Investigation of mecA, vanA and pvl genes in Staphylococcus aureus strains isolated from bovine mastitis in smallholder dairy farms

Yıl 2022, , 50 - 55, 15.06.2022

Müesser Yilmaz Esra Şeker

https://doi.org/10.35864/evmd.1008728

Cited By: 1

Öz

This study aimed to isolate the Staphylococcus aureus from bovine mastitis, investigate the presence of mecA, vanA and pvl genes in isolated strains and determine the antibiotic resistance of S. aureus strains to some antibiotics commonly used in veterinary field. In the present study, 602 mammary quarter milk samples belong to 170 lactating cows from 40 different smallholder dairy farms located center town and villages of Afyonkarahisar were used. After the CMT scoring, the milk samples were aseptically collected from each mammary quarter. In this study, a total of 23 (3.8%) S. aureus strains were isolated from 602 mammary quarter milk samples by using a commercial identification kit. The presence of mecA, vanA and pvl genes was found in none of the strains. Phenotypic resistances of all S. aureus strains to 12 antibiotics were investigated by using Kirby-Bauer disc diffusion method. The highest resistance rate in strains was against penicillin G (52.2%), followed by oxacillin (21.7%), erythromycin (21.7%) and amoxicillin+clavulanic acid (17.4%). In conclusion, it was noted that S. aureus was not a prevalent pathogen in the etiology of bovine mastitis for the smallholder farms sampled.

Anahtar Kelimeler

Mastitis, methicillin resistance, Panton-Valentine leukocidin, S. aureus, vancomycin resistance

Destekleyen Kurum

Afyon Kocatepe University Scientific Research Projects Coordination Unit

Proje Numarası

17.SAG.BIL.07

Küçük ölçekli süt işletmelerinde sığır mastitislerinden izole edilen Staphylococcus aureus suşlarında mecA, vanA ve pvl genlerinin araştırılması

Öz

Bu çalışmada sığır mastitislerinden Staphylococcus aureus'un izolasyonu, izole edilen suşlarda mecA, vanA ve pvl genlerinin varlığının araştırılması ve S. aureus suşlarının veteriner sahada yaygın olarak kullanılan bazı antibiyotiklere dirençliliklerinin belirlenmesi amaçlandı. Çalışmada, Afyonkarahisar'ın merkez kasaba ve köylerindeki 40 farklı küçük ölçekli süt işletmesinde bulunan 170 laktasyondaki ineğe ait 602 meme lobu süt örneği kullanıldı. CMT skorlamasının ardından, her bir meme lobundan aseptik olarak süt örnekleri toplandı. Çalışmada, ticari identifikasyon kiti kullanılarak 602 meme lobu süt örneğinden toplam 23 (%3,8) S. aureus suşu izole edildi. Suşların hiçbirinde mecA, vanA ve pvl genleri belirlenmedi. Tüm S. aureus suşlarının 12 antibiyotiğe karşı fenotipik dirençlilikleri Kirby-Bauer disk difüzyon yöntemi kullanılarak belirlendi. Suşlardaki en yüksek direnç oranı penisilin G'ye (%52,2) karşı iken, bunu oksasilin (%21,7), eritromisin (%21,7) ve amoksisilin+klavulanik aside (%17,4) karşı direnç oranları izledi. Sonuç olarak, S. aureus'un örneklemenin yapıldığı küçük ölçekli işletmeler için sığır mastitislerinin etiyolojisinde yaygın bir patojen olmadığı belirlendi.

