Tavuk Eti Örneklerinden Metisiline Dirençli (MR) Stafilokok İzolasyonu

Abstract

Bu çalışmanın amacı Hatay'da satılan 50 perakende çiğ tavuk eti örneğinde metisiline dirençli stafilokok (MRS) varlığının araştırılması idi. İzolatların antimikrobiyal duyarlılıkları disk difüzyon yöntemiyle 13 farklı antimikrobiyal için test edildi ve metisilin (mecA), tetrasiklin (tetM, tetK), penisilin (blaZ), makrolid (ermA, ermC), linkozamid (lnuA) ve aminoglikozid [aac(6′)/aph(2′′), aph(3′)‐IIIa, ant(4′)‐Ia] direncine aracılık eden genler yönünden polimeraz zincir reaksiyonu (PCR) ile incelendi. Ayrıca, stafilokokal enterotoksin (SE) genlerinin varlığı da PCR ile araştırıldı. İncelenen 50 perakende çiğ tavuk eti örneğinin 11'inden (%22) MRS izole edildi ve bu izolatların 8'i (%72.7) S. sciuri ve 3'ü (%27.3) de S. epidermidis olarak identifiye edildi. Tüm izolatlar oksasilin, penisilin ve ampisiline dirençli iken tetrasiklin (8, %72.7), klindamisin (3, %27.3), trimetoprim‐sülfametoksazol (2, %18.2), eritromisin (2, %18.2) ve rifampisine (1, %9.1) farklı direnç oranları gözlendi. Tüm MRS izolatları blaZ ile birlikte mecA genini taşıdı. Fenotipik olarak tetrasiklin dirençli izolatlarda ribozomal korumadan sorumlu tetM geni tespit edildi. Eritromisine dirençli izolatlarda ermC geni ve klindamisine dirençli izolatlarda lnuA geni saptandı. İzolatlar SE genlerinin varlığı açısından negatif bulundu. Bu çalışmanın sonuçları, perakende çiğ tavuk örneklerinde MRS'nin varlığının, antibiyotik direncinin insanlara olası yayılması nedeniyle halk sağlığı açısından önemli olabileceğini göstermiştir. Ayrıca bu çalışma, hayvansal kaynaklı gıdalarda antimikrobiyal dirençli bakterilerin izlenmesinin önemini de vurgulamaktadır.

Keywords

• Tavuk eti
• Metisilin direnci
• Stafilokok

Isolation of Methicillin Resistant (MR) Staphylococci from Chicken Meat Samples

Abstract

The aim of this study was to determine the presence of methicillin resistant staphylococci (MRS) in retail raw chicken meat samples sold in Hatay. The antimicrobial susceptibility of the isolates was tested for 13 different antimicrobials by disc diffusion method and investigated for resistance genes encoding methicillin (mecA), tetracycline (tetM, tetK), penicillin (blaZ), macrolide (ermA, ermC), lincosamide (lnuA) and aminoglycoside [aac(6′)/aph(2′′), aph(3′)‐IIIa, ant(4′)‐Ia] resistance via the use of polymerase chain reaction (PCR). In addition, the presence of staphylococcal enterotoxin (SE) genes was also searched by PCR. Out of 50 collected chicken meat samples, 11 (22%) MRS was isolated and the following species were determined: S. sciuri (72.7%, 8/11) and S. epidermidis (27.3%, 3/11). While all isolates were resistant to oxacillin, penicillin and ampicillin, various rates of resistance were observed for tetracycline (8, 72.7%), clindamycin (3, 27.3%), trimethoprim‐sulfamethoxazole (2, 18.2%), erythromycin (2, 18.2%) and rifampicin (1, 9.1%). All MRS harbored mecA gene together with blaZ. The tetM gene responsible for ribosomal protection was detected in all phenotypically tetracycline resistant isolates. ermC gene in erythromycin resistant isolates and lnuA gene in clindamycin resistant isolates were detected. None of the isolates was found to be positive for SE genes. The results of this study indicated that contamination of retail raw chicken meat samples with MRS poses a risk to public health due to transmission of these bacteria to humans. Additionally, this study also highlights the importance of monitoring antimicrobial resistant bacteria in animal originated foods.

