بررسی حضور استافیلوکوکوس اورئوس مقاوم به متی‌سیلین (MRSA) حامل انواع ژن mec در جدایه‌های مسبب ورم پستان تحت بالینی گاو

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش‌آموخته، گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران

2 استاد، گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران

3 استادیار، گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران

4 دانشیار، گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران

چکیده

استافیلوکوکوس اورئوس یکی از مهم‌ترین عوامل ایجادکننده ورم پستان در گاوهای شیری می‌باشد. بروز مقاومت به متی‌سیلین در این باکتری از نگرانی‌های عمده در دامپزشکی و بهداشت عمومی است. با توجه به شناسایی انواع جدیدی از ژن‌های کدکننده‌ی مقاومت به متی‌سیلین، هدف از این مطالعه بررسی حضور  استافیلوکوکوس اورئوس مقاوم به متی‌سیلین (MRSA) و انواع ژن mec برای نخستین بار در منطقه شمال شرق کشور می‌باشد. در مطالعه‌ی حاضر، هویت 54 جدایه‌ی استافیلوکوکوس اورئوس مسبب ورم پستان تحت بالینی گاو با استفاده از آزمون‌های بیوشیمیایی و مولکولی بررسی شد. از این تعداد، تنها در 44 سویه قسمت اختصاصی ژن nuc ردیابی شد. سپس، مقاومت جدایه‌ها به پنی‌سیلین، سفوکستین، انروفلوکساسین، تایلوزین و تری­متوپریم-سولفامتوکسازول بر اساس روش انتشار دیسک تعیین گردید. در انتها، بررسی مولکولی حضور ژن‌های mecA، mecB و mecC در جدایه‌ها صورت گرفت. طبق نتایج، بیشترین مقاومت مربوط به آنتی‌بیوتیک‌های پنی‌سیلین (54 درصد) و تری‌متوپریم-سولفامتوکسازول (23 درصد) و کمترین مقاومت مربوط به آنتی‌بیوتیک تایلوزین (7 درصد) بود. تنها در یک جدایه (3/2 درصد) حضور ژن mecA مورد تأیید قرار گرفت، در حالی که، این جدایه در تست فنوتیپی به‌عنوان MRSA شناسایی نگردید. هیچ یک از ژن‌های mecC وmecB  در این مطالعه شناسایی نشدند. نتایج این پژوهش نشان می‌دهد که در منطقه شمال شرق ایران، مقاومت به متی‌سیلین در سویه‌های ورم پستان گاو رخداد بالایی ندارد. همچنین، این مطالعه نشان می‌دهد که انواع کمتر شناخته شده ژن‌های مقاومت به متی‌سیلین در این منطقه شایع نیست. با این حال، توصیه می‌شود که مطالعات جامع‌تری در این خصوص صورت گیرد.

کلیدواژه‌ها


عنوان مقاله [English]

Investigation of the presence of methicillin resistant Staphylococcus aure-us (MRSA) carrying different mec types in isolates causing bovine sub-clinical mastitis

نویسندگان [English]

  • Mehri Amini Kamroodi 1
  • GholamReza HashemiTabar 2
  • Mahdi Askari Badouei 3
  • Babak Khoramian 4
  • Hamideh Kalateh Rahmani 1
1 Master of bacteriology, Department of Pathobiology, Faculty of veterinary, Ferdowsi University of Mashhad, Mash-had, Iran
2 PhD, Department of Pathobiology, Faculty of veterinary, Ferdowsi University of Mashhad, Mashhad, Iran
3 3 - PhD, Department of Pathobiology, Faculty of veterinary, Ferdowsi University of Mashhad, Mashhad, Iran
4 DVSc, Department of Clinical Sciences, Faculty of veterinary, Ferdowsi University of Mashhad, Mashhad, Iran
چکیده [English]

