| As the degree of intensive aquaculture gradually deepen and food safety, and public healthevents appear constantly, healthy farming has became the focus point.Because of the antibioticsand the long-term use as antibiotics, pathogenic bacteria and microbes appear constantly lead tothe transfer of resistance genes.So it needs to find an effective antibiotics substitutes.Lactobacillus (Lb) can adjust the animal intestinal microecological balance,can enhance thebody’s immune, also against pathogenic bacteria. So screening probiotic strains used for theprevention and treatment of swine diarrhea disease is a great significance to promote the healthydevelopment of pig industry.Through the pig small intestinal epithelial cells IPEC-J2model, we search the effect ofFaeG-DCpep recombinant lactic acid bacteria in vitro on cell adhesion, production enterotoxincolonize inhibitory effect, and its effects on intestinal physiological effect. Preliminary,weexplore the colonize of FaeG-DCpep recombinant lactic acid bacteria after entering the gut,induction of immune factors and the mechanism of immune regulation. Through experiment ofadhesion and scanning electron microscope, we study the attached form of FaeG-DCpeprecombinant lactic acid bacteria and ETEC to IPEC-J2, found that the closely adhesion ofFaeG-DCpep recombinant lactic acid bacteria and ETEC to IPEC-J2cell. The number of Lb(pSIP-409-FaeG) and Lb (pSIP-409-FaeG-DCpep) adhesion and the close degree is higherthan the control group, showed that the process of the adhesion between cells and bacteriaproduce the basis of biological effects.We researched attacks, competition, rejection andalternative experiment of FaeG-DCpep recombinant lactic acid bacteria on epithelial cell IPEC-J2. In invasive experiment, lactobacillus colony number higher than that of restructuring ofETEC group painted painted (P <0.01), indicating that ETEC invasive strongest ofcells.Competitive experiment, the maximum number of PBS+ETEC colonies, painted abovethe FaeG-DCpep recombinant lactic acid bacteria group (P <0.05), suggesting that theFaeG-DCpep recombinant lactic acid bacteria and ETEC competition and at the same time cannot effectively restrain ETEC for cell adhesion.Repelling experiment, PBS+ETEC colony inthe experimental group, the maximum number of its quantity and Lb (pSIP-409-FaeG-DCpep) painted painted+ETEC compared significant difference (P <0.01).This suggests thatthe Lb (pSIP-409-FaeG-DCpep) can be competitive inhibition ETEC on the cell surface bygumming the combination of effective inhibition of ETEC in cell adhesion on the surface of the quantity. In instead experiment, the experimental group between ETEC colony count nodifference (ns), suggested that cells with ETEC is not reversible.Immunofluorescence tests inthe Lb (pSIP-409-FaeG-DCpep) separate role amount of fluorescence expression in theexperimental group were better than the corresponding ETEC stimulating group.This suggeststhat the Lb (pSIP-409-FaeG-DCpep) stimulation can stimulate cell attached to the exocrineprotein (ZO-1), strengthen the connection strength and density between cells, strengthen theintestinal mechanical protection.ELISA experiment and quantitative PCR to detect thegeneration of recombinant cytokines induced by lactic acid bacteria, found that IL-8and TNFalpha with the increase of test time must be the time to peak, but after the peak expressionquantity of the two different development trend.Lb (pSIP-409-FaeG-DCpep)+ETECinflammatory molecules mRNA generation are lower than the other two groups, stimulatinggroup, showed that Lb (pSIP-409-FaeG-DCpep) has a certain immune stimulation... |
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