Keywords

• Chicken meat
• Methicillin resistance
• Staphylococci

References

1. References Abebe E, Gugsa G, Ahmed M, 2020: Review on major food-borne zoonotic bacterial pathogens. J Trop Med, 2020, 4674235.
2. Aslantaş Ö 2020: High occurence of methicillin resistant Staphylococcus sciuri (MRSS) and first detection of mecC from broiler flocks in Turkey. Isr J Vet Med, 75(4), 185-192.
3. Barbier F, Ruppé E, Hernandez D, Lebeaux D, Francois P, Felix B, Desprez A, Maiga A, Woerther PL, Gaillard K, Jeanrot C, Wolff M, Schrenzel J, Andremont A, Ruimy R 2010: Methicillin-resistant coagulase-negative staphylococci in the community: high homology of SCCmec IVa between Staphylococcus epidermidis and major clones of methicillin-resistant Staphylococcus aureus. J Infect Dis, 202, 270–281.
4. Becker K, Heilmann C, Peters G 2014: Coagulase-negative staphylococci. Clin Microbiol Rev, 27(4), 870-926.
5. Bhargava K, Zhang Y 2014: Characterization of methicillin-resistant coagulase-negative staphylococci (MRCoNS) in retail meat. Food Microbiol, 42, 56-60.
6. Bitrus AA, Peter OM, Abbas MA, Goni MD 2018: Staphylococcus aureus: A Review of Antimicrobial Resistance Mechanisms. Vet Scie Res Review, 4(2), 43-54.
7. Chajęcka-Wierzchowska W, Zadernowska A , Nalepa B, Sierpińska M, Łaniewska-Trokenheim Ł 2015: Coagulase-negative staphylococci (CoNS) isolated from ready-to-eat food of animal origin--phenotypic and genotypic antibiotic resistance. Food Microbiol, 46, 222-226.
8. Choi SM, Kim SH, Kim HJ, Lee DG, Choi JH, Yoo JH, Kang JH, Shin WS, Kang MW 2003: Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among staphylococcus species. J Korean Med Sci, 18, 631-636.

