Feeding Different Cellulolytic Bacteria Impact On Intestinal Microflora Of Geese

Geese intestinal microbial population is a complicated ecological environment, which plays an important role in digesting and absorbing nutrients and health of goose. This study was conducted to investigate the feeding different cellulolytic bacteria impact on intestinal microflora of geese by PCR-SSCP. Select three cellulolytic bacteria, namely: A cellulolytic strains from goose intestinal (referred to as A bacteria), and a cellulolytic strains from the termite gut (referred to as B bacteria), a cellulolytic strains in cow dung (referred to as C bacteria), composite bacteria were the mixture of A bacteria, B bacteria and C bacteria according to the ratio of1:1:1.80zero-day-old geese were randomly assigned five treatments with16geese each group according to similar weight as follows:Group A bacteria, Group B bacteria, Group C bacteria, the complex bacteria group and the control group. The corn stalks and alfalfa were the main fiber and the five treatments had the sane level of nutrition. Add cellulolytic bacteria in the feed since the geese were at their0days old, once every three days, the control group didn’t feed with cellulolytic bacteria. The geese were slaughtered at their70days old, and collect their duodenum, jejunum, ileum, cecum, and rectum, and stored at-80℃C refrigerator.This test was divided into two parts, in the first part the study was conducted to investigate the different pretreatments methods impact on the geese cecal chyme flora DNA quality. The geese cecal chyme samples were dealed whit the direct and indirect pre-treatments, then extracted the genomic DNA of pre-treatment samples by enzyme method and liquid nitrogen grinding method, and evaluated genomic DNA purity and PCR effect.In the second part, the study was conducted to analyze the flora diversity of the intestinal microflora of geese. The genomic DNA was extracted from the content of various intestinal segment of geese duodenum, jejunum, ileum, cecum, and rectum. The V1-V3regions of bacterial16S rRNA were amplified with universal primer, and then amplicons were separated directly by SSCP. Some common and special bands were identified, cloned and quantitated, in order to investigate the diversity of the predominant eubacteria in the duodenum, jejunum, ileum, cecum, and rectum digests. The results were showed as follows:The method combining the indirect pre-treatment with enzymatic was more suitable than other methods which laid the foundation for follow-up test. SSCP electrophoresis pattern showed that:different groups all had rich bands, which indicated that the intestinal had good flora diversity; Feeding cellulolytic bacteria had little impact on the geese preceding intestinal, there were more common bands and less specific bands in jejunum and ileum through different treatment groups; Feeding cellulolytic bacteria had larger impact on the geese latter segment of intestinal, there were more specific bands in ileum, cecum, and rectum through different treatment groups. Separation A bacteria and C bacteria in Group A and Group C, and the two cellulolytic bacteria were also detected in the other groups; B bacteria was not detected in Group B. The complex bacteria group had more bands than any other group. Clostridium, Bactericides, lively Ruminococcus, Ruminococcus were detected.