una investigación aplicada orientada a identificar la existencia de parásitos y bacterias del género Vibrio en los cultivos de camarón marino que se desarrollan en la zona de Salinas del Potrero y El Zompopero, así como evaluar su incidencia en el cultivo. Con la investigación se detectó la presencia de cinco géneros de parásitos que están presentes en branquias, intestinos y urópodos en el cultivo de camarón marino en grado 1 y 2; ambas categorías se consideran las etapas iniciales de propagación de los parásitos. También se evidenció a través del análisis bacteriológico en el medio de cultivo TCBS, la presencia de bacterias del genero Vibrio en agua, sedimento y camarón.
In El Salvador, 59.3% of the marine shrimp of the Litopenaeus vannamei species is produced in the Bay of Jiquilisco, department of Usulután, the culture lasts from 60 to 90 days and the harvest weight ranges between 7 and 12 grams. Low growth and high mortality are problems that concern this productive sector. For this reason, Escuela Espcializada en Ingeniería ITCA-FEPADE , through the major of Gestión Integral de Recursos Marino Costeros, developed an applied research aimed at identifying the existence of parasites and bacteria of the genus Vibrio in marine shrimp cultures that develop in the area of ââSalinas del Potrero and El Zompopero, as well as evaluating their impact on the crop. The investigation detected the presence of five types of parasites that are present in gills, intestines and uropods in marine shrimp culture in grade 1 and 2; Both categories are considered the initial stages of parasite spread. The presence of Vibrio bacteria in water, sediment and shrimp is also evidenced through bacteriological analysis in the TCBS culture medium.
Subject(s)
Vibrio , Astacoidea/parasitology , Parasites , Bacteria , Bacteriological Techniques , Aquaculture , BaysABSTRACT
The present study aimed to evaluate the mortality, reactive oxygen species production (ROS) and total hemocyte counts (THC) of the marine shrimp Litopenaeus vannamei infected with the white spot syndrome virus (WSSV) at three levels of oxygen saturation. For this, 360 shrimp (20±2g) were distributed in 24 tanks (60L), divided in two groups (infected and non-infected), which were subjected to 30, 60 and 100% of dissolved oxygen saturation (in quadruplicate). During 96 hours after infection, daily hemolymph samples were collected for hemato-immunological parameter evaluation (THC and ROS) and dead animals were removed and computed to assess cumulative mortality rates. In the infected group, animals subjected to 100% saturation showed higher ROS production (P<0.05) after 48 hours, while THC was significantly reduced (P<0.05), regardless of oxygen saturation. The hypoxia resulted in high mortality when compared to 100% saturation condition. In the uninfected group, no significant differences were observed in all evaluated parameters. Thus, the hypoxia condition increased the susceptibility of shrimp to the infection of WSSV, which may be partly related to the low ROS production showed by the animals subjected to 30% oxygen saturation.
O presente estudo teve por finalidade avaliar a mortalidade e a contagem total de hemócitos (CTH) e espécies reativas de oxigênio (EROs) de camarão Litopenaeus vannamei infectados com o vírus da mancha branca (WSSV) e submetidos a três níveis de saturação de oxigênio. Para tanto, 360 camarões (20±2g) foram distribuídos em 24 tanques (60L), divididos em dois grupos, infectados e não infectados e submetidos a 30, 60 e 100% de saturação de oxigênio (em quadruplicata). Após a infecção, diariamente foram coletadas amostras de hemolinfa dos animais para avaliação dos parâmetros hematoimunológicos (CTH e EROs) e foi estimada a mortalidade, por 96 horas. No grupo com infecção, os animais submetidos à saturação de 100% apresentaram um aumento na produção de EROs (P<0,05) após 48 horas, ao mesmo tempo em que a CTH demonstrou uma redução (P<0,05) independentemente da saturação do oxigênio, e a condição de hipóxia acarretou maiores mortalidades quando comparada à do grupo com 100% de saturação. No grupo sem infecção, não foram observadas diferenças significativas nos parâmetros avaliados nem mortalidade. Dessa forma, pode-se concluir que a hipóxia aumentou a susceptibilidade do camarão à infecção com o vírus da mancha branca, que pode estar, em parte, relacionada com a baixa contagem de hemócitos e produção de EROs observadas nos animais submetidos a essa condição.
