Survey on Methicillin Resistance and Panton Valentine Toxin of Staphylococcus aureus Strains Isolated from Raw Milk and Ice Cream: Molecular Study by Multiplex PCR

Yıl 2022, , 711 - 718, 31.08.2022

Erdoğan Güneş , Ahmet Uysal , Yusuf Durak

https://doi.org/10.35414/akufemubid.1115550

Cited By: 1

Öz

Staphylococcus aureus is a very important pathogenic bacterium that causes nosocomial and community-acquired infections in humans, and is also one of the leading pathogens that causes food-borne poisoning. The presence of S. aureus in raw milk and dairy products, and especially the presence of MRSA (Methicillin Resistance S. aureus) strains, poses a potential risk to public health. The aim of this study was to investigate the presence of methicillin resistance and Panton-Valentine Leucocidin (PVL) toxin in Staphylococcus aureus isolated and identified from raw milk and ice cream in Konya (Turkey) by multiplex PCR method. A total of 55 S. aureus were isolated 49 (18%) from 260 raw milk samples collected from various farms and 6 (4%) from 150 ice cream samples sold in patisseries. The obtained isolates were identified as S. aureus with conventional and genotypic methods. Multiplex PCR was performed to detect the 16S rRNA, mecA, femA and lukS genes. While no mecA gene was detected in any of the 49 S. aureus isolates obtained from raw milk samples, the presence of mecA gene was observed in one of the 6 S. aureus isolates isolated from ice cream samples. The PVL gene was not detected in any of the S. aureus isolates studied. S. aureus contamination is common in raw milk samples and ice cream samples. In order to avoid this, it is necessary to comply with the hygiene conditions and increase the precautions even more.

Anahtar Kelimeler

MRSA, Panton-Valentine Leucocidin, lukS, mecA, Dairy products

Destekleyen Kurum

Selcuk University Scientific Research Projects Coordinating Office (BAP)

Proje Numarası

14101021

Teşekkür

We would like to thank Selcuk University Scientific Research Projects Coordinating Office (BAP) for supporting this project financially (Project No: 14101021).

Çiğ Süt ve Dondurmadan İzole Edilen Staphylococcus aureus Suşlarının Metisilin Direnci ve Panton Valentine Toksini Üzerine Araştırma: Multiplex PCR ile Moleküler Çalışma

Öz

Staphylococcus aureus insanlarda hastane ve toplum kökenli enfeksiyonlara neden olan çok önemli bir patojen bakteridir ve aynı zamanda gıda kaynaklı zehirlenmelere neden olan patojenlerin başında gelmektedir. Çiğ süt ve süt ürünlerinde S. aureus ve özellikle MRSA (Metisilin Direnli S. aureus) suşlarının varlığı halk sağlığı için potansiyel bir risk oluşturmaktadır. Bu çalışmanın amacı, Multiplex PCR yöntemiyle, Konya'da (Türkiye) çiğ süt ve dondurmadan izole ve identifiye edilen Staphylococcus aureus'larda metisilin direnci ve Panton-Valentine Lökosidin (PVL) toksini varlığının araştırılmasıdır. Çeşitli çiftliklerden toplanan 260 çiğ süt örneğinden 49 (%18) ve pastanelerden toplanan 150 dondurma örneğinden 6 (%4) adet olmak üzere toplam 55 S. aureus izole edildi. Elde edilen izolatlar, konvansiyonel ve genotipik yöntemlerle S. aureus olarak tanımlandı. 16S rRNA, mecA, femA ve lukS genlerini saptamak için multipleks PCR yapıldı. Çiğ süt örneklerinden elde edilen 49 S. aureus izolatının hiçbirinde mecA geni saptanmazken, dondurma örneklerinden izole edilen 6 S. aureus izolatından birinde mecA geni varlığı belirlendi. PVL geni, çalışılan S. aureus izolatlarının hiçbirinde saptanmadı. S. aureus kontaminasyonu çiğ süt numunelerinde ve dondurma numunelerinde oldukça yaygındır. Bu kontaminasyonun önüne geçebilmek için hijyen koşullarına uyulması ve önlemlerin daha da artırılması gerekmektedir.

