Detection and Adhesion Gene Profiling of Biofilm-Producing Staphylococcus aureus from Chicken Meat and Raw Milk Samples
13 / 9
Authors
-
Dr Manikkavasagan Ilangopathy
College of Food and Dairy Technology, Tamil Nadu Veterinary and Animal Sciences University
-
Dr Vijayarani Kumanan
Tamil Nadu Veterinary and Animal Sciences University
-
Dr Murugan Balaraman
College of Food and Dairy Technology, Tamil Nadu Veterinary and Animal Sciences University
-
Dr Meignanalakshmi Sundaram
Madras Veterinary College, TANUVAS
-
Dr Eswari Soundian
Madras Veterinary College, TANUVAS
-
Dr Ragavendiran Venkatesan
College of Food and Dairy Technology, Tamil Nadu Veterinary and Animal Sciences University
Keywords:
PCR, adhesion genes, S. aureusAbstract
The present study, aimed to investigate the presence of S. aureus (Staphylococcus aureus) in food samples (chicken meat and raw milk), as well as the biofilm-forming ability of isolates, and to characterise different adhesion genes. Out of 20 chicken meat samples, five (25%) tested positive for S. aureus. Out of 20 raw milk samples, 6 (30%) tested positive for S. aureus. Microtitre plate assay revealed that all isolates were weak to moderate biofilm producers. The isolates were characterized and confirmed to be S.aureus by PCR using 16S rRNA. The adhesion genes were detected using PCR with specific primers. Among the isolates, nuclease gene nuc (60%), coagulase gene coa (80%), agr (80%), and sdrD (80%) were detected in S.aureus isolated from chicken meat. In S.aureus isolated from milk samples nuc (66%), coa (100%), agr (50%) and sdrD (50%) genes were detected. All isolates were negative for fnpB, bap, and icaD genes. This study identified biofilm-forming S. aureus in food samples, which serves as a contaminant that contributes to food spoilage. Furthermore, they pose risks to consumer health and safety.
Downloads
References
Achek, R., H. Hotzel, I. Nabi, S. Kechida, D. Mami, N. Didouh, H. Tomaso, H. Neubauer,
R. Ehricht, S. Monecke, and H. El-Adawy (2020), Phenotypic and molecular detection of biofilm formation in Staphylococcus aureus isolated from different sources in Algeria, Pathogens (Basel, Switzerland)., 9(2):153. https://doi.org/10.3390/pathogens9020153
Alshammari, M., A. Ahmad, M. AlKhulaifi, D. Al Farraj, S. Alsudir, M. Alarawi, G. Takashi, and E. Alyamani (2023), Reduction of biofilm formation of Escherichia coli by targeting quorum sensing and adhesion genes using the CRISPR/cas9-hdr approach, and its clinical application on urinary catheter, J. Infect. Public Health., 16(8): 1174–1183. https://doi.org/10.1016/j.jiph.2023.05.026
Azara, E., C. Longheu, G.Sanna, & S. Tola (2017). Biofilm formation and virulence factor analysis of Staphylococcus aureus isolates collected from ovine mastitis. J. Appl. Microbiol., 123(2): 372–379. https://doi.org/10.1111/jam.13502
Ballah, F. M., M. S. Islam, M. L. Rana, F. B. Ferdous, R. Ahmed, P. K.Pramanik, J. Karmoker, S. Ievy, M. A. Sobur, M. P. Siddique, M. M. Khatun, M.Rahman & M. T. Rahman, (2022). Phenotypic and genotypic detection of biofilm-forming Staphylococcus aureus from different food sources in Bangladesh. Biology, 11(7): 7. https://doi.org/10.3390/biology11070949
Bhardwaj, D. K., N. K.Taneja, S. Dp, A. Chakotiya, P. Patel, P. Taneja, D. Sachdev, S.Gupta & M. G. Sanal, (2021). Phenotypic and genotypic characterization of biofilm forming, antimicrobial resistant, pathogenic Escherichia coli isolated from Indian dairy and meat products. Int. J. Food Microbiol., 336: 108899. https://doi.org/10.1016/j.ijfoodmicro.2020.108899
Brakstad, O. G., K. Aasbakk & J. A. Maeland, (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J. Clin. Microbiol., 30(7), 1654–1660.
