Çiğ Sütte Stafilokokal Enterotoksinler ve Enterotoksijenik Staphylococcus aureus Varlığının Belirlenmesine Yönelik Bir Tarama Çalışması

Öz

Staphylococcus aureus, insanlarda gıda kaynaklı zehirlenmelerin başlıca nedenidir. Stafilokokal enterotoksinler (SE'ler) ile kontamine süt ve süt ürünleri tüketimi salgınlara neden olabilmektedir. Bu çalışmanın amacı, 120 çiğ süt örneğinde S. aureus izolatlarının stafilokokal enterotoksinlerin ve enterotoksijenik özelliklerinin analiz edilmesidir. SE'leri saptamak için enzim bağlantılı immünosorban testi (ELISA) yöntemi kullanılarak yüz yirmi çiğ süt örneği analiz edildi. Polimeraz zincir reaksiyonu (PCR) ile stafilokokkal entertoksin genleri (sea, seb, sec, sed, see) araştırıldı. Bu çalışmada, toplama tanklarından alınan 120 çiğ süt örneğinden 2’sinde SE tespit edilmiştir. Toplam 69 S. aureus izolatının 18'i (% 38.3) nuc ve coa genleri PCR yöntemi ile doğrulanmıştır. SE genleri, 18 S. aureus izolatının 3’ünde (%16,6) bulunmuştur. Süt ve süt ürünlerinde bulunan stafilokokal enterotoksinler halk sağlığı açısından potansiyel tehlikedir. Çiğ sütteki stafilokokal enterotoksinlerin kontrolü için sürveyans programları ve etkili izleme sistemleri gereklidir.

Anahtar Kelimeler

• Çiğ süt,Stafilokokal enterotoksinler,Staphylococcus aureus

Staphylococcal Enterotoxins and Enterotoxigenic Staphylococcus aureus in Raw Milk: A Screening Study

Abstract

Staphylococcus aureus is one of the most important cause of foodborne intoxications in human beings. Staphylococcal enterotoxins (SEs) may lead to outbreaks because of taking food such as milk and dairy products. The aims of this study were to analyze the presence of staphylococcal enterotoxins and enterotoxigenic properties of the S. aureus isolates in 120 raw milk samples. One hundred and twenty raw milk samples were analyzed to detect SEs using the enzim-linked immunosorbent assay (ELISA) method. Staphylococcal entertoxin genes (sea, seb, sec, sed, see) were analysed by polymerase chain reaction (PCR). In the current study, SEs were found 2 of 120 bulk tank milk samples. Totally 18 (38.3%) of 69 isolates were confirmed by PCR targeting nuc and coa genes in S. aureus. SEs genes were detected as 3 (16.6 %) of 18 S. aureus isolates. Staphylococcal enterotoxins in foods like milk and dairy products are the potential public health hazards. Surveillance programs and effective monitoring systems are required for controlling staphylococcal enterotoxins in raw milk.

Keywords

• Raw milk,Staphylococcal enterotoxins,Staphylococcus aureus

Kaynakça

1. Akineden Ö, Hassan AA, Schneider E, Usleber E. Enterotoxigenic properties of Staphylococcus aureus isolated from goats milk cheese. Int J Food Microbiol. 2008; 124(2):211-216.
2. Argudín MÁ, Mendoza MC, Rodio MR. Food poisoning and Staphylococcus aureus enterotoxins. Toxins. 2010; 2(7):1751–1773. Benkerroum N. Staphylococcal enterotoxins and enterotoxin-like toxins with special reference to dairy products: An overview. Crit Rev Food Sci Nutr. 2018; 58(12):1943-1970.
3. Bennett RW, Lancette GA. Staphylococcus aureus, In: BAM Bacteriological Analytical Manual. U.S. Food and Drug Administration, Centre for Food Safety & Applied Nutrition, AOAC International, Gaithersburg, MD. 2001; pp. 222–228.
4. Berdgoll M. Staphylococcus aureus, In: Foodborne Bacterial Pathogens, Ed; Doyle M, Marcel Dekker, New York, USA. 1989; pp. 464–523.
5. Boynukara B, Gulhan T, Alisarli M, Gurturk K, Solmaz H. Classical enterotoxigenic characteristics of Staphylococcus aureus strains isolated from bovine subclinical mastitis in Van, Turkey. Int J Food Microbiol. 2008; 125(2):209-211.
6. Cusato S, Gameiro AH, Sant'Ana AS, Corassin CH, Cruz AG, De Oliveira CAF. Assessing the costs involved in the implementation of GMP and HACCP in a small dairy factory. Qual Assur Saf of Crop. 2014; 6(2):135-139.
7. De Silva SASD, Kanugala KANP, Weerakkody NS. Microbiological quality of raw milk and effect on quality by implementing good management practices. Procedia Food Sci. 2016; 6:92-96.
8. Ding T, Yu YY, Schaffner DW, Chen SG, Ye XQ, Liu DH. Farm to consumption risk assessment for Staphylococcus aureus and staphylococcal enterotoxins in fluid milk in China. Food Control. 2016; 59:636-643.

