额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究

doi:10.12160/j.issn.1672-5190.2024.04.013

畜牧与饲料科学 ›› 2024, Vol. 45 ›› Issue (4): 103-108.doi: 10.12160/j.issn.1672-5190.2024.04.013

额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究

苏政¹, 于月通², 朱婷婷¹, 陀海欣², 李静², 吴静², 刘鹏¹, 彭健¹, 齐萌²

1.新疆生产建设兵团第九师农业科学研究所（畜牧科学研究所）,新疆额敏 834601;
2.塔里木大学动物科学与技术学院/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

收稿日期:2024-05-13 出版日期:2024-07-30 发布日期:2024-08-23
通讯作者: 齐萌（1985—）,男,教授,博士,硕士生导师,主要从事人兽共患病病原生物学研究工作。
作者简介:苏政（1993—）,男,硕士,主要研究方向为动物群发病防控。
基金资助:
新疆生产建设兵团科技创新人才计划（2021CB062）; 新疆生产建设兵团第九师科技计划项目（2021JS001,2022JS008）

Isolation,Identification,Antimicrobial Resistance and Pathogenicity of a Staphylococcus aureus Strain from Feces of Sheep in Emin County,China

SU Zheng¹, YU Yuetong², ZHU Tingting¹, TUO Haixin², LI Jing², WU Jing², LIU Peng¹, PENG Jian¹, QI Meng²

1. Agricultural Science Research Institute （Animal Husbandry Science Research Institute）of the Ninth Division of the Xinjiang Production and Construction Corps,Emin 834601,China;
2. College of Animal Science and Technology,Tarim University/Engineering Laboratory of Tarim Animal Diseases Diagnosis and Control of the Xinjiang Production and Construction Corps,Alar 843300,China

Received:2024-05-13 Online:2024-07-30 Published:2024-08-23

摘要/Abstract

摘要： [目的]对羔羊腹泻粪便样本中的金黄色葡萄球菌进行分离鉴定,了解其耐药性和致病性特征。[方法]从新疆维吾尔自治区塔城地区额敏县某养羊场采集腹泻羔羊肛拭子样本4份,使用高盐甘露醇培养基（MSA）选择性分离金黄色葡萄球菌;通过形态学观察和16S rDNA序列分析对分离菌株进行种属鉴定;采用纸片扩散法分析分离菌株对9类11种抗菌药物的敏感性;利用PCR法检测分离菌株毒力基因携带情况;建立昆明系小白鼠腹腔感染模型,开展分离菌株的致病性试验。[结果]从4份羔羊腹泻样本中分离到1株疑似金黄色葡萄球菌,该菌株在MSA培养基上形成有光泽、中度隆起、金黄色或柠檬色的菌落;革兰染色镜检可见分离菌株为革兰阳性,呈葡萄串状排列;16S rDNA序列分析显示,分离菌株的序列与我国湖南省长沙市猪肺源金黄色葡萄球菌分离株序列的相似性为100%;结合形态学观察和16S rDNA序列比对结果,鉴定该菌株为金黄色葡萄球菌。分离菌株对青霉素、克林霉素、头孢噻吩、利福平和红霉素耐药,对庆大霉素、阿奇霉素、氯霉素、诺氟沙星、头孢西丁和复方新诺明敏感。在检测的8类20种毒力基因中,该菌株携带sea、seg、sei、eta、clfA和clfB基因。该菌株对小鼠的半数致死量（LD₅₀）为2.24×10⁸ CFU/mL,死亡小鼠脏器病理变化明显。[结论]从额敏县某养羊场羔羊腹泻样本中分离鉴定到1株金黄色葡萄球菌,该菌株具有多重耐药表型,携带多种毒力基因,具有一定致病性。研究结果为该地区羊金黄色葡萄球菌病的合理用药提供了参考。

