原料乳和临床乳房炎金黄色葡萄球菌毒力基因检测及药敏分析

对临床乳房炎（57株）和原料乳（44株）金黄色葡萄球菌菌株,用PCR方法检测mecA基因、PVL基因、ETs基因、SEs基因和TSST-1基因;采用CLSI指导说明执行琼脂稀释法药敏性试验。结果显示原料乳菌株中,84.09%携带有毒素基因,其中PVL的检出率为84.09%,肠毒素的检出率为52.27%,主要流行的肠毒素基因为sea（56.82%）,均未检测到携带mecA、ETs、TSST-1、sei和sej基因的菌株;同时得到10种毒素基因型,其主要流行的毒素基因型为PVL＋sea（29.55%）和PVL（27.27%）。临床菌株中,78.95%携带有毒素基因,其中PVL的检出率为28.07%,肠毒素的检出率为77.19%,主要流行的肠毒素基因为sea（47.37%）,没有检测到携带ETs、TSST-1和seh基因菌株;同时得到25种毒素基因型,其主要流行的毒素基因型为sea（19.30%）,其次是seb（7.02%）,sea＋sed＋sej（3.51%）和PVL＋sea＋seb＋sec＋seg＋sei（3.51%）。6株（10.53%）携带有mecA基因菌株均含有较多毒素基因。原料乳分离株对甲氧苄啶和头孢西丁的耐药率较高,分别为100%和86.36%,其次对氯霉素、红霉素、苯唑西林、头孢哌酮和庆大霉素的耐药率分别为11.36%、4.55%、2.27%、2.27%和6.82%,所有原料乳菌株均对环丙沙星敏感,同时得到8种耐药谱,多重耐药率达22%;临床乳房炎菌株对红霉素和甲氧苄啶的耐药率较高,分别为100%和71.93%,其次对氯霉素、庆大霉素、环丙沙星、头孢西丁和苯唑西林的耐药率分别为28.07%、26.07%、24.56%、19.30%和7.02%,临床乳房炎菌株对头孢哌酮和四环素的敏感率为100%,同时得到13种耐药谱,多重耐药率达77.19%。所有原料乳和临床乳房炎菌株均对万古霉素和阿米卡星敏感。临床乳房炎菌株携带的毒素基因和多重耐药率比原料奶菌株高,同时在临床乳房炎乳中检测到MRSA菌株,提示我们应加强乳及其乳制品的管理,并对奶牛乳房炎加以重视。

Characterization of toxin genes and antimicrobial susceptibility of Staphylococcus aureus isolated from raw milk and milk of clinical mastitis

A total of 101 Staphylococcus aureus isolates from raw milk（44 isolates） and milk of clinical mastitis（57 isolates） were tested,for the prevalence of nine enterotoxin and four other exotoxin genes by polymerase chain reaction and antimicrobial susceptibility testing by the agar dilution method.It shows that in the 44 strains of S.aureus from raw milk,84.09% were positive for one or more toxin genes,84.09% isolates harbored PVL gene,52.27% isolates harbored one or more SE genes,and sea（56.82%） was the most common pattern,mecA,ETs,TSST-1 sei and sej genes were not detected.Resistance was most frequently observed to trimethoprim（100%）,followed by cefoxitin（86.36%）,chloramphenicol（11.36%）,erythromycin（4.55%）,oxacillin（2.27%）,cefoperazone（2.27%） and gentamicin（6.82%）.All S.aureus isolates from raw milk were susceptible to amikacin,ciprofloxacin and vancomycin.Of the 57 strains of S.aureus from milk of clinical mastitis,78.95% were positive for one or more toxin genes,28.07% isolates harbored PVL gene,77.19% isolates harbored one or more SE genes,and sea（47.37%） was the most common pattern,10.53% mecA gene was detected,ETs,TSST-1 and seh genes were not detected.Resistance was most frequently observed to erythromycin（100%）,followed by trimethoprim（71.93%）,chloramphenicol（28.07%）,gentamicin（26.07%）,ciprofloxacin（24.56%）,cefoxitin（19.30%） and oxacillin（7.02%）.All isolates of S.aureus from milk of clinical mastitis were susceptible to amikacin,cefoperazone,vancomycin and tetracycline.Many S.aureus isolates from raw milk and milk of clinical mastitis contained different multiple resistance and various toxin genes,MRSA was only isolated from cows with clinical mastitis.Our results may provide useful information for assessment of the possible risk posed to milk products.