Anahtar Kelimeler

MRSA, Panton-Valentine Lökosidin, lukS, mecA, Süt ürünleri

Proje Numarası

14101021

Kaynakça

• Ağaoğlu, S. and Alemdar, S., 2004. Van'da tüketime sunulan dondurmalarda bazı patojenlerin varlığının araştırılması, Yüzüncü Yıl Üniversitesi Veteriner Fakültesi Dergisi, 15 (1), 59-64.
• Al-Talib, H., Yean, C. Y., Al-Khateeb, A., Hassan, H., Singh, K.-K. B., Al-Jashamy, K. and Ravichandran, M., 2009. A pentaplex PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus and Panton-Valentine Leucocidin, BMC microbiology, 9 (1), 1-8.
• Alizadeh, S. and Amini, K., 2015. Determining the presence of virulence genes panton valentine leukocidin Pvl and methicillin resistance Gene mecA in Staphylococcus aureus strains isolated from food samples by multiplex PCR and antibiotic resistance, Journal of food microbiology, 2 (1), 49-58.
• Asiimwe, B. B., Baldan, R., Trovato, A. and Cirillo, D. M., 2017. Prevalence and molecular characteristics of Staphylococcus aureus, including methicillin resistant strains, isolated from bulk can milk and raw milk products in pastoral communities of South-West Uganda, BMC Infectious Diseases, 17 (1), 1-8.
• Aydin, A., Sudagidan, M. and Muratoglu, K., 2011. Prevalence of staphylococcal enterotoxins, toxin genes and genetic-relatedness of foodborne Staphylococcus aureus strains isolated in the Marmara Region of Turkey, International journal of food microbiology, 148 (2), 99-106.
• Basanisi, M., La Bella, G., Nobili, G., Franconieri, I. and La Salandra, G., 2017. Genotyping of methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in South Italy, Food Microbiology, 62, 141-146.
• Bonsaglia, E., Silva, N., Rossi, B., Camargo, C., Dantas, S., Langoni, H., Guimarães, F., Lima, F., Fitzgerald, J. and Júnior, A. F., 2018. Molecular epidemiology of methicillin-susceptible Staphylococcus aureus (MSSA) isolated from milk of cows with subclinical mastitis, Microbial pathogenesis, 124, 130-135.
• Can, H. Y., Elmalı, M. and Karagöz, A., 2017. Molecular typing and antimicrobial susceptibility of Staphylococcus aureus strains isolated from raw milk, cheese, minced meat, and chicken meat samples, Korean journal for food science of animal resources, 37 (2), 175.
• Duman, Y., Tekerekoğlu, M. S. and Otlu, B., 2013. Toplum ve hastane kökenli Staphylococcus aureus klinik izolatlarında Panton-Valentine lökosidin varlığının ve klonal ilişkinin araştırılması, Mikrobiyol Bul, 47 (3), 389-400.
• Ede, E., 2016. İstanbul ilinde açıkta satılan dondurmaların koagülaz pozitif staphylococcus aureus yönüyle incelenmesi, Yüksek lisans tezi, İstanbul Medipol Üniversitesi Sağlık Bilimleri Enstitüsü, İstanbul, 50.
• Ektik, N., Gökmen, M. and Çibik, R., 2017. The prevalence and antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) in milk and dairy products in Balikesir, Turkey, Journal of the Hellenic Veterinary Medical Society, 68 (4), 613-620.
• Ertaş, N. and Gönülalan, Z., 2010. Kayseri ilinde satılan çiğ sütlerde Staphylococcus aureus ve enterotoksinlerinin varlığı üzerine araştırmalar, Fırat Üniversitesi Sağlık Bilimleri Veteriner Dergisi, 24 (1), 11-15.
