Genotypic Assessment of Virulence Factors associated with Adhesion, Biofilm, and Quorum Sensing regulation in Staphylococcus aureus from Multi host Environments

Sandeep Kumar Sharma; Jyoti  Bishnoi; Vikas  Galav; Bincy  Joseph; Taruna  Bhati; Manish   Agrawal; Sharat   Mehta; Anil  Kataria

doi:10.56093/ijans.v96i8.169640

Authors

Sandeep Kumar Sharma Assistant Professor & PI, AMR Project , Post Graduate Institute of Veterinary Education & Research (PGIVER)
Jyoti Bishnoi
Vikas Galav
Bincy Joseph
Taruna Bhati
Manish Agrawal
Sharat Mehta
Anil Kataria

DOI:

https://doi.org/10.56093/ijans.v96i8.169640

Keywords:

Staphylococcus aureus, adhesion genes, biofilm formation, quorum sensing, agr typing

Abstract

Staphylococcus aureus is a versatile pathogen capable of infecting both humans and animals, largely due to its robust virulence factors. This study aimed to investigate the prevalence and diversity of key adhesion, colonization, slim production and quorum-sensing genes in S. aureus isolates from diverse animal and human host species. A total of 157 isolates were subjected to phenotypic and molecular assays. Biofilm production, assessed via Congo Red Agar (CRA), revealed that 92.4% of the isolates were slime producers. Molecular analysis demonstrated high prevalence of clumping factor genes, with clfA and clfB detected in 94.3% of isolates, and intercellular adhesion genes icaA and icaD present in 95.5% and 91.7%, respectively. The trap gene, a regulator of quorum sensing, was identified in 98.0% of isolates. Accessory gene regulator (agr) typing classified the majority into agr group I (40.1%), followed by agrII (26.8%), agrIII (17.2%), and agrIV (15.3%), with only one isolate being non-typable. All four agr types were detected among isolates from humans, camels, buffaloes, goats, and cattle, highlighting a broad agr diversity in these hosts. A small number of isolates (n=3) were deficient in three or more adherence-related genes, suggesting strain-specific variations in virulence mechanisms. No polymorphism was observed among the targeted virulence genes, indicating a high degree of genetic conservation. These findings underscore the widespread distribution and conservation of major virulence determinants in S. aureus, with implications for understanding host-pathogen interactions and developing targeted control strategies across species.

Downloads

Download data is not yet available.

References

Ando, E.; Monden, K.; Mitsuhata, R.; Kariyama, R. and Kumon, H. (2004). Biofilm formation among methicillin-resistant Staphylococcus aureus isolates from patients with Urinary tract infection. Acta Medica Okayama. 58 (4): 207-214.

Arciola, C.R.; Baldassarri, L. and Montanaro, L. (2001). Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. Journal of Clinical Microbiology. 39(6): 2151– 2156.

Atshan, S.S.; Shamsudin, M.N. and Sekawi, Z. (2012). Prevalence of adhesion and regulation of biofilm-related genes in different clones of Staphylococcus aureus. Journal of Biomedicine and Biotechnology. DOI: 10.1155/2012/976972.

Ballah, F. M., Islam, M. S., Rana, M. L., Ferdous, F. B., Ahmed, R., Pramanik, P. K., Karmoker, J., Ievy, S., Sobur, M. A., Siddique, M. P., Khatun, M. M., Rahman, M., & Rahman, M. T. (2022). Phenotypic and Genotypic Detection of Biofilm-Forming Staphylococcus aureus from Different Food Sources in Bangladesh. Biology, 11(7), 949.

Barbieri, R.; Pesce, M.; Franchelli, S.; Baldelli, I.; De Maria, A. and Marchese, A. (2015). Phenotypic and genotypic characterization of Staphylococci causing breast peri-implant infections in oncologic patients. BMC Microbiology. 15 (1): 26.

Bnyan, I. (2013). Detection of adhesion genes and slim production among Staphylococcus aureus and Staphylococcus epidermidis isolated from hemodialysis patients. Advances in Life Science and Technology. 15: 27-32.

Boles, B. R. and Horswill, A. R. (2008). "Agr-mediated dispersal of Staphylococcus aureus biofilms." PLoS Pathogens. 4(4): e1000052. doi:10.1371/journal.ppat.1000052.

Booth, M.C.; Pence, L.M.; Mahasreshti, P.; Callegan, M. and Gilmore, M. (2001). Clonal associations among Staphylococcus aureus isolates from various sites of infections. Infection and Immunity. 69 (1): 345–352.

Castelani, L.; Pilon, L.E.; Martins, T.; Pozzi, C.R. and Arcaro, J.R.P. (2015). Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis. Animal Science Journal. 86: 340–344.

Cheung, A.L.; Bayer, A.S.; Zhang, G.; Gresham, H. and Xiong Y-Q. (2004). Regulation of virulence determinants in vitro and in vivo in Staphylococcus aureus. FEMS Immunology & Medical Microbiology. 40 (1): 1-9.

Deepak SJ, Kannan P, Savariraj WR, Ayyasamy E, Tuticorin Maragatham Alagesan SK, Ravindran NB, Sundaram S, Mohanadasse NQ, Kang Q, Cull CA, Amachawadi RG. (2024). Characterization of Staphylococcus aureus isolated from milk samples for their virulence, biofilm, and antimicrobial resistance. (2024). Scientific Reports 14, 25635 (2024).

El-Sayed, A.; Alber, J.; Lammler, C.; Jager, S.; Wolter, W. and Castaneda-Vazquez, H. (2006). Comparative study on genotypic properties of Staphylococcus aureus isolated from clinical and subclinical mastitis in Mexico. Veterinaria México. 37(2): 165-179.