Anahtar Kelimeler

Mastitis, metisilin direnci, Panton-Valentine lökosidin, S. aureus, vankomisin direnci

Proje Numarası

17.SAG.BIL.07

Kaynakça

• Algammal AM, Enany ME, El-Tarabili RM, Ghobashy MOI, Helmy YA. (2020) Prevalence, antimicrobial resistance profiles, virulence and enterotoxins-determinant genes of MRSA isolated from subclinical bovine mastitis in Egypt. Pathogens. 9, 362 DOI: 10.3390/pathogens9050362
• Amin DHM, Guler E, Baddal B. (2020) Prevalence of Panton‑Valentine leukocidin in methicillin‑resistant Staphylococcus aureus clinical isolates at a university hospital in Northern Cyprus: a pilot study. BMC Res Notes. 13, 490. DOI: 10.1186/s13104-020-05339-0
• Balemi A, Gumi B, Amenu K, Girma S, Gebru M, Tekle M, Ríus AA, D’Souza DH, Agga GE, Kerro Dego O. (2021) Prevalence of mastitis and antibiotic resistance of bacterial isolates from CMT positive milk samples obtained from dairy cows, camels, and goats in two pastoral districts in Southern Ethiopia. Animals (Basel). 11(6), 1530. DOI: 10.3390/ani11061530
• Bhattacharyya D, Banerjee J, Bandyopadhyay S, Mondal B, Nanda PK, Samanta I, Mahanti A, Das AK, Das G, Dandapat P, Bandyopadhyay S. (2016) First report vancomycin resistant Staphylococcus aureus in bovine and caprine milk. Microb Drug Resist. 22(8): 675-681. DOI: 10.1089/mdr.2015.0330
• Bhutto AL, Murray RD, Woldehiwet Z. (2012) California mastitis test scores as indicators of subclinical intra-mammary infections at the end of lactation in dairy cows. Res Vet Sci. 92(1), 13-17. DOI: 10.1016/j.rvsc.2010.10.006
• Boyle-Vavra S, Daum RS. (2007) Community-acquired methicillin-resistant Staphylococcus aureus: the role of Panton-Valentine leukocidin. Lab Invest. 87(1), 3-9. DOI: 10.1038/labinvest.3700501
• Chang S, Sievert DM, Hageman JC, Boulton ML, Tenover FC, Downes FP, Shah S, Rudrik JT, Pupp GR, Brown WJ, Cardo D, Fridkin SK. (2003) Infection with vancomycin-resistant Staphylococcus aureus containing the vanA resistance gene. New Eng J Med. 348(14), 1342-1347. DOI: 10.1056/NEJMoa025025
• Choi SM, Kim SH, Kim HJ, Lee DG, Choi JH, Yoo JH, Kang JH, Shin WS, Kang MW. (2003) Multiplex PCR for detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species. J Korean Med Sci. 18(5), 631-636. DOI: 10.3346/jkms.2003.18.5.631
• Clinical and Laboratory Standards Institute (CLSI) (2007) Performance standards for antimicrobial susceptibility testing; 17th informational supplement. CLSI Document M100-S17. Wayne, PA.
• Clinical and Laboratory Standards Institute (CLSI) (2012) Performance standards for antimicrobial susceptibility testing; twenty-second informational supplement. CLSI Document M100-S22. Wayne, PA.
• Clinical and Laboratory Standards Institute (CLSI) (2013) Performance standard for antimicrobial susceptibility testing; twenty-third informational supplement. CLSI Document M100-S23. Wayne, PA.
• Clinical and Laboratory Standards Institute (CLSI) (2016) Performance standards for antimicrobial susceptibility testing; twenty. CLSI Document M100-S26. Wayne, PA.
• Clinical and Laboratory Standards Institute (CLSI) (2017) Performance standards for antimicrobial susceptibility testing; twenty. CLSI Document M100-S27. Wayne, PA.
