牦牛乳源产色葡萄球菌的分离鉴定及毒力与耐药性分析

doi:10.16431/j.cnki.1671-7236.2024.12.040

Abstract

Abstract: 【Objective】 The aim of this study was to investigate the drug resistance of Staphylococcus chromogenes derived from yak milk in Lhasa and Nagqu,Tibet,and explore the carriage of virulence and drug resistance genes.【Method】 Staphylococcus chromogenes was isolated and cultured from yak milk,and its biochemical characteristics were analyzed.Molecular biological identification was performed by PCR and virulence genes and drug resistance genes carried by the isolates were detected.The drug sensitivity test was carried out by Kirby-Baue (K-B) method.【Result】 11 strains of Staphylococcus chromogenes were isolated and identified from 66 yak milk,and the isolation rate was 16.67%.On 5% defibrated sheep blood agar medium,the isolates formed milky white colonies with neat edges,round,smooth and opaque,without hemolysis. Gram staining microscopy showed positive cocci in a moniliform arrangement.Biochemical identification showed that the isolates could ferment fructose,lactose,mannose and sucrose,and the nitrate reduction reaction was positive.The results of 16S rRNA amplification sequencing showed that the similarity between the isolates and Staphylococcus chromogenes in GenBank was more than 98%.2 biofilm-related regulatory genes,icaA and bap,were detected among 13 virulence genes,and the detection rates were 81.82%.Among the 14 drug resistance genes,the detection rate of aminoglycoside resistance gene aph(3')-Ⅰa,macrolide resistance gene ermB and sulfonamine resistance gene sul1 was 63.64%,27.27% and 9.09%,respectively. The resistance rates to penicillin,ampicillin and tetracycline was 100.00%,63.64% and 36.36%,respectively.【Conclusion】 In this study,11 strains of Staphylococcus chromogenes were isolated from 66 yak milk samples collected from Lhasa and Nagqu.The isolates had different degrees of resistance to penicillin,ampicillin,tetracycline and other common antimicrobials,and carried fewer types of virulence and resistance genes.The results provided a theoretical basis for the prevention and treatment of Staphylococcus chromogenes in yaks.

Key words: yak milk; Staphylococcus chromogens; isolation and identification; virulence genes; drug resistance genes

CLC Number:

S852.61

MENG Fanxing, ZENG Jiangyong, QIN Haofeng, ZHAO Congming, YUAN Zhenjie, WANG Dongjing, GESANG Zhuoga, ZHUO Ma, WU Qingxia, MA Hongcai. Isolation and Identification of Staphylococcus chromogenes from Yak Milk and Analysis of Virulence and Drug Resistance[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(12): 5539-5548.

