【摘要】 为了明确致猫呼吸道症状的病原菌及该菌的病原特征及其基因组功能信息，本研究对采集的3例发病猫的鼻腔拭子进行病原菌的分离、纯化及生化和16S rRNA基因的PCR鉴定；采用纸片法检测分离菌的药物敏感性及其对小鼠的致病性。并对其中一株分离菌进行基因组的生物信息学分析，利用不同的数据库对基因组编码的蛋白进行功能注释以及耐药和毒力相关基因的预测。细菌的分离及鉴定结果显示：分离出3株支气管败血波氏杆菌（Bb）。药敏试验结果显示，分离菌对丁胺卡那、卡那霉素、四环素、氟苯尼考、恩诺沙星、环丙沙星、诺氟沙星和氯霉素敏感，对氨苄西林、链霉素、利福平、磷霉素和呋喃唑酮耐药；致病性试验结果显示，3株分离的Bb对小鼠均有一定的致病性，且浓度越高致病性越强。基因组分析结果显示，其中一株菌基因组大小为4 520 328 bp,GC%含量为68.12%，含有4 690个蛋白质编码基因，87个tRNA基因，25个rRNA基因，1个tmRNA基因等信息；分离菌基因组编码蛋白功能的注释结果显示该菌的功能蛋白主要表现在细菌的复制增殖和生物代谢方面；分离菌携带多种耐药基因，包括主动外排系统相关基因、多粘菌素抗性基因、磷霉素抗性基因等，并基本与耐药表型一致；携带66种毒力基因，主要是分泌系统类毒力基因以及内外毒素、黏附侵袭类毒力因子基因。上述结果表明，本研究分离的Bb耐药性不强，但对小鼠呈强致病力，且含多种毒力相关基因。本研究为对该菌的研究奠定基础，并为该菌感染的防控研究提供参考。更多还原

【Abstract】 In order to study the pathogen characteristics and genomic function information of Bordetella bronchiseptica(Bb),which can infect many animals and cause respiratory diseases. In this study, the nasal swabs of 3 infected cats were isolated,purified, identified, tested for drug sensitivity and pathogenicity in mice, and the genome of one isolate was sequenced and analyzed. Different databases were used for functional annotation of gene sequences, as well as gene prediction of drug resistance and virulence. The results showed that all the three isolates were Bordetella bronchiseptica, and were highly sensitive to butanicana, kanamycin, tetracycline, fluorobenicol, enrofloxacin, ciprofloxacin, norfloxacin and chloramphenicol, but resistant to ampicillin, streptomycin, rifampicin, fosfomycin and furazolidone. The results of pathogenicity experiments showed that the three isolated Bb strains had certain pathogenicity to mice, and the higher the concentration, the stronger the pathogenicity. The genome size of this bacterium was 4 520 328 bp, and GC content was 68.12%. It contained 4690 protein-coding genes(CDS), 87 tRNA, 25 rRNA, 1 tmRNA, among other information. The CDS shows that the bacteria’s functional proteins are mainly involved in the replication and proliferation of bacteria and biological metabolism. Additionally, it had multiple resistance genes: active efflux systems related genes, the polymyxin resistance gene, the fosfomycin resistance gene, and so on, which were consistent with the drug resistance phenotype. There are 66 virulence genes, mainly including secretion systems, endotoxin and exotoxin, adhesion and invasion virulence factors. The above results indicate that the Bb isolates in this study has weak drug resistance but display strong pathogenicity to mice, and contains a variety of virulence-related genes. These results expand our knowledge on the pathogens of cat disease in China, and will provide a basis for control and prevention of Bb-caused diseases.更多还原