肠炎沙门氏菌感染对鸭肠道菌群结构和呼吸道6种菌数量的影响
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摘要
肠炎沙门氏菌(Salmonella enteritidis,S.enteritidis)是一种常见的泛嗜性人畜共患病原菌,为无宿主特异性而有侵害性的病原菌之一。本研究对肠炎沙门氏菌经不同途径感染鸭呼吸道中的大肠杆菌、葡萄球菌、乳酸杆菌、双歧杆菌、芽孢杆菌和肠球菌进行定量研究并应用ERIC-PCR技术对感染鸭肠道菌群结构的多样性和动态变化进行了初步研究和分析,获得了以下结果:
1.应用FQ-PCR方法对致病性肠炎沙门氏菌经不同途径感染7日龄鸭呼吸道内六种菌属数量的动态变化进行检测。结果表明:各攻毒途径感染鸭呼吸道内这六种菌群除乳酸杆菌在个别检测时间段呈阴性,其他五种菌均能在呼吸道内检测到;而双歧杆菌属细菌含量(2.83±0.3—4.446±0.49)为检测的六中菌属中细菌含量最低的。大肠杆菌属细菌数量出现峰值的时间顺序为皮下组(攻毒后8 h)＞口服组(攻毒后24 h)＞滴鼻组(攻毒后48 h);葡萄球菌属细菌整体含量高低顺序为滴鼻组(8.37±0.29—10.54±0.23)＞口服组(7.41±0.31—9.66±0.24)＞皮下组(7.06±0.41-8.76±0.45);其他四种菌属细菌受攻毒途径影响的动态变化趋势相似,且波动剧烈程度依次为皮下组＞口服组＞滴鼻组。表明细菌失调程度与肠炎沙门氏菌感染途径密切相关。
2.应用ERIC-PCR方法对临床健康对照鸭和不同途径感染致病性肠炎沙门氏菌鸭各肠段中细菌菌群结构的多样性和动态变化进行检测。结果表明:临床健康对照鸭肠道菌群结构相对稳定,ERIC-PCR条带数量以盲肠最多,其次是十二指肠、空肠和回肠,直肠最少。感染鸭肠道菌群失调程度与其感染途径密切相关,其中以皮下感染鸭最为明显,其次是口服感染鸭,而滴鼻感染鸭没有明显变化。需氧菌或兼性厌氧菌(大肠杆菌、志贺氏菌、沙门氏菌等)成为感染鸭的优势菌群。
S.enteritidis is a common pathogenic bacterium that can result in zoonosis.In this study,FQ-PCR was used to detect the quantity of six kind of bacterium in respiratory tract,ERIC-PCR was used to fingerprint the kinetics of microbial community of fecal samples of S.enteritidis-infected ducklings.The contents are summarized as follows:
1.FQ-PCR was used to detect the quantity of six kind of bacterium in respiratory tract. The results indicated that five of the six kind of bacteria in respiratory tract of all ducklings can be well detected except Lactobacillus and the quantity of Bifidobacterium is the least(3.18±0.3—5.06±0.25).The chronologicaol order of Eschenchia coli's quantity crest value is hypodermically inoculated ducklings(8 h p.i.)＞orally inoculated duckling s (24 h p.i.)＞nasally inoculated ducklings(48 h p.i.).The quantity of StaphyLococcus is nasally inoculated ducklings(8.37±0.29—10.54±0.23)＞orally inoculated ducklings (7.41±0.31—9.66±0.24)＞hypodermically inoculated ducklings(7.06±0.41-8.76±0.45).The dynamic changes of the other four kind of bacterium are also hypodermically inoculated ducklings＞orally inoculated ducklings＞nasally inoculated ducklings.So the maladjusted degree of microbial community in respiratory tract was correlative with the infected routes.
2.Enterobacterial Repetitive Intergenic Consensus(ERIC)-PCR was applied to analyze the intestinal microbial community diversity and dynamic change of healthy ducklings and 7-day-old ducklings infected with S.enteritidis in different routes.The results indicated that the intestinal microbial community of the control healthy ducklings was steady and the ERIC-PCR band numbers of the control healthy ducklings were the least with rectum and were the most with caecum.The maladjusted degree of intestinal microbial community was correlative with the infected routes and it successively abated in hypodermically inoculated ducklings,orally inoculated ducklings and nasally inoculated ducklings.The intestinal microbial community dynamic change of hypodermically inoculated ducklings was severely maladjusted while it did not change in nasally infected ducklings.The prominent bacteria had changed because of S.enteritidis infection and the DNAstar of staple of ERIC-PCR showed that aerobe and facultative aerobe(Escherichia coli,Shigella,Salmonella) became preponderant bacilli in the intestine of all infected ducklings with S.enteritidis.
