Methicillin Resistance Coagulase Positive Staphylococci and Staphylococcus aureus Isolated from Raw Milk and Cheese, Samsun Province-TURKEY

Today, MRSA is among the most important causes of antimicrobial-resistant health care-associated infections worldwide. The aim of the present study, 100 coagulase positive staphylococci (CPS) isolates recovered from raw cow milk and different kind of cheese samples, collected from different part of Samsun province, Turkey, using classic culture technique were determined for Staphylococcus aureus (S. aureus) and methicillin-resistant (MR) properties. For S. aureus determination of the isolates, nuc gene (S. aureus specific gene) was detected, and to determination of methicillin resistance properties of the isolates, mecA gene was determined in the isolates using PCR assay technique. As a result; nuc gene was detected in 72 (72%) (44 of which milk and 28 of which cheese origin) out of 100 CPS. In the 100 CPS isolates, mecA gene was detected 15 (15%, n=15; 4 milk and 11 cheese origin) CPS isolates, and the isolates were evaluated as MRCPS.  The number of both nuc and mecA present isolates were 5 (4 milk and 1 cheese origin). These 5 isolates were evaluated as MRSA (5%). The remaining 10 (%10) containing mecA isolates were evaluated as MRCPS. In conclusion, raw cow milk and cheese samples have a potential health risk for mainly S. aureus and then coagulase positive staphylococci. In addition to this, analyzed samples also contaminated with MRSA or MRCPS isolates, become a big challenge for human health. Günümüzde MRSA, dünya genelinde antimikrobiyel dirençlilik ile ilgili sağlık problemlerinin en önemli nedenleri arasında yer almaktadır. Bu çalışmada, çiğ inek sütü (n=50) ve çeşitli tip peynirlerden (n=50) klasik kültür tekniği ile izole edilen toplam 100 koagulaz pozitif staphylococcus (KPS) izolatlarında S. aureus’un belirlenmesi ve metisiline dirençlilik (MR) özelliklerinin moleküler yöntemle saptanması amacıyla analiz edildi.  S.aureus’un saptanması için nuc, metisiline dirençlilik özelliklerini saptanması ise mecA gen varlıkları PZR tekniği kullanılarak saptandı. Sonuç olarak, nuc geni toplam 100 KPS izolatının 72 (%72)’isinde saptandı ve bu izolatlar S.aureus (%72, n=72; 44’ü süt orjinli, 28’i ise peynir orjinli) olarak değerlendirildi.100 KPS izolatında mecA geni ise 15 izolatta (%15, n=15; 4 süt ve 11 peynir orjinli) saptandı. mecA geni saptanan bu 15 izolat MRKPS olarak belirlendi. Çalışmada nuc ve mecA genlerinin her ikisinin de bulunduğu izolat sayısı ise 5 (4 süt, 1 peynir orjinli) olup, bu izolatlar MRSA (%5) olarak değerlendirilirken, geriye kalan 10 izolat ise MRKPS (%10) olarak değerlendirildi. Bu çalışma bulguları çerçevesinde, Samsun ilinde tüketime sunulan peynirlerin ve çiğ inek sütlerinin başta S. aureus olmak üzere koagulaz pozitif stafilokoklarla kontamine olduğu, ayrıca analiz edilen örneklerin halk sağlığı yönünden çok önemli bir tehdit olan MRSA ve MRCPS ile de kontamine olduğu saptandı.

Methicillin Resistance Coagulase Positive Staphylococci and Staphylococcus aureus Isolated from Raw Milk and Cheese, Samsun Province-TURKEY

Today, MRSA is among the most important causes of antimicrobial-resistant health care-associated infections worldwide. The aim of the present study, 100 coagulase positive staphylococci (CPS) isolates recovered from raw cow milk and different kind of cheese samples, collected from different part of Samsun province, Turkey, using classic culture technique were determined for Staphylococcus aureus (S. aureus) and methicillin-resistant (MR) properties. For S. aureus determination of the isolates, nuc gene (S. aureus specific gene) was detected, and to determination of methicillin resistance properties of the isolates, mecA gene was determined in the isolates using PCR assay technique. As a result; nuc gene was detected in 72 (72%) (44 of which milk and 28 of which cheese origin) out of 100 CPS. In the 100 CPS isolates, mecA gene was detected 15 (15%, n=15; 4 milk and 11 cheese origin) CPS isolates, and the isolates were evaluated as MRCPS.  The number of both nuc and mecA present isolates were 5 (4 milk and 1 cheese origin). These 5 isolates were evaluated as MRSA (5%). The remaining 10 (%10) containing mecA isolates were evaluated as MRCPS. In conclusion, raw cow milk and cheese samples have a potential health risk for mainly S. aureus and then coagulase positive staphylococci. In addition to this, analyzed samples also contaminated with MRSA or MRCPS isolates, become a big challenge for human health.
