家兔肠道微生物ERIC-PCR指纹图谱构建及分析

Construction and analysis of ERIC-PCR fingerprinting of rabbit intestinal microbes

  • 摘要: 目的构建不同周龄健康家兔及腹泻家兔肠道微生物的ERIC-PCR指纹图谱,分析二者肠道菌群结构的多样性及其差异性,为优化家兔饲养和疾病防治打下基础.方法收集l~8周龄健康家兔与腹泻家兔粪便样本,分离肠道微生物并提取其基因组DNA,PCR扩增其16S rDNA与ERIC,构建ERIC-PCR指纹图谱,对不同同龄健康家兔及腹泻家兔肠道菌群结构进行分析.结果肠道微生物迅速在1周龄仔兔肠道中定植且总量达最高值,其后不断减少,5周龄后总量趋于稳定,且l周龄仔兔与5周龄后(断奶后)家兔的肠道微生物总量间均存在显著差异(P<0.05).随着周龄的增加,肠道微生物ERIC条带特征发生明显变化,各周龄出现了不同的特征性ERIC条带.与同一周龄健康家兔相比,腹泻家兔肠道微生物ERIC条带明显减少,条带特征存在明显差异,菌群多样性低于健康家兔.结论随着周龄的增加,健康家兔的肠道微生物菌群结构和多样性发生明显变化.与同一周龄健康家兔相比,腹泻家兔肠道微生物菌群结构明显改变,多样性明显降低,且出现健康家兔中未出现过的优势菌群,进一步证明肠道微生物菌群结构与家兔生长发育和腹泻存在关联性.

     

    Abstract: ObjectiveThe present study was conducted to construct intestinal microbial ERIC-PCR fingerprint of healthy rabbits and diarrheal rabbits and analyze structural difference between their intestinal microbial communities,in order to lay a foundation for optimizing rabbit feeding and disease control.Method Intestinal microbes were isolated from fecal samples which was collected from 1-8 week-old healthy rabbits and diarrheal rabbits,and their genomic DNA was extracted.Then 16S rDNA and ERIC were amplified by PCR technique.ERIC-PCR fingerprint was further constructed to analyze structure of intestinal microbial community in different week-old healthy rabbits and diarrheal rabbits.Result Intestinal microbes colonized quickly in 1 week-old rabbits,and total quantity of intestinal microbes reached the highest level,thereafter total quantity of intestinal microbes began to reduce,and became stable after 5 weeks old.And there were some significant differences in total quantity of intestinal microbes between 1 week-old rabbits and more than 5 week-old (after weaning) rabbits (P<0.05).Characteristic ERIC bands changed obviously with increase of week age,different characteristic ERIC bands were found in different week-old rabbits.Compared with age-matched healthy rabbits,number of ERIC bands in diarrheal rabbits was reduced markedly,and characteristics of ERIC bands were also markedly different,and diversity of intestinal microbial communities in healthy rabbits were lower than that in diarrheal rabbits.Conclusion With increase of week age,structure and diversity of intestinal flora in healthy rabbits change obviously.Compared with age-matched healthy rabbits,structure of intestinal microbial communities of healthy rabbit changes obviously,and diversity is also reduced,but dominant bacterial communities which do not appear in healthy rabbits are found in diarrheal rabbits.It is suggested that structure of intestinal microbial communities is significantly correlated with development and diarrhea of rabbits.

     

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