Phenotypic and Genotypic Antibiotic Resistance of Staphylococcus warneri and Staphylococcus pasteuri Isolated from Stuffed Mussels

Abstract

This study aimed to assess the presence of Staphylococcus species in stuffed mussel samples sold in Çanakkale province and to determine the antibiotic resistance of the isolates. A total of 246 stuffed mussel samples were examined, and two different Staphylococcus (S. warneri and S. pasteuri) were isolated from 12.19% of the samples. Among the Staphylococcus isolates, S. warneri was detected at a rate of 73.33% and S. pasteuri at 26.67%. Bacterial isolates (N=30) were examined for their resistance to amoxicillin/clavulanic acid (30 µg), ampicillin (10 μg), erythromycin (15 µg), clindamycin (10 µg), vancomycin (30 µg), oxacillin (5 µg), tetracycline (30 µg), doxycycline hydrochloride (30 µg), chloramphenicol (30 µg), streptomycin (10 µg), gentamicin (10 µg), kanamycin (30 µg), nalidixic acid (30 µg), ciprofloxacin (5 µg), meropenem (10 µg), imipenem (10 µg), sulfamethox/ trimethoprim (25 µg), cefotaxime (30 µg), cephalothin (30 µg), ceftriaxone (30 µg), and levofloxacin (5 µg) antibiotics. Resistance to amoxicillin/clavulanic acid and erythromycin was found in all S. warneri isolates. In addition, all S. pasteuri isolates were found to be resistant to amoxicillin/ clavulanic acid. In S. warneri isolates, at least three resistance genes (BlaTEM, tetB-6, tetK-8) and up to eight resistance genes (BlaTEM, tetB-6, tetK-8, strA-strB, aphAI-IAB, ermC) were identified. All S. pasteuri isolates exhibited blaTEM, strA-strB, and aphAI-IAB resistance genes. In conclusion, it was determined that antibiotic-resistant S. warneri and/or S. pasteuri contaminates stuffed mussel samples. This study will serve as a valuable resource for enhancing monitoring strategies in stuffed mussel production. Further studies should be conducted to determine whether the products are suitable for food safety. Therefore, it is necessary to clarify the disease effects and mechanisms of the pathogens identified in stuffed mussels.

Keywords

• Stuffed mussel
• Mytilus galloprovincialis
• S. warneri
• S. pasteuri
• Public health
• antibiotic resistance

References

1. Bauer, A. W., Kirby, W. M. M., Sherris, J. C., & Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45(4_ts), 493-496. https://doi. org/10.1093/ajcp/45.4_ts.493 google scholar
2. Becker, K., Heilmann, C., & Peters, G. (2014). Coagulase-negative staphylococci. Clinical Microbiology Reviews, 27(4), 870-926. https:// doi.org/10.1128/CMR.00109-13 google scholar
3. Bhargava, K., & Zhang, Y. (2014). Characterization of methicillin-resistant coagulase-negative staphylococci (MRCoNS) in retail meat. Food Microbiology, 42, 56-60. https://doi.org/10.1016/j.fm.2014.02.019 google scholar
4. Bingol, E. B., Colak, H., Hampikyan, H., & Muratoglu, K. (2008). The microbiological quality of stuffed mussels (Midye Dolma) sold in Istanbul. British Food Journal, 110, 1079-1087. https://doi. org/10.1108/00070700810917992 google scholar
5. Çavdar, G., Köroğlu, M., Ölmez, M., Şahin, E. Ö., & Altindiş, M. (2022). Hastane İçme ve Kullanma Sularının Mikrobiyolojik Analizi; Sakarya. Online Türk Sağlık Bilimleri Dergisi, 7(3), 346-355. google scholar
6. Chesneau, O., Morvan, A., Grimont, F., Labischinski, H., & El Solh, N. (1993). Staphylococcus pasteuri sp. nov., isolated from human, animal, and food specimens. International Journal of Systematic and Evolutionary Microbiology, 43(2), 237-244. https://doi.org/10.1099/00207713-43-2-237 google scholar
7. CLSI (2015). Clinical and Laboratory Standards Institute (CLSI). Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard—Tenth Edition. CLSI document M07-A10 (ISBN 1-56238-987-4 [Print]; ISBN 1-56238-988-2 [Electronic]). Clinical and Laboratory Standards Institute, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087 USA, 2015. google scholar
8. CLSI (2017). Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing. 27th ed. CLSI supplement M100 (ISBN 1-56238-804-5 [Print]; ISBN 1-56238-805-3 [Electronic]). Clinical and Laboratory Standards Institute, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087 USA, 2017. google scholar

