KANATLI ETLERİNDE STAPHYLOCOCCUS AUREUS YAYGINLIĞI VE ANTİBİYOTİK DİRENÇ PROFİLLERİNİN, ANTİBİYOTİK DİRENÇ VE ENTEROTOKSİN GENLERİNİN BELİRLENMESİ

Year 2021, Volume: 46 Issue: 3, 692 - 706, 15.05.2021

Çilem KISA Yasin TUNCER

https://doi.org/10.15237/gida.GD21048

Abstract

Bu çalışmada, Isparta ve Antalya illerinde perakende satışı yapılan kanatlı etlerinde (32 piliç, 9 hindi, 9 bıldırcın) Staphylococcus aureus yaygınlığının araştırılması, antibiyotik direnç profilleri ve direnç genlerinin yanı sıra enterotoksin genleri varlığının belirlenmesi amaçlanmıştır. Kanatlı etlerinden muhtemel S. aureus izolasyonunda Baird Parker agar besiyeri kullanılmıştır. İzolatların tür düzeyinde tanısı S. aureus’da termostabil nükleaz genine (nuc) özgü primer çifti kullanılarak polimeraz zincir reaksiyonu (PZR) ile yapılmıştır. 130 muhtemel S. aureus izolatının 16’sında nuc genine özgü 458 bç büyüklüğünde amplikonlar elde edilmiştir. Kanatlı eti örneklerinde S. aureus bulunma sıklığı % 20 (10/50) bulunmuştur. Koagülaz testi, S. aureus izolatlarının tamamının koagülaz pozitif olduğunu göstermiştir. Disk difüzyon testi, izolatların tamamının (% 100) kloramfenikole ve teikoplanine duyarlı, penisilin G’ye ise dirençli olduğunu göstermiştir. İzolatların % 81.25’inin (13/16) metisilin dirençli S. aureus (MRSA) olduğu tespit edilmiştir. PZR işlemi sonucu S. aureus suşlarında en sık rastlanan antibiyotik direnç geninin blaZ (% 62.5, 10/16) olduğu tespit edilmiştir. S. aureus suşlarının hiçbirinde sea, seb, sec, sed ve see geni varlığı belirlenmemiştir.

Keywords

Kanatlı eti, Staphylococcus aureus, antibiyotik direnç, antibiyotik direnç geni, enterotoksin geni

Supporting Institution

Süleyman Demirel Üniversitesi

Thanks

Bu çalışmayı FYL-2019-6994 nolu proje ile maddi olarak destekleyen Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Yönetim Birimi’ne teşekkür ederiz.

DETERMINATION OF STAPHYLOCOCCUS AUREUS PREVALENCE AND ANTIBIOTIC RESISTANCE PROFILES, ANTIBIOTIC RESISTANCE AND ENTEROTOXIN GENES IN POULTRY MEATS

Abstract

In this study, it was aimed to investigate the prevalence of Staphylococcus aureus in poultry meats (32 chickens, 9 turkeys, 9 quails) sold in Isparta and Antalya provinces, and to determine the presence of enterotoxin genes as well as antibiotic resistance profiles and resistance genes. Baird Parker agar medium was used to isolate presumptive S. aureus from poultry meats. The species-level identification of isolates was done by polymerase chain reaction (PCR) using primer pair specific to the thermostable nuclease gene (nuc) in S. aureus. As a result of the PCR analysis, amplicons of 458 bp specific to the nuc gene were obtained in 16 of 130 presumptive S. aureus isolates. The frequency of S. aureus in poultry meat samples was found to be 20% (10/50). Coagulase test showed that all the S. aureus isolates were coagulase positive. Disk diffusion test showed that all of the isolates (100 %) were susceptible to chloramphenicol and teicoplanin and resistant to penicillin G. It was determined that 81.25 % (13/16) of the isolates were methicillin resistant S. aureus (MRSA). The most common antibiotic resistance gene in S. aureus strains was found to be blaZ (62.5 %, 10/16) as a result of PCR. The presence of sea, seb, sec, sed and see genes was not detected in any of the S. aureus strains.

