RESUMO
Abstract Background: Shrimp farming is evolving from semi-intensive to hyper-intensive systems with biofloc technology and water recirculation systems. Objective: To evaluate the transcriptional response promoted by biofloc on shrimp (Litopenaeus vannamei) under a recirculating aquaculture system (RAS). Methods: Quantitative real-time RT-PCR was used to monitor seven key genes related to the immune system in shrimp post-larvae, reared in a RAS with and without biofloc (BF and no- BF). In addition, we present for the first time nucleotide sequences of ADP-ribosylation factor 4 (LvArf4) from Litopenaeus vannamei. Results: Transcripts for penaeidin3 (Pen3), penaeidin4 (Pen4), crustin, and Toll receptor (LvToll) genes were up-regulated between 3 and 24 h in both systems, and tumor necrosis factor receptor-associated factor 6 (TRAF6) in no-BF as an early response. Regarding differential expression between treatments, 13 occurrences were encountered. Nine that were higher in BF than in no-BF and four higher in no-BF than in BF. In some sample times, expression of Pen3, crustin, LvToll, TRAF6, IMD, and LvArf4 was higher in BF than in no-BF and in others, expression of Pen3, Pen4, and TRAF6 was higher in no-BF than in BF. Conclusions: BF modulates the transcription of genes related to the immune response in shrimp as an early response. However, the RAS with no-BF promotes a similar response.
Resumen Antecedentes: Los cultivos de camarón están evolucionando de sistemas semi-intensivos a hiper-intensivos con biofloc y con recirculación. Objetivo: Evaluar la respuesta transcripcional promovida por el biofloc en un sistema acuícola con recirculación (SAR). Métodos: Monitoreamos mediante RT-PCR cuantitativo siete genes relacionados con el sistema inmune en postlarvas de camarón cultivadas en un SAR con y sin biofloc (BF y no-BF). Además, presentamos por primera vez la secuencia de nucleótidos del factor de ribosilación 4 de ADP (LvArf4) de Litopenaeus vannamei. Resultados: Los genes penaeidina3 (Pen3), penaeidina4 (Pen4), Crustina y Toll (LvToll) se sobre-expresaron entre las 3 y 24 h en ambos sistemas, y el factor 6 asociado al factor de necrosis tumoral (TRAF6) en BF como una respuesta temprana. Con respecto a la expresión diferencial entre los tratamientos, se presentaron 13 ocurrencias. Nueve donde el BF fue mayor que sin-BF y cuatro donde el no-BF fue mayor que el BF. La expresión fue más alta en BF que en no-BF en Pen3, Crustin, LvToll, TRAF6, IMD y LvArf4. En contraste, la expresión fue mayor en no-BF en Pen3, Pen4 y TRAF6. Conclusión: el BF modula la transcripción de los genes relacionados con la respuesta inmune en camarón como una respuesta temprana. Sin embargo, el SAR sin-BF promueve una respuesta similar.
Resumo Antecedentes: A criação de camarões está evoluindo de sistemas semi-intensivos para hiper-intensivos como tecnologia de bioflocos e sistemas de recirculação. Objetivo: Avaliar a resposta transcricional promovida pelo biofloco em um sistema de aquicultura recirculante (SAR). Métodos: Utilizamos RT-PCR quantitativo em tempo real para monitorar sete genes-chave relacionados ao sistema imune em pós-larvas de camarão, criados em SAR com e sem bioflocos (BF e no-BF). Além disso, apresentamos pela primeira vez sequências nucleotídicas do fator de ribosilação do ADP 4 (LvArf4) de Litopenaeus vannamei. Resultados: Os resultados mostraram que o Penaeidina3 (PEN3), Penaeidina4 (Pen4), Crustina e Toll genes (LvToll) foram sobre-expressos entre 3 e 24 h em ambos os sistemas, e o Factor de Necrose do Receptor 6 associado e protuberância (TRAF6) no BF como uma resposta precoce. Com relação à expressão diferencial entre tratamentos, 13 ocorrências foram apresentadas. Nove onde o BF foi maior do que os não-BF e quatro onde o não-BF foi maior do que o BF. A expressão foi maior do que em BF não-BF em Pen3, Crustin, LvToll, TRAF6, IMD e LvArf4. Em contraste, a expressão foi mais elevada no não-BF em Pen3, Pen4 e TRAF6. Conclusões: O BF modula a transcrição de resposta imune relacionada no camarão como um genes de resposta precoce. No entanto, o SAR não BF promove uma resposta semelhante.
RESUMO
Molecular analysis of the 16S rDNA of the intestinal microbiota of whiteleg shrimp Litopenaeus vannamei was examined to investigate the effect of a Bacillus mix (Bacillus endophyticus YC3-b, Bacillus endophyticus C2-2, Bacillus tequilensisYC5-2) and the commercial probiotic (Alibio(®)) on intestinal bacterial communities and resistance to Vibrio infection. PCR and single strain conformation polymorphism (SSCP) analyses were then performed on DNA extracted directly from guts. Injection of shrimp with V. parahaemolyticus at 2.5 × 10(5) CFU g(-1) per shrimp followed 168 h after inoculation with Bacillus mix or the Alibio probiotic or the positive control. Diversity analyses showed that the bacterial community resulting from the Bacillus mix had the highest diversity and evenness and the bacterial community of the control had the lowest diversity. The bacterial community treated with probiotics mainly consisted of α- and γ-proteobacteria, fusobacteria, sphingobacteria, and flavobacteria, while the control mainly consisted of α-proteobacteria and flavobacteria. Differences were grouped using principal component analyses of PCR-SSCP of the microbiota, according to the time of inoculation. In Vibrio parahaemolyticus-infected shrimp, the Bacillus mix (~33 %) induced a significant increase in survival compared to Alibio (~21 %) and the control (~9 %). We conclude that administration of the Bacillus mix induced modulation of the intestinal microbiota of L. vannamei and increased its resistance to V. parahaemolyticus.