Subject(s)
Animals , Artemia , Hypoxia/veterinary , Hemocytes , White spot syndrome virus 1/pathogenicity , Hemolymph/immunology , Oxygen ConsumptionABSTRACT
El sector El Zompopero ubicado en la Bahía de Jiquilisco Usulután, es un núcleo productivo integrado por cinco cooperativas que se dedican al cultivo de camarón blanco de la especie Litopenaeus vannamei. Los miembros que integran las cooperativas son familias desmovilizadas de la guerra civil de El Salvador, a quienes se les repartieron tierras en esta zona costera como estrategia de reinserción a la vida productiva. Cuentan con un total de 21 estanques de tierra, que cubren un área de cultivo de 84.3 hectáreas y realizan tres ciclos de cultivo de camarón al año. Actualmente este sector productivo enfrenta serios problemas debido a la alta mortalidad que se registra en cada ciclo productivo. Por esta razón, la Escuela de Ciencias del Mar del Centro Regional MEGATEC La Unión realizó un proyecto de investigación aplicada para diagnosticar las principales causas que generan la mortalidad en los cultivos, así como el impacto económico y social que perciben las familias que se dedican a la producción de camarón. A través del diagnóstico se identificaron 5 causas externas y 15 internas que afectaron la producción en las granjas camaroneras en el año 2014. Las causas están asociadas al manejo del cultivo y a las condiciones de infraestructura de los estanques en las cooperativas
El Zompopero zone located in Jiquilisco Bay in Usulután, is a productive center formed by five farm cooperatives dedicated to the white shrimp farming, Litopenaeus vannamei species. The members forming the cooperatives are demobilized families from the civil conflict in El Salvador who were distributed lands in this coastal area as a reinsertion into productive life strategy. They currently have a total of 21 earth ponds with a farming area of 84.3 hectares that allow three shrimp farming cycles a year. At the moment, this productive area is facing serious problems due to the high mortality rate registered every productive cycle. Thy is why, the Marine Science School at the regional Centre MEGATEC in La Unión, implemented an applied research project to identify the main reasons causing farming mortality, as well as the economic and social impact in the families dedicated to shrimp farm production. The research identified 5 external causes and 15 internal ones that affected shrimp farm production in the year 2014. These causes are related to farming handling and to the conditions of the ponds infrastructure in the cooperatives.
Subject(s)
Coasts , Astacoidea/growth & development , Vibrio Infections , Aquaculture , Bays , Applied ResearchABSTRACT
O objetivo do presente trabalho foi avaliar métodos de controle e tratamento de bacterioses na larvicultura de Litopenaeus vannamei, avaliando o uso contínuo de probiótico e o uso pontual de enrofloxacino 15mg.L-1 e propionato de sódio 0,5 mM.L-1 nos momentos de metamorfose sobre os, parâmetros zootécnicos e microbiológicos (larvas e da água). Foram utilizadas 16 unidades de 60L, povoadas na densidade de 325 náuplios.5L-1, divididos em três tratamentos: enrofloxacino, probiótico, propionato de sódio e controle. O enrofloxacino e o propionato foram ministrados em protozoea 3, misis 3 e pós-larva 4 e o probiótico foi ministrado na ração ao longo de todo o experimento. O probiótico aumentou as contagens de bactérias ácido-láticas em relação aos demais tratamentos na água de cultivo (p=0,00001) e em relação ao enrofloxacino e o proprionato nas larvas (p=0,0048). A água do tratamento com probiótico apresentou menor contagem de Vibrio spp. que o enrofloxacino (p=0,0011) e as larvas tratadas com probiótico apresentaram menor contagem de Vibrio spp. que o propionato (p=0,0158). Contudo, não foi observada diferença nos índices zootécnicos avaliados. Assim, os aditivos na dose utilizada não alteram parâmetros zootécnicos da larvicultura do camarão L. vannamei.
The purpose of this study was to assess control and treatment methods for bacterial diseases in Litopenaeus vannamei, evaluating the continuous use of probiotics and occasional use of 15mg-L-1 enrofloxacin and 0.5-mM.L-1 sodium propionate at the morphological change moments on the performance and microbiological parameters of larvae and water. A total of 16 60-L units were used, stocked with 325 nauplii/5L-1, divided into three treatments and one control. Enrofloxacin and propionate were administered into protozoea 3, misis 3 and 4, and post-larvae 4, while the probiotic was administered in the feed throughout the experiment. The probiotic increased the counts of lactic acid bacteria in relation to the other treatments in the culture water (p = 0.00001) and in relation to enrofloxacin and propionate in larvae (p = 0.0048). The treatment water with probiotic had lower counts of Vibrio ssp. than enrofloxacin (p = 0.0011) and larvae treated with probiotic showed lower counts of Vibrio ssp. that propionate (p = 0.0158). However, no difference was observed in the performance indexes assessed. Thus, it can be concluded that additives in the assessed doses did not influence the performance parameters of L. vannamei.