Keywords

Poultry meat, Staphylococcus aureus, antibiotic resistance, antibiotic resistance gene, enterotoxin gene

References

• Abolghait, S.K., Fathi, A.G., Youssef, F.M., Algammal, A.M. (2020). Methicillin-resistant Staphylococcus aureus (MRSA) isolated from chicken meat and giblets often produces staphylococcal enterotoxin B (SEB) in non-refrigerated raw chicken livers. Int J Food Microbiol, 328: 1-9.
• Anand, K.B., Agrawal, P., Kummar S., Kapila K. (2009). Comparison of sefoxitin disc diffusion test, oxacillin screen agar, and PCR for mecA gene for detection of MRSA. Indian J Med Microbiol, 27(1): 27-29.
• Anonymous (2020). UK standarts for microbiology investigations. Identification of Staphylococcus species, Micrococcus species and Rothia species. UK SMI ID: 7, Issue no: 4, Issue date: 26.05.20, England.
• Argudín, M., Mendoza, M., González-Hevia, M., Bances, M., Guerra, B., Rodicioa, M. (2012). Genotypes, exotoxin gene content, and antimicrobial resistance of Staphylococcus aureus strains recovered from foods and food handlers. Appl Environ Microbiol, 78(8): 2930-2935.
• Asfour, H.A.E., Darwish, S.F. (2011). Phenotypic and genotypic detection of both mecA- and blaZ- genes mediated β-lactam resistance in Staphylococcus strains isolates from bovine mastitis. Glob Vet, 6(1): 39-50.
• Bacigil, A.F., Taponen S., Koort, J., Bengtsson, B., Myllyniemi, A.L., Pyörἃlἃ, S. (2012). Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis. Acta Vet Scand, 54(69): 1-7.
• Bayston, R. (2006). Coagulase-negative methicillin-resistant Staphylococcus aureus. J Hosp Infect, 62(1): 127.
• Brakstad, O.G., Aasbakk, K., Maeland, J.A. (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J Clin Microbiol, 30(7): 1654-1660.
• Cancilla, M.R., Powell, I.B., Hillier, A.J., Davidson, B.E. (1992). Rapid genomic fingerprinting of Lactococcus lactis strains by arbitrarily primed polymerase chain reaction with 32P and fluorescent labels. Appl Environ Microbiol, 58(5): 1772-1775.
• Cariolato, D., Andrighetto, C., Lombardi, A. (2008). Occurrence of virulence factors and antibiotic resistances in Enterococcus faecalis and Enterococcus faecium collected from dairy and human samples in North Italy. Food Control, 19(9): 886-892.
• Clinical and Laboratory Standards Institute, (CLSI 2016). Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fifth Informational Supplement M100-S26 CLSI, Wayne, PA.
• Depardieu, F., Perichon, B., Courvalin, P. (2004). Detection of van alphabet and identification of enterococci and staphylococci at the species level by multiplex PCR. J Clin Microbiol, 42(12): 5857-5860.
• Donham, K.J. (2010). Community and occupational health concerns in pork production. J Anim Sci, 88: 102-111.
• Dutka M.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(1): 24-27.
• Frey, Y., Rodriguez, J.P., Thomann, A., Schwendener, S., Perreten, V. (2013). Genetic characterization of antimicrobial resistance in coagulase-negative staphylococci from bovine mastitis Milk. J Dairy Sci, 96: 2247-2257.
• Gandra, E., Fernandez, M.A., Sılva, J.A., Sılva, W.P. (2011). Standardization of a multiplex PCR for the identification of coagulase-positive Staphylococcus. Ciênc Tecnol Aliment, 31(4): 946-949.
• Gencay, Y.E., Ayaz, N.D., Doğru, A.K. (2010). Enterotoxin gene profiles of Staphylococcus aureus and other staphylococcal isolates from various foods and food ingredients, Erciyes Üniv Vet Fak, 7(2): 75-80.
• Hanson, B.M., Dressler, A.E., Harper A.L., Scheibel R.P., Wardyn, S.E., Roberts, L.K., Kroeger J.S., Smith, T.C. (2011). Prevalence of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) on retail meat in lowa. J Infect Public Health, 4: 169-174.
• Hauschild, T., Sacha, P., Wieczorek, P., Zalewska, M., Kaczyǹska, K., Tryniszewska, E. (2008). Aminoglycosides resistance in clinical isolates of Staphylococcus aureus from a university hospital in Bialystok, Poland. Follia Histochem Cytobiol, 46(2): 225-228.
• John, J., George, S., Nori, S.R.C. Nelson-Sathi, S. (2019). Phylogenomic analysis reveals the evolutionary route of resistant genes in Staphylococcus aureus. Genome Biol Evol, 11(10): 2917-2926.
