新疆南疆地区乳源金黄色葡萄球菌的分离鉴定及对氟苯尼考纳米凝胶的药物敏感性研究  

Isolation and identification of milk-derived Staphylococcus aureus from the southern Xinjiang region and study on its drug sensitivity to florfenicol nanogel

作　　者：冷楠楠 刘燕 窦蕊 鞠牧洁 刘金环 陈伟[1,2,3] 罗万和 LENG Nannan;LIU Yan;DOU Rui;JU Mujie;LIU Jinhuan;CHEN Wei;LUO Wanhe(School of Animal Science and Technology,Tarim University,Aral 843300,China;Tarim Animal Disease Diagnosis and Prevention and Control Engineering Laboratory of Xinjiang Production and Construction Corps,Tarim University,Aral 843300,China;Tarim Basin Key Laboratory of Conservation and Utilization of Biological Resources,Xinjiang Production and Construction Corps,Tarim University,Aral 843300,China)

机构地区：[1]塔里木大学动物科学与技术学院,新疆阿拉尔843300 [2]塔里木大学新疆生产建设兵团塔里木动物疫病诊断与防控工程实验室,新疆阿拉尔843300 [3]塔里木大学新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,新疆阿拉尔843300

出　　处：《黑龙江畜牧兽医》2024年第6期59-65,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：塔里木大学大学生创新创业训练计划资助项目(202310757001);塔里木大学校长基金项目(TDZKSS202144);新疆生产建设兵团中青年科技创新领军人才项目(2019CB029)。

摘　　要：为了研究氟苯尼考纳米凝胶对新疆南疆地区乳源金黄色葡萄球菌的抑菌效果,试验从新疆南疆地区某奶牛场的乳房炎患病奶牛乳样中分离纯化金黄色葡萄球菌,采用生化试验、PCR扩增及测序等方法进行细菌鉴定,并通过比较氟苯尼考原料药、氟苯尼考纳米凝胶和氟苯尼考注射液对金黄色葡萄球菌分离株的抑菌圈直径(DIZ)、最小抑菌浓度(MIC)和抑菌曲线分析氟苯尼考纳米凝胶对金黄色葡萄球菌分离株的抑菌效果。结果表明:从乳样中分离到一株细菌,该分离株革兰氏染色、镜检后呈蓝紫色并排列成葡萄串状,触酶、血浆凝固酶和甘露醇发酵试验结果均为阳性,其基因组DNA中能扩增出金黄色葡萄球菌耐热核酸酶(nuc)基因,16S rDNA基因序列与金黄色葡萄球菌的相似性为99.7%,判断该分离菌为金黄色葡萄球菌。氟苯尼考原料药、氟苯尼考纳米凝胶和氟苯尼考注射液对金黄色葡萄球菌分离株和金黄色葡萄球菌ATCC 29213株均产生抑菌作用,DIZ大小为氟苯尼考纳米凝胶>氟苯尼考注射液>氟苯尼考原料药,均表现为高度敏感,其中氟苯尼考纳米凝胶对金黄色葡萄球菌分离株和金黄色葡萄球菌ATCC 29213株的DIZ均显著高于氟苯尼考原料药和氟苯尼考注射液(P<0.05)。3种药物对金黄色葡萄球菌分离株的MIC分别为4μg/mL、2μg/mL和4μg/mL。1μg/mL氟苯尼考纳米凝胶对金黄色葡萄球菌分离株的抑菌效果与2μg/mL氟苯尼考原料药和氟苯尼考注射液相当,当氟苯尼考纳米凝胶的浓度为4μg/mL时金黄色葡萄球菌分离株不再生长。说明从新疆南疆地区成功分离出一株乳源金黄色葡萄球菌;相比于氟苯尼考原料药和氟苯尼考注射液,氟苯尼考纳米凝胶对该分离株具有较好的抑菌效果。In order to study the bacteriostatic effects of florfenicol nanogel on milk-derived Staphylococcus aureus from the southern Xinjiang region, in the experiment, milk-derived Staphylococcus aureus was isolated and purified from the cows suffering from mastitis in a dairy farm in the southern Xinjiang region. Bacteria were identified by biochemical test, PCR amplification and sequencing.The bacteriostatic effects of florfenicol nanogel against Staphylococcus aureus isolate were also analyzed by comparing the diameters of the inhibition zone(DIZ), the minimum inhibitory concentrations(MIC) and the inhibitory curves of florfenicol raw material, florfenicol nanogel and florfenicol injection against Staphylococcus aureus isolate. The results showed that one bacterial strain was isolated from the milk sample;the isolate was blue-purple and arranged in grape bunches after Gram staining microscopy. The catalase, plasma coagulase and mannitol fermentation were positive;the Staphylococcus aureus heat-resistant nuclease(nuc) gene was amplified in its genome DNA;the 16S rDNA gene sequence showed 99.7% similarity to that of Staphylococcus aureus. Florfenicol API, florfenicol nanogel and florfenicol injection all showed bacteriostatic effect against Staphylococcus aureus isolate and Staphylococcus aureus ATCC 29213 strain. The DIZ sizes were florfenicol nanogel > florfenicol injection > florfenicol raw material, all of which showed high sensitivity. The DIZ of flufenicol nanogel on Staphylococcus aureus isolate and Staphylococcus aureus ATCC 29213 strain was significantly higher than that of florfenicol API and florfenicol injection(P < 0.05).The MICs of the three drugs against Staphylococcus aureus isolates were 4 μg/mL, 2 μg/mL, and 4 μg/mL, respectively. The inhibitory effect of 1 μg/mL florfenicol nanogel on the Staphylococcus aureus isolate was comparable to that of 2 μg/mL florfenicol API and commercially available florfenicol injection;when the concentration of florfenicol nanogel was 4 μg/mL, the Staphylococcus aur

关 键 词：金黄色葡萄球菌 氟苯尼考纳米凝胶 抑菌活性 奶牛乳房炎 菌株分离 

分 类 号：S859.79[农业科学—临床兽医学]