• Darboe S, Dobreniecki S, Jarju S, Jallow M, Mohammed NI, Wathuo M, Ceesay B, Tweed S, Basu Roy R, Okomo U, Kwambana-Adams B, Antonio M, Bradbury RS, de Silva TI, Forrest K, Roca A, Lawal BJ, Nwakanma D, Secka O. (2019) Prevalence of Panton-Valentine Leukocidin (PVL) and antimicrobial resistance in community-acquired clinical Staphylococcus aureus in an Urban Gambian Hospital: A 11-year period retrospective pilot study. Front Cell Infect Microbiol. 9, 170. DOI: 10.3389/fcimb.2019.00170
• Dutka-Malen S, Evers S, Courvalin P. (1995) Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant Enterococci by PCR. J Clin Microbiol. 33, 24-27 (Erratum 33:1434). DOI: 10.1128/jcm.33.5.1434-1434.1995.
• Elias L, Balasubramanyam AS, Ayshpur OY, Mushtuk IU, Sheremet NO, Gumeniuk VV, Musser JMB, Rogovskyy AS. (2020) Antimicrobial susceptibility of Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli isolated from mastitic dairy cattle in Ukraine. Antibiotics. 9, 469. DOI:10.3390/antibiotics9080469
• Gezgen C, Seker E. (2016) Investigation of methicillin resistance and Panton-Valentine leukocidin in staphylococci isolated from bovine mastitis. Acta Sci Vet. 44(1), 1373. DOI: 10.22456/1679-9216.81080
• Guimarães FF, Manzi MP, Joaquim SF, Richini-Pereira VB, Langoni H. (2017) Outbreak of methicillin-resistant Staphylococcus aureus (MRSA)-associated mastitis in a closed dairy herd. J Dairy Sci. 100, 726-730. DOI: 10.3168/jds.2016-11700
• Hiramatsu K, Hanaki H, Ino T, Yabuta K, Oguri T, Tenover FC. (1997) Methicillin-resistant Staphylococcus aureus clinical strain with reduced vancomycin susceptibility. J Antimicrob Chemother. 40(1), 135-146. DOI: 10.1093/jac/40.1.135
• Hogan JS. (1999) Laboratory handbook on bovine mastitis. Revised edition. Madison, WI: National Mastitis Council Inc., p.222.
• Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. (2000) Bergey’s Manual of Determinative Bacteriology. Philadelphia: Lippincott Williams and Wilkins, p. 787.
• Hoque MN, Das ZC, Rahman ANMA, Haider MG, Islam MA. (2018) Molecular characterization of Staphylococcus aureus strains in bovine mastitis milk in Bangladesh. Int J Vet Sci Med. 6, 53-60. DOI: 10.1016/j.ijvsm.2018.03.008
• Igbinosa EO, Beshiru A, Akporehe LU, Ogofure AG. (2016) Detection of methicillin-resistant Staphylococci isolated from food producing animals: A public health implication. Vet Sci. 3(3), 14. DOI: 10.3390/vetsci3030014.
• Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter M, Gauduchon V, Vandenesch F, Etienne J. (1999) Involvement of Panton–Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 29(5), 1128-1132. DOI: 10.1086/313461.
• Lo W, Wang C. (2011) Panton-Valentine Leukocidin in the pathogenesis of community-associated methicillin-resistant Staphylococcus aureus infection. Pediatr Neonatol. 52(2), 59-65. DOI: 10.1016/j.pedneo.2011.02.008
• Lowy FD. (2003) Antimicrobial resistance: the example of Staphylococcus aureus. J Clin Invest. 111(9), 1265-1273. DOI: 10.1172/JCI18535
• Pajić MJ, Rašić ZB, Velebit BM, Boboš F, Mihajlović-Ukropina MM, Radinović MŽ, Galfi AL, Petković JM, Trojaćanec SI. (2014) The prevalence of methicillin resistance and Panton-Valentine leukocidin synthesis genes in Staphylococcus aureus isolates of bovine and human origin. Vet Arhiv. 84(3), 205-214.