• Fagundes, H., Barchesi, L., Nader Filho, A., Ferreira, L. M. and Oliveira, C. A. F., 2010. Occurrence of Staphylococcus aureus in raw milk produced in dairy farms in São Paulo state, Brazil, Brazilian Journal of Microbiology, 41, 376-380.
• Giacinti, G., Carfora, V., Caprioli, A., Sagrafoli, D., Marri, N., Giangolini, G., Amoruso, R., Iurescia, M., Stravino, F. and Dottarelli, S., 2017. Prevalence and characterization of methicillin-resistant Staphylococcus aureus carrying mecA or mecC and methicillin-susceptible Staphylococcus aureus in dairy sheep farms in central Italy, Journal of Dairy Science, 100 (10), 7857-7863.
• Guven, K., Mutlu, M. B., Gulbandilar, A. and Cakir, P., 2010. Occurrence and characterization of Staphylococcus aureus isolated from meat and dairy products consumed in Turkey, Journal of Food Safety, 30 (1), 196-212.
• Gücükoğlu, A., Onur Kevenk, T., Uyanik, T., Çadirci, Ö., Terzi, G. and Alişarli, M., 2012. Detection of enterotoxigenic Staphylococcus aureus in raw milk and dairy products by multiplex PCR, Journal of food science, 77 (11), M620-M623.
• Ichiyama, S., Ohta, M., Shimokata, K., Kato, N. and Takeuchi, J., 1991. Genomic DNA fingerprinting by pulsed-field gel electrophoresis as an epidemiological marker for study of nosocomial infections caused by methicillin-resistant Staphylococcus aureus, Journal of clinical microbiology, 29 (12), 2690-2695.
• Issa, G. and Aksu, H., 2020. Detection of Methicillin-Resistant Staphylococcus aureus in Milk by PCR-Based Phenotyping and Genotyping, Acta Veterinaria Eurasia, 46 (3), 120-125.
• Keskin, Y., Başkaya, R., Özyaral, O. and Kıyan, P., 2007. Sade dondurmaların mikrobiyolojik incelenmesi, Türk Mikrobiyoloji Cemiyeti Dergisi, 37 (1), 51-58.
• Keyvan, E., Yurdakul, O., Demirtas, A., Yalcin, H. and Bilgen, N., 2020. Identification of methicillin-resistant Staphylococcus aureus in bulk tank milk, Food Science and Technology, 40, 150-156.
• Korpysa-Dzirba, W. and Osek, J., 2011. Identification of genes encoding classical staphylococcal enterotoxins in Staphylococcus aureus isolated from raw milk, Bulletin of the Veterinary Institute in Puławy, 1 (55).
• Kramer, J., Gilbert, R. and Doyle, M., 1989, Foodborne bacterial pathogens, by MP Doyle, Marcel Dekker, Inc., New York and Basel, 22-70.
• Löffler, B., Hussain, M., Grundmeier, M., Brück, M., Holzinger, D., Varga, G., Roth, J., Kahl, B. C., Proctor, R. A. and Peters, G., 2010. Staphylococcus aureus panton-valentine leukocidin is a very potent cytotoxic factor for human neutrophils, PLoS pathogens, 6 (1), e1000715.
• Martins, K. B., Faccioli-Martins, P. Y., Riboli, D. F. M., Pereira, V. C., Fernandes, S., Oliveira, A. A., Dantas, A., Zafalon, L. F. and Cunha, M. d. L. R. d. S. d., 2015. Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk, Brazilian Journal of Microbiology, 46, 535-543.
• McKay, A. M., 2008. Antimicrobial resistance and heat sensitivity of oxacillin-resistant, mecA-positive Staphylococcus spp. from unpasteurized milk, Journal of Food Protection, 71 (1), 186-190.
• Nazari, R., Godarzi, H., Baghi, F. R. and Moeinrad, M., 2014. Enterotoxin gene profiles among Staphylococcus aureus isolated from raw milk, Iranian journal of veterinary research, 15 (4), 409.
