山羊伪结核棒状杆菌的分离与鉴定

Isolation and identification of a goat Corynebacterium parvum

近年来陕西省规模化羊场中病羊躯干出现脓包的情况不断出现。为了获得引起陕西某山羊场病羊脓包的病原菌,无菌采集病羊脓包液,进行细菌分离、革兰染色、生化试验、16S rDNA序列分析及小鼠致病性试验。结果：获得1株具有β溶血特性、菌落直径大小为1~2 mm、呈苍白色、表面粗糙的革兰阳性球杆菌;16S rDNA扩增大小为1 412 bp,比对结果显示与伪结核棒状杆菌ATCC 19410菌株的核苷酸相似性为99%;用0.2 mL/只（1×10~8cfu/m L）的菌液剂量接种昆明小鼠,死亡率为100%（10/10）,最终确定分离纯化病原菌为具有致病性的伪结核棒状杆菌。药敏试验结果显示,分离菌对环丙沙星、头孢噻肟、氧氟沙星等多种抗生素敏感,对复方新诺明、杆菌肽等多种抗生素表现为耐药,药敏结果为该病的防治提供了依据。

In recent years,the incidence of pustule on sheep torso continues to appear in large-scale sheep farms in Shaanxi Province. In order to obtain the pathogenic bacteria that cause pustules in sick goats,we collected samples of pustules liquid from the sick goats and biochemical experiments,16 S r DNA sequence analysis and pathogenicity test with the samples in mice. The result was that a strain of Grampositive cocci was obtained and was of β-hemolytic character,colony size of 1 to 2 mm in diameter,of pale color,and rough surface. The Gram-positive cocci was amplified by 16 S rDNA to 1412 bp in size and showed 99% nucleotide similarity to Corynebacterium parvum. Then,a bacterial volume of 0. 2 mL/mouse（1 × 10~8 cfu/m L） was inoculated to the Kunming mice resulting in a mortality of 100%（10/10）. Finally,the pathogenic bacterium was isolated and identified as Pseudomonas Corynebacterium with pathogenicity. The result of the drug susceptibility test showed that the isolates of Corynebacterium tuberculosis were resistant to various antibiotics such as ciprofloxacin,cefotaxime and ofloxacin,was sensitive to compound cotinine,bacitracin and other antibiotics. The drug susceptibility results here provides a basis for prevention and treatment of goat pustules.