• Pehlivanoğlu F, Yardımcı H. (2012) Detection of methicillin and vancomycin resistance in Staphylococcus strains isolated from bovine milk samples with mastitis. Kafkas Univ Vet Fak Derg. 18(5): 849-855.
• Pinho MG, Filipe SR, de Lencastre H, Tomasz A. (2001) Complementation of the essential peptidoglycan transpeptidase function of penicillin-binding protein 2 (PBP2) by the drug resistance protein PBP2A in Staphylococcus aureus. J Bacteriol. 183(22), 6525- 6531. DOI: 10.1128/JB.183.22.6525-6531.2001.
• Pu W, Su Y, Li J, Li C, Yang Z, Deng H, Ni C. (2014) High incidence of oxacillin-susceptible mecA-positive Staphylococcus aureus (OS-MRSA) associated with bovine mastitis in China. PLoS One. 9(2), e88134. DOI: 10.1371/journal.pone.0088134
• Schalm OW, Carroll EJ, Jain NC. (1971) Bovine mastitis. Philadelphia: Lea&Febiger, pp. 136-157.
• Shrestha A, Bhattarai RK, Luitel H, Karki S, Basnet HB. (2021) Prevalence of methicillin-resistant Staphylococcus aureus and pattern of antimicrobial resistance in mastitis milk of cattle in Chitwan, Nepal. BMC Vet Res. 17, 239. DOI: 10.1186/s12917-021-02942-6
• Sievert DM, Rudrik JT, Patel JB, McDonald LC, Wilkins MJ, Hageman JC. (2008) Vancomycin-resistant Staphylococcus aureus in the United States, 2002-2006. Clin Infect Dis. 46(5), 668-674. DOI: 10.1086/527392
• Strommenger B, Kettlitz C, Werner G, Witte W. (2003) Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol. 41(9), 4089-4094. DOI: 10.1128/JCM.41.9.4089-4094.2003
• Szweda P, Schielmann M, Frankowska A, Kot B, Zalewska M. (2014) Antibiotic resistance in Staphylococcus aureus strains isolated from cows with mastitis in Eastern Poland and analysis of susceptibility of resistant strains to alternative nonantibiotic agents: lysostaphin, nisin and polymyxin B. J Vet Med Sci. 76(3), 355-362. DOI: 10.1292/jvms.13-0177
• Tel OY, Bayraktar M, Keskin O. (2012) Investigation of antibiotic resistance among Staphylococcus aureus strains of human and bovine origin. Ankara Univ Vet Fak Derg. 59(3), 191-196. DOI: 10.1501/Vetfak_0000002524
• Tenhagen B-A, Köster G, Wallmann J, Heuwieser W. (2006) Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg, Germany. J Dairy Sci. 89, 2542-2551. DOI: 10.3168/jds.S0022-0302(06)72330-X
• Tiwari HK, Sen MR. (2006) Emergence of vancomycin resistant Staphylococcus aureus (VRSA) from a tertiary care hospital from northern part of India. BMC Infect Dis. 6, 156. DOI: 10.1186/1471-2334-6-156
• Türkyılmaz S, Tekbıyık S, Oryasin E, Bozdogan B. (2010) Molecular epidemiology and antimicrobial resistance mechanisms of methicillin-resistant Staphylococcus aureus isolated from bovine milk. Zoonoses Public Health. 57, 197-203. DOI: 10.1111/j.1863-2378.2009.01257.x
• Wang D, Wang Z, Yan Z, Wu J, Ali T, Li J, Lv Y, Han B. (2015) Bovine mastitis Staphylococcus aureus: Antibiotic susceptibility profile, resistance genes and molecular typing of methicillin-resistant and methicillin-sensitive strains in China. Infect Genet Evol. 31, 9-16. DOI: 10.1016/j.meegid.2014.12.039
• Zecconi A, Cesaris L, Liandris E, Daprá V, Piccinini R. (2006) Role of several Staphylococcus aureus virulence factors on the inflammatory response in bovine mammary gland. Microb Pathog. 40(4), 177-183. DOI: 10.1016/j.micpath.2006.01.001