Keywords:

milk, cheese, Coagulase,

___

  • Abdou, A.M., Ibrahım, S.K., Mohamed, E. and Tomohiro, T., 2016. Prevalence, molecular characterization, and antimicrobial susceptibility of Methicillin-Resistant Staphylococcus aureus isolated from milk and dairy products. Foodborne Pathogens and Disease, 13 (3): 156-162.
  • Aghazadeh, M., Ghotaslou, R., Rezaee, M.A., Moshafi, M.H., Hojabri, Z. and Saffari, F., 2015. Determination of antimicrobial resistance profile and inducible clindamycin resistance of coagulase negative staphylococci in pediatric patients: the first report from Iran. World Journal of Pediatrics, 11(3): 250-254.
  • André, M.C.D.P.B., Campos, M.R.H., Borges, L.J., Kipnis, A., Pimenta, F.C. and Serafini, Á.B.C., 2008. Comparison of Staphylococcus aureus isolates from food handlers, raw bovine milk and Minas Frescal cheese by antibiogram and pulsed-field gel electrophoresis following SmaI digestion. Food Control, 19: 200–207.
  • Anonymous, 2002. Use of Antimicrobials Outside Human Medicine and Resultant Antimicrobial Resistance in Humans (Fact Sheet No. 268). Geneva, Switzerland.
  • Anonymous, 2009. European Food Safety Authority (EFSA). Analysis of the baseline survey on the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in holdings withbreeding pigs in the EU, Part A: MRSA prevalence estimates. EFSA Journal, 7:1376.
  • Anonymous, 2015. Eurosurveillance Editorial Team. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2013. EFSA Journal, 13:3991.
  • Anonymous, 2017. EU summary report on zoonoses, zoonotic agents and food-borne outbreaks 2016. EFSA Journal 2017, 15 (12):5077.
  • Anonymous, 2017a. Global consumption per capita. Canadian Dairy Information Centre. http://www.dairyinfo.gc.ca/index_e. php?s1=dff-fcil&s2= cons&s3= consglo&s4=tc-ft. Accesed on May 24 2018.
  • Antunes, P., Machado, J. and Peixe, L., 2006. Characterization of antimicrobial resistanceand class 1and 2 integron in Salmonella enteric isolates from different sources in Portugal. Journal of Antimicrobial Chemotheraphy, 58: 297-304.
  • Aras, Z., Aydin, I. and Kav, K., 2012. Isolation of methicillin-resistant Staphylococcus aureus from caprinemastitis cases. Small Ruminant Research, 102: 68-73.
  • Argudín, M.A., Tenhagen, B.A., Fetsch, A., Sachsenröder, J., Käsbohrer, A., Schroeter, J., Hammerl, A., Hertwig, S., Helmuth, R., Bräning, J., Mendoza, M.C., Appel, B., Rodicio, M.R. and Guerra, B., 2011. Virulence and resistance determinants of German Staphylococcus aureus ST398 isolates from nonhuman sources. Applied and Environmental Microbiology, 77(9): 3052–3060.
  • Asao, T., Kumeda, Y., Kawa, T., Shibata, T., Oda, H., Haruk, K., Nakazawa, H. and Kozak, S., 2003. An extensive outbreak of staphylococcal food poisoning due to low-fat milk in Japan:estimation of enterotoxin A in the incriminated milk and powdered skim milk. Epidemiology and Infection, 130: 33–40.
  • Bardiau, M., Yamazaki, K., Duprez, J-N., Taminiau, B., Mainil, J.G. and Ote, I., 2013. Genotypic and phenotypic characterization of methicillin resistant Staphylococcus aureus (MRSA) isolated from milk of bovine mastitis. Letters in Applied Microbiology, 57:181-186.
  • Basanisi, landra, M.G., La Bella, G., Nobili, G., Franconieri, I. and La Salandra, G., 2017. Genotyping ofmethicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in South Italy. Food Microbiology, 62:141-146.
  • Bennett, R.W., Lancette, G.A., 2001. Staphylococcus aureus. Chap 12. In Bacteriological analytical manual. Available at: http.//www.fda.gov/Food/oodScienceResearch/Laboratory Methods/ ucm071429.htm. Accessed 15 November 2016.