References

[1] HU H, FANG Z, MU T, et al.Application of metabolomics in diagnosis of cow mastitis:A review[J].Frontiers in Veterinary Science, 2021, 8:747519.
[2] TAPONEN S, PYÖRÄLÄ S.Coagulase-negative staphylococci as cause of bovine mastitis—Not so different from Staphylococcus aureus?[J].Veterinary Microbiology, 2009, 134(1):29-36.
[3] PYORALA S, TAPONEN S.Coagulase-negative Staphylococci-emerging mastitis pathogens[J].Veterinary Microbiology, 2009, 134(1-2):3-8.
[4] DE VISSCHER A, PIEPERS S, HAESEBROUCK F, et al.Coagulase-negative Staphylococcus species in bulk milk:Prevalence, distribution, and associated subgroup- and species-specific risk factors[J].Journal of Dairy Science, 2017, 100(1):629-642.
[5] GREEN L E, SCHUKKEN Y H, GREEN M J.On distinguishing cause and consequence:Do high somatic cell counts lead to lower milk yield or does high milk yield lead to lower somatic cell count?[J].Preventive Veterinary Medicine, 2006, 76(1-2):74-89.
[6] GROHN Y T, WILSON D J, GONZALEZ R N, et al.Effect of pathogen-specific clinical mastitis on milk yield in dairy cows[J]. Journal of Dairy Science, 2004, 87(10):3358-3374.
[7] ANDRESEN L O, AHRENS P, DAUGAARD L, et al.Exudative epidermitis in pigs caused by toxigenic Staphylococcus chromogenes[J]. Veterinary Microbiology, 2005, 105(3-4):291-300.
[8] 马玲, 谭义俊, 邵帅, 等.可形成生物膜的禽源产色葡萄球菌的分离鉴定及耐药性研究[J].西南农业学报, 2012, 25(2):703-707.MA L, TAN Y Y, SHAO S, et al.Identification of biofilm-forming Staphylococcus chromogenes isolated from poultry and its drug-resistance experiment[J].Southwest China Journal of Agricultural Sciences, 2012, 25(2):703-707.(in Chinese)
[9] WHITE D G, HARMON R J, MATOS J E, et al.Isolation and identification of coagulase-negative Staphylococcus species from bovine body sites and streak canals of nulliparous heifers[J].Journal of Dairy Science, 1989, 72(7):1886-1892.
[10] DE VISSCHER A, PIEPERS S, HAESEBROUCK F, et al.Teat apex colonization with coagulase-negative Staphylococcus species before parturition:Distribution and species-specific risk factors[J]. Journal of Dairy Science, 2016, 99(2):1427-1439.
[11] ADKINS P R F, DUFOUR S, SPAIN J N, et al.Molecular characterization of non-aureus Staphylococcus spp.from heifer intramammary infections and body sites[J]. Journal of Dairy Science, 2018, 101(6):5388-5403.
[12] TAPONEN S, BJÖRKROTH J, PYÖRÄLÄ S.Coagulase-negative Staphylococci isolated from bovine extramammary sites and intramammary infections in a single dairy herd[J].Journal of Dairy Research, 2008, 75(4):422-429.
[13] GILLESPIE B E, HEADRICK S I, BOONYAYATRA S, et al.Prevalence and persistence of coagulase-negative Staphylococcus species in three dairy research herds[J]. Veterinary Microbiology, 2009, 134(1-2):65-72.
[14] MATTHEWS K R, HARMON R J, SMITH B A.Protective effect of Staphylococcus chromogenes infection against Staphylococcus aureus infection in the lactating bovine mammary gland[J].Journal of Dairy Science, 1990, 73(12):3457-3462.
[15] VANDER ELST N, BELLEMANS J, STEENBRUGGE J, et al.Priming of the murine mammary gland with Staphylococcus chromogenes IM reduces bacterial growth of Streptococcus uberis:A proof-of-concept study[J]. Veterinary Research, 2023, 54(1)：28.
[16] 张鹏飞, 王印, 德西措姆, 等.豪猪源福氏志贺菌的分离鉴定及喹诺酮类耐药基因检测[J].中国预防兽医学报, 2019, 41(2):204-208.ZHANG P F, WANG Y, DEXI C M, et al.Isolation and identification of Shigella flexneri from Hystrix hodgsoni and detection of quinolone resistance genes[J].Chinese Journal of Preventive Veterinary Medicine, 2019, 41(2):204-208.(in Chinese)
[17] 梁成孔.新疆阿克苏地区蛋鸡场鸡群产色葡萄球菌的感染情况调查及耐药与毒力基因检测[D].阿拉尔：塔里木大学, 2023.LIANG C K.Investigation of Staphylococcus chromogenes infection and detection of drug resistance and virulence genes in laying hens in Aksu area of Xinjiang province[D].Alaer:Tarim University, 2023.(in Chinese)
[18] 胡慧, 李田美, 姜艳芬.临床型乳房炎乳样中主要致病菌的分离鉴定及部分分离菌株的药物敏感性[J].中国兽医学报, 2022, 42(12):2529-2533.HU H, LI T M, JIANG Y F.Isolation and identification of main pathogenic bacteriafrom clinical mastitis milk samples and detection of antibiotic susceptivity of some isolated strains[J].Chinese Journal of Veterinary Science, 2022, 42(12):2529-2533.(in Chinese)
[19] 邹明昊, 马弘财, 王冰艺, 等.牦牛产色葡萄球菌的分离鉴定及耐药性分析[J].中国畜牧兽医, 2023, 50(7):3017-3025.ZOU M H, MA H C, WANG B Y, et al.Isolation, identification and drug resistance analysis of Staphylococcus chromogenes in yak[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(7):3017-3025.(in Chinese)