El objetivo del presente trabajo fue evaluar métodos de control y tratamiento de enfermedades bacterianas en larvicultura de Litopenaeus vannamei, evaluando el uso continuo de probiótico y el uso puntual de 15 mg.L-1 de enrofloxacino y 0,5 mM.L-1 de propionato de sodio en momentos de metamorfosis sobre los parámetros zootécnicos y microbiológicos (larvas y del agua). Se han utilizado 16 unidades de 60L, pobladas en la densidad de 325 nauplios.5L-1, divididos en tres tratamientos: enrofloxacino, probiótico, propionato de sodio y control. El enrofloxacino y el propionato fueron suministrados en protozoea 3, misis 3 y postlarva 4 y el probiótico suministrado en alimento durante el transcurso del experimento. El probiótico aumentó el contaje de bacterias ácido-lácticas en correlación a los demás tratamientos en agua de cultivo (p=0,0001) y las larvas en relación al enrofloxacino y el propionato en las larvas (p=0,0048). El agua del tratamiento con probiótico presentó menor contaje de Vibrio spp. que el enrofloxacino (p=0,0011) y las larvas tratadas con probiótico presentaron menor contaje de Vibrio spp. que el propionato (p=0,0158). Sin embargo, no se ha observado diferencia en los índices zootécnicos de larvicultura de camarón L. vannamei.
Subject(s)
Animals , Penaeidae/microbiology , Penaeidae/virology , Vibrio Infections/prevention & control , Disease Prevention , PropionatesABSTRACT
Objective:To identify enolase,47 kD allergen,from Litopenaeus vannamei by Mass spectrometry. Methods: The proteins were extracted from Litopenaeus vannamei tissue with acetone precipitation method. The protein components were analyzed by SDS-PAGE and Western blot. By using Matrix-Assisted Laser Desorption/Ionization time of flight mass spectrometry ( MALDI-TOF/TOF-MS) ,the 47 kD allergen from Litopenaeus vannamei was identified as enolase. Results:By SDS-PAGE,we proved that the native protein components from Litopenaeus vannamei were completely. According to the Western blot result more than 14 components could react with the positive serum. MALDI-TOF/TOF analysis results showed that the suspected proteins were enolase. A sensitization frequency was 55%. Conclusion:Enolase was identified as a new allergen of Litopenaeus vannamei.
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In shrimp farming, screening for economically significant viral pathogens in nucleic acids of shrimps is vital for disease surveillance programmes and further, to take necessary precautions to ensure the sustainability of the farms and thereby the shrimp industry. Different preservatives, temperature and storage durations of the pleopod tissues of Penaeus vannamei broodstock were tested to investigate its effect on the quality and quantity of the nucleic acids. The pleopods were subjected to two preservation regimes and the yield and stability of the extracted nucleic acids were monitored over a time period of 12 months. Stability of the nucleic acids was assessed with nested polymerase chain reaction, and the yield was checked spectrophotometrically. Data was analysed by performing two way ANOVA and Tukeys Paired test. Preservation treatments included storage at −20°C and 5°C in RNAlaterTM and in 70 % ethanol. Significant variation (P<0.05) was observed in both DNA and RNA yield and stability from ethanol and RNAlaterTM stored pleopods at 5°C. However, the yield and stability did not differ (P >0.05) in both the preservatives at −20°C. The RNA was degraded and yielded lesser quantity when pleopod tissues were stored in ethanol at −20°C than when stored in RNAlaterTM during storage duration of 9 months. This study would help the shrimp farmers and researchers to adopt better preservation strategy, vital for shrimp disease surveillance programmes and for traceability studies in the event of any disease outbreak.