Cappitelli, F., A. Polo & F. Villa, (2014). Biofilm formation in food processing environments is still poorly understood and controlled. Food Eng. Rev., 6: 29–42. https://doi.org/10.1007/s12393-014-9077-8
Carrascosa, C., D. Raheem, F. Ramos, A. Saraiva & A. Raposo, (2021). Microbial biofilms in the food industry- A comprehensive review. Int. J. Environ. Res. Public Health., 18(4): 2014. https://doi.org/10.3390/ijerph18042014
Chen, Q., S. Xie, X. Lou, S. Cheng, X. Liu, W. Zheng, Z. Zheng & H. Wang, (2020). Biofilm formation and prevalence of adhesion genes among Staphylococcus aureus isolates from different food sources. MicrobiologyOpen, 9(1): e00946. https://doi.org/10.1002/mbo3.946
Cucarella, C., M. A. Tormo, C. Ubeda, M. P. Trotonda, M. Monzón, C. Peris, B. Amorena, I.Lasa & J. R. Penadés, (2004). Role of biofilm-associated protein bap in the pathogenesis of bovine Staphylococcus aureus. Infect. Immun., 72(4): 2177–2185. https://doi.org/10.1128/IAI.72.4.2177-2185.2004
Ganesh, P. S., K. Veena, R. Senthil, K. Iswamy, E. M. Ponmalar, V. Mariappan, A. S. S. Girija, J. Vadivelu, S. Nagarajan, D. Challabathula & E. M. Shankar, (2022). Biofilm-associated agr and sar quorum sensing systems of Staphylococcus aureus are inhibited by 3-Hydroxybenzoic acid derived from illicium verum. ACS Omega, 7(17): 14653–14665. https://doi.org/10.1021/acsomega.1c07178
Herve, T. & G. Kumar, (2017). Prevalence of Staphylococcus aureus in retail chicken meat samples in jalandhar, punjab. Res. J. Pharm. Technol., 10: 1–5. https://doi.org/10.5958/0974-360X.2017.00058.0
Kumar, R. (2010). Detection of E.coli and Staphylococcus in milk and milk products in and around pantnagar. Vet. World., 3. https://doi.org/10.5455/vetworld.2010.495-496
Lakshminarayanan, S., & R. Jayalakshmy, (2015). Diarrheal diseases among children in India: Current scenario and future perspectives. J. Nat. Sci. Biol. Med., 6(1): 24–28. https://doi.org/10.4103/0976-9668.149073
Liang, J., T. Y. Huang, Y. Mao & X. Li, (2023). Biofilm formation of two genetically diverse Staphylococcus aureus isolates under beta-lactam antibiotics. Front. Microbiol., 14: 1139753. https://doi.org/10.3389/fmicb.2023.1139753
Liang, T., Z. Liang, S. Wu, Y. Ding, Q. Wu & B. Gu, (2023). Global prevalence of Staphylococcus aureus in food products and its relationship with the occurrence and development of diabetes mellitus. https://doi.org/10.1002/med4.6
Ma, Y., Y. Xu, B. D. Yestrepsky, R. J. Sorenson, M. Chen, S. D. Larsen & H. Sun, (2012). Novel inhibitors of Staphylococcus aureus virulence gene expression and biofilm formation. PloS One, 7(10): e47255. https://doi.org/10.1371/journal.pone.0047255
Neelam, V. K. Jain, M. Singh, V. G. Joshi, R. Chhabra, K.Singh & Y. S. Rana, (2022). Virulence and antimicrobial resistance gene profiles of Staphylococcus aureus associated with clinical mastitis in cattle. PloS One, 17(5), e0264762. https://doi.org/10.1371/journal.pone.0264762
