Salmonella Serotiplerinde Biofilm ve Virülans İlişkisinin Araştırılması

Abstract

Salmonella gıda kaynaklı bir patojendir ve çiftlik hayvanlarında, kümes hayvanlarında ve insanlarda gastroenterite neden olur. Salmonella'nın patojenitesi, biyofilm oluşumu, motilite, invazyon, adezyon ve tip III sekresyon sistemi gibi virülans faktörleriyle ilişkilidir. Bu çalışmada, farklı kaynaklardan toplanan Salmonella serotiplerinin biyofilm oluşumu ile virülans genleri arasındaki ilişki incelenmiştir. Salmonella serotiplerinin adrA, agfD, invA, sdiA, bapA genlerinin biyofilm ile ilişkisi Polimeraz Zincir Reaksiyonu (PCR) yöntemi ile araştırıldı. Bu amaçla farklı kaynaklardan 44 adet Salmonella izolatı alınmıştır, bunların 10 adeti Salmonella Typhimurium, 10 adeti Salmonella Enteritidis ve 24 adeti Salmonella Infantis'ten oluşmaktadır. Bu izolatların sırasıyla, 38 (%86,36), 26 (%59), 44 (%100), 39 (%88,64), 35 (%79,55) oranında bapA, sdiA, invA, agfD ve adrA genlerini içerdiği belirlendi. Fenotipik olarak örneklerde biyofilm oluşumu gözlenmedi. Fenotipik ve moleküler yöntemlerin birlikte kullanılması da daha güvenilir sonuçlar elde etmek için gereklidir. Elde edilen bulgular, Salmonella’da biyofilm oluşumunun çok bileşenli bir düzenleme mekanizmasına bağlı olduğunu, ayrıca biyofilmle ilişkili genlerin varlığının tek başına yeterli olmayabileceğini göstermektedir. Bu nedenle, gelecekte yapılacak çalışmaların biyofilm gelişimini etkileyen çevresel ve genetik faktörlerin daha ayrıntılı olarak incelenmesine odaklanılması, hem Salmonella’nın patojenitesinin daha iyi anlaşılması hem de insan sağlığı açısından korunma stratejilerinin geliştirilmesi bakımından büyük önem taşımaktadır.

Keywords

• Biyofilm
• PCR
• Salmonella
• Virülans

Supporting Institution

ADÜ-BAP

Project Number

VTF15042

Ethical Statement

Çalışmamızda Etik Kurul Kararına gerek yoktur.

Investigation of Biofilm Production and Virulence Genes in Salmonella Serotypes

Abstract

Salmonella is a foodborne pathogen and cause of gastroenteritis in livestock, poultry and in humans. Pathogenicity of Salmonella is related to its virulence factors like formation of biofilm, motility, invasion, adhesion and its type III secretion system. In this study, the relation between biofilm formation and virulence genes of Salmonella serotypes which were collected from different sources was examined. The presence of biofilm-associated genes (adrA, agfD, invA, sdiA, bapA) in Salmonella serotypes was analyzed using the Polymerase Chain Reaction (PCR) method. For this aim, the 44 Salmonella isolates were taken from different sources, those consist of 10 Salmonella Typhimurium, 10 Salmonella Enteritidis and 24 Salmonella Infantis. It was determined that these isolates contained the genes bapA, sdiA, invA, agfD and adrA at the rates of 38 (86.36%), 26 (59%), 44 (100%), 39 (88.64%), and 35 (79.55%) respectively. Phenotypically, biofilm formation was not observed in the samples. Integration of phenotypic and molecular methods is crucial for obtaining more reliable results. The findings suggest that biofilm formation in Salmonella is governed by a multicomponent regulatory mechanism, and that the presence of biofilm-associated genes alone may not be sufficient for biofilm formation. Therefore, future studies focusing on a more detailed investigation of the environmental and genetic factors influencing biofilm development are of great importance for a better understanding of Salmonella pathogenicity and for the development of protection strategies in terms of human health.

Keywords

• Biofilm
• PCR
• Salmonella
• Virulence

Supporting Institution

ADU-BAP

Project Number

VTF15042

Ethical Statement

This study does not present any ethical concerns.

References

1. Afshari, A., Baratpour, A., Khanzade, S., & Jamshidi, A. (2018). Salmonella Enteritidis and Salmonella Typhimurium identification in poultry carcasses. Iranian Journal of Microbiology, 10(1), 45-50.
2. Akçelik, M., & Akkoç, N. (2010). Salmonella Patojenitesinin Molekuler Mekanizmaları. Türkiye Klinikleri Journal of Medical Sciences, 30(1), 312-22. https://doi.org/10.5336/medsci.2009-13658
3. Amrutha, B., Sundar, K., & Shetty, P.H. (2017). Study on E. coli and Salmonella biofilms from fresh fruits and vegetables. Journal of Food Science and Technology, 54, 1091–1097. https://doi.org/10.1007/s13197-017-2555-2
4. Barilli, E., Bacci, C., StellaVilla, Z., Merialdi, G., D’Incau, M., Brindani, F., & Vismarra, A. (2018). Antimicrobial resistance, biofilm synthesis and virulence genes in Salmonella isolated from pigs bred on intensive farms. Italian Journal of Food Safety, 7(2). 7223. https://doi.org/10.4081/ijfs.2018.7223.
5. Bhatta, D.R., Bangtrakulnonth, A., Tishyadhigama, P., Saroj, S.D., Bandekar, J.R., Hendriksen R.S., & Kapadnis, B.P. (2007). Serotyping, PCR, phage‐typing and antibiotic sensitivity testing of Salmonella serovars isolated from urban drinking water supply systems of Nepal. Letters in Applied Microbiology, 44(6), 588–594. https://doi.org/10.1111/j.1472-765X.2007.02133.x
6. Biswas, R., Agarwal, R.K., Bhilegaonkar, K.N., Kumar, A., Nambiar, P., Rawat, S., & Singh, M. (2011). Cloning and sequencing of biofilmassociated protein (bapA) gene and its occurrence in different serotypes of Salmonella. Letters in Applied Microbiology, 52(2), 138–143. https://doi.org/10.1111/j.1472-765X.2010.02975.x
7. Çiftci, A., Fındık, A., Onuk, E.E., & Savaşan, S. (2009). Detection of methicillin resistance and slime factor production of Staphylococcus aureus in bovine mastitis. Brazilian Journal of Microbiology, 40, 254-261. https://doi.org/10.1590/S1517-83822009000200009
8. De Oliveira D.C.V, Júnior, A.F., Kaneno, R., Silva, G.M., Júnior, J.P.A., Silva, N.C.C., & Rall, V.L.M. (2014). Ability of Salmonella spp. to Produce Biyofilm Is Dependent on Temperature and Surface Material. Foodborne Pathogens and Disease, 11, 478-483. https://doi.org/10.1089/fpd.2013.1710

