Mikotik Mastitislerde Non-Albicans Candida Türlerinin Etkinliğinin ve Antifungal İlaç Dirençlerinin Araştırılması

Year 2022, , 113 - 119, 15.05.2022

Hafize Tuğba Yüksel Dolgun , Şükrü Kırkan , Volkan Özavcı

https://doi.org/10.31196/huvfd.1117714

Abstract

Bu çalışmanın amacı, sığır klinik mastitislerinden izole edilen Candida türlerinin konvansiyonel ve moleküler yöntemlerle tanımlanması ve antifungal duyarlılıklarının belirlenmesidir. Bu araştırmada İzmir'in beş farklı ilçesindeki sığır çiftliklerinden 100 mastitisli süt örneği alınmıştır. Mastitisli süt örneklerinden 23 Candida izolatı izole edildi ve kromojenik agar ile 20 (%87.0) izolat C. krusei, iki (%8.6) izolat C. albicans ve bir (%4.3) izolat C. tropicalis olarak tanımlandı. Tüm Candida izolatları PCR analizi ile C. krusei, C. albicans ve C.tropicalis olarak doğrulandı. Sekans analizine göre C. krusei izolatlarından 11 tanesinin “Pichia kudriavzeii ZKI117 5.8S ribosomal gene” ile; altı tanesinin “Pichia kudriavzeii isolate L-012 small subunit ribosomal RNA gene” ile; iki tanesinin “Pichia kudriavzeii isolate 3 internal transcribed spacer 1”, bir tanesinin “Pichia kudriavzeii LL11_078 18S ribosomal gene” ile; 2 C. albicans izolatının “Candida albicans isolate B02 5.8S ribosomal gene” ile; bir C.tropicalis izolatının “Candida tropicalis isolate CTR1201 18S ribosomal gene” ile >%97 benzerlik gösterdiği belirlendi. Tüm Candida izolatlarının antifungal duyarlılık testi sonuçlarında metronidazol ve flusitozine karşı %100 dirençli olduğu saptandı. Sonuç olarak, mikotik mastitisli çiftliklerde Candida etkenlerinin identifikasyonu ve antifungal duyarlılıklarının belirlenmesinin doğru tedavi protokollerinin uygulanmasını sağlayacağı ve mikotik mastitise bağlı ekonomik kayıpları önleyeceği kanaatine varıldı.

Keywords

Antifungal İlaç Direnci, Candida, DNA SEKANSI, İNEK, MASTİTİS

Supporting Institution

Bulunmamaktadır

Project Number

Bulunmamaktadır

Investigation of the Efficacy and Antifungal Drug Resistance of Non-Albicans Candida Species in Mycotic Mastitis

Abstract

This research aimed to use a conventional and molecular approach to determine Candida species isolated from cattle clinical mastitis and examine their antifungal susceptibility. In this research, 100 milk samples with mastitis were collected from dairy farms in five different Izmir districts. On chromogenic agar, 23 Candida isolates were isolated from milk samples with mastitis, and 20 (87.0%) isolates were identified as C. krusei, two (8.6%) isolates as C. albicans, and one (4.3%) isolate as C. tropicalis. PCR analysis verified that all Candida isolates were C. krusei, C. albicans, and C. tropicalis. According to sequence analysis, it was determined that 11 of the C. krusei isolates had >97% similarity to “Pichia kudriavzeii ZKI117 5.8S ribosomal gene”; six of the C. krusei isolates to “Pichia kudriavzeii isolate L-012 small subunit ribosomal RNA gene”; two C. krusei isolates to “Pichia kudriavzeii isolate 3 internal transcribed spacer 1”; and one C. krusei isolates to “Pichia kudriavzeii LL11_078 18S ribosomal gene”; two C. albicans isolates to “Candida albicans isolate B02 5.8S ribosomal gene”; one C. tropicalis isolate to “Candida tropicalis isolate CTR1201 18S ribosomal gene”. In the antifungal susceptibility test results, all Candida isolates (%100) were resistant to metronidazole and flucytosine. Consequently, it was determined that identifying Candida agents and determining antifungal susceptibility in farms with mycotic mastitis will ensure the application of correct treatment protocols and reduce economic losses due to fungal mastitis.