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AbstractThe aim of this study was to evaluate the influence of different levels of alkalinity for the superintensive cultivation of marine shrimp Litopenaeus vannamei in biofloc system. A total of 12 experimental circular units of 1000L were used supplied with 850L water from a nursery, populated at a density of 165 shrimps.m–3 and average weight of 5.6 g. The treatments, in triplicate, consisted in four levels of alkalinity in the water: 40, 80, 120 and 160 mg.L–1 of calcium carbonate. To correct the alkalinity was used calcium hydroxide (CaOH). It was observed a decrease in pH of the water in the treatments with lower alkalinity (p<0.05). The total suspended settleable solids were also lower in the treatment of low alkalinity. No significant difference was observed in other physico-chemical and biological parameters in the water quality assessed, as well as the zootechnical parameters of cultivation between treatments (p≥0.05). The results of survival and growth rate of shrimps were considered suitable for the cultivation system used in the different treatments. The cultivation of marine shrimp Litopenaeus vannamei in biofloc at density of 165 shrimps.m–3 can be performed in waters with alkalinity between 40 and 160 mg.L–1 of CaCO3, without compromising the zootechnical indexes of cultivation.
ResumoO objetivo deste trabalho foi avaliar a influência de diferentes níveis de alcalinidade para o cultivo do camarão marinho Litopenaeus vannamei em sistema superintensivo em bioflocos. Foram utilizadas 12 unidades experimentais circulares de 1000L abastecido com 850L de água provenientes de um berçário intensivo, povoadas a uma densidade de 165 camarões.m-3 e peso médio 5,6g. Os tratamentos em triplicata consistiram de quatro níveis de alcalinidade na água: 40, 80, 120 e 160 mg.L–1 de carbonato de cálcio. Para correção da alcalinidade, foi utilizado cal hidratada (CaOH). Foi observado um decréscimo no pH da água nos tratamentos de menor alcalinidade (p<0,05). Os sólidos suspensos sedimentáveis totais também foram menores nos tratamentos de menor alcalinidade. Não foi observada diferença significativa nos demais parâmetros físico-químicos e biológicos de qualidade de água avaliados, assim como nos parâmetros zootécnicos do cultivo entre os tratamentos (p≥0,05). Os resultados de sobrevivência e taxa de crescimento dos camarões foram considerados adequados para o sistema de cultivo utilizado nos distintos tratamentos. O cultivo do camarão marinho Litopenaeus vannamei em bioflocos na densidade de 165 camarões.m–3 pode ser realizado em águas com alcalinidade entre 40 a 160 mg.L–1 de CaCO3, sem comprometer os índices zootécnicos do cultivo.
Subject(s)
Animals , Calcium Carbonate/analysis , Penaeidae/growth & development , Seawater/chemistry , Water Quality , AquacultureABSTRACT
The objective of this study was to identify the toxigenic mycobiota and the occurrence of aflatoxins in shrimp feed products intended for shrimp cultivated in the coastal area of the state of Piauí, Brazil, in three farms ("A", "B" and "C"). The toxigenic capacity of the fungal species isolated was tested for aflatoxins (AF) and ochratoxin A production. The fungal counts of shrimp feed were similar for the "A" and "B" farms at all cultivation phases, collection sites, in closed and opened packages (1.33 to 2.66CFU g-1 log10 -1). The lowest fungal counts were found in feed from "C" farm (0.65CFU g-1 log10 -1) from closed packages. Thirty-four strains of Aspergillus were detected with a greater prevalence of A. flavus. Two strains produced B1, B2, G1 and G2 aflatoxins at concentrations from 0.39 to 0.42ng g-1; 0.18 to 0.27ng g-1; 1.78ng g-1 and 0.09ng g-1 respectively and were classified as atypical A. flavus. Two strains of A. niger aggregate were OTA producers. Fifteen samples (13.88%) presented AFB1 contamination at levels ranging from 0.25ng to 360ng g-1. This study demonstrates the presence of toxigenic fungi in shrimp feed used at different phases of cultivation and farms. Atypical strains of A. flavus were isolated which produced AF B1, B2, G1 and G2 in shrimp feeds. Only AFB1 was detected in the analyzed feed.