Staphylococcus aureus is one of the most important agents causing mastitis in dairy cattle. The emergence of methicillin resistance in S. aureus is a concern in veterinary medicine and public health. Due to the identification of new types of genes encoding methicillin resistance in recent years, the aim of this study was to investigate the presence of methicillin resistant (MRSA) and types of mec gene for the first time in the northeast of Iran. In the present study, the identity of 54 staphylococcus aureus isolates causing bovine subclinical mastitis were investigated using biochemical and molecular tests. Among them, the species-specific nuc gene was detected in 44 strains. Then, resistance of isolates to penicillin, cefoxitin, enrofloxacin, tylosin and trimethoprim-sulfamethoxazole were determined using the disk diffusion method. Finally, the molecular investigations for detection of the mecA, mecC, mecB genes were conducted. According to the results, the highest resistance were related to penicillin (54%) and trimethoprim-sulfamethoxazole (23%), and the lowest resistance was related to taylosin (7%). The mecA gene was observed in only one isolate (2.3%), while the isolate was not identified as MRSA in phenotypic test. The results of the present study show that the incidence of methicillin resistance in bovine mastitis strains of the area is not high. This study also showed that less known types of mec genes were not common in this region. However, further studies must be perform in this regard.

کلیدواژه‌ها [English]

  • Staphylococcus aureus
  • mec
  • subclinical mastitis
1- El-Tawab A, Ashraf A, El Hofy AI, Amaar AM, Sleim MA, Salem HS. Molecular characterization for some virulence and antibiotic resistance genes of Staphylococcus aureus isolated from dairy cattle's subclinical mastitis in EL-Sharkia Governorate. Benha Veterinary Medical Journal. 2016; 30(1): 219-30.
2- Abera M, Demie B, Aragaw K, Regassa F, Regassa A. Isolation and identification of Staphylococcus aureus from bovine mastitic milk and their drug resistance patterns in Adama town, Ethiopia. Journal of Veterinary Medicine and Animal Health. 2010; 2(3): 29-34.
3- Shrivastava N, Sharma V, Nayak A, Shrivastava AB, Sarkhel BC, Shukla PC, et al. Prevalence and Characterization of Methicillin-Resistant Staphylococcus aureus (MRSA) Mastitis in Dairy Cattle in Jabalpur, Madhya Pradesh. Journal of Animal Research. 2017; 7(1): 77.
4- Havaei SA, Assadbeigi B, Esfahani BN, Hoseini NS, Rezaei N, Havaei SR. Detection of mecA and enterotoxin genes in Staphylococcus aureus isolates associated with bovine mastitis and characterization of Staphylococcal cassette chromosome mec (SCCmec) in MRSA strains. Iranian journal of microbiology. 2015; 7(3): 161. [In Persian]
5- Abebe R, Hatiya H, Abera M, Megersa B, Asmare K. Bovine mastitis: prevalence, risk factors and isolation of Staphylococcus aureus in dairy herds at Hawassa milk shed, South Ethiopia. BMC Veterinary Research. 2016; 12(1): 1-1.
6- Turlej AG AT A, Hryniewic W A L E R I A, Empel J. Staphylococcal cassette chromosome mec (Sccmec) classification and typing methods: an overview. Pol J Microbiol. 2011; 60(2): 95-103.
7- Devries LA, VanDamme LR, Fameree L. Methicillin (cloxacillin)‐resistant Staphylococcus aureus strains isolated from bovine mastitis cases. Zentralblatt für Veterinärmedizin Reihe B. 1972; 19(7): 598-605.
