池塘养殖牙鲆肠道和环境菌群结构对益生菌制剂的响应

Responses of Microbiota Structure in the Intestinal Tract and Pond Culture Environment of Japanese Flounder (Paralichthys olivaceus) to Probiotics

中文关键词: 高通量测序  肠道菌群  牙鲆  池塘养殖  益生菌制剂

英文关键词: High throughput sequencing  Intestinal tract microbiota  Paralichthys olivaceus  Pond culture  Probiotics

中文摘要:

      为研究益生菌制剂对池塘养殖牙鲆(Paralichthys olivaceus)肠道及环境菌群结构的调控效果，采用高通量测序技术和生物信息学分析手段构建牙鲆肠道、养殖水体、饵料和池塘底泥的16S rDNA基因测序文库，分析不同样品中菌群组成和多样性在益生菌制剂调控过程中的变化趋势。结果显示，添加益生菌制剂后，池塘底泥和牙鲆肠道的菌群多样性升高，且池塘底泥的菌群多样性依然最高；而养殖水体的菌群多样性明显下降，并低于牙鲆肠道的。牙鲆肠道中的肠杆菌属(Enterobacter)、芽孢杆菌属(Bacillus)相对丰度呈上升趋势，不动杆菌属(Acinetobacter)、发光杆菌属(Photobacterium)相对丰度先上升后下降；池塘养殖水体中NS3a_marine_group代表的菌属相对丰度先下降后上升；底泥中芽孢杆菌属相对丰度变化最为明显，由最初的3.78%增加到33.64%。养殖牙鲆肠道、养殖水体和底泥中的弧菌属(Vibrio)相对丰度在益生菌制剂添加后出现不同程度的降低。而在水产养殖中，不动杆菌属和弧菌属中的部分菌株通常被认为是重要病原菌。说明益生菌制剂的添加能在一定程度上优化鱼体肠道和环境(养殖水体和池塘底泥)的菌群结构。相似性分析发现，在饵料不变的条件下，牙鲆肠道菌群结构与底泥的更相近；且益生菌产品对池塘底泥和牙鲆肠道菌群的影响较为明显。本研究结果可为池塘养殖过程中微生态制剂筛选和使用提供参考。

英文摘要:

      The deterioration of the aquaculture environment and disease outbreaks occur often in high-density pond cultures of fish. Probiotics are the main and efficient method to regulate the microbiota of the fish intestinal tract and pond water environments, which will improve aquaculture conditions and reduce the incidence of diseases. To investigate the effects of probiotics on the composition and diversity of microbiota in the intestinal tract and pond culture environment of the Japanese flounder (Paralichthys olivaceus), samples from the Japanese flounder intestinal tract and environmental media (culture water, feed, and pond sediment) were collected. High-throughput sequencing of the 16S rDNA genes was performed to analyze the regulation of the microbiota of the intestinal tract and environmental samples by probiotics. Our results showed that the diversity of microbiota in the pond sediment and intestinal tract of Japanese flounder increased after the addition of probiotics, and was still in the pond sediment. Inversely, the diversity of the microbiota in the culture water declined, which was lower than that in the intestinal tract. Firmicutes, Proteobacteria, and Fusobacteria were dominant in the Japanese flounder intestinal tract, and the relative abundances of Firmicutes and Proteobacteria increased, whereas Fusobacteria decreased after the addition of probiotics. At the genus level, the relative abundances of Enterobacter and Bacillus increased, whereas Acinetobacter and Photobacterium first increased and then decreased in the intestinal tract. The relative abundance of genus represented by NS3a_marine_group firstly decreased and then increased in the Japanese flounder culture water. In the pond sediment, the change in the relative abundance of Bacillus was obvious, increasing from 3.78% to 33.64%. The relative abundances of Vibrio in the fish intestinal tract, culture water, and pond sediment decreased after the addition of probiotics. Some strains of Acinetobacter and Vibrio are important pathogens in aquaculture. The microbiota structure in the intestinal tract of Japanese flounder was similar to that in the pond sediment. Changes in the relative abundances of these main microbiota indicated that the addition of probiotics could optimize the microbiota structure of the Japanese flounder intestinal tract and environment. However, it is impossible that Lactobacillus, as the main microbiota of the probiotics, was able to colonize and persist in the fish intestinal tract, water, and pond sediment, which indicated that probiotics did not deliver bacteria but their probiotic function. Hence, the addition of probiotics could affect the microbiota structure of the Japanese flounder intestinal tract, water, and pond sediment.