关键词: 羊, 金黄色葡萄球菌, 分离, 鉴定, 耐药性, 致病性

Abstract: [Objective] The aims of the present study were to isolate and identify the Staphylococcus aureus （S. aureus） strain from fecal samples of lambs with diarrhoea, and to characterize the antimicrobial resistance and pathogenicity of the isolate. [Method] Four anal swab samples of diarrheic lambs were collected from a sheep farm in Emin County, Tacheng Prefecture, Xinjiang Uygur Autonomous Region, and S. aureus strain was selectively isolated with mannitol salt agar （MSA）. The isolate was identified by morphological observation and 16S rDNA sequence analysis. The in vitro susceptibility of the isolate to 11 antimicrobial agents belonging to 9 categories was analyzed by paper diffusion method. The presence of virulence genes of the isolate was detected by PCR assay. A model of abdominal infection in Kunming mice was established to evaluate the pathogenicity of the isolate. [Result] One suspected S. aureus strain was isolated from the four fecal samples of diarrheic lambs. It formed glossy, moderately raised, golden or lemon coloured colonies on MSA. Gram staining and morphological observation revealed that the isolate was Gram-positive and arranged in the form of grape bunches. The 16S rDNA sequence of the isolate was 100% similar to that of a S. aureus isolate of porcineluny origin from Changsha City, Hunan Province, China. Morphological observation and 16S rDNA sequence analysis confirmed that the isolate was S. aureus. The isolate was resistant to penicillin, clindamycin, cefotiamine, rifampicin and erythromycin, and was sensitive to gentamicin, azithromycin, chloramphenicol, norfloxacin, cefoxitin and cotrimoxazole. Out of the 20 virulence genes belonging to 8 classes tested, the presence of sea, seg, sei, eta, clfA and clfB genes was observed. The LD₅₀ of the isolate for Kunming mice was 2.24×10⁸ CFU/mL, with observable pathological changes in the organs of the dead mice. [Conclusion] One S. aureus strain was isolated and identified from the fecal samples of diarrheic lambs in a sheep farm in Emin County, which exhibited a multi-drug resistant phenotype, harbored multiple virulence genes and had certain pathogenicity. The results of this study provide references for the rational use of medication for the diseases caused by S. aureus infection in sheep in this region.

Key words: sheep, Staphylococcus aureus, isolation, identification, antimicrobial resistance, pathogenicity

中图分类号:

S852.611

苏政, 于月通, 朱婷婷, 陀海欣, 李静, 吴静, 刘鹏, 彭健, 齐萌. 额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究[J]. 畜牧与饲料科学, 2024, 45(4): 103-108.

SU Zheng, YU Yuetong, ZHU Tingting, TUO Haixin, LI Jing, WU Jing, LIU Peng, PENG Jian, QI Meng. Isolation,Identification,Antimicrobial Resistance and Pathogenicity of a Staphylococcus aureus Strain from Feces of Sheep in Emin County,China[J]. Animal Husbandry and Feed Science, 2024, 45(4): 103-108.

