| Based on the isolation and identification technologies of rabbit DiplococcusPneumoniae, the studies were done from2013to2014. the rabbit DiplococcusPneumoniae was investigated in North-Shandong Veterinary Clinic of ShandongBinzhou animal science and veterinary medicine academy. Five rabbit farms weresurveyed and analyzed that if a single rabbit Diplococcus Pneumococcal had occurred.Physiological and biochemical test, hemolytic test and the animal pathogenicity testwere done. Meanwhile nasal swabs were collected from healthy rabbit to analyze thehealth rabbit carrier Diplococcus Pneumococcus, to provide important reference forthe development of rational and effective rabbit Diplococcus pneumococcal diseaseprevention and control measures. One isolated rabbit Diplococcus pneumococcus wastested to screen medium. Single-factor test components and ratios were optimized onrabbit Diplococcus pneumococcus growth medium, ultimately appropriate enrichmentthe bacteria growth medium was got. The foundation lied for rabbit Diplococcuspneumococcal vaccine-scale production.1. Isolation and identification of Diplococcus pneumoniae in rabbitsShandong Binzhou City, a rabbit farm has over1000rabbits. February2013,pups sporadic deaths occurred. A week after, the sudden death of rabbit pupsincreased and an increase in the case of pregnant female miscarriage and stillbirth lowearners. Some females appear otitis media and rhinitis, mostly cold symptoms, andrabbit mortality rate was low. To mid-March, the incidence rate of40%of pups, themortality rate as high as70%. The incidence and mortality rate of females was20%and10%, respectively. There are less disease onset and death in commercial rabbit.Pathogens were collected and isolated form dead rabbit lungs. A strain ofgram-positive cocci was successfully isolated. A series of Diplococcus pneumococcustraditional identification test results showed that isolates haveα-hemolytic, Ao Botuonew test positive, bile lysis test positive and PCR test showed that the16S rRNAsequence homology was99%. Animal regression test successfully replicate the typical symptoms of Diplococcus pneumococcal disease in rabbits and pathological change.Pathogens were confirmed as rabbit Diplococcus pneumococcus. Isolation andidentification supported technical for the investigation of rabbit pneumococcal.2. Investigation of rabbit Diplococcus pneumococcal disease in BinzhoudistrictFrom2013to2014, the North-Shandong Veterinary Clinic selected223suspectedrabbits samples from417cases for rabbit Diplococcus pneumococcal diseaseinvestigation and pathogenic bacteria isolation. The respiratory system samples wereused to isolate the bacteria and identification. The information such as morbidity rate,clinical symptoms, breeding aquatic and rabbit numbers was all collected andanalyzed. Nasal swabs from healthy rabbits in6large scale farms and10householdswere collected, of which the pathogenic bacteria were isolated and identified forevaluating the bacteria-bring rate. All the studies could provide us important data andinformation for the control and prevent of rabbit Diplococcus pneumococcal disease.3. Screening and optimizing of enrichment liquid medium for DiplococcuspneumoniaeDuring this study, four medium including general broth, buffered Martin broth,brain-heart infusion broth, and BYGPglur broth were firstly used for evaluating theirbacteria enriching capacity. The result indicated that BYGPglur broth was the bestmedium for Diplococcus pneumonia growth. Secondly, different formulas of theBYGP-glur broth ingredients were optimized, and the eventually broth formula was:beef infusion powder5.0g/L, soy peptone10.0g/L, yeast extract50.0ml/L, glucose1.0g/L, NaCl5.0g/L, glutamine1.0g/L,K2HPO43.68g/L,KH2PO41.32g/L, rabbitserum5%. The OD value of optimized broth was remarkably higher than generalbroth（124.5%）,indicating that it can be useful to improving the production of rabbitDiplococcus pneumococcal vaccine. |
PDF Full Text Request