Fabres-Klein, M.H.; Santos, M.J.C.; Klein, R.C.; Nunes de Souza, G. and Ribon, A.O.B. (2015). An association between milk and slime increases biofilm production by bovine Staphylococcus aureus. BMC Veterinary Research. 11: 3.

Fitzpatrick, F.; Humphreys, H. and O’Gara, J.P. (2005) The genetics of staphylococcal biofilm formation will a greater understanding of pathogenesis lead to better management of device-related infection? Clinical Microbiology and Infection. 11 (12): 967–973.

Gerke, C.; Kraft, A.; Sussmuth, R.; Schweitzer, O. and Gotz, F. (1998). Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin. The Journal of Biological Chemistry. 273 (29): 18586–18593.

Gilot, P.; Lina, G.; Cochard, T. and Poutrel, B. (2002). Analysis of the genetic variability of genes encoding the RNA III-activating components Agr and TRAP in a Population of Staphylococcus aureus strains isolated from cows with mastitis. Journal of Clinical Microbiology. 40(11): 4060–4067.

Jarraud, S.; Mougel, C.; Thioulouse, J.; Lina, G.; Meugnier, H.; Forey, F.; Nesme, X.; Etienne, J. and Vandenesch, F. (2002). Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease. Infection and Immunity. 70 (2): 631–641.

Keller, L. and Surette, M.G. (2006). "Communication in bacteria: an ecological and evolutionary perspective." Nature Reviews, Microbiology. 4 (4): 249-258.

Khoramian, B.; Jabalameli, F.; Niasari-Naslaji, A.; Taherikalani, M. and Emaneini M. (2015). Comparison of virulence factors and biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections. Microbial Pathogenesis. 88: 73-77.

Li-li Yang, H.J.; Liu, D.C.; He, H.B.; Wang, C.F.; Zhong, J.F.; Gao, Y.D. and Yanjun, Z. (2012). Analysis of biofilms formation and associated genes detection in staphylococcal isolates from bovine mastitis. International Journal of Applied Research and Veterinary Medicine. 10 (1): 62-68.

Melo, P.C.; Ferreira, L.M.; Filho, A.N.; Zafalon, L.N.; Vicente, H.I.G. and Souza, V. (2013). Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis. Brazilian Journal of Microbiology. 44 (1): 119-124.

Momtaz, H.; Rahimi, E. and Tajbakhsh, E. (2010). Detection of some virulence factors in Staphylococcus aureus isolated from clinical and subclinical bovine mastitis in Iran. African Journal of Biotechnology. 9 (25): 3753-3758.

Peerayeh, S.N.; Azimian, A.; Nejad, Q.B.; Kashi, M. (2009). Prevalence of agr specificity groups among Staphylococcus aureus isolates from University Hospitals in Tehran. Science. 40 (1): 27-29.

Salgueiro, V., Manageiro, V., Bandarra, N. M., Ferreira, E., Clemente, L., & Caniça, M. (2020). Genetic Relatedness and Diversity of Staphylococcus aureus from Different Reservoirs: Humans and Animals of Livestock, Poultry, Zoo, and Aquaculture. Microorganisms, 8(9), 1345.

Sharma, S.K., Yadav, R, Mehta, S.C. and Kataria, A.K. (2020). Detection and analysis of antibiotic resistance variability among Staphylococcus aureus isolates from animal and human sources Veterinarski arhiV., 90 (5), 493-508.

Sharma, S.K.; Mehta S.C. and Kataria A.K. (2016). Capsular typing of Staphylococcus aureus isolates from camel and other domestic animals using duplex polymerase chain reaction. Journal of Camel Practice and Research. 23(1): 81-84.

Stephan, R.; Annemuller, C.; Hassan, A. and Lammler, C. (2001). Characterization of enterotoxigenic Staphylococcus aureus strains isolated from bovine mastitis in north-east Switzerland. Veterinary Microbiology. 78 (4): 373–382.

Tang, J.; Chen, J.; Li, H.; Zeng, P. and Li. J. (2013). Characterization of adhesion genes, Staphylococcal nuclease, hemolysis, and biofilm formation among Staphylococcus aureus strains isolated from different sources. Foodborne Pathogens and Disease. 10 (9): 757-763.

Tristan, A.; Ying, L.; Bes, M.; Etienne, J.; Vandenesch, F. and Lina, G. (2003). Use of multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. Journal of Clinical Microbiology. 41: 4465- 4467.

Vasudevan, P.; Nair, M.K.M.; Annamalai, T. and Venkitanarayanan, K.S. (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for bioﬁlm formation. Veterinary Microbiology. 92 (1-2): 179–185.

Xu, J.; Tan, X.; Zhang, X.; Xia, X. and Sun, H. (2015). The diversities of staphylococcal species, virulence and antibiotic resistance genes in the subclinical mastitis milk from a single Chinese cow herd. Microbial Pathogens. 88: 29-38.

Yadav, J.A.; Hertel, C. and Hammes, W.P. (1999). A 23S rRNA target polymerase chain reaction based system for detection of Staphylococcus aureus in meat starter cultures and dairy products. Journal of Food Protection. 62 (10): 1150-1156.

Yazdani, R.; Oshaghi, M.; Havayi, A.; Pishva, E.; Salehi, R.; Sadeghizadeh, M. and Foroohesh, H. (2006). Detection of icaAD gene and biofilm formation in Staphylococcus aureus isolates from wound infections. Iranian Journal of Public Health. 35 (2): 25-28.