• Kaçmaz, B., Gül, S., Öztürk, D.B., Ecemiş, E. (2015). Staphylococcus aureus suşlarında koagulaz testi için uygun sıcaklığın araştırılması. Turk Hij Den Biyol Derg, 75(2): 99-102.
• Kadariya, J., Smith, T., Thapaliya, D. (2014). Staphylococcus aureus and staphylococcal food-borne disease: an ongoing challenge in public health. BioMed Res Int, 2014: 827965.
• Kadiroğlu, P., Korel, F., Ceylan, Ç. (2019). Identification of Staphylococcus aureus cheese isolates with respect to virulence properties, genetic relatedness and antibiotic resistance profiles. Food and Health, 5(3): 149-159.
• Karmakar, A., Dua, P., Ghosh, C. (2016). Biochemical and molecular analysis of Staphylococcus aureus clinical isolates from hospitalized patients. Can J Infect Dıs Med, 2016: 9041636.
• Kaya, H., Onmaz, N.E., Gönülalan, Z., Al, S. (2015). Kayseri ilinde tüketime sunulan tavuk etlerinde Staphylococcus aureus ve enterotoksin varlığının araştırılması. Erciyes Üniv Vet Fak, 12(2): 93-98.
• Lee, J.H. (2003). Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol, 69(11): 6489-6494 .
• Lina G., Quaglia A., Reverdy M.E., 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.
• Loir, Y., Baron, F., Gautier, M. (2003). Staphylococcal food poisoning. Genetic and Molecular Research, 2(1): 7-28.
• Martineau, F., Picard, F., Lansac, N., Menard, C., Roy, P.H., Ouellette, M., Bergeron, M.G. (2000). Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother, 44(2): 231-238.
• Niu H., Yu H., Hu T., Tian G., Zhang L., Guo X., Hu H., Wang Z. (2016). The prevalence of aminoglycoside-modifying enzyme and virulence genes among enterococci with high-level aminoglycoside resistance in Inner Mongolia, China. Braz J Microbiol, 47: 691-696.
• Olsen, J.E., Christensen, H., Aarestrup F.M. (2006). Diversity and evolution of blaZ from Staphylococcus aureus and coagulase-negative staphylococci. J Antimicrob Chemother, 57: 450-460.
• Omoe, K., Hu, D.L., Takahashi-Omoe, H., Nakane, A., Shinagawa, K. (2005). Comprehensive analysis of clasical and newly described staphylococcal superantihenic toxin genes in Staphylococcus aureus isolates. FEMS Microbiol Lett, 246: 191-198.
• Omoe, K., Ishikawa, M., Shimoda, Y., Hu, D.L., Ueda, S., Shinagawa, K. (2002). Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes. J Clin Microbiol, 40(3): 857-862.
• Ouoba, L.I.I., Lei, V., Jensen, L.B. (2008). Resistance of potential probiotic lactic acid bacteria and bifidobacteria of African and European origin to antimicrobials: determination and transferability of the resistance genes to other bacteria. Int J Food Microbiol, 121: 217-224.
• Özdemir H., Keyvan, E. (2016). Isolation and characterisation of Staphylococcus aureus strains isolated from beef, sheep and chicken meat. Ankara Üniv Vet Fak Derg, 63: 333-338.
• Pal, M., Kerorsa, G.B., Marami, L.M., Kandi, V. (2020). Epidemiology, pathogenicity, animal infections, antibiotic resistance, public health significance, and economic impact of Staphylococcus aureus: a comprehensive review. Am J Public Health Res, 8(1): 14-21.
• Pereira, V., Lopes, C., Castro, A., Silva, J., Gibbs, P., Teixeria, P. (2009). Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from serious foods in Portugal. Food Microbiol, 26: 278-282.
• Pyzik, E., Marek, A., Pysniak, D.S., Chmiel, R.U., Jarosz, L.S., Podebska, I.J. (2019). Detection of antibiotic resistance and classical enterotoxin genes in coagulase negative staphylococci isolated from poultry in Poland. J Vet Res, 63: 183-190.
• Reyes, J., Hidalgo, M., Diaz, L., Rincon, S., Moreno, J., Vanegas, N., Castaneda, E., Arias, C.A. (2007). Characterization of macrolide resistance in Gram-positive cocci from Colombian hospitals: a countrwide surveillance. Int J Infect Dis, 11: 329-336.
• Saad, M.S., A, N., Elroos, A., Abdel-Fadeel, S.R. (2018). Incidence and characterization of S. aureus in broiler carcasses. Benha Veterinary Medical Journal, 34(2): 191-200.
• Sadiq, A., Samad, M., Saddam, Basharat, N., Ali, S., Roohullah, Saad, Z., Khan, A.N., Ahmad, Y., Khan, A., Khan, J. (2020). Methicillin-resistant Staphylococcus aureus (MRSA) in slaughter houses and meat shops in capital territory of Pakistan during 2018-2019. Front Microbiol, 11: 1-12.