9. Çoban, E. P., & Yaman, F. (2023). Biofilm Formation and Prevention of Bacteria Isolated from Fish and Fish Stalls. Acta Aquatica Turcica, 19(1), 21-34. https://doi.org/10.22392/actaquatr.1116548 google scholar Demir, Ş., Akdağ, G., Sormaz, Ü. & Özata, E. (2018). Sokak lezzetlerinin gastronomik değeri: İstanbul sokak lezzetleri. Güncel Turizm Araştırmaları Dergisi, 2(Ek.1), 589-601. google scholar
10. Diler, Ö., Yilmaz, H. E., Çağatay, İ. T., Naziroğlu, M., Öznur, Ö. Z. İ. L., & Şeydanur, K. A. N. (2023). Gökkuşağı Alabalıklarında (Oncorhynchus mykiss) Staphylococcus warneri’nin Teşhis ve Histopatolojisi. Acta Aquatica Turcica, 19(3), 277-288. https://doi.org/10.26453/otjhs.1006312 google scholar
11. Dimitriadi, D., Charitidou, C., & Charvalos, E. (2014). Urinary tract infection due to beta-lactams-resistant Staphylococcus warneri: a case report. The International Arabic Journal of Antimicrobial Agents, 4(4). google scholar
12. Durgun, S. (2013). İzmir’de açıkta satılan midye dolmaların mikrobiyolojik açıdan incelenmesi. Celal Bayar Üniversitesi / Fen Bilimleri Enstitüsü / Gıda Mühendisliği Anabilim Dalı, Yüksek Lisans Tezi, Manisa. google scholar
13. FAO, 1997. Food and Agriculture Organization of the United Nations. Street foods. FAO food and nutrition paper 63. Report of an FAO technical meeting on street foods, Calcutta, India, 6 to 9 November 1995. Food and Agriculture Organization of the United Nations, Rome. google scholar
14. Faria, C., Vaz-Moreira, I., Serapicos, E., Nunes, O. C., & Manaia, C. M. (2009). Antibiotic resistance in coagulase negative staphylococci isolated from wastewater and drinking water. Science of the Total Environment, 407(12), 3876-3882. https://doi.org/10.1016/j.scitotenv.2009.02.034 google scholar
15. Gelman, R., Potruch, A., Oster, Y., Ishay, Y., Gur, C., Beeri, R., & Strahilevitz, J. (2022). Native aortic valve Staphylococcus warneri endocarditis after COVID-19 infection: a case report and a review of literature. Apmis, 130(5), 270-275. google scholar
16. Geniş, B., & Tuncer, Y. (2018). Determination of antibiotic susceptibility and decarboxylase activity of coagulase-negative Staphylococcus and Macrococcus caseolyticus strains isolated from fermented Turkish sausage (sucuk). Journal of Food Processing and Preservation, 42(1), e13329. https://doi.org/10.1111/jfpp.13329 google scholar
17. Güngörür, N.M. (2019). İstanbul’da satışa sunulan midye dolmaların mikrobiyolojik kalitesinin incelenmesi. İstanbul Aydın Üniversitesi / Fen Bilimleri Enstitüsü / Gıda Güvenliği ve Beslenme Anabilim Yazım alanları gerektiği kadar uzatılabilir Dalı, Yüksek Lisans Tezi, İstanbul. google scholar
18. Hoque, M. N., Faisal, G. M., Das, Z. C., Sakif, T. I., Al Mahtab, M., Hossain, M. A., & Islam, T. (2023). Genomic Features and Pathophysiological Impact of a Multidrug-Resistant Staphylococcus warneri Variant in Murine Mastitis. Microbes and Infection, 105285. google scholar
19. Hoque, M. N., Moyna, Z., Faisal, G. M., & Das, Z. C. (2023). Whole-Genome Sequence of the Multidrug-Resistant Staphylococcus warneri Strain G1M1F, Isolated from Mice with Mastitis. Microbiology Resource Announcements, 12(5), e00275-23. google scholar
