Occurrence, enterotoxigenic properties and antimicrobial resistance of Staphylococcus aureus on beef carcasses

Abstract

The objectives of this study were to determine the incidence, enterotoxigenic properties and antimicrobial resistance of Staphylococcus aureus. For this purpose, 120 beef carcass samples, obtained from 2 different slaughterhouses in Ankara. All samples were collected to each of carcass regions of rump, flank and brisket per 100 cm2 (10 × 10 cm, a total of 300 cm2) under aseptic conditions with sponge swab technique. Results of microbiological examinations, 15 (12.5%) of the 120 beef carcasses samples were confirmed as S. auerus. 19 (86,3 %) out of 22 S. aureus isolates determined as enteotoxigenic. It was determined that these enterotoxins contained 8 (36.3%) seh, 5 (22.7%) sea, 2 (9%) seg+sei, 1 (4.5%) sei, 1 (4.5%) sed+sej, 1 (4.5%) sea+tst, 1 (4.5%) tst gene distribution, respectively. According to the disc diffusion test results, 72.7%, 54.5%, 40.9%, 22.7% of S. aureus isolates were high level resistant to ampicillin, tetracycline, erythromycin, sulphamethoxazole- trimethoprim, respectively. All of the isolates were sensitive to vancomycin. As a conclusion, it was found that beef carcasses contaminated with S. aureus 15 (12.5%) and enterotoxigenic S. aureus level is relatively high 19 (86.3%). It is therefore recommended that slaughterhouses introduce internal hygiene measures such as the HACCP system in order to produce beef meat of good quality and thus protect public health

Keywords

• Antimicrobial resistance
• beef carcass
• PCR
• staphylococcal enterotoxin
• S. aureus

Sığır karkaslarında Staphylococcus aureus’un varlığı, enterotoksijenik özellikleri ve antimikrobiyal dirençliliği

Abstract

Bu çalışma, sığır karkaslarında Staphylococcus aureus’un varlığı ile izolatların enterotoksijenik özellikleri ve antimikrobiyal dirençliliğini belirlemek için yapılmıştır. Bu amaçla, Ankara’da bulunan 2 farklı mezbahadaki 120 adet sığır karkasından alınan örnekler materyal olarak kullanılmıştır. Örnekler aseptik koşullarda her bir sığır karkasının but, kavram ve döş bölgelerinin 100 cm2’ lik (10×10 cm) alanından (toplam 300 cm2) sünger swab tekniği ile alınmıştır. Yapılan mikrobiyolojik analizler sonucu, 120 sığır karkas örneğinin 15’i (%12,5) S. aureus yönünden pozitif bulunmuştur. S. aureus olarak belirlenen 22 izolattan 19’unun (%86,3) ise enterotoksijenik özellikte olduğu belirlenmiştir. Enterotoksijenik izolatlardan 8’inin (% 36,3) seh, 5’inin ( %22,7) sea, 2’sinin (%9) seg+sei, 1’inin (%4,5) sei, 1’inin (%4,5) sed+sej, 1’inin (%4,5) sea+tst ve 1’ininde (%4,5) tst tipi enterotoksin genine sahip olduğu saptanmıştır. Yapılan antibiyotik dirençlilik testleri sonucunda S. aureus izolatlarından %72,7’sinin ampisiline, %54,5’inin tetrasikline, %40,9’unun eritromisine ve %22,7’sinin sülfametaksazol-trimetoprime yüksek oranda dirençli olduğu, vankomisine ise tümünün duyarlı olduğu saptanmıştır. Sonuç olarak, sığır karkaslarında S. aureus, 15 örnekte (%12.5) bulunmuş olup, izolatların 19’u (%86,3) enterotoksijenik özelliktedir. Bu nedenle sağlıklı sığır eti üretimi ve halk sağlığının korunması için mezbahalarda etkin HACCP (Hazard Analysis Critical Control Point) sisteminin uygulanması önemlidir

Keywords

• Antimikrobiyal direnç
• PCR
• sığır karkas
• stafilokokal enterotoksin
• S. aureus

References

1. Akineden Ö, Hassan AA, Schneider E ve ark. (2008): Enterotoxigenic properties of Staphylococcus aureus isolated from goats’milk cheese. Int J Food Microbiol, 124, 211-216.
2. Anon (1996): USDA/FSIS (United States Department of Agriculture/Food Safety and Inspection Service). Federal Register, 61, 38931-38937.
3. Anon (2003): ISO (International Standard Office, 6888-1). Microbiology of food and animal feedingstuffs-horizontal method for the enumeration of coagulase-positive staphylococci (S. aureus and other species).
4. Anon (2011): CLSI (The Clinical and Laboratory Standards Institute). The Clinical and Laboratory Performance Standards for Antimicrobial Susceptibility Testing, 21th Informational Supplement. M100-S21. Wayne Pa.
5. Arcuri EF, Angelo FF, Guimarães MF ve ark. (2010): Toxigenic status of Staphylococcus aureus isolated from bovine raw milk and Minas frescal cheese in Brazil. J Food Prot, 73, 2225–2231.
6. Aydın A, Sudagıdan M, Muratoğlu K (2011): Prevalence of Staphylococcal enterotoxins, toxin genes and genetic- relatedness of foodborne S. aureus strains isolated in the Marmara Region of Turkey. Int J Food Microbiol, 148, 99- 106.
7. Bania J, Dabrowska A, Bystron J ve ark. (2006): Distribution of newly described enterotoxin like genes in Staphylococcus aureus from food. Int J Food Microbiol, 108, 36-41.
8. Bergdoll MS, Lee Wong AC (2006): Staphylococcal intoxications. 523-562. In: HP Rieman, DO Cliver (Eds), Foodborne Infections and Intoxications, Academic Press, Elsevier Inc., San Diego, California.