9. Clinical and Laboratory Standard Institute. 2012. Performance standards for antimicrobial susceptibility testing; nineteenth informational supplement. CLSI Document M100-S19. Wayne, PA, USA. Dekker D, Eibach D, Boahen KG, Akenten CW, Pfeifer Y, Zautner AE, Mertens E, Krumkamp R, Jaeger A, Flieger A, Owusu-Dabo E, May J 2019: Fluoroquinolone-Resistant Salmonella enterica, Campylobacter spp., and Arcobacter butzleri from Local and Imported Poultry Meat in Kumasi, Ghana. Foodborne Pathog Dis, 16(5), 352-358.
10. EFSA 2021: The European Union Summary Report on Antimicrobial Resistance in zoonotic and indicator bacteria from humans, animals and food in 2018/2019. EFSA J, 19(4), 6490.
11. Fessler AT, Billerbeck C, Kadlec K, Schwarz S 2010: Identification and characterization of methicillin-resistant coagulase-negative staphylococci from bovine mastitis. J Antimicrob Chemother, 65(8), 1576-82.
12. França A, Gaio V, Lopes N, Melo LDR 2021: Virulence factors in coagulase-negative staphylococci. Pathogens, 10(2), 170.
13. Hanssen AM, Kjeldsen G, Sollid JUE 2004. Local variants of staphylococcal cassette chromosome mec in sporadic methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci: evidence of horizontal gene transfer ? Antimicrob Agents Chemother, 48, 285-296. Hanssen AM, Ericson Sollid JU 2006: SCCmec in staphylococci: genes on the move. FEMS Immunol Med Microbiol, 46(1), 8-20.
14. Huber H, Ziegler D, Pflüger V, Vogel G, Zweifel C, Stephan R 2011: Prevalence and characteristics of methicillin-resistant coagulase-negative staphylococci from livestock, chicken carcasses, bulk tank milk, minced meat, and contact persons. BMC Vet Res, 7, 6.
15. Huys G, D'Haene K, Collard JM, Swings J 2004: Prevalence and molecular characterization of tetracycline resistance in Enterococcus isolates from food. Appl Environ Microbiol, 70, 1555-1562.
16. Jamaluddin TZ, Kuwahara-Arai K, Hisata K, Terasawa M, Cui L, Baba T, Sotozono C, Kinoshita S, Ito T, Hiramatsu K. Extreme genetic diversity of methicillin-resistant Staphylococcus epidermidis strains disseminated among healthy Japanese children. J Clin Microbiol, 46, 3778–3783.
17. Kim CH, Khan M, Morin DE, Hurley WL, Tripathy DN, Kehrli M Jr, Oluoch AO, Kakoma I 2001: Optimization of the PCR for detection of Staphylococcus aureus nuc gene in bovine milk. J Dairy Sci, 84(1), 74-83.
18. Kondo Y, Ito T, Ma XX, Watanabe S, Kreiswirth BN, Etienne J, Hiramatsu K, 2007: Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment: rapid identification system for mec, ccr, and major differences in junkyard regions. Antimicrob Agents Chemother, 51, 264-274.
19. Koneman EW, Allen SD, Janda WM, Schreckenberger PC, Winn WC, Washington MD. Staphylococci and Related Gram- Positive Cocci. In Koneman's Color Atlas and Textbook of Diagnostic Microbiology 5th Ed. USA: Lippincott Williams and Wilkins. 1997. p. 539-565.
20. Lina G, Quaglia A, Reverdy ME, Leclercq R, Vandenesch F, Etienne J 1999: Distribution of genes encoding resistance to macrolides, lincosamides, and streptogramins among staphylococci. Antimicrob Agents Chemother, 43, 1062-1066.
21. Martins PD, de Almeida TT, Basso AP, de Moura TM, Frazzon J, Tondo EC, Frazzon APG 2013: Coagulase-positive staphylococci isolated from chicken meat: Pathogenic potential and vancomycin resistance. Foodborne Pathog Dis, 10, 771-776.
22. Nemeghaire S, Vanderhaeghen W, Argudín MA, Haesebrouck F, Butaye P 2014: Characterization of methicillin-resistant Staphylococcus sciuri isolates from industrially raised pigs, cattle and broiler chickens. J Antimicrob Chemother, 69, 2928-2934.
23. Olsen JE, Christensen H, Aarestrup FM 2006: Diversity andevolution of blaZ from Staphylococcus aureus and coagulase-negative staphylococci. J Antimicrob Chemother, 57, 450-460.
24. Omoe K, Hu DL, Takahashi-Omoe H, Nakane A, Shinagawa K 2005: Comprehensive analysis of classical and newly described staphylococcal superantigenic toxin genes in Staphylococcus aureus isolates. FEMS Microbiol Lett, 246(2), 191-198.
25. Osman K, Alvarez-Ordóñez A, Ruiz L, Badr J, ElHofy F, Al-Maary KS, Moussa IMI, Hessain AM, Orabi A, Saad A, Elhadidy M 2017: Antimicrobial resistance and virulence characterization of Staphylococcus aureus and coagulase-negative staphylococci from imported beef meat. Ann Clin Microbiol Antimicrob, 16, 35.
26. Otto M 2013: Coagulase-negative staphylococci as reservoirs of genes facilitating MRSA infection: Staphylococcal commensal species such as Staphylococcus epidermidis are being recognized as important sources of genes promoting MRSA colonization and virulence. Bioessays, 35(1), 4-11.
27. Pehlivanlar Önen S, Aslantaş Ö, Şebnem Yılmaz E, Kürekci C 2015: Prevalence of β-Lactamase Producing Escherichia coli from Retail Meat in Turkey. J Food Sci, 80(9), M2023-9.
28. Preethirani PL, Isloor S, Sundareshan S, Nuthanalakshmi V, Deepthikiran K, Sinha AY, Rathnamma D, Nithin Prabhu K, Sharada R, Mukkur TK, Hegde NR 2015: Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India. PLoS One, 10(11), e0142717.
29. Prestinaci F, Pezzotti P, Pantosti A 2015: Antimicrobial resistance: a global multifaceted phenomenon. Pathog Glob Health, 109(7), 309-318.
30. Rolo J, Worning P, Nielsen JB, BowdenR, BouchamiO, DamborgP, GuardabassiL, Perreten V, Tomasz A, Westh H, de Lencastre H, Miragaia M 2017: Evolutionary origin of the staphylococcal cassette chromosome (SCCmec). Antimicrob Agents Chemother, 6, e02302-16.
31. Silva NC, Guimarães FF, de P Manzi M, Gómez-Sanz E, Gómez P, Araújo-Júnior JP, Langoni H, Rall VL, Torres C 2014: Characterization of methicillin-resistant coagulase-negative staphylococci in milk from cows with mastitis in Brazil. Antonie Van Leeuwenhoek, 106(2), 227-33.
32. 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, 4089–4094.
33. Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG 2015: Staphylococcus aureus infections: Epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev, 28, 603-661.
34. Tsubakishita S, Kuwahara-Arai K, Sasaki T, Hiramatsu K 2010: Origin and molecular evolution of the determinant of methicillin resistance in staphylococci. Antimicrob Agents Chemother, 54(10), 4352-4359.
35. Vanderhaeghen W, Vandendriessche S, Crombe F, Dispas M, Denis O, Hermans K, Haesebrouck F, Butaye P 2012: Species and staphylococcal cassette chromosome mec (SCCmec) diversity among methicillin-resistant non-Staphylococcus aureus staphylococci isolated from pigs. Vet Microbiol, 158, 123-128.
36. Udo E, Al-BustanM, JacobL, Chugh T 1999: Enterotoxin production by coagulase-negative staphylococci in restaurant workers from Kuwait city may be a potential cause of food poisoning. J Med Microbiol, 48, 819-823.
37. Ünal S 2006: Toplumda kazanılmış metisilin dirençli Staphylococcus aureus. Genetik özellikler. Ankem Derg, 20, 100–101.
38. Wisplinghoff H, Rosato AE, Enright MC, Noto M, Craig W, Archer GL 2003: Related clones containing SCCmec type IV predominate among clinically significant Staphylococcus epidermidis isolates. Antimicrob Agents Chemother, 47, 3574–3579.
39. Yılmaz EŞ, Aslantaş Ö, Pehlivanlar Önen S, Türkyılmaz S, Kürekci C 2016: Prevalence, antimicrobial resistance and virulence traits in enterococci from food of animal origin in Turkey. LWT- Food Sci Technol, 66, 20-26.
40. Zhang Y, Agidi S, LeJeune J 2009: Diversity of staphylococcal cassette chromosome in coagulase‐negative staphylococci from animal sources. J Appl Microbiol, 107(4), 1375-1383.