Keywords

Antifungal drug resistance, Candida, Cattle, DNA Sequence, Mastitis

Project Number

Bulunmamaktadır

References

• Bakr EM, El-Tawab A, Elshemey TM, Abd-Elrhman AH, 2015: Diagnostic and Therapeutic Studies on Mycotic Mastitis in Cattle. Alex J Vet Sci, 46 (1), 138-145.
• Cilvez P, Turkyilmaz S, 2019: Molecular Diagnosis of Candida Species Isolated from Cases of Subclinical Bovine Mastitis. Israel J Veterinary Med, 74 (3), 134-140.
• Clinical Laboratory Standards Institute (CLSI). Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts: Approved Guidelines. Second Edition. CLSI Document M44-A2. Wayne, PA, USA: Clinical Laboratory Standard Institute; 2009.
• Du J, Wang X, Luo H, Wang Y, Liu X, Zhou X, 2018: Epidemiological investigation of non-albicans Candida species recovered from mycotic mastitis of cows in Yinchuan, Ningxia of China. BMC Veterinary Research, 14 (251), 1-9.
• Dworecka-Kaszak B, Krutkiewicz A, Szopa D, Kleczkowski M, Biegańska M, 2012: High prevalence of Candida yeast in milk samples from cows suffering from mastitis in poland. The Scientific World Journal, 2012, 5.
• Eldesouky I, Mohamed N, Khalaf D, Salama A, Elsify A, Ombarak R, El-Ballal S, Effat M, Alshabrawy M, 2016: Candida mastitis in dairy cattle with molecular detection of Candida albicans. Kafkas Univ Vet Fak Derg, 22(3), 461-464.
• Erbas G, Parin U, Kirkan S, Savasan S, Ozavci V, Yuksel HT, 2017: Identification of Candida strains with nested PCR in bovinemastitis and determination of antifungal susceptibilities. Turk J Vet Anim Sci, 41(6), 757-763.
• Fujita SI, Senda Y, Nakaguchi S, Hashimoto T, 2001: Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains. Journal of clinical microbiology, 39 (10), 3617-3622.
• Hizlisoy H, Ertas Onmaz N, Al S, Karadal F, Yildirim Y, Gonulalan Z, Gumussoy KS, Aydemir GD, Tekinsen FK, Dinc G, 2020: Clonal diversity and antifungal susceptibility of Candida spp. recovered from cow milk. Mljekarstvo: časopis za unaprjeđenje proizvodnje i prerade mlijeka, 70(1), 40-49.
• Khalaf DD, Soliman MMH, Mansour AS, 2021: Conventional and molecular identification of mycotic mastitis caused by Candida in farm animals. Int J Vet Sci, 10(1), 64-68.
• Khalaf RA, Fattouh N, Medvecky M, Hrabak J, 2021: Whole genome sequencing of a clinical drug resistant Candida albicans isolate reveals known and novel mutations in genes involved in resistance acquisition mechanisms. J Med Microbiol, 70 (4), 001351.
• Milanov D, Prunic BO, Velhner MA, Bojkovski J, 2014: Diagnosis of yeast mastitis in dairy cows. Lucrari Stiintifice Medicina Veterinara, 47(1), 56-64.
• Sanger F, Nicklen S, Coulson R, 1977: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA, 74, 5463-5467.
• Sartori LCA, Santos RC, Marin JM, 2014: Identification of Candida species isolated from cows suffering mastitis in four Brazilian states. Arq. Bras. Med. Vet. Zootec, 66 (5), 1615-1617.
• Seker E, 2010: Identification of Candida species isolated from bovine mastitic milk and their in vitro hemolytic activity in Western Turkey. Mycopathologia, 169 (4), 303-308.
• Sonmez M, Erbas G, 2017: Isolation and identification of Candida spp. from mastitis cattle milk and determination of antifungal susceptibilities. Int J Vet Sci, 6 (2), 104-107.
• Turkyilmaz S and Kaynarca S, 2010: The slime production by Yeast Isolated from Subclinical Mastitic Cows, Acta Vet Brno, 79, 581-586.
• Wawron W, Bochniarz M, Piech T, 2010: Yeast mastitis in dairy cows in the middleeastern part of Poland. Bull. Vet. Inst. Pulawy, 54, 201-204.
• Zaragoza CS, Olivares RA, Watty AE, Moctezuma Ade L, Tanaca LV, 2011: Yeasts isolation from bovine mammary glands under different mastitis status in the Mexican High Plateu. Rev Iberoam Micol, 28(2), 79-82.