9. Bhargava K, Wang X, Donabedian ve ark. (2011): Methicillin-resistant Staphylococcus aureus in retail meat, Detroit, MI, USA. Emerg Infect Dis, 17, 1135–1137.
10. Brakstad OG, Aasbakk K, Maeland J A (1992): Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J Clin Microbiol, 30, 1654-1660.
11. Gündoğan N, Çıtak S, Yücel N ve ark. (2005): A note on the incidence and antibiotic resistance of Staphylococcus aureus isolated from meat and chicken samples. Meat Sci, 69, 807-810.
12. Hanson BM, Dressler AE, Harper AL ve ark. (2011): Prevalence of Staphylococcus aureus and methicillin- resistant Staphylococcus aureus (MRSA) on retail meat in Iowa. J Infect Public Health, 4, 169–174.
13. Jackson CR, Davis JA, Barrett JB (2013): Prevalence and characterization of methicillin-resistant Staphylococ- cus aureus isolates from retail meat and humans in Geor- gia. J Clin Microbiol, 51, 1199-1207.
14. Jarraud S, Cozon G, Vandenesch FBM ve ark. (1999): Involvement of enterotoxins G and I in staphylococcal toxic shock syndrome and staphylococcal scarlet fever. J Clin Microbiol, 37, 2446–49.
15. Johnson WM, Tyler SD, Ewan, EP ve ark. (1991): Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in Staphylococcus aureus by the polymerase chain reaction. J Clin Microbiol, 29, 426– 430.
16. Kitai S, Shimizu A, Kawano J, ve ark. (2005): Characterization of methicillin-resistant S. aureus isolated from retail raw chicken meat in Japan. J Vet Med Sci, 67, 107-110.
17. Lim SK, Nam HM, Park HJ ve ark. (2010): Prevalence and characterization of methicillin resistant Staphylococcus aureus in raw meat in Korea. J Microbiol Biotech, 20, 775–778.
18. Lowy, F. D (1998): Staphylococcus aureus infections. The New England J Med. 339, 520-532.
19. Maes N, Magdalena J, Rottiers S ve ark. (2002): Evaluation of a triplex PCR assay to discriminate Staphylococcus staphylococci and determine methicillin resistance from blood Cultures. J Clin Microbiol, 40, 1514–1517.
20. Mehrotra M, Wang G, Johnson WM (2000): Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1 and methicillin resistance. J Clin Microbiol, 38, 1032- 1035.
21. Monday SR, Bohach GA (1999): Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates. J Clin Microbiol, 37, 3411- 3414.
22. Nitzsche S, Zweifel C, Stephan R (2007): Phenotypic and genotypic traits of Staphylococcus aureus strains isolated from pig carcasses. Vet Microbiol, 120, 292–299.
23. Normanno G, Corrente M, La Salandra G ve ark. (2007): Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products. Int J Food Microbiol, 115, 290-296.
24. Pelisser MR, Klein CS, Ascoli KR ve ark. (2009): Ocurrence of Staphylococcus aureus and multiplex PCR detection of classic enterotoxin genes in cheese and meat products. Braz J Microbiol, 40, 145–148.
25. Pereira V, Lopes C, Castro A ve ark. (2009): Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of staphylococcus aureus isolates from various foods in portugal. Food Microbiol, 26, 278-282.
26. Pesavento G, Duccib B, Comodo N ve ark. (2007): Antimicrobial resistance profile of Staphylococcus aureus isolated from raw meat: A research for methicilin resistant Staphylococcus aureus (MRSA). Food Control, 18, 196- 200.
27. Pu S, Han F, Ge B (2009): Isolation and characterization of methicillin-resistant Staphylococcus aureus strains from Louisiana retail meats. Appl Environ Microbiol, 75, 265- 267.
28. Rosec JP, Gigaud O. (2002): Staphylococcal enterotoxin genes of classical and New types detected by PCR in France. Int J Food Microbiol, 77, 61-70.
29. Schneewind O, Missiakas D (2009): Staphylococcus aureus and related Staphylococci. 275-294. In: E Goldman, LH Green (Eds), Practical Handbook of Microbiology. CRC Press, USA.
30. Van Loo IHM, Diederen BMW, Savelkoul PHM ve ark. (2007): Methicilin-resistant Staphylococcus aureus in meat products, the Netherlands. Emerg Infect Dis, 13, 1753- 1755.
31. Witte, W, Cuny, C, Strommenger B, ve ark. (2004): Emergence of community-acquired MRSA in Germany. Eurosurveillance, 9, 16-18.
32. Yılmaz İ, Gümüş T (2004): Sığır karkaslarının mikrobi- yolojik kalitesinin belirlenmesi. Türkiye 10. Gıda Kongre- si, 21-23 Mayıs. Erzurum.
33. Zhang S, Iandolo J.J, Stewart GC (1998): The enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotox in determinant (sej). FEMS Microbiol Lett, 168, 227–233.