Study On The Effect Of Combined Application Of Dog-derived Probiotics And Purslane On Dog Diarrhea

Canine diarrhea is a clinical disease characterized by increased defecation frequency and increased water in feces caused by multiple pathogens and multiple factors,which seriously endangers the health of dogs.How to prevent and control canine diarrhea has become an urgent problem in veterinary clinic.Traditional Chinese medicine is widely used clinically because of its naturalness,low residue and drug resistance,and it is a good substitute for antibiotics;probiotics are a type of active microorganisms that are beneficial to the host and are often used to treat gastrointestinal-related diseases.However,there is no relevant report about the combined application of canine-derived probiotics and traditional Chinese medicine.To this end,this study provided experimental evidence for the prevention of canine diarrhea by isolating,identifying,and screening probiotics in dog feces,and combining them with purslane for clinical application.In this study,probiotics bacteria were isolated from feces of healthy dogs;the isolated probiotics were identified by morphological observation and molecular biology methods;acid and bile salt-resistantstrains with inhibitory effect on pathogenic Escherichia coli were screened,while purslane water extract showed no inhibitory effect on them.Single factor test and orthogonal test were used to determine the best combined application conditions of probiotics and purslane.Taking the model of dog diarrhea caused by senna as the research object,to observe the effect of combined application of probiotics and purslane on dog diarrhea.Results:8 strains of probiotics were isolated from the feces of healthy dogs;morphological observation revealed that 3 strains of Bacillus formed round or oval,convex or concave,milky white,smooth or rough edged colonies in LB solid medium;5 strains of lactic acid bacteria formed round,convex,milky white,smooth-edge colonies in MRS solid medium;the results of molecular biology identification:1 strain of Bacillus subtilis(DY001),1 strain of Bacillus subtilis(DY002),1 strain of Bacillus licheniformis(DY003),2 strains of Enterococcus faecalis(DR001 and DR005),1 strain of Enterococcus hayali(DR002),1 strain of Enterococcus lactis(DR003)and,l strain of Lactobacillus reuteri(DR004).The results of acid and bile salt resistance test showed that 2 strains of Bacillus(DY002,DY003)and 2 strains of lactic acid bacteria(DR003,DR004)showed strong acid and bile salt resistance;the results of bacteriostatic experiment of pathogenic Escherichia coli showed 3 strains of lactic acid bacteria(DR001,DR003,DR004)had inhibitory effect on Escherichia coli;therefore,Lactobacillus reuteri(DR004)as the strain to use in combined application;the results of in vitro bacteriostasis test of purslane showed that the water extract of purslane had no inhibitory effect on the 8 strains but has an inhibitory effect on Escherichia coli;the single factor test results showed that the optimal fermentation time was 36 h,the purslane addition amount was 25 g,and the inoculation amount was 20%;the L9(33)orthogonal experiment showed the best combined conditions are:fermentation time 36 h,purslane addition amount 20 g,inoculation amount 20%,fermented purslane polysaccharide content reached the maximum of 0.066±0.02 mg/mL,and the amount of viable bacteria was 6.84×107 cfu/mL.Through animal experiment,it is found that the combined application of probiotics and purslane is better than probiotics or purslane alone in preventing and treating canine diarrhea.Conclusion:the combined of probiotics isolated from healthy dog feces and purslane can increase the purslane polysaccharide content and the amount of viable probiotics,and has a certain preventive effect on dog diarrhea caused by senna leaves.