采用微生物组学方法研究自然养猪法仔猪肠道菌群变化规律

目的通过在垫料中添加微生态制剂,研究使用微生态垫料与水泥地饲养条件影响仔猪结肠菌群变化的程度,揭示微生态制剂对猪肠道菌群的调控规律。方法无菌采取仔猪结肠黏膜,提取DNA,采用微生物组学方法 454测序法测定细菌种类。结果使用微生态垫料21、42 d后,21天试验组(J111)有效序列分别为5 435个,OTU为757个,21天试验组(J212)有效序列为5 298个,OTU为655个,42天对照组(J121)有效序列为5 480个,OTU为937,42天对照组(J222)有效序列为6 514个,OTU为907个。占主导地位的细菌有弯曲杆菌、螺杆菌、毛螺菌、乳酸杆菌、RC9肠道菌、瘤胃球菌、螺旋体以及未确定种属的细菌。通过比较21 d、42 d仔猪结肠菌群数量,发现弯曲杆菌比例试验组中增加,而对照组降低;乳酸杆菌、螺杆菌比例试验组降低,而对照组升高;瘤胃球菌比例试验组基本不变,而对照组略有升高;毛螺菌、RC9肠道菌群、螺旋体、未确定菌比例试验组与对照组均下降。埃希-志贺菌比例在试验组降低,而对照组基本不变。结论微生物制剂垫料影响仔猪结肠菌群变化,细菌多样性降低,常见细菌比例与传统方法研究结果一致,但部分细菌呈现与传统方法研究结果不一致,且尚未确定的未培养细菌种类比例较高,有待于进一步深入研究。

Variation of intestinal microflora of piglets raised on fermentation bed： a study by microbial genomics method

Objective To explore the alteration of the colonic flora of piglets raised on probiotic-fermentation bed com- pared with those raised on concrete floor, and reveal the mechanfism of probiotics regulating pig＇ s intestinal flora. Methods The colonic mucosa samples of the piglets, after breeding for 21 and 42 days repectively, were collected aseptically to extract the DNA. Microbial genomies method, 454 Sequencing, was used to determine the bacterialspecies. Results 5,435 effective sequences and 757 OTUs were detected in the 21 -day test group J111, while 5,298 sequences and 655 OTUs were detected in the 42-day test group J212. There were 5,480 effective sequences and 937 OTUs detected in the 21-day control group J121, while 6,514 effective sequences and 907 OTUs in the 42- day control group J222. Campylobacter, Helicobacter, Lachnospiraceae, Lactobacillus, RC9-gut-bacteria, Rumino- coccus, Spirochetes and unidentified bacteria were dominant bacteria at the genus level. The proportion of Campy- lobacter increased in the test groups, while decreased in the control groups; the proportions of Lactobacilli and Heli- cobacter reduced in test groups, while increased in the control groups. The ratio of Ruminococcus remained un- changed in the test groups while slightly increased in the control groups. The ratios of Lachnospiraceae, the RC9- gut-flora, Spirochetes, and unidentified bacteria decreased in both test groups and control groups. The proportion of Escherichia-Shigella reduced in the test groups, but remained unchanged in the control groups. Conclusion Litter added with microbial agents can affect the colonic flora of piglets and reduce bacterial diversity. The proportions of common bacteria are similar to the results from traditional method. Some bacteria present inconsistent findings with those of traditional methods, while the higher proportion of unidentified uncultured bacteria needs to be further studied.