20. Incani, R. N., Hernândez, M., Cortez, J., Gonzâlez, M. E., & Dorel Salazar, Y. (2010). Staphylococcus warneri meningitis in a patient with Strongyloides stercoralis hyperinfection and lymphoma: first report of a case. Revista do Instituto de Medicina Tropical de Sâo Paulo, 52, 169-170. google scholar
21. Jeong, D. W., & Lee, J. H. (2015). Safety assessment of coagulase-negative staphylococci from jeotgal, a Korean high-salt-fermented seafood. Microbiology and Biotechnology Letters, 43(1), 84-90. https://doi.org/10.4014/mbl.1501.01001 google scholar
22. Karademir, F. (2018). İstanbul’da tüketime sunulan midye dolmaların mikrobiyolojik kalitesinin belirlenmesi. İstanbul Üniversitesi / Sağlık Bilimleri Enstitüsü / Besin Hijyeni ve Teknolojisi Anabilim Dalı, Yüksek Lisans Tezi, İstanbul. google scholar
23. Kim, T. Y., Lee, J. J., Kim, B. S., & Choi, S. H. (2017). Whole-body microbiota of sea cucumber (Apostichopus japonicus) from South Korea for improved seafood management. Journal of Microbiology and Biotechnology, 27(10), 1753-1762. https://doi.org/10.4014/jmb.1707.07067 google scholar
24. Kişla, D., & Üzgün, Y. (2008). Microbiological evaluation of stuffed mussels. Journal of Food Protection, 71(3), 616-620. https://doi. org/10.4315/0362-028X-71.3.616 google scholar
25. Lane, DJ. (1991). 16S/23S rRNA sequencing. In: Stackebrant E., Goodfellow M. (eds.) Nucleic acid techniques in bacterial systematics. John Wiley & Sons Ltd. London, UK, pp 115-75. google scholar
26. Louail, R., Florin, F., Bernard, S., Michaud, J. B., Breton, J., Achamrah, N., ... & Ribet, D. (2023). Invasion of intestinal cells by Staphylococcus warneri, a member of the human gut microbiota. Gut Pathogens, 15(1), 4. google scholar
27. Lucca, A., & da Silva Torres, E. A. F. (2006). Street-food: the hygiene conditions of hot-dogs sold in Sâo Paulo, Brazil. Food Control, 17(4), 312-316. https://doi.org/10.1016/j.foodcont.2004.11.005 google scholar
28. Marino, M., Frigo, F., Bartolomeoli, I., & Maifreni, M. (2011). Safety-related properties of staphylococci isolated from food and food environments. Journal of Applied Microbiology, 110(2), 550-561. https://doi.org/10.1111/j.1365-2672.2010.04909.x google scholar
29. Mepba, H. D., Achinewhu, S. C., Aso, S. N., & Wachukwu, C. K. (2007). Microbiological quality of selected street foods in Port Harcourt, Nigeria. Journal of Food Safety, 27(2), 208-218. https://doi. org/10.1111/j.1745-4565.2007.00073.x google scholar
30. Morfin-Otero, R., Martınez-Vâzquez, M. A., Lopez, D., Rodrîguez-Noriega, E., & Garza-Gonzâlez, E. (2012). Isolation of rare coagulase-negative isolates in immunocompromised patients: Staphylococcus gallinarum, Staphylococcus pettenkoferi and Staphylococcus pasteuri. Annals of Clinical & Laboratory Science, 42(2), 182-185. google scholar
31. Mosupye, F. & Von Holy, A. (1999). Microbiological quality and safety of ready to eat street-vended foods in Johannesburg, South Africa. Journal of Food Protection, 62, 1278-1284. https://doi.org/10.4315/0362-028x-62.11.1278 google scholar