  • Beuvier, E. and Buchin, S., 2004. Raw milk cheeses. 319–345. In: Fox PF, McSweene PLH, Cogan, TM, Guinee TP (Eds.) Cheese: chemistry, physics and microbiology. Elsevier Academic Press, Amsterdam.
  • Can, H.Y. and Çelik, T.H., 2012. Detection of enterotoxigenic and antimicrobial resistant S. aureus in Turkish cheeses. Food Control, 24:100-103.
  • Chambers, H.F., 1997. Methicillin resistance in staphylococci: molecular and biochemical basis and clinical implications. Clinical Microbiology Review, 10: 781-91.
  • Cho, J., Joo, I., Choi, S.I., Jung, K.H., Choi, E., Son, N.R., Han, M.K., Jeong, S.J., Lee, H., Hwang, I.G., 2014. Distribution of methicillin-resistant Staphylococcus aureus (MRSA) in raw meat and fish samples in Korea. Food Science Biotechnology, 23:999-1003.
  • Choi, K-H., Lee, H., Lee S., Kim, S. and Yoon, Y., 2016. Cheese Microbial Risk Assessments- A Review. Asian-Australasian Journal of Animal Sciences, 29(3):307-314.
  • David, M. Z. and Daum, R.S., 2010. Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. Clinical Microbial Review, 23: 616–687.
  • Denayer, S., Delbrassinne, L., Nia Y., Botteldoorn, N., 2017. Food-Borne Outbreak Investigation and Molecular Typing: High Diversity of Staphylococcus aureus Strains and Importance of Toxin Detection. Toxins, 9: 407.
  • Doyle, M.E., Hartmann, F.A. and Lee Wong, A.C., 2012. Methicillin-resistant staphylococci: implications for our food supply? Animal Health Research Review, 13(2):157-80.
  • Enright, M. C., 2003. The evolution of a resistant pathogen-The case of MRSA. Current Opinion in Pharmacology, 3:474–479.
  • Ercoli, L., Gallina, S., Nia, Y., Auvray, F., Primavilla, S., Guidi, F., Pierucci, B., Graziotti, C., Decastelli, L. and Scuota, S., 2017. Investigation of a Staphylococcal Food Poisoning Outbreak from a Chantilly Cream Dessert, in Umbria (Italy). Foodborne Pathogens and Diseases, 14: 407-413.
  • Erol İ., 2007. Gıda Hijyeni ve Kontrolü. Pozitif Matbaacılık Ltd. Şti., Yenimahalle, Ankara. 135-144.
  • Gorbach, S.L. and Goldin, B.R., 1992. Nutriton and gastrointestinal microflora. Nutrition Reviews, 50(12):378-81.
  • Grundmann, H., Aires-de-Sousa, M., Boyce, J. and Tiemersma, E., 2006. Emergence and resurgence of meticillin-resistant Staphylococcus aureus as a public-health threat. Lancet, 368(9538):874-85.
  • Haran, K.P., Godden, S.M., Boxrud, D., Jaeahir, S., Bender, J.B. and Sreevatsan, S., 2012. Prevalence and Characterization of Staphylococcus aureus, Including Methicillin-Resistant Staphylococcus aureus, Isolated from Bulk Tank Milk from Minnesota Dairy Farms. Journal of Clincal Microbiology, 50 (3): 688-695.
  • Hummerjohann, J., Naskova, J., Baumgartner, A. and Graber, H.U., 2014. Enterotoxin-producing Staphylococcus aureus genotype B as a major contaminant in Swiss raw milk cheese. Journal of Dairy Science, 97(3):1305-12.
  • Jay, J., Loessner, M.J. and Golden, D.A., 2005. Staphylococal Gastroenteritis. Seven Edition. Springer Science+Business Media Inc. In: Modern Food Microbiology (Chapter 23).
  • Jevons, M.P., Coe, A.W. and Parker, M.T., 1963. Methicillin resistance in staphylococci. Lancet, 7287:904–907.
  • Johler, S., Weder, D., Bridy, C., Huguenin ,M-C., Robert, L., Hummerjohann, J. and Stephan, R., 2015. Outbreak of staphylococcal food poisoning among children and staff at a Swiss boarding school due to soft cheese made from raw milk. Journal of Food Science, 98 (5): 2944-2948.
  • Kamal, R.M., Bayoumi, M.A. and Abd El, S.F.A., 2013. MRSA detection in raw milk, some dairy products and hands of dairy workers in Egypt, a mini-survey. Food Control, 33:49–53.
  • Louie, L., Goodfellow,J., Mathieu,A., Glatt,M., Louie, M. and Simor, A.E., 2002. Rapid detection of methicillin-resistant staphylococci from blood culture bottles by using a multiplex PCR assay. Journal Clinical Microbiology, 40: 2786-2790.
