零换水条件下养殖水体中碳氮比对生物絮团形成及团头鲂肠道菌群结构的影响

孙盛明, 朱健, 戈贤平, 江晓浚

doi:10.3969/j.issn.1006-267x.2015.03.036

动物营养学报 >

2015 , Vol. 27 >Issue 3: 948 - 955

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.03.036

研究简报 Short Communications

零换水条件下养殖水体中碳氮比对生物絮团形成及团头鲂肠道菌群结构的影响

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1. 中国水产科学研究院淡水渔业研究中心, 农业部淡水渔业和种质资源利用重点实验室, 无锡 214081;
2. 南京农业大学无锡渔业学院, 无锡 214081

孙盛明(1983—),男,山东莱州人,助理研究员,博士,研究方向为水产动物营养与饲料研究。E-mail:sunsm@ffrc.cn

收稿日期: 2014-09-22

网络出版日期: 2015-03-18

基金资助

中央级公益性科研院所基本科研业务费专项资金(2014A08XK01);十二五国家科技支撑计划"长江下游池塘高效生态养殖技术集成与示范"(2012BAD25B07);国家大宗淡水鱼类产业技术体系华东养殖岗位(CARS-46-14)

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Effects of Different Carbon/Nitrogen Ratios in Cultured Water under Zero-Water Exchange Condition on Bioflocs Formation and Intestinal Microflora Structure of Blunt Anout Bream (Megalobrama amblycephala)

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1. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China;
2. WuXi Fishery College, Nanjing Agricultural University, Wuxi 214081, China

Received date: 2014-09-22

Online published: 2015-03-18

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摘要

本试验旨在运用454焦磷酸测序技术同时结合PCR-变性梯度凝胶电泳(DGGE)技术分析零换水条件下养殖水体中碳氮比对生物絮团形成及团头鲂(Megalobrama amblycephala)肠道菌群结构的影响。试验共设置5组,其养殖水体中碳氮比分别为8(CN8组,设为对照组)、12(CN12组)、16(CN16组)、20(CN20组)、24(CN24组),每组设置3个水泥池(重复),每个水泥池投放团头鲂幼鱼20尾。养殖期为8周,每隔1周采集水样1次,检测养殖水体中氨氮、亚硝酸盐氮及生物絮团含量。养殖8周后,每个水泥池分别随机挑选3尾鱼无菌采集盲肠内容物,运用454焦磷酸测序技术分析不同碳氮比零换水养殖条件下团头鲂肠道菌群结构的变化。结果表明:当碳氮比≥16时,形成的生物絮团可以有效调节水质,降低水体中的氨氮和亚硝酸盐氮含量。随着碳氮比由8升高到16和20,肠道中放线菌门(Actinobateria)含量变化不显著(P>0.05),厚壁菌门(Firmicutes)含量显著增加(P<0.05),梭杆菌门(Fusobateria)和变形菌门(Proteobacteria)含量显著减少(P<0.05)。对厚壁菌门进行深入分析发现,在纲水平上,与对照组相比,CN16、CN20和CN24组团头鲂肠道中梭菌纲(Clostridia)含量显著减少(P>0.05),而芽孢杆菌纲(Baccilli)含量显著增加(P>0.05)。DGGE指纹图谱分析表明,养殖水体中碳氮比能够影响团头鲂的肠道菌群的多样性。CN8组和其他组的相似性系数最低,仅为0.43,而CN20和CN24组的相似性系数最高,达到0.96。由此可见,零换水条件下养殖水体的碳氮比影响了生物絮团的形成量,改变了团头鲂肠道菌群结构和多样性。以团头鲂肠道菌群结构为评价指标,零换水条件下团头鲂养殖水体中适宜碳氮比为16~20。

关键词： 团头鲂; 肠道菌群; 454焦磷酸测序; 碳氮比

本文引用格式

孙盛明, 朱健, 戈贤平, 江晓浚 . 零换水条件下养殖水体中碳氮比对生物絮团形成及团头鲂肠道菌群结构的影响[J]. 动物营养学报, 2015 , 27(3) : 948 -955 . DOI: 10.3969/j.issn.1006-267x.2015.03.036

Abstract

This present study was conducted to investigate the effects of different carbon (C)/nitrogen (N) ratios in cultured water under zero-water exchange condition on bioflocs formation and intestinal microflora structure of blunt anout bream (Megalobrama amblycephala) using by 454 pyrosequencing technology and PCR-denaturing gradient gel electrophoresis (DGGE) technology. Five groups were designed, and the C/N ratios in those groups were 8(CN8 group, as control group), 12(CN12 group), 16(CN16 group), 20(CN20 group) and 24(CN24 group), respectively. Each group had 3 tanks (replicates), and each tank cultured 20 juvenile blunt anout bream. Culture period was 8 weeks, and water samples were collected every other week to detect the contents of ammonia nitrogen, nitrite nitrogen and bioflocs in cultured water. After 8 weeks culture, three fish were selected in each tank, and the cecal contents of those fish were collected to analyze the change of intestinal microflora structure of blunt anout bream cultured in different C/N ratios under zero-water exchange tanks using by 454 pyrosequencing technology. The results showed that when the C/N ratio was over 16, the biofloc could effectively regulate water quality, and reduce ammonia nitrogen and nitrite nitrogen contents in water. With the C/N ratio increased from 8 to 16 and 20, intestinal Actinobateria content had no significant change (P>0.05), and Firmicutes content was significantly increased (P<0.05), while the contents of Fusobateria and Proteobacteria were significantly decreased (P<0.05). For Firmicutes, a further analysis was made, at class level, the content of intestinal Clostridia in CN16, CN20 and CN24 groups was significantly decresed compared with control group (P<0.05), while the content of intestinal Bacilli significantly increased (P<0.05). The DGGE fingerprints also demonstrated that the intestinal microflora community diversity of blunt anout bream was affected by C/N ratio in cultured water. The similarity coefficient between CN8 group and other groups was 0.43, which was the lowest. CN20 and CN24 groups were related to each other with the similarity coefficient was 0.96, which was the higest. It is concluded that the formation of bioflocs is affected and the diversity and structure of intestinal microflora of blunt anout bream are altered by C/N ratio in cultured water under zero-water exchange condition. Using the intestinal microflora structure of blunt anout bream as an evaluation index, the suitable C/N ratio in cultured water under zero-water exchange condition is 16 to 20.

Key words： blunt anout bream (Megalobrama amblycephala); intestinal microflora; 454 pyrosequencing; C/N ratio

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