9. Duquenne M, Fleurot I, Aigle M, Darrigo C, Borezée-Durant E, Derzelle S, Bouix M, Deperrois-Lafarge V, Delacroix-Buchet AS. Tool for quantification of staphylococcal enterotoxin gene expression in cheese. Appl Environ Microbiol. 2010; 76(5):1367-1374.
10. Fagundes H, Barchesi L, Filho AN, Ferreira LM, Oliveira CAF. Occurrence of Staphylococcus aureus in raw milk produced in dairy farms in São Paulo state, Brazil. Brazilian J Microbiol. 2010; 41(2):376-380.
11. International Standart Office, 6888-1-ISO (2003). Microbiology of food and animal feedingstuffs – Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species).
12. Jørgensen HJ, Mørk T, Høgåsen HR, Rørvik LM. Enterotoxigenic Staphylococcus aureus in bulk milk in Norway. J Appl Microbiol. 2005a; 99(1):1589-166. Jørgensen HJ, Mørk T, Rørvik LM. The Occurrence of Staphylococcus aureus on a farm with small-scale production of raw milk cheese. J Dairy Sci. 2005b; 88(11):3810-3817.
13. Kandpal SD, Srivastava AK, Negi KS. Estimation of quality of raw milk (Open & branded) by milk adulteration testing kit. Indian J Community Heal. 2012; 24:188-192. Kearns AM, Seiders PR, Wheeler, J., Freeman, R., Steward, M. Rapid detection of methicillin-resistant staphylococci by multiplex PCR. J Hosp Infect 1999; 43(1):33-37.
14. Keyvan E, Özdemir H. Occurrence, enterotoxigenic properties and antimicrobial resistance of Staphylococcus aureus on beef carcasses. Ankara Univ Vet Fak Derg. 2016; 63(1):17-23.
15. Kluytmans J, van Belkum A, Verbrugh H. Nasal carriage of Staphylococcus aureus: epidemiology, underlying mechanisms, and associated risks. Clin Microbiol Rev. 1997; 10:505-520.
16. Korpysa-Dzirba W, Osek J. Molecular characterization of enterotoxigenic Staphylococcus aureus isolated from raw cow milk in Poland. Foodborne Pathog Dis. 2019; 16(2):114-118.
17. Kuzma K, Malinowski E, Lassa H, Klossowska A. Detection of genes for enterotoxins and toxic shock syndrome toxin-1 in Staphylococcus aureus isolated from bovine mastitis. B Vet I Pulawy. 2003; 47(2):419-426.
18. Le Loir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genet Mol Res. 2003; 2:7-28.
19. Lee SHI, Camargo CH, Gonçalves JL, Cruz AG, Sartori BT, Machado MB, Oliveira CAF. Characterization of Staphylococcus aureus isolates in milk and the milking environment from small-scale dairy farms of São Paulo, Brazil, using pulsed-field gel electrophoresis. J Dairy Sci. 2012; 95(12):7377-7383.
20. Lee SHI, Cappato LP, Corassin CH, Cruz AGD, Oliveira CAFD. Effect of peracetic acid on biofilms formed by Staphylococcus aureus and Listeria monocytogenes isolated from dairy plants. J Dairy Sci. 2016; 99(3):2384-2390.
21. Lee SHI, Mangolin BLC, Gonçalves JL, Neeff DV, Silva MP, Cruz AG, Oliveira CAF. Biofilm-producing ability of Staphylococcus aureus isolates from Brazilian dairy farms. J Dairy Sci. 2014; 97(3):1812-1816.
22. Lem P, Spiegelman J, Toye B, Ramotar K. Direct detection of mecA, nuc and 16S rRNA genes in BacT/Alert blood culture bottles. Diagn Microbiol Infect Dis. 2001; 41(3):165-168.
23. Mansour AS, Wagih GES, Morgan SD, Elhariri M, El-Shabrawy MA, Abuelnaga ASM, Elgabry EA. Detection of Staphylococcus aureus enterotoxigenic strains in bovine raw milk by reversed passive latex agglutination and multiplex polymerase chain reaction. Vet World. 2017; 10(8):843–847.
24. Mehrotra M, Wang G, Johnson WM. Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1 and methicillin resistance. J Clin Microbiol. 2000; 38(3):1032-1035.
25. Morandi S, Brasca M, Lodi R, Cremonesi P, Castiglioni B. Detection of classical enterotoxins and identification of enterotoxin genes in Staphylococcus aureus from milk and dairy products. Vet Microbiol. 2007; 124(1-2):66-72.