8- Devriese LA, Hommez J. Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds. Research in veterinary science. 1975; 19(1): 23-27.
9- Paul I, Isore DP, Joardar SN, Mukhopadhayay SK, Jana C, Ganguly S. Investigation on methicillin resistant gene of Staphylococcus aureus for causing bovine mastitis. Indian J Comp Microbiol Immunol. Infect Dis. 2015; 36(1): 35-8.
10- Bardiau M, Yamazaki K, Duprez JN, Taminiau B, Mainil JG, Ote I. Genotypic and phenotypi ccharacterization of methicillin‐resist Staphylococcus aureus (MRSA) isolated from milk of bovine mastitis. Letters in Applied Microbiology. 2013; 57(3): 181-6.
11- Srednik ME, Crespi E, Testorelli MF, Puigdevall T, Pereyra AM, Rumi MV, et al. First isolation of a methicillin-resistant Staphylococcus aureus from bovine mastitis in Argentina. Veterinary and Animal Science. 2019; 7: 100043.
12- Becker K, van Alen S, Idelevich EA, Schleimer N, Seggewiß J, Mellmann A, et al. Plasmid-encoded transferable mecB-mediated methicillin resistance in Staphylococcus aureus. Emerging infectious diseases. 2018; 24(2): 242.
13- García-Álvarez L, Holden MT, Lindsay H, Webb CR, Brown DF, Curran MD, et al. Meticillin-resistant Staphylococcus aureus with a novel mecA homologue in human and bovine populations in the UK and Denmark: a descriptive study. The Lancet infectious diseases. 2011; 11(8): 595-603.
14- Cikman A, Aydin M, Gulhan B, Karakecili F, Kurtoglu MG, Yuksekkaya S, et al. Absence of the mecC gene in methicillin-resistant Staphylococcus aureus isolated from various clinical samples: The first multi-centered study in Turkey. Journal of infection and public health. 2019; Feb 7.
15- Turlej AGATA, Hryniewicz WALERIA, Empel J. Staphylococcal cassette chromosome mec (Sccmec) classification and typing methods:an overview. Pol J Microbiol. 2011; 60(2): 95-103.
16- Juuti K. Surface protein Pls of methicillin-resistant Staphylococcus aureus-role in adhesion, invasion and pathogenesis, and evolutionary aspects. 2004.
17- Peacock S J, Paterson G K. Mechanisms of methicillin resistance in Staphylococcus aureus. Annual review of biochemistry. 2015; 84: 577-601.
18- Macfadyen AC, Fisher EA, Costa B, Cullen C, Paterson GK. Genome analysis of methicillin resistance in Macrococcus caselyticus from dairy cattle in England and Wales. Microbial genomics. 2018; 4(8).
19- Chambers HF. Methicillin-resistant staphylococci. Clinical microbiology reviews. 1988; 1(2): 173-86
20- Dweba CC, Zishiri OT, El Zowalaty ME. Isolation and Molecular Identification of Virulence, Antimicrobial and Heavy Metal Resistance Genes in Livestock-Associated Methicillin-Resistant Staphylococcus aureus. Pathogens. 2019; 8(2):79.
21- Misiura A, Pigli YZ, BoyleVavra S, Daum RS, Boocock MR, Rice PA. Roles of two large serine recombinases in mobilizing the methicillin‐resistance cassette SCCmec. Molecular microbiology. 2013; 88(6): 1218-29.
22- Kim SH, Jeong HS, Kim YH, Song SA, Lee JY, Oh SH, et al. Evaluation of DNA extraction methods and their clinical application for direct detection of causative bacteria in continuous ambulatory peritoneal dialysis culture fluids from patients with peritonitis by using broad-range PCR. Annals of laboratory medicine. 2012; 32(2): 119-25.
23- Shebekhtiari N, Nochi Z, Eslampour M, Dabiri M, Bolifion M, Teherikalani M, et al. Characterization of staphylococcus aureus strains isolated from raw milk of bovine subclinical mastitis in Tehran and Mashhad. Acta Microbiologica et immunologica Hungarica. 2011; 58(2): 113-21.