9. Elfaky, M.A., Thabit, A.K., Eljaaly, K., Zawawi, A., Abdelkhalek, A.S., Almalki, A.J., & Hegazy, W.A.H. (2022). Controlling of Bacterial Virulence: Evaluation of Anti-Virulence Activities of Prazosin against Salmonella enterica. Antibiotics, 11, 1585. https://doi.org/10.3390/antibiotics11111585
10. Guillen, S., Marcén, M., Fau, E., Mañas, P., & Cebrián, G. (2022). Relationship between growth ability, virulence, and resistance to food-processing related stresses in non-typhoidal Salmonellae. International Journal of Food Microbiology, 361, 109462. https://doi.org/10.1016/j.ijfoodmicro.2021.109462
11. Halatsi, K., Oikonomou, I., Lambiri, M., Mandilara, G., Vatopoulos, A., & Kyriacou, A. (2006). PCR detection of Salmonella spp. using primers targeting the quorum sensing gene sdiA. FEMS Microbiology Letters, 259(2), 201–207. https://doi.org/10.1111/j.1574-6968.2006.00266.x
12. Hidalgo, G.S., Valenzuela, L.M., Aréchiga, C.N., Guajardo, C.S., Salazar, L.M., & Vázquez, M.E. (2016). Identification of bapA in Strains of Salmonella enterica subsp. enterica Isolated from Wild Animals Kept in Captivity in Sinaloa, Mexico. Veterinary Medicine International, 2016(1), 4. https://doi.org/10.1155/2016/3478746
13. Horton, R.A., Card, R.R., Randall, R., & Teale, C.J. (2016). Differentiation of UK endemic strains of Salmonella enterica serovar Newport from epidemic North American strains by PCR detection of a truncated bapA chromosomal gene. Research in Veterinary Science, 104, 113-116. https://doi.org/10.1016/j.rvsc.2015.12.008
14. Jahan, F., Chinni, S.V., Samuggam, S., Reddy, L.V., Solayappan, M., & Yin, S.L. (2022). The Complex Mechanism of the Salmonella Typhi Biofilm Formation That Facilitates Pathogenicity: A Review. International Journal of Molecular Sciences, 23, 6462. https://doi.org/10.3390/ijms23126462
15. Lapierre, A.L., Cornejo, J., Zavala, S., Galarce, N., Sánchez, F., Benavides, M.B.,…&, Sáenz, L. (2020). Phenotypic and Genotypic Characterization of Virulence Factors and Susceptibility to Antibiotics in Salmonella Infantis Strains Isolated from Chicken Meat: First Findings in Chile. Animals, 10, 1049. https://doi.org/10.3390/ani10061049
16. Pantu, K.R., Angela, J.W.H., Mizan, M.F.R., Hossain, M.I., Ashrafudoulla, M., Toushik, S.H.,…&, Yu Kyung Kim, S.D.H. (2021). Effects of environmental conditions (temperature, pH, and glucose) on biofilm formation of Salmonella enterica serotype Kentucky and virulence gene expression. Poultry Science, 100, 101209. https://doi.org/10.1016/j.psj.2021.101209
17. Rana, K., Nayak, S.R., Bihary, A., Sahoo, A.K., Mohanty, K.C., Palo, S.,…&, Dash, P. (2021). Association of quorum sensing and biofilm formation with Salmonella virulence: story beyond gathering and cross-talk. Archives of Microbiology, 203, 5887–5897. https://doi.org/10.1007/s00203-021-02594-y
18. Sharma, İ., & Das, K. (2016). Detection of invA Gene in Isolated Salmonella from Marketed Poultry Meat by PCR Assay. Journal of Food Processing and Technology, 7(3). http://dx.doi.org/10.4172/2157-7110.1000564
19. Sunar, N.M., Stentiford, E.I., Stewart, D.I., & Fletcher, L.A. (2014). Molecular techniques to characterize the invA genes of Salmonella spp. for pathogen inactivation study in composting. ArXiv Preprint Archive, 1404.5208. https://doi.org/10.48550/arXiv.1404.5208
20. Wang, H., Dong, Y., Wang, G., Xu, X., & Zhou, G. (2016). Effect of growth media on gene expression levels in Salmonella Typhimurium biyofilm formed on stainless steel surface. Food Control, 59, 546- 552. https://doi.org/10.1016/j.foodcont.2015.06.026s