O objetivo deste estudo foi identificar a micobiota toxigênica e a incidência de aflatoxinas em rações comerciais para camarão cultivado no litoral do Estado do Piauí, Brasil, em três fazendas ("A", "B" e "C"). Foi realizada a capacidade toxigênica das espécies de fungos isolados e a produção de aflatoxinas (AF) e ocratoxina A (OTA). As contagens fúngicas da ração foram semelhantes nas fazendas "A" e "B" e em todas as fases de cultivo, locais de coleta e de embalagens fechadas e abertas (1,33-2,66UFC g-1 log10 -1). As mais baixas contagens de fungos foram encontradas nas rações de embalagens fechadas da fazenda "C" (0,65UFC g-1 log10 -1). Foram isoladas trinta e quatro cepas de Aspergillus com maior prevalência de A. flavus e duas linhagens eram produtoras de aflatoxinas B1, B2, G1 e G2 em concentrações 0,39-0,42ng g-1; 0,18-0,27ng g-1; 1,78ng g-1, e 0,09ng g-1, respectivamente, e foi classificado como A. flavus atípico, sendo necessária posteriormente a classificação filogenética desta cepa. Duas cepas de A. niger agregados eram produtoras de OTA. Quinze amostras de ração (13,88%) apresentaram contaminação AFB1 em níveis que variam de 0,25ng a 360ng g-1. Este estudo demonstra a presença de fungos toxigênicos em rações de camarão nas fazendas analisadas e nas diferentes fases de cultivo. Foram isoladas, em rações de camarões, cepas atípicas de A. flavus, produzindo AF B1, B2, G1 e G2. Apenas AFB1 foi detectada na ração analisada.
ABSTRACT
Objective:To obtain purified recombinant Litopenaeus vannamei allergen protein Lit v 1.2.Methods: The target gene of Lit v 1.2 was inserted into clone vector pGEM-T and then ligated to the expression vector pET 44a.The pET44a-Liv 1.2 was transformed into Rosetta and screened by ampicillin resistance .The recombinant protein was expressed by IPTG induction .The protein was purified by 6-His tag affinity chromatography and the purification was analyzed by SDS-PAGE gel electrophoresis .Results:The ex-pression plasmid pET44a-Lit v 1.2 was constructed.SDS-PAGE showed that expressed Lit v 1.2 was efficient and soluble in E.coli Rosetta.The protein molecular weight was consistent with the theoretical value .The highly purified target protein was obtained.Conclusion:In this study ,we successfully gained highly purified recombinant allergen protein Lit v 1.2 which was expressed in prokaryotic system and purified by affinity chromatography column .The purified Lit v 1.2 protein will facilitate us to further study its role in immunological responses .
ABSTRACT
Objective: To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host–pathogen interaction model.Methods:ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug.Results:Thirty species of marine plants were subjected to Soxhlet extraction using water, formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection.Conclusions:Nine plant isolates exhibited significant survivability in host. The drug MP07X thus The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug.
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Objective: To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host–pathogen interaction model.Methods:Thirty plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti–WSSV property in Litopenaeus vannamei. The best anti–WSSV plant isolate, TP22C was isolated and further analyzed. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug.Results: Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection.Conclusions:The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug.
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<p><b>OBJECTIVE</b>To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host-pathogen interaction model.</p><p><b>METHODS</b>Thirty species of marine plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug.</p><p><b>RESULTS</b>Nine plant isolates exhibited significant survivability in host. The drug MP07X thus formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1 000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection.</p><p><b>CONCLUSIONS</b>The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug.</p>
ABSTRACT
<p><b>OBJECTIVE</b>To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host-pathogen interaction model.</p><p><b>METHODS</b>Thirty plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. The best anti-WSSV plant isolate, TP22C was isolated and further analyzed. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug.</p><p><b>RESULTS</b>Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection.</p><p><b>CONCLUSIONS</b>The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug.</p>
ABSTRACT
White spot syndrome virus (WSSV), is the most contagious pathogen of cultured shrimp that causes mass mortality, leading to huge economic loss to the shrimp industry. The lack of effective therapeutic or prophylactic measures has aggravated the situation, necessitating the development of antiviral drugs. With this objective, the antiviral activity of the drug, (MP07X -derived from the marine plant) in the host, Litopenaeus vannamei was evaluated. The biochemical changes aggravated by WSSV in the host, and the in vivo efficacy of the drug in the host – pathogen interaction were analyzed. The survival percentage of the treated (with MP07X) WSSV infected host was 85 %. Significant results were obtained from the cytotoxicity assays of the drug in both the brine shrimp and host. A total of 9 biochemical parameters such as, total protein, total carbohydrate, total glucose, total free amino acid, total fatty acid, fructose 1, 6 diphosphatase, aldolase, glucose 6 phosphatase and glucose 6 phosphate dehydrogenase were examined for healthy (NEG), WSSV infected (POS) and test sample (TS) shrimps. Significant differences (p < 0.01) were observed between the POS, NEG and TS in the biochemical variables at different time intervals post infection with WSSV. In the case of POS, significantly (p < 0.01) reduced variables were observed when compared to the NEG. In contrast, significant (p < 0.01) elevations were observed in the TS after a certain time interval due to the anti-WSSV activity of MP07X. Neither the VP 28 gene nor the immediate early genes (ie 1) were expressed in the host at the 42nd and 84th hrs. Thus, in accordance with the above results it can be concluded that acute WSSV infection triggers alterations in biochemical parameters in L. vannamei and at the same time the drug is efficient enough to combat the deadly virus and can increase the survivability of the host.
