摘要
为了解某奶牛养殖场奶牛临床乳房炎的致病菌及酸性电解水对奶牛乳房炎致病菌的体外杀菌效果,本试验采集该奶牛养殖场临床乳房炎患病奶牛乳样,对乳样中的病原菌进行分离后通过革兰氏染色和镜检、生化试验、16S rDNA基因序列分析鉴定病原菌种类,并对分离菌株进行药敏试验和动物致病性试验,通过体外杀菌试验分析酸性电解水对分离菌株的杀菌效果。结果表明:分离菌株在Baird-Parker(BP)培养基上生长良好,培养24 h后可见菌落表面光滑,呈灰黑色,无透明圈。革兰氏染色、镜检可见分离菌株呈蓝紫色并排列成葡萄串状,将该分离菌株命名为QSPT2023。分离菌株QSPT2023与葡萄球菌属的生化特性基本一致;16S rDNA基因序列与GenBank收录的产色葡萄球菌OP068076.1的基因序列相似性最高,为99.93%;与产色葡萄球菌OP068076.1位于系统发育树的同一分支,亲缘关系最近;与产色葡萄球菌KJ783397.1、LC317309.1、ON715882.1的亲缘关系较近,与金黄色葡萄球菌、腐生葡萄球菌、表皮葡萄球菌等的亲缘关系较远;对氯霉素、头孢噻肟、头孢呋辛、头孢氨苄、头孢他啶、头孢唑啉、卡那霉素、米诺环素、氧氟沙星、多西环素、氟苯尼考、复方新诺明敏感,对大观霉素、恩诺沙星、罗红霉素、氨苄西林、阿莫西林、两性霉素B、磺胺嘧啶耐药。小鼠腹腔注射0.2 mL分离菌株菌液5 h后出现紧闭双眼、眼周有渗出物、畏寒抱团等症状,3 d内有死亡小鼠,死亡小鼠剖检后有明显的病理特征。用酸性电解水作用产色葡萄球菌分离菌株菌液5 s后杀菌率为99.21%,作用10 s后杀菌率为99.99%,作用15 s后杀菌率为100%,杀菌效果优于传统消毒剂5%聚维酮碘。说明从奶牛乳房炎患病奶牛乳样中成功分离到一株产色葡萄球菌,酸性电解水对该分离菌株具有较好的杀菌效果。
In order to understand the pathogen of the clinical mastitis in dairy cows in a dairy farm and the in vitro bactericidal effects of acid electrolytic water on the pathogen of mastitis in dairy cows, in this experiment, milk samples were collected from cows with clinical mastitis in a dairy farm, and the pathogenic bacteria were isolated from the milk samples;the species of the pathogenic bacteria were identified by Gram staining, microscopic examination, biochemical test, and 16S rDNA gene sequence analysis. The strain was also subjected to drug sensitivity test and animal pathogenicity test, and the bactericidal effect of acid electrolytic water on the strain was analyzed by in vitro bactericidal test. The results showed that the strain grew well on Baird-Parker(BP) medium, and the surface of the colonies was smooth, gray-black and opaque without hyaline ring after 24 h of incubation. Gram staining and microscopic examination showed that the strain was bluish-purple in color and arranged in the shape of grape bunches, and the strain was named QSPT2023. The biochemical characteristics of isolate QSPT2023 were basically consistent with those of Staphylococcus spp. The 16S rDNA gene sequence showed the highest similarity of 99.93% with that of Staphylococcus chromogenes OP068076.1 in GenBank. It was located in the same branch of the phylogenetic tree with Staphylococcus chromogenes OP068076.1 and had the closest affinity;it was closer to Staphylococcus chromogenes KJ783397.1, LC317309.1 and ON715882.1, and was more distant from Staphylococcus aureus, and Staphylococcus saprophyticus, Staphylococcus epidermidis. It was sensitive to chloramphenicol, cefotaxime, cefadroxil, ceftazidime, cefazolin, kanamycin, minocycline, oxfloxacin, doxycycline, florfenicol, and cotrimoxazole;and was resistant to daunorubicin, enrofloxacin, roxithromycin, ampicillin, amoxicillin, amphotericin B, and sulfadiazine. After the mice were injected intraperitoneally with 0.2 mL of bacterial solution of the isolate strain for 5 h, the symptoms such as tightly closed eyes, ooze around the eyes, and chilling and cuddling appeared;there were dead mice within 3 d, and the dead mice had obvious pathological characteristics after autopsy. The bactericidal rates of Staphylococcus chromogenes isolate treated with acid electrolytic water were 99.21% after 5 s, 99.99% after 10 s, and 100% after 15 s. The bactericidal effect was better than that of the traditional disinfectant 5% povidone-iodine. The results suggested that a strain of Staphylococcus chromogenes was successfully isolated from the milk samples of cows suffering from mastitis in a dairy farm, and that acid electrolytic water had a good bactericidal effect on this isolated strain.
作者
田秋丰
尹珺伊
蒋志平
唐旭银
龚有谋
白长胜
刘秋瑾
王欢
张红
赵金波
张蕾
钟鹏
李红宇
TIAN Qiufeng;YIN Juny;JIANG Zhiping;TANG Xuyin;CONG Youmou;BAI Changsheng;LIU Qiujin;WANG Huan;ZHANG Hong;ZHAO Jinbo;ZHANG Lei;ZHONG Peng;LI Hongyu(Animal Husbandry and Veterinary Branch,Heilongjiang Academy of Agricultural Sciences,Qiqihar 161005,China;Heilongjiang Provincial Engineering and Technology Research Center for Forage Herbs Fermentation,Qiqihar 161005,China;Wuhan Luoge Purunsi Eco-Technology Co.,Ltd.,Wuhan 430074,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2024年第6期66-71,共6页
Heilongjiang Animal Science And veterinary Medicine
基金
黑龙江省重点研发计划项目(GA21B006)
黑龙江省农业科学院畜牧兽医分院横向课题“电解水在规模化奶牛场消杀有害微生物的研究与应用”。
关键词
产色葡萄球菌
奶牛乳房炎
酸性电解水
体外杀菌试验
杀菌率
Staphylococcus chromogenes
cow mastitis
acid electrolytic water
in vitro bactericidal test
bactericidal rate
