Rungrassamee Wanilada, Klanchui Amornpan, Maibunkaew Sawarot, Chaiyapechara Sage, Jiravanichpaisal Pikul, Karoonuthaisiri Nitsara
Microarray Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Khlong Luang, Pathum Thani, Thailand.
Aquatic Molecular Genetics and Biotechnology Laboratory, National Center for Genetic Engineering and Biotechnology (BIOTEC), Khlong Luang, Pathum Thani, Thailand.
PLoS One. 2014 Mar 11;9(3):e91853. doi: 10.1371/journal.pone.0091853. eCollection 2014.
The black tiger shrimp (Penaeus monodon) is a marine crustacean of economic importance in the world market. To ensure sustainability of the shrimp industry, production capacity and disease outbreak prevention must be improved. Understanding healthy microbial balance inside the shrimp intestine can provide an initial step toward better farming practice and probiotic applications. In this study, we employed a barcode pyrosequencing analysis of V3-4 regions of 16S rRNA genes to examine intestinal bacteria communities in wild-caught and domesticated P. monodon broodstock. Shrimp faeces were removed from intestines prior to further analysis in attempt to identify mucosal bacterial population. Five phyla, Actinobacteria, Fusobacteria, Proteobacteria, Firmicutes and Bacteroidetes, were found in all shrimp from both wild and domesticated environments. The operational taxonomic unit (OTU) was assigned at 97% sequence identity, and our pyrosequencing results identified 18 OTUs commonly found in both groups. Sequences of the shared OTUs were similar to bacteria in three phyla, namely i) Proteobacteria (Vibrio, Photobacterium, Novosphingobium, Pseudomonas, Sphingomonas and Undibacterium), ii) Firmicutes (Fusibacter), and iii) Bacteroidetes (Cloacibacterium). The shared bacterial members in P. monodon from two different habitats provide evidence that the internal environments within the host shrimp also exerts selective pressure on bacterial members. Intestinal bacterial profiles were compared using denaturing gradient gel electrophoresis (DGGE). The sequences from DGGE bands were similar to those of Vibrio and Photobacterium in all shrimp, consistent with pyrosequencing results. This work provides the first comprehensive report on bacterial populations in the intestine of adult black tiger shrimp and reveals some similar bacterial members between the intestine of wild-caught and domesticated shrimp.
黑虎虾（斑节对虾）是世界市场上具有重要经济价值的海洋甲壳类动物。为确保对虾产业的可持续性，必须提高生产能力并预防疾病爆发。了解对虾肠道内健康的微生物平衡可为更好的养殖实践和益生菌应用提供初步步骤。在本研究中，我们采用对16S rRNA基因V3 - 4区域的条形码焦磷酸测序分析，来检测野生捕捞和养殖的斑节对虾亲虾的肠道细菌群落。在进一步分析之前，从肠道中取出虾粪便，以试图鉴定黏膜细菌种群。在来自野生和养殖环境的所有虾中均发现了五个门，即放线菌门、梭杆菌门、变形菌门、厚壁菌门和拟杆菌门。以97%的序列同一性分配操作分类单元（OTU），我们的焦磷酸测序结果确定了两组中常见的18个OTU。共享OTU的序列与三个门中的细菌相似，即i）变形菌门（弧菌属、发光杆菌属、新鞘氨醇菌属、假单胞菌属、鞘氨醇单胞菌属和无定形杆菌属），ii）厚壁菌门（梭杆菌属），以及iii）拟杆菌门（泄殖腔杆菌属）。来自两个不同栖息地的斑节对虾中共享的细菌成员提供了证据，表明宿主对虾体内的环境也对细菌成员施加选择压力。使用变性梯度凝胶电泳（DGGE）比较肠道细菌谱。DGGE条带的序列在所有虾中均与弧菌属和发光杆菌属的序列相似，与焦磷酸测序结果一致。这项工作提供了关于成年黑虎虾肠道细菌种群的第一份综合报告，并揭示了野生捕捞和养殖虾肠道之间一些相似的细菌成员。