32. Petti, C. A., Simmon, K. E., Miro, J. M., Hoen, B., Marco, F., & Chu, V. H. (2008). International collaboration on endocarditis-microbiology investigators genotypic diversity of coagulase-negative staphylococci causing endocarditis: a global perspective. J Clin Microbiol, 46(5), 1780-1784. google scholar
33. Ramnarain, J., Yoon, J., & Runnegar, N. (2019). Staphylococcus pasteuri infective endocarditis: A case report. IDCases, 18, e00656. google scholar
34. Rantsiou, K., Iacumin, L., Cantoni, C., Comi, G., & Cocolin, L. (2005). Ecology and characterization by molecular methods of Staphylococcus species isolated from fresh sausages. International Journal of Food Microbiology, 97(3), 277-284. https://doi. org/10.1016/j.ijfoodmicro.2004.02.018 google scholar
35. Regecovâ, I., Pipovâ, M., Jevinovâ, P., Maruskovâ, K., Kmet', V., & Popelka, P. (2014). Species identification and antimicrobial resistance of coagulase-negative staphylococci isolated from the meat of sea fish. Journal of Food Science, 79(5), M898-M902. https://doi. org/10.1111/1750-3841.12429 google scholar
36. Sânchez, E., Donat, E., Ribes-Koninckx, C., Fernândez-Murga, M. L., & Sanz, Y. (2013). Duodenal-mucosal bacteria associated with celiac disease in children. Applied and environmental microbiology, 79(18), 5472-5479. google scholar
37. Savini, V., Bianco, A., Catavitello, C., Balbinot, A., Pompilio, A., Piccolomini, R., & D’Antonio, D. (2009a). Meticillin-heteroresistant Staphylococcus pasteuri from an apheresis platelet product. Journal of medical microbiology, 58(11), 1527-1528. google scholar
38. Savini, V., Catavitello, C., Carlino, D., Pompilio, A., Piccolomini, R., & Di Bonaventura, G. (2009b). Staphylococcus pasteuri bacteraemia in a leukemic patient. Journal of Clinical Pathology. google scholar
39. Savini, V., Catavitello, C., Pompetti, F., Passeri, C., Di Zacomo, S., Esattore, F., & D’Antonio, D. (2008). Contamination of a donated platelet unit by Staphylococcus pasteuri. Journal of Infection, 57(6), 494-496. google scholar
40. Si, W., Li, M., Wang, K., Li, J., Xu, M., Zhou, X., ... & Tian, Z. (2024). Staphylococcus warneri strain XSB102 exacerbates psoriasis and promotes keratinocyte proliferation in imiquimod-induced psoriasis-like dermatitis mice. Archives of Microbiology, 206(1), 3. google scholar
41. US FDA. (2001). Bacteriological analytical manual, 8th edition, revision A. In R. W. Benett & G. A. Lancett (Eds.). Rockville, MD, USA: AOAC International. google scholar
42. Üzgün, Y (2005). İzmir>in çeşitli semtlerinde satışa sunulan midye dolmaların mikrobiyolojik kalite kontrolü. Ege Üniversitesi / Fen Bilimleri Enstitüsü / Su Ürünleri Avlama ve İşleme Teknolojisi Anabilim Dalı, Yüksek Lisans Tezi, İzmir. google scholar
43. WHO (1996). World Health Organisation. Essential safety requirements for street-vended foods (Revised Edition). WHO/FAO Food Safety Unit - Division of Food and Nutrition, Geneva. google scholar
44. WHO (2017). World Health Organization guidelines on use of medically important antimicrobials in food-producing animals. google scholar