Kedi ve köpek kökenli stafilokok türlerinde beta-laktam direncinin fenotipik ve genotipik karakterizasyonu
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Date
2022-03-01
Authors
Kurnaz, Havva
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Bu çalışmada sağlıklı kedi ve köpeklerde Staphylococcus spp. taşıyıcılığı ile hasta kedi ve köpeklerde Staphylococcus spp. kaynaklı enfeksiyon prevalansının belirlenmesi hedeflendi. Ayrıca Staphylococcus spp.’de beta-laktam grubu antimikrobiyallere direnç profilinin fenotipik ve genotipik yöntemler ile araştırılması amaçlandı. Bu amaçla sağlıklı hayvanlardan (165 kedi ve 85 köpek) sırası ile burun, ağız, kasık ve perineal bölgeden olacak şekilde tek svab örneği, hasta hayvanlardan (56 kedi ve 95 köpek) ise hastalıkla ilişkili bölgelerden (kulak, göz, deri, ağız, burun ve yara) alınan svab örnekleri incelendi. Tüm svablar ön zenginleştirme için %6,5 sodyum klorid içeren Mueller Hinton Broth’a (MHB + %6,5 NaCI) inoküle edildi, sonrasında selektif besiyeri olarak Egg Yolk Tellurite içeren Baird Parker (BP) agar izolasyonda kullanıldı. BD Phoenix 100 otomatize cihazı kullanılarak izolatların identifikasyonu ve antimikrobiyal duyarlılık testleri yapıldı. Staphylococcus intermedius Grup (SIG) olarak belirlenen türlerin ayrımı MALDI-TOF MS ile gerçekleştirildi. Tüm Staphylococcus spp. blaZ, mecA ve mecC genleri yönünden multiplex PCR ile analiz edildi. Sağlıklı kedilerin %53,9 ve sağlıklı köpeklerin %78,8 oranında Staphylococcus spp. taşıyıcılığı bulunurken, Staphylococcus spp. prevalansı hasta kedi ve köpeklerde sırasıyla %83,9 ve %60 idi. Kedilerde izole edilen en yaygın türler S. felis, S. epidermidis ve S. aureus iken, köpeklerde S. pseudintermedius idi. İzole edilen Staphylococcus spp.’de beta-laktamaz aktivitesi fenotipik yöntemler (penisilin direnci ve nitrosefin test sonucu) ile %53,8 ve genotipik yöntemler (blaZ) ile %50,4 bulundu. Metisilin direnci fenotipik yöntemler (sefoksitin ve/veya oksasilin direnci) ile %15,8 genotipik yöntemler (mecA) ile %13,5 bulundu. İzolatlarda mecC geni tespit edilmedi. Metisilin dirençli Staphylococcus spp’nin (MRS) dağılımı S. pseudintermedius (n:18), S. epidermidis (n:8), S. aureus (n:4), S. hominis (n:2), S. cohnii subsp. cohnii (n:1), S. haemolyticus (n:1) ve S. capitis (n:1) olarak saptandı. Kedi ve köpeklerde Staphylococcus spp.’nin güncel prevalansının ve beta-laktam grubu antimikrobiyallere güncel direnç profilinin bilinmesi, hem laboratuvarlarda tanısal yaklaşımlara hem de klinisyen hekimlerin bilinçli terapotik yaklaşımlarına fayda sağlayacağı düşünülmektedir.