[20] 王秋东, 刘琪, 董志民, 等.内蒙古部分地区致奶牛乳房炎金黄色葡萄球菌和产色葡萄球菌的分子流行病学及耐药性研究[J].中国预防兽医学报, 2017, 39(4):257-261.WANG Q D, LIU Q, DONG Z M, et al.Molecular epidemiology and antimicrobial resistance of Staphylococcus aureus and Staphylococcus chromogenes from dairy cow in the Inner Mongolia[J]. Chinese Journal of Preventive Veterinary Medicine, 2017, 39(4):257-261.(in Chinese)
[21] 赵华男, 瞿玥, 韩博, 等.奶牛乳房炎源性产色葡萄球菌的肠毒素基因检测[J].中国兽医杂志, 2018, 54(5):24-27.ZHAO H N, QU Y, HAN B, et al.Detection of enterotoxin genes in Staphylococcus chromogenes isolated from bovine mastitis[J].Chinese Journal of Veterinary Medicine, 2018, 54(5):24-27.(in Chinese)
[22] 赵霞玲, 肖琦, 钱雯娴, 等.藏猪产色葡萄球菌分离鉴定及生物学特性研究[J].中国畜牧兽医, 2020, 47(9):3014-3021.ZHAO X L, XIAO Q, QIAN W X, et al.Isolation, identification and biological characteristics of Staphylococcus chromogenes from Tibetan pigs[J].China Animal Husbandry & Veterinary Medicine, 2020, 47(9):3014-3021.(in Chinese)
[23] PARSEK M R, SINGH P K.Bacterial biofilms:An emerging link to disease pathogenesis[J].Annual Review of Microbiology, 2003, 57(1):677-701.
[24] MARTINS K B, FACCIOLI P Y, BONESSO M F, et al.Characteristics of resistance and virulence factors in different species of coagulase-negative Staphylococci isolated from milk of healthy sheep and animals with subclinical mastitis[J]. Journal of Dairy Science, 2017, 100(3):2184-2195.
[25] 汤琦, 袁兵, 黄云超, 等.乳腺外科表皮葡萄球菌icaA、icaD及聚集相关蛋白基因对细菌生物膜形成的影响[J].中国修复重建外科杂志, 2014, 28(2):244-249.TANG Q, YUAN B, HUANG Y C, et al.Biofilm and role of icaA, icaD and accumulation-associated protein[J].Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(2):244-249.(in Chinese)
[26] VANDERHAEGHEN W, PIEPERS S, LEROY F, et al.Invited review:Effect, persistence, and virulence of coagulase-negative Staphylococcus species associated with ruminant udder health[J].Journal of Dairy Science, 2014, 97(9):5275-5293.
[27] 徐结.甘、青、宁地区奶牛乳房炎葡萄球菌分子特性研究[D].北京：中国农业科学院, 2019.XU J.Molecular charactrtistics of Staphylococcus from bovine mastitis in Gansu, Qinghai and Ningxia province[D].Beijing：Chinese Academy of Agricultural Sciences, 2019.(in Chinese)
[28] 裘鹏, 郭玉茹, 张泽辉, 等.基于转录组学技术分析细菌耐药机制的研究进展[J].中国家禽, 2019, 4:45-49.QIU P, GUO Y R, ZHANG Z H, et al.Progress in drug resistance mechanism of bacteria based on transcriptome technology[J].China Poultry, 2019, 4:45-49.(in Chinese)
[29] REGECOVÁ I, V AY＇G ROSTKOVÁ J, ZIGO F, et al.Detection of antimicrobial resistance of bacteria Staphylococcus chromogenes isolated from sheep’s milk and cheese[J].Antibiotics (Basel), 2021, 10(5):570.
[30] 赵德军, 张碧霞, 曹雁, 等.乳腺分泌物中检出产色葡萄球菌1例[J].西南军医, 2007, 9(6):36.ZHAO D J, ZHANG B X, CAO Y, et al.One case of Staphylococcus chromogenes was detected in breast secretions[J].Journal of Military Surgeon in Southwest China, 2007, 9(6):36.(in Chinese)
[31] 傅冬梅, 明德松.产色葡萄球菌临床分离株耐药性及耐药机制分析[J].国际检验医学杂志, 2012, 33(18):2265-2266.FU D M, MING D S.Analysis of drug resistance and drug resistance mechanism of clinical isolates of Staphylococcus chromogenes[J].International Journal of Laboratory Medicine, 2012, 33(18):2265-2266.(in Chinese)
[32] 陈广, 温贵兰, 张喜懿, 等.禽源产色葡萄球菌的分离鉴定及其耐药性分析[J].中国畜牧兽医, 2020, 47(7):2215-2222.CHEN G, WEN G L, ZHANG X Y, et al.Isolation and identification of avian-origin Staphylococcus chromogenes and its drug resistance analysis[J].China Animal Husbandry & Veterinary Medicine, 2020, 47(7):2215-2222.(in Chinese)
[33] 李文杰.奶牛乳房炎凝固酶阴性葡萄球菌的分离鉴定与耐药性分析[D].杨凌：西北农林科技大学, 2017.LI W J.The separation and identification of dairy cow mastitis and drug resistance analysis in coagulase negative Staphylococcus[D].Yangling:Northwest A&F University, 2017.(in Chinese)
[34] 李一鸣, 王少林.肠杆菌科细菌耐药基因表达的遗传和环境调控[J].生物工程学报, 2021, 37(4):1092-1106.LI Y M, WANG S L.The impact of genetic and environmental regulation on the expression of antibiotic resistance genes in Enterobacteriaceae[J].Chinese Journal of Biotechnology, 2021, 37(4):1092-1106.(in Chinese)
[35] WANG X, YU D, CHEN L.Antimicrobial resistance and mechanisms of epigenetic regulation[J].Frontiers in Cellular and Infection Microbiology, 2023, 13:1199646.

Isolation and Identification of Staphylococcus chromogenes from Yak Milk and Analysis of Virulence and Drug Resistance

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