  • Ma, X. X., T. Ito, C., Tiensasitorn, M., Jamklang, P., Chongtrakool, S., Boyle-Vavra, R., Daum, R.S. and Hiramatsu, K., 2002. Novel type of staphylococcal cassette chromosome mec identified in community acquired methicillin-resistant Staphylococcus aureus strains. Antimicrobial Agents and Chemotherapy, 46:1147–1152.
  • McClure, J.A, Conly, J.M., Lau, V., Elsayed, S., Louie, T., Hutchins, W. and Zhang, K., 2006. Novel Multiplex PCR Assay for Detection of the Staphylococcal Virulence Marker Panton-Valentine Leukocidin Genes and Simultaneous Discrimination of Methicillin-Susceptible from- Resistant Staphylococci. Journal of Clinical Microbiology, 44: 1141-1144.
  • Normanno, G., Corrente, M., La Salandra, G., Dambrosio, A., Quaglia, N.C., Parisi, A., Greco, G., Bellacicco, A.L., Virgilio, S. and Celano, G.V., 2007. Methicillin-resistant Staphylococcus aureus (MRSA) in foods of animal origin product in Italy. International Journal of Food Microbiology, 117 (2): 219-222.
  • Rabello, R.F., Moreira BM., Lopes RM., Teixeira, L.M., Riley LW. and Castro, A.C., 2007. Multilocus sequence typing of Staphylococcus aureus isolates recovered from cows withmastitis in Brazilian dairy herds. Journal of Medical Microbiology, 56 (1): 1505–1511.
  • Riva, A., Borghi, E., Cirasola, D., Colmegna,S., Borgo,F., Amato,E., Pontello, M.M. and Morace, B., 2015. Methicilli-Resistant Staphylococcus aureus in in Raw Milk: Prevalence, SCCmec Typing, Enterotoxin Characterization, and Antimicrobial Resistance Patterns. Journal of Food Protection, 78 (6): 1142-1146.
  • Rodríguez-Lázaro, D., Oniciuc, E-A., García, P.G., Fernández-Natal, I., Dominquez-Gil, M., Eiros-Bouza, J.M., Wagner, M., Nicolau, A. and Hernández, M., 2017. Detection and Characterization of Staphylococcus aureus and Methicillin-Resistant S.aureus in Foods Confiscated in EU Borders. Front Microbiology, 8:1344.
  • Rossolini, G.M., Arena, F., Pecile, P. and Pollini, S., 2014. Update on the antibiotic resistance crisis. Current Opinion Pharmacology,18: 56-60.
  • Scallan, E., Hoekstra, R.M., Angulo, F.J., Tauxe, R.V., Widdowson, M.A., Roy, S.L., Jones, J.L. and Griffin, P.M., 2011. Foodborne illness acquired in the United States-Major pathogens. Emerging Infectious Diseases, 17: 7-15.
  • Schmid, D., Fretz, R., Winter, P., Mann, M., Höger, G., Stöger, A., Ruppitsch, W., Ladstätter, J., Mayer, N., Martin, A. and Allerberger, F., 2009. Outbreak of staphylococca food intoxication after consumption of pasteurized milk products, June 2007, Austria. Wiener Klinische Wochenschrift, 121(3-4): 125-131.
  • Spohr, M., Rau, J., Friedrich, A.W. and Tenhagen, B.A., 2011. Methicillin-Resistant Staphylococcus aureus (MRSA) in Three Dairy Herds in Southwest Germany. Zoonoses and Public Health, 58 (4): 252-61.
  • Sorum, H. and L’Abee-Lund, T. M., 2000. Antibiotic resistance in food-related bacteria-A result of interfering with global web of bacterial genetics. International Journal of Food Microbiology, 78: 43-56.
  • Tong, S.Y., Davis, J.S., Eichenberger, E., Holland, T.L. and Fowler, V.G. Jr., 2015. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clinical Microbiological Review, 28 (3): 603-661.
  • Zhang, K., Sarling, J., Chow, B.L., Elsayed, S., Hussain, Z., Church, D.L., Gregson, D.B., Louie, T. and Conly, J.M., 2004. New quadriplex PCR assay for detection of methicillin resistance and simultaneous discrimination of S.aureus from coagulase-negative staphylococci. Journal of Clinical Microbiology, 2: 4947–4955.
  • Zinke, C., Winter, M., Mohr, E. and Krömker, V., 2012. Occurrence of Methicillin-Resistant Staphylococcus aureus in Cheese Produced in German Farm-Dairies. Advances in Microbiology, 2: 629-633.