Antibiotic Resistance and Virulence Gene Profiles in Staphylococci Isolated from Cattle with Mastitis

Year 2021, Volume: 6 Issue: 3, 395 - 402, 28.09.2021

Mehmet Onur Gökdağ Alper Çiftci

https://doi.org/10.35229/jaes.954156

Abstract

Staphylococcus spp. are the important bacterial agents of subclinical and clinical mastitis cases. This study was aimed to determine the vancomycin resistances, antibacterial resistance profiles, some virulence genes, and pheno- and genotyping of staphylococci from mastitis. For this aim, 121 staphylococcal isolates were analyzed. The identifications of isolates were confirmed with PCR for being Staphylococcus spp. and Staphylococcus aureus. The antibiotic resistance patterns were determined by Kirby bauer Disc Diffusion Tests and according to the resistance profiles, the isolates were antibiotyped. The vancomycin resistance genes were determined by PCR for investigating the vanA, vanH, vanR, vanS, vanZ, vanY and vanX genes. The vancomycin resistant isolates were genotyped with RAPD-PCR. The nuc gene was detected in 86 of 121 staphylococcal isolates examined and named as Staphylococcus aureus. The remaining 35 isolates were defined as Staphylococcus spp. S. aureus isolates were found to be resistant to penicillin G, amoxicillin/clavulanic acid, oxacillin, tetracycline, cefaperazone, teicoplanin, vancomycin and trimethoprim-sulfomethaxazole at 50% (43/86), 40.7% (35/86), 34.9% (30/86), 23.3% (20/86), 22.1% (19/86), 18.6% (16/86) 10.5% (9/86) and 8.1% (7/86) respectively. On the other hand, 4 (11.4%) of 35 Staphylococcus spp. isolates were resistant to penicillin G, 3 (8.6%) to amoxicillin/clavulanic acid, 2 (5.7%) to trimethoprim-sulfometaxazole, 1 (2.9%) to oxacillin, cefaperazone, teicoplanin, while all isolates were sensitive to vancomycin and tetracycline. Of the 9 S. aureus isolates that were phenotypically resistant to vancomycin, vanA gene was detected in 2 and vanR gene in 3 isolates. Multiple resistances to three or more antibiotics were determined in 42 of 86 S. aureus isolates. In addition, coa gene was detected in 61 (70.9%) of 86 S. aureus isolates. 10 different gene polymorphisms were detected in coa gene positive isolates. While the spa gene was determined in all S. aureus isolates, it was revealed that they had 4 spa gene polymorphisms. Nine different genotypes with a similarity between 51-75% were detected in the genotyping of vancomycin resistant 9 isolates. In conclusion, multiple antibiotic resistance rates in S. aureus isolates investigated were found to be important for mastitis treatment. The results obtained from this study show that milk and dairy products containing these factors pose a public health risk due to the determination of vancomycin resistance in mastitis-derived Staphylococcus strains.

Keywords

Antibiotic resistance, bovine, coa, mastitis, RAPD-PCR, spa, Staphylococcus

Supporting Institution

Scientific Research Projects Commission of Ondokuz Mayis University

Project Number

PYO.VET.1904.14.005

Mastitisli Sığırlardan İzole Edilen Stafilokoklarda Antibiyotik Direnci ve Virülens Gen Profilleri

Abstract

Staphylococcus spp. subklinik ve klinik mastitis olgularının önemli bakteriyel etkenleridir. Bu çalışmanın amacı, mastitis kaynaklı stafilokokların vankomisin dirençlerini, antibakteriyel direnç profillerini, bazı virülans genlerini ve feno- ve genotiplendirmesini belirlemektir. Bu amaçla 121 stafilokok izolatının analizi yapılmıştır. İzolatların Staphylococcus spp. ve Staphylococcus aureus olmak üzere identifikasyonları PCR ile ve antibiyotik direnç paternleri de Kirby Bauer Disk Difüzyon Testleri ile belirlendi. İzolatlar direnç profillerine göre antibiyotiplendirildi. VanA, vanH, vanR, vanS, vanZ, vanY ve vanX genlerinin PCR ile araştırılması sonucunda vankomisin direnç genleri belirlendi. Vankomisine dirençli izolatlar, RAPD-PCR ile genotiplendirildi. İncelenen 121 stafilokok izolatının 86'sında nuc geni saptandı ve Staphylococcus aureus olarak identifiye edildi. S. aureus izolatlarının penisilin G, amoksisilin/klavulanik asit, okzasilin, tetrasiklin, sefaperazon, teikoplanin, vankomisin ve trimetoprim-sulfometaksazole sırasıyla %50 (43/86), %40.7 (35/86), %34,9 (30/86), %23,3 (20/86), %22.1 (19/86), %18,6 (16/86) %10,5 (9/86) ve %8,1 (7/86) oranında dirençli bulundu. 35 adet Staphylococcus spp.'nin 4'ü (%11,4) penisilin G'ye, 3'ü (%8,6) amoksisilin/klavulanik aside, 2'si (%5,7) trimetoprim-sülfometaksazole, 1'i (%2,9) oksasilin, sefaperazon, teikoplanine direnç gösterirken, tüm izolatlar vankomisin ve tetrasikline duyarlıydı. Vankomisine fenotipik olarak dirençli olan 9 S. aureus izolatından 2'sinde vanA, 3'ünde vanR geni saptandı. 86 S. aureus izolatının 42'sinde üç veya daha fazla antibiyotiğe karşı çoklu direnç belirlendi. Ayrıca 86 S. aureus izolatının 61'inde (%70.9) coa geni tespit edildi. coa geni pozitif izolatlarda 10 farklı gen polimorfizmi tespit edildi. Tüm S. aureus izolatlarında spa geni belirlenirken, 4 spa gen polimorfizmi görüldü. Vankomisine dirençli 9 izolatın genotiplendirilmesinde %51-75 arasında benzerlik gösteren dokuz farklı genotip tespit edildi. Sonuç olarak, araştırılan S. aureus izolatlarında çoklu antibiyotik direnç oranları mastitis tedavisi için önemli bulunmuştur. Bu çalışmadan elde edilen sonuçlar, bu faktörleri içeren süt ve süt ürünlerinin mastitis kaynaklı Staphylococcus suşlarında vankomisin direncinin belirlenmesi nedeniyle halk sağlığı açısından risk oluşturduğunu göstermektedir.

Keywords

Antibiyotik direnci, coa, inek, mastitis, RAPD-PCR, spa, Staphylococcus

Project Number

PYO.VET.1904.14.005

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