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应用 ERIC-PCR 技术分析健康兔与经口感染溶组织内阿米巴兔肠道微生物群落多样性的比较。

Intestinal microbial community diversity between healthy and orally infected rabbit with Entamoeba histolytica by ERIC-PCR.

机构信息

Guiyang College of Traditional Chinese Medicine, Guiyang, 550002, Guizhou Province, China.

出版信息

Parasitol Res. 2012 Sep;111(3):1123-6. doi: 10.1007/s00436-012-2942-6. Epub 2012 May 5.

Abstract

Enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to analyze the difference of intestinal microbial community diversity between healthy and orally infected rabbits with Entamoeba histolytica. The dynamic changes in different parts of digestive system including the duodenum, jejunum, ileum, caecum, and rectum in healthy and infected rabbits at different time points were also tested. The intestinal microbial community of the control healthy rabbits was steady, and the total number of ERIC-PCR bands in the control healthy rabbit was the least in the rectum and the most in the caecum. ERIC-PCR bands of orally inoculated rabbits did not obviously change until 24 h after postinoculation (p.i.). The numbers of the ERIC-PCR bands gradually decreased from 24 to 72 h p.i., and then, with the development of disease, the band numbers gradually increased until 6 days p.i. Sequence analysis showed that the nucleotide sequence homologies of the fragment about 1,200 bp of infected ileum sampled at 32 h p.i. were above 95 % with Sinorhizobium meliloti enterobacterial, Erwinia amylovora and Salmonella typhimurium, and the nucleotide sequence homologies of the fragment about 300 bp of infected ileum sampled at 48 h p.i. were more than 90 % with Xanthomonas campestris enterobacterial, Yersinia enterocolitica subsp., Shigella flexneri, S. meliloti enterobacterial, Yersinia pestis, Klebsiella pneumoniae subsp., and Escherichia coli. The prominent bacteria had changed after E. histolytica infection. The DNAstar of staple of ERIC-PCR showed that aerobe and facultative aerobe (E. coli, Shigella, and Salmonella) became preponderant bacilli in the intestine of orally infected rabbits with E. histolytica.

摘要

应用肠道细菌重复基因间一致性(ERIC)-PCR 分析分析健康兔和经口感染溶组织内阿米巴兔肠道微生物群落多样性的差异。还检测了不同时间点健康和感染兔不同消化部位(十二指肠、空肠、回肠、盲肠和直肠)的动态变化。对照组健康兔的肠道微生物群落稳定,对照组健康兔的 ERIC-PCR 总条带数在直肠中最少,在盲肠中最多。经口接种兔的 ERIC-PCR 条带在接种后 24 小时(p.i.)内没有明显变化。从 24 小时到 72 小时 p.i.,条带数量逐渐减少,然后随着疾病的发展,条带数量逐渐增加,直到 6 天 p.i.。序列分析表明,32 小时 p.i.感染回肠样本中约 1200bp 片段的核苷酸序列同源性与根瘤菌属、梨火疫病菌和鼠伤寒沙门氏菌均在 95%以上,48 小时 p.i.感染回肠样本中约 300bp 片段的核苷酸序列同源性与黄单胞菌属、肠侵袭性大肠杆菌亚种、福氏志贺菌、根瘤菌属、鼠疫耶尔森菌、肺炎克雷伯菌亚种和大肠埃希菌均在 90%以上。溶组织内阿米巴感染后优势菌发生了变化。ERIC-PCR 的 staple DNAstar 显示,需氧菌和兼性需氧菌(大肠杆菌、志贺菌和沙门氏菌)成为经口感染溶组织内阿米巴兔肠道中的优势杆菌。

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