1.应用FQ-PCR方法对致病性肠炎沙门氏菌经不同途径感染7日龄鸭呼吸道内六种菌属数量的动态变化进行检测。结果表明:各攻毒途径感染鸭呼吸道内这六种菌群除乳酸杆菌在个别检测时间段呈阴性,其他五种菌均能在呼吸道内检测到;而双歧杆菌属细菌含量(2.83±0.3—4.446±0.49)为检测的六中菌属中细菌含量最低的。大肠杆菌属细菌数量出现峰值的时间顺序为皮下组(攻毒后8 h)＞口服组(攻毒后24 h)＞滴鼻组(攻毒后48 h);葡萄球菌属细菌整体含量高低顺序为滴鼻组(8.37±0.29—10.54±0.23)＞口服组(7.41±0.31—9.66±0.24)＞皮下组(7.06±0.41-8.76±0.45);其他四种菌属细菌受攻毒途径影响的动态变化趋势相似,且波动剧烈程度依次为皮下组＞口服组＞滴鼻组。表明细菌失调程度与肠炎沙门氏菌感染途径密切相关。
2.应用ERIC-PCR方法对临床健康对照鸭和不同途径感染致病性肠炎沙门氏菌鸭各肠段中细菌菌群结构的多样性和动态变化进行检测。结果表明:临床健康对照鸭肠道菌群结构相对稳定,ERIC-PCR条带数量以盲肠最多,其次是十二指肠、空肠和回肠,直肠最少。感染鸭肠道菌群失调程度与其感染途径密切相关,其中以皮下感染鸭最为明显,其次是口服感染鸭,而滴鼻感染鸭没有明显变化。需氧菌或兼性厌氧菌(大肠杆菌、志贺氏菌、沙门氏菌等)成为感染鸭的优势菌群。
S.enteritidis is a common pathogenic bacterium that can result in zoonosis.In this study,FQ-PCR was used to detect the quantity of six kind of bacterium in respiratory tract,ERIC-PCR was used to fingerprint the kinetics of microbial community of fecal samples of S.enteritidis-infected ducklings.The contents are summarized as follows:
1.FQ-PCR was used to detect the quantity of six kind of bacterium in respiratory tract. The results indicated that five of the six kind of bacteria in respiratory tract of all ducklings can be well detected except Lactobacillus and the quantity of Bifidobacterium is the least(3.18±0.3—5.06±0.25).The chronologicaol order of Eschenchia coli's quantity crest value is hypodermically inoculated ducklings(8 h p.i.)＞orally inoculated duckling s (24 h p.i.)＞nasally inoculated ducklings(48 h p.i.).The quantity of StaphyLococcus is nasally inoculated ducklings(8.37±0.29—10.54±0.23)＞orally inoculated ducklings (7.41±0.31—9.66±0.24)＞hypodermically inoculated ducklings(7.06±0.41-8.76±0.45).The dynamic changes of the other four kind of bacterium are also hypodermically inoculated ducklings＞orally inoculated ducklings＞nasally inoculated ducklings.So the maladjusted degree of microbial community in respiratory tract was correlative with the infected routes.
2.Enterobacterial Repetitive Intergenic Consensus(ERIC)-PCR was applied to analyze the intestinal microbial community diversity and dynamic change of healthy ducklings and 7-day-old ducklings infected with S.enteritidis in different routes.The results indicated that the intestinal microbial community of the control healthy ducklings was steady and the ERIC-PCR band numbers of the control healthy ducklings were the least with rectum and were the most with caecum.The maladjusted degree of intestinal microbial community was correlative with the infected routes and it successively abated in hypodermically inoculated ducklings,orally inoculated ducklings and nasally inoculated ducklings.The intestinal microbial community dynamic change of hypodermically inoculated ducklings was severely maladjusted while it did not change in nasally infected ducklings.The prominent bacteria had changed because of S.enteritidis infection and the DNAstar of staple of ERIC-PCR showed that aerobe and facultative aerobe(Escherichia coli,Shigella,Salmonella) became preponderant bacilli in the intestine of all infected ducklings with S.enteritidis.
引文
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