Effect Of Lactobacillus Plantarum BS22 On Intestinal Flora Of Broilers Fed With Dietary Containing AFB1

This study is aim to study the effect of Lactobacillus plantarum BS22 (BS22) on digestive tract flora of broilers fed with dietary containing aflatoxin B1(AFB1). Test choosed one-day-old Avian broiler 120 which male and female were randomly divided into three groups with four repeat often. Three groups were control group I(basal diet), test group Ⅱ (basal diet+50μg/kgAFB1)and test group III(basal diet+50μg/kgAFB1+1% L.plantarum BS22). The trial lasted for 28 days. The digestive tract flora structure in the crop, glandular stomach, duodenum, jejunum, ileum and cecum at the age of 4 weeks were studied by 16S rDNA-based denaturing gradient gel electrophoresis (DGGE), cloning and sequencing, viable counting and quantitative real-time polymerase chain reaction (qRT-PCR). The test results were as follows:1.PCR-DGGE:The results of DGGE showed that number of DGGE bands in group Ⅲ were higher than that in group Ⅱ and group Ⅰ of contents in gastrointestinal except ileum. Cluster analysis showed that group Ⅱ and group Ⅰ stayed together, but group Ⅲ was separated in gastrointestinal except glandular stomach and cecum. Number of DGGE bands in group Ⅱ was higher than that in group Ⅰ and group Ⅲ of mucosa except ileum and cecum. At the same time, number of DGGE bands in group Ⅰ and group Ⅲ were similar or same. Except ileum and cecum, group Ⅰ and group Ⅲ stayed together, but group Ⅱ was separated in mucosa.2.Clone sequencing:Advantage bacterium groups were Lactobacillus crispatus, Lactobacillus salivarius, Lactobacillus acidophilus et al. Specific bands were mostly Uncultured bacterium.3.Viable counting: In contents, amounts of Ecoli in craw showed group Ⅲ significantly lower than group Ⅰ and group Ⅱ. Amounts of Ehterococcus and Ecoli showed detoxification group significantly lower than group Ⅰ and group Il in glandular stomach. Amounts of same kind of bacteria showed unconspicuous between groups in duodenum, jejunum and caecum. Amounts of Lactobacilli showed group Ⅲ higher than that of group Ⅰ and group Ⅱ in all parts except cecum. Amounts of Enterococcus and Ecoli showed group Ⅰ and group Ⅱ higher than group Ⅲ. At the same time, group Ⅲ was different with group Ⅰ and group Ⅱ. In mucosa, amounts of Enterococcus showed group Ⅱ very significantly higher than group Ⅰ and group Ⅲ in glandular stomach. Amounts of Ecoli showed group Ⅱ very significantly higher than group Ⅰ but significantly higher than group Ⅲ. Amounts of Lactobacilli in duodenum showed group Ⅰ significantly higher than group Ⅱ and amounts of Ecoli showed group Ⅱ significantly higher than group Ⅰ or group Ⅲ. Amounts of Enterococcus showed group Ⅱ very significantly higher than group Ⅰ in jejunum. Amounts of Lactobacilli showed group Ⅰ significantly higher than group Ⅱ in ileum. Amounts of the same kinds of flora in crop and cecum showed indistinctive result in three groups.4. Real-time PCR: The results of DNA copy number of microbiota by Real-time PCR in digesta were as follows. The DNA copy number of Enterobacteriaceae family showed group Ⅰ and group Ⅱ significantly higher than group Ⅲ in crop. In glandular stomach, the DNA copy number of Lactobacillus spp. showed group Ⅲ very significantly higher than group Ⅰ and group Ⅱ; Enterococcus spp. showed group I significantly higher than group Ⅲ; Enterobacteriaceae family showed group Ⅱ significantly higher than group Ⅲ. The results of DNA copy number of Enterobacteriaceae family showed group Ⅰ very significantly higher than group Ⅱ and group Ⅲ in ileum. Lactobacillus spp．and Enterococcus spp．had no significant difference between group Ⅰ and group Ⅱ from whole contents. Further more, Lactobacillus spp. of group Ⅲ increased to group Ⅰ and group Ⅱ but Enterobacteriaceae family decreased to group Ⅰ and group Ⅱ. In mucosa, Enterococcus spp. and Enterobacteriaceae family of group Ⅱ were very significantly higher than group Ⅰ and group Ⅲ in glandular stomach. Lactobacillus spp. of group Ⅰ showed significantly high- er than group Ⅱ and group Ⅲ in duodenum. Meanwhile, Enterobacteriaceae family of group Ⅱ showed very significantly higher than group Ⅰ and group Ⅲ and group Ⅲ showed significantly higher than group Ⅰ. Lactobacillus spp. of group Ⅱ and group Ⅲ reduced to group Ⅰ but Enterococcus spp. and Enterobacteriaceae family increased to group Ⅰ.In conclusion, AFB1 had no obvious effect on flora of digesta but had more effect on flora of mucosa especially on duodenum. At the same time, L.plantarum BS22 can adjust flora of mucosa to balance by AFB1.