Phenotypic and Genotypic Characterization of Klebsiella pneumoniae isolated from Clinical Mastitis in Buffaloes

W. Mousa, H. Abdelkhalek, Hanan E. Nagib, Randa S. Elias, S. Mansour
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Abstract

Mastitis is the most common serious and economical disease affecting the dairy industry. The current study aimed to determine the molecular characterization of K. pneumoniae isolated from buffaloes with clinical mastitis as well as determine the antimicrobial pattern and virulence associated genes. Out of examination one hundred and fifty dairy buffaloes at Beheria governorate, 57 (38%) showed clinical mastitis signs. All samples are submitted to bacteriological examination and confirmed by biochemical tests. The result showed that K.pneumoniae was identified in only five (8.7%). Regarding to the antibiogram profile, K. pneumoniae isolates exhibited high resistance pattern against Carbenicillin, Cefotaxime, Flumequine, Gentamicin, Kanamycin (100%) followed by Neomycin, Nitrofurantoin, Penicillin (80%) and Oxytetracycline, Sulfamethoxazole / trimethoprim (60%). Meanwhile, higher susceptibility to Ampicillin (100%), followed by Chloramphenicol, Colistin Sulphate, Erythromycin, and Streptomycin (80%). Molecular identification using the mPCR approach applied efficiently to detect the K. pneumoniae virulence genes (mrkA, mrkD, iutA) at 475, 226, 300bp respectively. In addition, the successfully detection of blaTEM and blaSHV for β lactams resistance genes at 516 and 392 bp. In conclusion, the phenotypic and genotypic resistance pattern of K. pneumoniae isolates indicates the portability of transmission of resistance genes through milk and food chain with public health concern.
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水牛临床乳腺炎肺炎克雷伯菌的表型和基因型分析
乳腺炎是影响乳制品行业最常见的严重和经济疾病。本研究旨在确定临床乳腺炎水牛中分离的肺炎克雷伯菌的分子特征,并确定其抗菌模式和毒力相关基因。在贝赫里亚省对150头奶牛进行的检查中,57头(38%)表现出临床乳腺炎症状。所有样品均经细菌学检查和生化试验确认。结果显示,检出肺炎克雷伯菌5株(8.7%)。在抗生素谱方面,肺炎克雷伯菌分离株对卡霉素、头孢噻肟、氟喹、庆大霉素、卡那霉素的耐药率为100%,其次是新霉素、呋喃妥英、青霉素(80%)和土霉素、磺胺甲恶唑/甲氧苄啶(60%)。对氨苄西林的敏感性较高(100%),其次是氯霉素、硫酸粘菌素、红霉素和链霉素(80%)。分子鉴定方法能有效地检测到肺炎克雷伯菌毒力基因(mrkA、mrkD、iutA),分别为475,226,300 bp。此外,blaTEM和blaSHV分别在516和392 bp处成功检测到β内酰胺耐药基因。综上所述,肺炎克雷伯菌分离株的表型和基因型耐药模式表明耐药基因通过牛奶和食物链传播的可移植性,具有公共卫生问题。
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