ABSTRACT
Whiteleg shrimp, Litopenaeus vannamei is one of the most commercially farmed species worldwide because of its fast growth, good survival rate at high farming densities, and osmoregulatory capacity, which makes it an excellent candidate for cultures at different salinities. The knowledge of shrimp nutritional requirements is critical in the formulation of diets to allow optimal growth at different environmental conditions and development stages. The effect of salinity on apparent digestibility of shrimp feed is not well known, and this information is required in shrimp diet formulation. For this purpose, the apparent digestibility coefficients of carbohydrates (ACD) and lipids (ALD) were determined for juvenile whiteleg shrimps under controlled culture conditions. We evaluated the apparent digestibility of six commercial (D1:37CP, D2:38CP, D3:39CP, D4:34CP, D5:35CP, and D6:37CP) and two experimental (E1:33CP and E2:33CP) diets for juvenile whiteleg shrimp cultivated at three salinities (5, 35 and 50psu) in 60L aquariums. ACD and ALD were determined in vivo using chromic oxide as an inert marker. Our results showed that ALD in most cases was over 80%, independent of salinity, except the E1:33CPdiet which had 74.0% at 50psu. Diet D3:39CP showed the highest ALD coefficient (90.1 and 90.6% at 5 and 35psu, respectively). For ACD, differences were detected between commercial and experimental diets at every salinity level, although salinity effect on ACD was not significant. Diet D4:34CP had the highest coefficient (92.4%) at 5psu, and E2:33CP at 35 and 50psu (97.3 and 94.7%). This study demonstrated that there is no significant effect of saline variations on carbohydrate and lipid digestibility by juvenile whiteleg shrimp, under the experimental conditions. Rev. Biol. Trop. 61 (3): 1201-1213. Epub 2013 September 01.
El camarón blanco, Litopenaeus vannamei, es una de las especies más cultivadas comercialmente en el mundo, debido a su velocidad de crecimiento y tasa de supervivencia en altas densidades de cultivo, y su capacidad de osmoregulación, que lo hacen un excelente candidato para cultivo en diferente salinidades. El conocimiento de los requerimientos nutricionales del camarón es fundamental en la formulación de dietas que permita el crecimiento óptimo en diferentes condiciones ambientales y fases de desarrollo del animal. El efecto de la salinidad sobre la digestibilidad aparente de alimentos comerciales para camarones no está documentado. Esta información es necesaria en el cultivo de camarón para la formulación de los alimentos comerciales. Se determinó la digestibilidad aparente de los carbohidratos (ACD) y lípidos (ALD), en juveniles del camarón blanco L. vannamei cultivado en condiciones controladas. Los tratamientos fueron seis dietas comerciales (D1:37CP, D2:38CP, D3:39CP, D4:34CP, D5:35CP y D6:37CP) y dos dietas experimentales (E1:33CP y E2:33CP) para juveniles de camarón blanco en cultivo, a tres salinidades (5, 35 y 50ups) en acuarios de 60L. Los coeficientes de digestibilidad aparente de los carbohidratos (ACD) y los lípidos (ALD) fueron determinados in vivo utilizando óxido crómico como marcador inerte en la dieta. ALD en la mayoría de los tratamientos fue superior al 80%, independientemente de la salinidad, con excepción de la dieta E1:33CP que presentó 74.0% a salinidad de 50ups. La dieta D3:39CP presentó el mayor coeficiente ALD (90.1 y 90.6 en 5 y 35ups, respectivamente). En ACD, se presentaron diferencias entre dietas comerciales y experimentales en cada nivel de salinidad, aunque la salinidad no presentó un efecto significativo. La dieta D4:34CP presentó el coeficiente de digestibilidad más alto (92.4%) en la salinidad de 5ups, mientras que E2:33CP en salinidades de 35 y 50ups fue de 97.3 y 94.7%, respectivamente. En general, el presente estudio demuestra que en juveniles de camarón blanco la variación de salinidad no afecta significativamente la digestibilidad de lípidos y carbohidratos en las condiciones experimentales de este estudio.
