Research On Adherence And Anti-bacteria Property Of Clostridium Butyricum And Its Effect On Miichthys Miiuy Intestinal Physiology

With the development of aquaculture cultivation, aquatic animal heath faces greatchanllenges. The long-term use of antibiotics and anti-bacteria substance causes muchharmful effect, such as anti-medicine ability of bacteria, translocation of anti-medicinegenes and medicine remnants in muscle. Probiotics, the same meaning of microbialecology products are various live bacteria, which can modulate intestinal microbialecology of aquatic animals, improve water environment, enhance the body immunity,advance the food digestion and inhibit the pathogenic bacteria. It is proved that use ofprobiotics is safe and effective to animal body, so it appears valuable significance tostudy and develop probiotic for aquaculture.Clostridium butyricum which is usually used as human and animal medicine, cancure diarrhea and enteritis. Our research focuses on the cell adhesion and anti-pathogenof Clostridium butyricum, and effect to intestinal physiology of Miichthys miiuy. Theresearch contains three parts: (Ⅰ) Separate and cultivate intestinal epithelial cell(IEC),develop the research model of cell; (Ⅱ) Study Clostridium butyricum adhesion to cellmodel, and inhibition to pathogenic invasion and cell safety evaluation; (Ⅲ) By take ofClostridium butyricum, we observe the fish intestine morpha, gut digestion enzymeactivity, intestinal short chain fatty acid (SCFA) and the ecology of intestinal innerwallThe main results are as follows:1. We can separate the IEC withⅣcollagen enzyme,Ⅰcollagen enzyme and EDTA.Purifying the IEC in line with the time difference of adhesion to wall and reducingthe treatment time. In this way, IEC can be obtained lively, and appear monolayergrowth.2. Clostridium butyric can adhere to IEC, adhesion rate 8.03%, and will not invade orharm cells. It can reduce the adhesion of Vibrio anguillarum in different treatmentgroups. On competition experiment, adhesion rate of Vibrio anguillarum candecrease 6.71%; repulsion experiment, decrease 8.27%; substitution experiment,4.47%. 3. Feeding Miichthys miiuy with Clostridium butyric, it can increase its intestinaldigest enzyme activity and improve SCFA quantity, especially butyric acid withthe increase of 2.09mmol/kg in mid-gut and 2.12mmol/kg in hinder gut. Intestinalvillus height increases 28.81μm in mid-gut and 34.23μm in hinder gut; microvillusheight increases 1.89μm, 1.09μm separately. With the electronic lens, we canobserve bacteria adhering to intestinal inner wall.Conclusion:Clostridium butyricum has cell adhesion ability, and has no pathogenic property.It can inhibit intestinal adhesion of pathogenic bacteria, and improve the quantity ofintestinal SCFA, digest enzyme activity, intestinal villus height. In vitro, the researchon bacteria adherence and inhibition can reflect the mutual action of fish intestinalbacteria in certain extent.