摘要
为了查明河北省唐山市某豚鼠养殖场发病豚鼠感染的病原菌,本试验采集发病豚鼠的眼角脓性分泌物,采用细菌分离纯化、革兰染色、生化鉴定、16S rRNA基因PCR扩增方法对分离菌进行鉴定,通过K-B试纸法检测分离菌株对24种抗菌药的敏感性,试管二倍稀释法测定19种中药对该分离菌株的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),并通过动物回归试验检测该分离菌株的致病性。结果显示,分离菌株在血琼脂培养基培养24 h生长为针尖状透明菌落、呈β溶血,革兰染色为阳性圆球状、3~5个连接呈短链状,生化特性与马链球菌兽疫亚种相符,将其命名为MLQJ-1;16S rRNA序列比对结果显示,MLQJ-1与马链球菌兽疫亚种同源性高达99.72%,在系统进化树中处于同一分支,鉴定为马链球菌兽疫亚种;MLQJ-1对24种抗菌药的敏感性不同,对青霉素、头孢曲松和阿莫西林等8种抗菌药敏感,对阿米卡星、新霉素和卡那霉素等10种抗菌药耐药;苦地丁、苦参、黄芩和千里光对MLQJ-1的MIC值介于1.9~15.6 mg/mL,MBC值介于1.9~31.2 mg/mL;小鼠腹腔接种MLQJ-1菌液后24 h内全部死亡。结果表明,马链球菌兽疫亚种为本次发病豚鼠的病原菌,青霉素、头孢曲松和苦地丁等药物对该菌具有较强的抑菌效果。
To investigate the pathogenic bacteria infecting diseased guinea pigs in a breeding farm in Tangshan,Hebei Province,purulent secretion from the eye corners of affected guinea pigs was collected.The isolated strains were identified using bacterial isolation and purification,Gram staining,biochemical identification,and 16S rRNA gene PCR amplification.The sensitivity of the isolated strains to 24 antibiotics was determined by the K-B paper method.The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of 19 traditional Chinese medicines against the isolated strain were determined using the tube two-fold dilution method.The pathogenicity of the isolated strain was evaluated through animal regression experiments.The results showed that the isolated strain,named MLQJ-1,grew as needle-like transparent colonies withβ-hemolysis on blood agar after 24 hours of culture.Gram staining revealed positive round cocci,connected in short chains of 3-5.Biochemical characteristics were consistent with Streptococcus equi subsp.zooepidemicus.The 16S rRNA sequence alignment indicated a high homology of 99.72%with Streptococcus equi subsp.zooepidemicus,placing MLQJ-1 in the same branch on the phylogenetic tree,confirming its identification as Streptococcus equi subsp. zooepidemicus . MLQJ-1 showed varying sensitivity to 24 antibiotics, being sensitive to eight, including penicillin, ceftriaxone, and amoxicillin, and resistant to ten, including amikacin, neomycin, and kanamycin. Traditional Chinese medicines such as Corydalis bungeana , Sophora flavescens , Baikal skullcap, and Senecio scandens had MIC values ranging from 1.9 to 15.6 mg/mL and MBC values ranging from 1.9 to 31.2 mg/mL against MLQJ-1. Mice intraperitoneally injected with MLQJ-1 died within 24 hours. The results indicate that Streptococcus equi subsp. zooepidemicus is the pathogenic bacterium in the diseased guinea pigs, and drugs like penicillin, ceftriaxone, and Corydalis bungeana exhibit strong bacteriostatic effects against this bacterium.
作者
李艳红
白和平
信玉春
郭柯宇
王金锋
夏力亮
吴同垒
史秋梅
LI Yanhong;BAI Heping;XIN Yuchun;GUO Keyu;WANG Jinfeng;XIA Liliang;WU Tonglei;SHI Qiumei(Leting County Agriculture and Rural Bureau,Tangshan 063699,China;Key Laboratory of Preventive Veterinary Medicine in Hebei Province,Hebei Normal University of Science and Technology,Qinhuangdao 066604,China;Agriculture and Rural Bureau of Yongqing County,Langfang City,Hebei Province,Langfang 065600,China;Hebei Shuobo Animal Husbandry Service Co.,Ltd.,Tangshan 063500,China)
出处
《中国兽医杂志》
CAS
北大核心
2024年第4期86-91,共6页
Chinese Journal of Veterinary Medicine
基金
中央引导地方科技发展资金项目(236Z6604G)
河北省现代农业产业技术体系肉牛产业创新团队疫病防控与减抗岗位(HBCT2023190201)。
关键词
豚鼠
马链球菌兽疫亚种
分离鉴定
药敏试验
guinea pigs
Streptococcus equi subsp.zooepidemicus
isolation and identification
drug sensitivity test