24- Ishihara k, Shimokubo N, Sakagami A, Ueno H, Muramatsu Y, Kadosawa T, et al. Occurrence and Molecular Characteristics Of Methicillin-Resistant staphylococcus aureus and Methicillin-Resistant staphylococcus pseudintermedius in an Academic Veterinary Hospital. Applied and environmental microbiology. 2010; 76 (15): 5165-74.
25- CaLSI CL. Performance Standards for Antimicrobial Susceptibility Testing: Approved Twenty: Document M100-S28. Wayne, PA, USA: CLSI.2018; 2018 -19
26- Khoei F, Mobaiyen H, Nahaei MR, Sedeghi Mohammadi S. Antibiotic resistance pattern and Frequency of mecA gene in Staphylococcus aureus isolated from shohada hospital, Tabriz. Journal of medical Microbiology and Infectious Diseases. 2014; 2(3): 105-8. [In Persian]
27- Ahangari Z, Ghorbanpoor M, Shapouri MR, Gharibi D, Ghazvini K. Methicillin resistance and selective genetic determinants of Staphylococcus aureus isolates with bovine mastitis milk origin. Iranian journal of microbiology. 2017; 9(3): 152.
28- Sangarifar Samira, Hassan Ghajavand, Behrooz Johari, and Abazar Yari. Frequency of clf-A, mecA, and mecC Genes in Staphylococcus Aureus Strain isolated From Nosocomial Infections and Cows Milk. 2016; 233-238.
29- Hashemi M. Prevalence of antibiotic resistance among bacterial pathogens isolated from dairy cows with mastitis in Far province. Veterinary Researches & Biological Products. 2016; 29(3): 85-93. [In Persian]
30- Paterson GK, Harrison EM, Holmes MA. The emergence of mecC methicillin-resistant Staphylococcus aureus. Trends in microbiology. 2014; 22(1): 42-7.
31- Dastmalchi HS, Panahi M. Genotyping and antimicrobial resistance of Staphylococcus aureus isolates from dairy ruminants: differences in the distribution of clonal types between cattle and small ruminants. Archives of microbiology. 2019; Sep.
32- Haenni M, Châtre P, Tasse J, Nowak N, Bes M, Madec JY, et al. Geographical clustering of mecC-positive Staphylococcus aureus from bovine mastitis in France. Journal of Antimicrobial Chemotherapy. 2014; 69(8): 2292-2293.
33- Aklilu E, Chia HY. First mecC and mecA positive livestock-associated methicillin resistant Staphylococcu saureus (mecC MRSA/LA-MRSA) from dairy cattle in Malaysia. Microorganisms. 2020; 8(2): 147.
34- Solgi S, Razavi S, Nateghian A, Irajian G, Pournajaf A, Haasnnejad-Bibalan M, et al. Resistance-related determinants in clinically relevant Staphylococcus aureus isolated from teaching therapeutic centers, Tehran, Iran. Reviews in Medical Microbiology. 2019; May 31. [In Persian]
35- Youssif NH, Hafiz NM, Halawa MA, Aziz HM. Genes conferring antimicrobial resistance in cattle with Subclinical mastitis. Bulgarian J Vet Med, DOI. 2019; 10: 2019-0028.
36- Akkou M, Antri K, Bachtarzi MA, Bes M, Tristan A, Dauwalder O, et al. Phenotypic and genotypic characterization of Staphylococcus aureus strains associated with bovine mastitis and nasal carriage of workers in contact to animals in Algeria. Pak. Vet. J. 2016; 36(2): 184-8.
37- Pourtaghi H, Aziz AG, Sodagari H. Antimicrobial resistance patterns of Staphylococcus aureus isolated from bovine subclinical mastitis in Alborz province, Iran. Bulg J vet Med. 2016; 19(2): 169-74
38- Bahraminia F, Emadi SR, Emaneini M, Farzaneh N, Rad M, Khoramian B. Ahigh prevalence of tylosin resistance among staphylococcus aureus strains isolated from bovine mastitis. In Veterinary Research Forum. 2017; 8(2): 121. Faculty of Veterinary Medicine, Urmia University, Urmia, Iran. [In Persian]