• Schelin, J., Wallin-Carlquist, N., Cohn, M., Lindqvist, R., Barker, G. (2011). The formation of Staphylococcus aureus enterotoxin in food environments and advances in risk assessment. Virulence, 2(6): 580-592.
• Sırıken, B., İnat, G., Yıldırım, T., Yavuz, C., Çiftci, A. (2018). Methicillin resistance coagulase positive staphylococci and Staphylococcus aureus isolated from raw milk and cheese, Samsun province-Turkey. Gıda Yem Bilimi-Teknolojisi Dergisi, 20: 55-63.
• Swenson, J.M. (2002). News tests for the detection of oxacillin-resistant Staphylococcus aureus. Clin Microbiol Newslett, 24(21): 159-163.
• Takayama, Y., Tanaka, T., Oikawa, K., Fukano, N., Goto, M., Takahashi, T. (2015). Prevalence of blaZ gene and performance of phenotypic tests to detect penicillinase in Staphylococcus aureus isolates from Japan. Ann Lab Med, 38: 155-159.
• Temiz, A. (1994). Genel Mikrobiyoloji Uygulama Teknikleri. Şafak Matbaacılık Ltd. Şti., Ankara, Türkiye, 266 s. ISBN:975-95834-0-2
• Tomasz, A., Drugeon H.B., Lencastre, H.M., Jabes, D., McDougal, L., Bille, J. (1989). New mechanism for methicillin resistance in Staphylococcus aureus: clinical isolates that lack the PBP 2a gene and contain modified penicillin-binding proteins with modified penicillin-binding capacity. Antimicrob Agents Chemother, 33: 1869-1874.
• Ullah, F., Malik, S.A., Ahmed, J., Ullah, F., Shah, S.M., Ayaz, M., Hussain, S., Khatoon, L. (2012). Investigation of the genetic basis of tetracycline resistance in Staphylococcus aureus from Pakistan. Trop J Pharm Res, 11(6): 925-931.
• Vandenesch, F., Lebeau, C., Bes, M., Mcdevitt, D., Greenland, T., Novickj, R.P., Etienne, J. (1994). Coagulase deficiency in clinical isolates of Staphyococcus aureus invoIves both transcriptional and post-transcriptional defects. J Med Microbiol, 40: 344-349.
• Vakulenko, S.B., Donabedian, S.M., Voskresenskiy, A.M., Zervos, M.J., Lerner, A., Chow, J.W. (2003). Multiplex PCR for detection of aminoglycoside resistance genes in enterococci. Antimicrob Agents Chemother, 47(4): 1423-1426.
• Wang, W., Baloch, Z., Jiang, T., Zhang, C., Peng, Z., Li, F., Fanning, S., Ma, A. Xu J. (2017). Enterotoxigenicity and antimicrobial resistance of Staphylococcus aureus isolated from retail food in China. Front Microbiol, 8: 1-11.
• Woegerbauer, M., Zeinzinger, J., Springer, B., Hufnagl, P., Indra, A., Korschineck, I., Hofrichter, J., Kopacka, I., Fuchs, R., Steinwider, J., Fuchs, K., Nielsen, K.M., Allerberg, F. (2014). Prevalence of the aminoglycoside phosphotransferase genes aph(3’)-IIIa and aph(3’)-IIa in Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica and Staphylococcus aureus isolates in Austria. J Med Microbiol, 63: 210-217.
• Wu, S., Huang, J., Wu, Q., Zhang, J., Zhang, F., Yang, X., Wu, H., Zeng, H., Chen, M., Ding, Y., Wang, J., Lei, T., Zhang, S. Xue L. (2018). Staphylococcus aureus isolated from retail meat and meat products in China: incidence, antibiotic resistance and genetic diversity. Front Microbiol, 9: 1-14.
• Wu, S., Zhang, F., Huang, J., Wu, Q., Zhang, Y., Xue, L., Wang, J., Ding, Y. (2019). Phenotypic and genotypic characterization of PVL-positive Staphylococcus aureus isolated from retail foods in China. Int J Food Microbiol, 304: 119-126.
• Yang, F., Wang, O., Wang, X., Wang, L., Xiao, M., Lı, X., Luo, J., Zhang, S., Lı., H. (2015). Prevalence of blaZ Gene and other virulence genes in penicillin-resistant Staphylococcus aureus isolated from bovine mastitis cases in Gansu, China. Turk J Vet Anim Sci, 39: 634-636.
• Yogurtcu, N.N., Tuncer, Y. (2013). Antibiotic susceptibility patterns of Enterococcus strains isolated from Turkish tulum cheese. Int J Dairy Technol, 66: 236-242.
• Zhang, K., Sparling, J., Chow., B.L., Elsayed, S., Hussain, Z., Church, D.L., Gregson, D.B., Louie, T., Conly, J.M. (2004). New quadriplex PCR assay for detection of methicillin and mupirocin resistance, and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci. J Clin Microbiol, 42: 4947-4955.