26. Normanno G, La Salandra G, Dambrosio A, Quaglia NC, Corrente M, Parisi A, Santagada G, Firinu A, Crisetti E, Celano GV. Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products. Int J Food Microbiol. 2007; 115(3):290-296.
27. Obaidat MM, Salman AEB, Roess AA. High prevalence and antimicrobial resistance of mecA Staphylococcus aureus in dairy cattle, sheep, and goat bulk tank milk in Jordan. Trop Anim Health Prod. 2018; 50(2):405-412.
28. Ote I, Taminiau B, Duprez JN, Dizier I, Mainil JG. Genotypic characterization by polymerase chain reaction of Staphylococcus aureus isolates associated with bovine mastitis. Vet Microbiol. 2011; 153:285-292.
29. Özdemir H, Keyvan E. Isolation and characterisation of Staphylococcus aureus strains isolated from beef, sheep and chicken meat. Ankara Univ Vet. Fak. Derg. 2016; 63:333-338.
30. Painter JA, Hoekstra RM, Ayers T, Tauxe RV, Braden CR, Angulo FJ, Griffin PM. Attribution of foodborne illnesses, hospitalizations, and deaths to food commodities by using outbreak data, United States, 1998–2008. Emerg Infect Dis. 2013; 19:407-415.
31. Parisi A, Caruso M, Normanno G, L. Latorre R, Sottili A, Miccolupo R, Fraccalvieri G. Prevalence, antimicrobial susceptibility and molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) in bulk tank milk from southern Italy. Food Microbiol. 2016; 58: 36-42.
32. Peles F, Wagner M, Varga L, Hein I, Rieck P, Gutser K, Keresztúri P, Kardos G, Turcsányi I, Béri B, Szabó A. Characterization of Staphylococcus aureus strains isolated from bovine milk in Hungary. Int J Food Microbiol. 2007; 118(2):186-193.
33. Pereira V, Lopes C, Castro A, Silva J, Gibbs P, Teixeira P. Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from various foods in Portugal. Food Microbiol. 2009; 26(3):278-282.
34. Rall VLM, Miranda ES, Castilho IG, Camargo CH, Langoni H, Guimarães FF, Araújo Júnior JP, Fernandes Júnior A. Diversity of Staphylococcus species and prevalence of enterotoxin genes isolated from milk of healthy cows and cows with subclinical mastitis. J Dairy Sci. 2014; 97(2):829-837.
35. Rall VLM, Vieira FP, Rall R, Vieitis RL, Fernandes A, Candeias JMG, Cardoso KFG, Araújo JP. PCR detection of staphylococcal enterotoxin genes in Staphylococcus aureus strains isolated from raw and pasteurized milk. Vet Microbiol. 2008; 132(3-4):408-413.
36. Riva A, Borghi E, Cirasols D, Colmegna S, Borgo F, Amato E, Pontello MM, Morace G. Methicillin-resistant Staphylococcus aureus in raw milk: Prevalence, SCC mec typing, enterotoxin characterization and antimicrobial resistance patterns. J Food Prot. 2015; 78(6):1142-1146.
37. Schmid D, Fretz R, Winter P, Mann M, Höger G, Stöger A, Ruppitsch W, Ladstätter J, Mayer N, De Martin A, Allerberger F. Outbreak of staphylococcal food intoxication after consumption of pasteurized milk products, June 2007, Austria. Wien Klin Wochenschr. 2009; 121(3-4):125-131.
38. Türkyılmaz S, Tekbıyık S, Oryasin E, Bozdogan B. Molecular epidemiology and antimicrobial resistance mechanisms of methicillin-resistant Staphylococcus aureus isolated from bovine milk. Zoonoses Public Health. 2010; 57(3):197-203.
39. Umeda K, Nakamura H, Yamamoto K, Nishina N, Yasufuku K, Hirai Y, Hirayama T, Goto K, Hase A, Ogasawara J. Molecular and epidemiological characterization of staphylococcal foodborne outbreak of Staphylococcus aureus harboring seg, sei, sem, sen, seo, and selu genes without production of classical enterotoxins. Int J Food Microbiol. 2017; 256:30-35.
40. Zouharova M, Rysanek D. Multiplex PCR and RPLA identification of Staphylococcus aureus enterotoxigenic strains from bulk tank milk. Zoonoses Public Health 2008; 55(6):313-319.