参考文献

[1] 董炜,李军.奶牛乳房炎源金黄色葡萄球菌血清型鉴定、毒力基因及耐药性检测与分析[J].中国动物传染病学报,2022,30(6):113-117.
[2] JARRAUD S, MOUGEL C, THIOULOUSE J, et al.Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease[J]. Infection and Immunity,2002,70(2):631-641.
[3] THOMAS D Y, JARRAUD S, LEMERCIER B, et al.Staphylococcal enterotoxin-like toxins U2 and V, two new staphylococcal superantigens arising from recombination within the enterotoxin gene cluster[J]. Infection and Immunity,2006,74(8):4724-4734.
[4] LØVSETH A, LONCAREVIC S, BERDAL K G. Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates[J]. Journal of Clinical Microbiology,2004,42(8):3869-3872.
[5] 马弘财,董海龙,樊世杰,等.日喀则藏羊源金黄色葡萄球菌的分离鉴定和药物敏感性分析[J].中国兽医杂志,2023,59(8):77-81.
[6] RAHBARNIA L, KHOSRAVI R R, DEHNAD A R, et al.The examination of some virulence factors in S. aureus isolates obtained from the healthy human population, sheep mastitis, and cheese[J]. Iranian Journal of Veterinary Research,2023,24(2):110-115.
[7] 杨雪娇,王一涵,项勋,等.奶山羊隐性乳房炎金黄色葡萄球菌的分离鉴定及毒力基因的检测[J].中国兽医科学,2024,54(5):650-657.
[8] 陈泽世. 羊源金黄色葡萄球菌的耐药和分型检测及金黄色葡萄球菌ndhF缺失株耐药相关功能研究[D].海口:海南大学,2022.
[9] 马木尔·阿克木汉,姚晓慧,刘英玉,等.新疆焉耆县不同动物源葡萄球菌耐药性及其亲缘关系分析[J].动物医学进展,2020,41(7):27-33.
[10] 刘肖利,刘璐瑶,李镔罡,等.金黄色葡萄球菌性奶牛乳房炎调查及菌株耐药性和毒力分析[J].西北农业学报,2021,30(10):1452-1460.
[11] LI X H, FANG F, ZHAO J, et al.Molecular characteristics and virulence gene profiles of Staphylococcus aureus causing bloodstream infection[J]. The Brazilian Journal of Infectious Diseases,2018,22(6):487-494.
[12] SONOWAL D, GHATAK S, BARUA A G, et al.Livestock, pets and humans as carriers of methicillin-resistant Staphylococcus aureus and comparative evaluation of two PCR protocols for detection[J]. Veterinary Research Forum,2023,14(7):351-358.
[13] SABIR M J, IJAZ M, AHMED A, et al.First report on genotypic estimation of MRSA load in udder of nomadic sheep flocks affected with subclinical mastitis in Pakistan[J]. Research in Veterinary Science,2024,166:105107.
[14] 李永光,简莹娜,贺胜,等.一起羊皮肤脓肿病例的病原诊断及其防治[J].青海畜牧兽医杂志,2022,52(3):26-29.
[15] SPANU V, SCARANO C, COSSU F, et al.Antibiotic resistance traits and molecular subtyping of Staphylococcus aureus isolated from raw sheep milk cheese[J]. Journal of Food Science,2014,79(10):2066-2071.
[16] 王琦,宋衍燕.金黄色葡萄球菌肠毒素与多位点序列分型分子分型研究进展[J].食品安全质量检测学报,2021,12(12):4714-4720.
[17] AUNG M S, URUSHIBARA N, KAWAGUCHIYA M, et al.Prevalence and genetic diversity of staphylococcal enterotoxin (-like) genes sey, selw, selx, selz, sel26 and sel27 in community-acquired methicillin-resistant Staphylococcus aureus[J]. Toxins,2020,12(5):347.
[18] KATSAROU I, PARASKEVOPOULOU N M, PAPADIMITRIOU-OLIVGERIS M, et al.Fatality of Staphylococcus aureus infections in a Greek university hospital:Role of inappropriate empiric treatment, methicillin resistance, and toxin genes′ presence[J]. European Journal of Clinical Microbiology and Infectious Diseases,2020,39(3):443-450.
[19] CHAN Y L, CHEE C F, TANG S N, et al.Unveilling genetic profiles and correlations of biofilm-associated genes, quorum sensing, and antibiotic resistance in Staphylococcus aureus isolated from a Malaysian Teaching Hospital[J]. European Journal of Medical Research,2024,29(1):246.

额敏县1株羊粪源金黄色葡萄球菌的分离鉴定及其耐药性与致病性研究

Isolation,Identification,Antimicrobial Resistance and Pathogenicity of a Staphylococcus aureus Strain from Feces of Sheep in Emin County,China

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