• Nhatsave, N., Garrine, M., Messa, A., Massinga, A. J., Cossa, A., Vaz, R., Ombi, A., Zimba, T. F., Alfredo, H. and Mandomando, I., 2021. Molecular Characterization of Staphylococcus aureus Isolated from Raw Milk Samples of Dairy Cows in Manhiça District, Southern Mozambique, Microorganisms, 9 (8), 1684.
• Paterson, G., Larsen, J., Harrison, E. M., Larsen, A. R., Morgan, F., Peacock, S. J., Parkhill, J., Zadoks, R. and Holmes, M. A., 2012. First detection of livestock-associated meticillin-resistant Staphylococcus aureus CC398 in bulk tank milk in the United Kingdom, January to July 2012, Eurosurveillance, 17 (50), 20337.
• Riva, A., Borghi, E., Cirasola, D., Colmegna, S., Borgo, F., Amato, E., Pontello, M. M. and Morace, G., 2015. Methicillin-resistant Staphylococcus aureus in raw milk: Prevalence, SCC mec typing, enterotoxin characterization, and antimicrobial resistance patterns, Journal of Food Protection, 78 (6), 1142-1146.
• Samir, H., Younis, W., Sultan, S. and Abd El-Azeem, M., 2018. Isolation of Staphylococcus aureus from ice-cream samples, J Vet Ani Res, 1, 204.
• Sharma, V., Sharma, S., Dahiya, D. K., Khan, A., Mathur, M. and Sharma, A., 2017. Coagulase gene polymorphism, enterotoxigenecity, biofilm production, and antibiotic resistance in Staphylococcus aureus isolated from bovine raw milk in North West India, Annals of clinical microbiology and antimicrobials, 16 (1), 1-14.
• Titouche, Y., Hakem, A., Houali, K., Meheut, T., Vingadassalon, N., Ruiz-Ripa, L., Salmi, D., Chergui, A., Chenouf, N. and Hennekinne, J., 2019. Emergence of methicillin-resistant Staphylococcus aureus (MRSA) ST8 in raw milk and traditional dairy products in the Tizi Ouzou area of Algeria, Journal of Dairy Science, 102 (8), 6876-6884.
• Visciano, P., Pomilio, F., Tofalo, R., Sacchini, L., Saletti, M. A., Tieri, E., Schirone, M. and Suzzi, G., 2014. Detection of methicillin-resistant Staphylococcus aureus in dairy cow farms, Food Control, 46, 532-538.
• Wang, W., Lin, X., Jiang, T., Peng, Z., Xu, J., Yi, L., Li, F., Fanning, S. and Baloch, Z., 2018, Prevalence and characterization of Staphylococcus aureus cultured from raw milk taken from dairy cows with mastitis in Beijing, China, Frontiers in Microbiology, 9, 1123.
• Yildirim, T., Sadati, F., Kocaman, B. and Siriken, B., 2019. Staphylococcus aureus and Staphylococcal enterotoxin detection in raw milk and cheese origin coagulase positive isolates, International Journal of Science Letters, 1 (1), 30-41.
• Yücel, N. and Anıl, Y., 2011. Çiğ süt ve peynir örneklerinden Staphylococcus aureus ve koagülaz negatif stafilokokların identifikasyonu ve antibiyotik duyarlılığı, Türk Hijyen ve Deneysel Biyoloji Dergisi, 68 (2), 73-78.
• Zecconi, A. and Hahn, G., 1999, Staphylococcus aureus in raw milk and human health risk, International Dairy Federation, 345, 15-18
• Zhang, P., Liu, X., Zhang, M., Kou, M., Chang, G., Wan, Y., Xu, X., Ruan, F., Wang, Y. and Wang, X., 2021. Prevalence, Antimicrobial Resistance, and Molecular Characteristics of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus from Retail Ice Cream in Shaanxi Province, China, Foodborne Pathogens and Disease, 19 (3), 217-225.