Epidemiological Investigation And Probiotic Treatment On Helicobacter Pylori Infection Of Dogs In Nanjing

Helicobacter pylori(H.pylori)is a gram-negative and microaerobic bacteria.It was isolated successfully for the first time by Marshsll and Warren in 1983,and is recognized the higher infection rate and one of the important pathogenic bacteria of upper gastrointestinal diseases in the world now.Since 1994 it has been considered as class I carcinogen of human by WHO(World Health Organisation).The natural reservoir of H.pylori is human,and the infection rate is 40%?80%,which is generally higher in developing countries than in developed countries.It has reported that H.pylori can be spread between pet and its owners,and mutual infection rates are as high as 100%.So far,there has not epidemiological investigation of pet infected with H.pylori and relevant study report of cases.It has an important clinical value to carry out the detailed investigation of epidemiological characteristics and comparative research on the preventive and treatment effects of the drug used in order to provide the guidance for clinical veterinary treatment of H.pylori infection effectively.Objective:(1)The present study was designed to investigate the infection rate of H.pylori of dogs in Nanjing,to analyze the corresponding relationship between their living environment and the infection rate,and to provide the foundation data for pet clinical;(2)To isolate and identify H.pylori to make drug sensitivity test and assess antibiotic resistance status of different H.pylori strains in order to provide reference for reasonable selection of antibiotics;(3)To compare the eradication effect between the standard triple therapy and standard triple therapy combined with probiotics for H.pylori in dogs and provide the data for pet clinical to select a better therapeutic regimen of H.pylori.Method:(1)We selected 518 dogs living in different environment in Nanjing,collected the fresh feces from rectum,recorded the basic information of dogs,detected feces samples of dogs rapidly using colloidal gold test plate,and analysed the positive rate of H.pylori.The feces samples were selected randomly,extracted the genomic DNA,proceed PCR amplification and compared the infection situation of H.pylori and Hlicobacter spp.(H spp.);(2)24 fresh feces sample which from eligible positive dogs were selected to isolate culture under the condition of 37 ? and microaerobic.The morphology of bacterial colonies and bacterial were identified when the number of baterial colonies met the requirement,then gram stain,urease,oxidase and catalase were determined.Antibiotic susceptibility test was made to determine the strains of H.pylori and evaluate the antibiotic resistance status of H.pylori in local dogs;(3)41 H.pylori positive dogs were selected as group A receiving the triple therapy(omeprazole 0.4 mg.kg-1+clarithromycin 7.5 mg.kg-1+amoxicillin 15 mg.kg-1,twice a day for 14 days as a course).34 H.pylori positive dogs were selected as group B which received probiotics treatment as well as triple therapy(probiotics 0.8 mg.kg-1 after 2 h of antibiotic administration).Colloidal gold method was used to detecte the feces samples of dogs at 4 weeks after the medicine withdrawal and to compare the eradication rate of H.pylori of various regimens.Results:(1)There were 393(75.87%)positive of H.pylori among the 518 dog feces samples in Nanjing,the range of H.pylori positive rate was from 57.14%to 80.65%.The highest infection rate was found in stray-dog shelters(80.65%),the lowest was the hospitalized dogs(57.14%).Total positive rate of dogs at pet hospital clinic was 72.64%in Nanjing.The results of PCR amplification showd that the positive rate of H spp.was 87.10%(108/124),and H.pylori was 76.61%(95/124),H.pylori accounted for 87.96%(95/108)of H spp.;(2)The H.pylori from the fresh feces of 24 dogs were isolated and identified and then 15 ascertain H.pylori strains were obtained.The results of antibiotic sensitivity test showed that the resistance rate of quinolones(e.g.,levofloxacin)and furazolidone(e.g.,furazolidone)were the lowest as 20%,and the clarithromycin,amoxicillin and metronidazole was 26.67%,33.33%and 33.33%,respectively.The rest of the antibiotic resistant rate was more higher,even up to 53.33%.(3)In the animal experiment of H.pylori eradication,20 cases were recorded in A group,the rate was 48.78%,H.pylori eradication rate at the end of first treatment course was 40%,and that at the end of the second course was 80%.20 cases were recorded in B group,and the rate was 58.82%,H.pylori eradication rate at the end of the first course was 70%,and that at the end of the second courses was 100%.Conclusion:(1)The infection rate of H.pylori of dogs in Nanjing was consistent with previous reports in various regions,and the proportion of H.pylori accounted for H spp.was the highest.H.pylori infection in dogs was closely related to living habits and environment hygiene status.(2)It was important to reduce the prophylactic medication of antibiotic in order to decrease the problems of bacterial resistance.Levofloxacin can be used to replace the clarithromycin in standard triple therapy to improve therapeutic effect.(3)It suggested that standard triple therapy was combined with probiotics adjuvant therapy or prolonged treatment so as to improve the eradication rates of H.pylori effectively.