Subject(s)
Animals , Animal Feed/analysis , Aquaculture/methods , Dietary Carbohydrates/metabolism , Lipid Metabolism , Penaeidae/physiology , Penaeidae/classification , SalinityABSTRACT
The use of probiotics has gained acceptance in aquaculture, particularly in maintaining water quality and enhancing growth in organisms. This study analyzed the effect of the commercial (EM TM, Japan) natural product composed by (Rhodopseudomonas palustris, Lactobacillus plantarum, Lactobacillus casei and Saccharomyces cerevisiae) added to the water, in order to determine its effect in water quality, sediment and growth of L. vannamei under intensive culture. The evaluation included three treatments with a weekly addition of EM: i) tanks without probiotics (C), ii) tanks with a dose of 4L/ha (EM1) and iii) tanks with a dose of 10L/ha (EM2). The treatment C was carried out three times, while treatments EM1 and EM2 were carried out four times. A total of 4 350 shrimps were measured for total length and weight, to calculate total and porcentual weight gain, daily weight gain, specific growth rate (TCE), and food conversion factor (FCA); besides, the survival rate was estimated. The use of probiotics allowed a shorter harvest time in treatments EM1 (90d) and EM2 (105d) with relation to the treatment C (120d). Treatments EM1 and EM2 were within the recommended intervals for culture, with respect to treatment C. The use of probiotic bacteria significantly regulated pH (EM1, 8.03±0.33; EM2, 7.77±0.22; C, 9.08±0.35) and reduced nitrate concentration (EM1, 0.64±0.25mg/L; EM2, 0.39±0.26mg/L; C, 0.71mg/L). Water pH mostly explained the variance with respect to the treatments. Treatment EM2 presented the greatest removal of organic matter (1.77±0.45%), whereas the contents of extractable phosphorus increased significantly in treatment EM1 with 21.6±7.99mg/kg and in treatment EM2 with 21.6±8.45mg/kg with control relation (14.3±5.47). The shrimp growth was influenced by dissolved oxygen, salinity and pH in the sediment, establishing that salinity was the most important variable in the weight with a negative association. Treatment EM1 recorded an improved TCE (2.69±0.35%/d) and FCA (1.46±0.20) with relation to the control treatment (TCE, 1.88±0.25%/d; FCA, 2.13±0.48). Survival was significantly greater in treatments containing probiotics with 61±8.76% and 60±10.5% for EM1 and EM2, respectively. This study indicated the positive effect obtained with the use of this commercial probiotic, to improve culture conditions and growth parameters in an intensive culture of L. vannamei.
Los probióticos han ganado aceptación en la acuicultura para mantener la calidad del agua y aumentar el crecimiento de los organismos. En este estudio se analizó el efecto de una mezcla comercial de microorganismos eficientes (EM) (Rhodopseudomonas palustris, Lactobacillus plantarum, Lactobacillus casei y Saccharomyces cerevisiae) sobre la calidad del agua, sedimento y el crecimiento en un cultivo intensivo de camarón L. vannamei. La evaluación consistió en tres tratamientos: i) estanques sin EM (C), ii) estanques con dosis de 4L/ha (EM1) y iii) estanques con dosis de 10L/ha (EM2). Los resultados demostraron menor tiempo de cosecha en los tratamientos EM1 (90d) y EM2 (105d). Los tratamientos EM1 y EM2 mantuvieron significativamente regulados los valores del pH (EM1, 8.03±0.33; EM2, 7.77±0.22) y redujeron las concentraciones de nitrato (EM1, 0.64±0.25mg/L; EM2, 0.39±0.26mg/L). El tratamiento EM2 presentó la mayor remoción de materia orgánica (1.77±0.45%). El tratamiento EM1 mejoró la TCE (2.69±0.35%/d) y FCA (1.46±0.20). Los tratamientos EM1 y EM2 presentaron mayor supervivencia con 61±8.76% y 60±10.5%, respectivamente. Este estudio demostró el efecto benéfico del uso de la mezcla comercial en los parámetros ambientales y de crecimiento en un cultivo intensivo de L. vannamei.