In this study, it was aimed to determine the prevalence of Staphylococcus spp. carriage in healthy cats and dogs and the prevalence of Staphylococcus spp-induced infection in sick cats and dogs. It was also intended to investigate their resistance profile of beta-lactam group of antimicrobials in Staphlococcus spp. isolates with phenotypic and genotypic methods. For this purpose, a single swab sample has taken from the nose, mouth, groin and perineal regions, respectively, from healthy animals (n:165 cats and n:85 dogs). Sample collection was applied to sick animals (n:56 cats and n:95 dogs) from disease-related regions (ear, eye, skin, mouth, nose and wound). All swabs were inoculated in to Mueller Hinton Broth (MHB + 6.5% NaCl) containing 6.5% sodium chloride for pre-enrichment, this followed by Baird Parker (BP) agar containing Egg Yolk Tellurite as a selective medium for the isolation. BD Phoenix 100 Automatic Identification System, was used for the identification and the detection of antimicrobial susceptibility of the isolates. Species differentiation of isolates identified as a Staphylococcus intermedius Group (SIG) were analysed with MALDI-TOF MS. All Staphylococcus spp.,isolates were analyzed by multiplex PCR for blaZ, mecA and mecC genes. The results showed that 53,9% of healthy cats and 78,8% of healthy dogs were carrying Staphylococcus spp. however the prevelance of Staphylococcus spp. was 83.9% and 60% in sick cats and dogs respectively. The most common Staphylococcus species in cats were S. felis, S. epidermidis and S. aureus, while S. pseudintermedius was predominant in dogs. Beta-lactamase activity was found to be 53,8% by phenotypical methods (penicillin resistance and nitrocefin test results) and 50,4% by genotypic methods (blaZ) in all Staphylococcus spp. isolates examined. Methicillin resistance was found to be 15,8% with phenotypic methods (cefoxitin and/or oxacillin resistance) and 13,5% with genotypic methods (mecA). The mecC gene was not detected in any isolate. The distribution of methicillin-resistant Staphylococcus species (MRS) was determined as S. pseudintermedius (n:18), S. epidermidis (n:8), S. aureus (n:4), S. hominis (n:2), S. cohnii subsp. cohnii (n:1), S. haemolyticus (n:1) and S. capitis (n:1). Knowing the current prevalence of Staphylococcus spp. in cats and dogs and their resistance profile to beta-lactam group of antimicrobials may help both diagnostic approaches in laboratories and conscious therapeutic to clinicians.
In this study, it was aimed to determine the prevalence of Staphylococcus spp. carriage in healthy cats and dogs and the prevalence of Staphylococcus spp-induced infection in sick cats and dogs. It was also intended to investigate their resistance profile of beta-lactam group of antimicrobials in Staphlococcus spp. isolates with phenotypic and genotypic methods. For this purpose, a single swab sample has taken from the nose, mouth, groin and perineal regions, respectively, from healthy animals (n:165 cats and n:85 dogs). Sample collection was applied to sick animals (n:56 cats and n:95 dogs) from disease-related regions (ear, eye, skin, mouth, nose and wound). All swabs were inoculated in to Mueller Hinton Broth (MHB + 6.5% NaCl) containing 6.5% sodium chloride for pre-enrichment, this followed by Baird Parker (BP) agar containing Egg Yolk Tellurite as a selective medium for the isolation. BD Phoenix 100 Automatic Identification System, was used for the identification and the detection of antimicrobial susceptibility of the isolates. Species differentiation of isolates identified as a Staphylococcus intermedius Group (SIG) were analysed with MALDI-TOF MS. All Staphylococcus spp.,isolates were analyzed by multiplex PCR for blaZ, mecA and mecC genes. The results showed that 53,9% of healthy cats and 78,8% of healthy dogs were carrying Staphylococcus spp. however the prevelance of Staphylococcus spp. was 83.9% and 60% in sick cats and dogs respectively. The most common Staphylococcus species in cats were S. felis, S. epidermidis and S. aureus, while S. pseudintermedius was predominant in dogs. Beta-lactamase activity was found to be 53,8% by phenotypical methods (penicillin resistance and nitrocefin test results) and 50,4% by genotypic methods (blaZ) in all Staphylococcus spp. isolates examined. Methicillin resistance was found to be 15,8% with phenotypic methods (cefoxitin and/or oxacillin resistance) and 13,5% with genotypic methods (mecA). The mecC gene was not detected in any isolate. The distribution of methicillin-resistant Staphylococcus species (MRS) was determined as S. pseudintermedius (n:18), S. epidermidis (n:8), S. aureus (n:4), S. hominis (n:2), S. cohnii subsp. cohnii (n:1), S. haemolyticus (n:1) and S. capitis (n:1). Knowing the current prevalence of Staphylococcus spp. in cats and dogs and their resistance profile to beta-lactam group of antimicrobials may help both diagnostic approaches in laboratories and conscious therapeutic to clinicians.
Description
Keywords
Beta-laktam direnci, Staphylococcus spp, MecA, MecC, Kedi, Köpek, MRS, Beta-lactam resistance, Cat, Dog
Citation
Kurnaz, H. (2022). Kedi ve köpek kökenli stafilokok türlerinde beta-laktam direncinin fenotipik ve genotipik karakterizasyonu. Yayınlanmamış doktora tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.