| The Klebsiella disease is an acute zoonotic infectious disease which is caused by Klebsiella, Enterobacter Koch Bacillus genus. The disease occurred in a variety of poultry, livestock and wildlife frequently. Klebsiella pneumoniae, belonging to Enterobacter Cork Klebsiella genus, is a common condition pathogens and widely distributed in the animal’s digestive tract, digestive tract, water, soil and grain. It can cause pneumonia, peritonitis, metritis, diarrhea, and even sepsisis in humans and animals.But for a long time, Klebsiella pneumoniae disease as a condition of pathogenic bacteria was considered to be a little risk on livestock and poultry, thus it has not been paid enough attention. In recent years, Modern aquaculture production methods has developed rapidly, but there are several reports about infected cases in different livestock, poultry with sheep due to the management level of the farmers, farm hardware and software facilities Lagging behind the scale and other reasons. However, the relevant rabbit group infected by Klebsiella was rarely reported. With the development of the rabbit industry in China, Rabbit manner was changed from fragmented and small amount breeding to the regional farming and large-scale breeding, large-scale, high density and ventilation led to an upward trend of the rabbit respiratory diseases. Moreover, part of the females and pups died of diarrheal disease and clinical symptoms suspected it to be Klebsiella disease in Qindao after the winter of2011. Thus it was significant to carry out studies on the Klebsiella disease in order to prevent and control the disease. In this study, Isolation and identification of Klebsiella and diversity analysis of the disease provide basic information for effective treatment and prevention and control of the disease.In this study, thirty cases of dead rabbits in the incidence of rabbit house were detected. Samples of liver, spleen, lung efforts are collected and divided into two parts. One was isolated, cultured and made into smear. Bacterial examination and physiological and biochemical identification of bacterial were conducted. Then animal regression testing and drug sensitivity test were carried out to determine the types of bacterial pathogens and filter effective antibiotic treatment of the disease. And the other materials are used for DNA extraction using ERIC-PCR technology to get ERIC-PCR fingerprints of Klebsiella. Then the molecular biology of Klebsiella diversity were analyzed using NTSYS-pc.The results showed:Bacillus was found in24heart-blood smears of the total30samples. The bacillus dyed strongly in amphi with Methylene Blue was Gram-negative bacterium. It had capsule but without gemma. The capsule was easily observed after staining with Indian ink. The experimental result of cultural character and biochemical characteristics of pathogenic bacterium showed that the strain was K. peumoniae of rabbit. We cultured the bacteria purely and inculated mice intraperitoneally with pure culture bacteria diluted with PBS. The result showed that the inoculation of0.3ml/mouse died firstly, and all died after three to seven hours inoculation. And inoculum of0.2ml/mouse died after five to twelve hours inoculation. The nectropsy of the dead mice shows similar pathological changes to the rabbits died for the disease. Animals regression experimental results showed that the isolates were strong pathogenic, and the cause of the dead clinical rabbits was Klebsiella pneumoniae infection. Susceptibility test results showed that this bacteria was only sensitive to norfloxacin and doxycycline drugs and resistant to most antimicrobial agents.24clinical isolates were amplified using ERIC-PCR method and the bands were obvious. The smallest band was about280bp and the biggest band was3000bp. The different fingerprints got were analyzed by NTSYS-pc software to construct the cluster tree. Twenty-four clinical isolates of Klebsiella were divided into fourteen genotypes, then denoted as A-N. Among these genotypes, genotype A accounted for16.67%and was designated as dominant bacteria; secondly, genotype B、C、D、F、G、 J and K accounted for8.33%; thirdly genotype E、G、I、L、M and N accounted for4.17%. It could be seen from the results of the analysis that genotype A Klebsiella bacteria was the dominant bacteria causing the rabbit infected. Then were the genotypeB, C, D, F, G, J and K. The results of this study provided a scientific basis for the prevention and control of Klebsiella disease.The conclusions of this study:in this research conventional bacterial tests, biochemical identification tests and ERIC-PCR technique were used to study the cause of the rabbits death in Qingdao. The bacterial pathogens Klebsiella were confirmed and sensitive drugs such as norfloxacin and doxycycline filtered out. this research provided a favorable theoretical basis for the effective prevention and control of the disease and treatment; the molecular diversity of the Klebsiella in the region was analyzed by ERIC-PCR method, thus it provided a new theoretical basis for further study of the bacteria. |
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