Obe, T., R. Nannapaneni, W. Schilling, L. Zhang & A. Kiess, (2021). Antimicrobial tolerance, biofilm formation, and molecular characterization of Salmonella isolates from poultry processing equipment. J. Appl. Poult. Res., 30(4): 100195. https://doi.org/10.1016/j.japr.2021.100195
Peng, Q., X. Tang, W. Dong, N. Sun & W. Yuan, (2022). A review of biofilm formation of Staphylococcus aureus and its regulation mechanism. Antibiotics, 12(1): 12. https://doi.org/10.3390/antibiotics12010012
Ryu, S., M. Shin, B. Yun, W. Lee, H. Choi, M. Kang, S. Oh & Y. Kim, (2021). Bacterial quality, prevalence of pathogens, and molecular characterization of biofilm-producing Staphylococcus aureus from korean dairy farm environments. Animals: An Open Access Journal from MDPI, 11(5): 1306. https://doi.org/10.3390/ani11051306
Selem, E., A. F Mekky, W. A. Hassanein, F. M. Reda & Y. A. Selim, (2022). Antibacterial and antibiofilm effects of silver nanoparticles against the uropathogen Escherichia coli U12. Saudi J. Biol. Sci., 29(11): 103457. https://doi.org/10.1016/j.sjbs.2022.103457
Sharma, V., S. Sharma, D. K. Dahiya, A. Khan, M. Mathur & A. Sharma, (2017). Coagulase gene polymorphism, enterotoxigenecity, biofilm production, and antibiotic resistance in Staphylococcus aureus isolated from bovine raw milk in north west India. Ann. Clin. Microbiol. Antimicrob., 16(1): 65. https://doi.org/10.1186/s12941-017-0242-9
Speziale, P. & G. Pietrocola, (2020). The Multivalent role of fibronectin-binding proteins A and B (FnBPA and FnBPB) of Staphylococcus aureus in host infections. Front. Microbiol., 11: 2054. https://doi.org/10.3389/fmicb.2020.02054
Tang, J., J. Chen, H. Li, P. Zeng & J. Li, (2013). Characterization of adhesin genes, staphylococcal nuclease, hemolysis, and biofilm formation among Staphylococcus aureus strains isolated from different sources. Foodborne Pathog., Dis., 10(9): 757–763. https://doi.org/10.1089/fpd.2012.1474
Tang, J.N., X.M. Shi, C.L. Shi & H. C Chen, (2006). Characterization of a duplex polymerase chain reaction assay for the detection of enterotoxigenic Strains of Staphylococcus Aureus. J. Rapid Methods Autom. Microbiol., 14(3): 201–217. https://doi.org/10.1111/j.1745-4581.2006.00047.x
Thakur, P., R. Chawla, A. Tanwar, A. S. Chakotiya, A. Narula, R. Goel, R. Arora & R. K. Sharma, (2016). Attenuation of adhesion, quorum sensing and biofilm mediated virulence of carbapenem resistant Escherichia coli by selected natural plant products. Microb. Pathog., 92: 76–85. https://doi.org/10.1016/j.micpath.2016.01.001
Van Houdt, R. & C. W. Michiels, (2010). Biofilm formation and the food industry, a focus on the bacterial outer surface. J. Appl. Microbiol., 109(4): 1117–1131. https://doi.org/10.1111/j.1365-2672.2010.04756.x
Yu, J., F. Jiang, F. Zhang, M. Hamushan, J. Du, Y. Mao, Q. Wang, P. Han, J.Tang & H. Shen, (2021). Thermonucleases contribute to staphylococcus aureus biofilm formation in implant-associated infections–A redundant and complementary story. Front. Microbiol., 12, 687888. https://doi.org/10.3389/fmicb.2021.687888
