Evaluation of the Role of Staphylococci in the Pathomechanism of Conjunctivitis

Overview

Journal Int Ophthalmol

Specialty Ophthalmology

Date 2021 Mar 29

PMID 33778922

Authors

Ewa Jasinska

Agnieszka Bogut

Agnieszka Magrys

Alina Olender

Affiliations

Soon will be listed here.

Abstract

Purpose: Determination of the association between ica genes and phenotypic biofilm formation in staphylococcal isolates involved in conjunctivitis, their antibiotic resistance as well as detection of selected virulence characteristics: adhesion to epithelial cells and in vitro cytotoxicity.

Methods: The study included 26 Staphylococcus aureus (SA) and 26 Staphylococcus epidermidis (SE) isolates. The presence of icaAD genes and ica operon was determined by the PCR assay. Phenotypic biofilm formation was verified using the microtitre plate assay. Antibiotic resistance was performed using the disc diffusion method. Staphylococcal ability to attach to host cells was assessed by flow cytometry. Cytotoxicity on epithelial cells was evaluated by LDH assay.

Results: The ica genes were detected in 26.9% of SE and in 42.3% of SA isolates. Only 15.3% of isolates (SE) were positive for both the icaAD and the ica operon. Phenotypically, 19.2% of SE isolates were strong biofilm producers, among which three were both icaAD- and ica operon-positive. About 26.9% of SA isolates were strong biofilm producers. Methicillin resistance (MR) was detected in 34.6% of SE and 26.9% of SA isolates. About 75% of MR isolates were multidrug resistant. SA isolates adhered to host cells more extensively than SE. SA isolates released higher level of LDH than SE.

Conclusions: Adherence abilities were commonly observed in staphylococci associated with conjunctivitis. However, low prevalence of isolates positive for a complete and functional ica locus and low prevalence of strong biofilm producers was detected. SA adhered to a greater extent to eukaryotic cells than SE and were more cytotoxic.

References

Arciola C, Campoccia D, Ravaioli S, Montanaro L . Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects. Front Cell Infect Microbiol. 2015; 5:7. PMC: 4322838. DOI: 10.3389/fcimb.2015.00007. View

Josse J, Laurent F, Diot A . Staphylococcal Adhesion and Host Cell Invasion: Fibronectin-Binding and Other Mechanisms. Front Microbiol. 2017; 8:2433. PMC: 5723312. DOI: 10.3389/fmicb.2017.02433. View

Otto M . Staphylococcus epidermidis--the 'accidental' pathogen. Nat Rev Microbiol. 2009; 7(8):555-67. PMC: 2807625. DOI: 10.1038/nrmicro2182. View

McKenney D, Pouliot K, Wang Y, Murthy V, Ulrich M, DORING G . Broadly protective vaccine for Staphylococcus aureus based on an in vivo-expressed antigen. Science. 1999; 284(5419):1523-7. DOI: 10.1126/science.284.5419.1523. View

Schlievert P, Strandberg K, Lin Y, Peterson M, Leung D . Secreted virulence factor comparison between methicillin-resistant and methicillin-sensitive Staphylococcus aureus, and its relevance to atopic dermatitis. J Allergy Clin Immunol. 2010; 125(1):39-49. PMC: 2814367. DOI: 10.1016/j.jaci.2009.10.039. View

Sabate Bresco M, Harris L, Thompson K, Stanic B, Morgenstern M, OMahony L . Pathogenic Mechanisms and Host Interactions in Device-Related Infection. Front Microbiol. 2017; 8:1401. PMC: 5539136. DOI: 10.3389/fmicb.2017.01401. View

Ziebuhr W, Krimmer V, Rachid S, Lossner I, Gotz F, Hacker J . A novel mechanism of phase variation of virulence in Staphylococcus epidermidis: evidence for control of the polysaccharide intercellular adhesin synthesis by alternating insertion and excision of the insertion sequence element IS256. Mol Microbiol. 1999; 32(2):345-56. DOI: 10.1046/j.1365-2958.1999.01353.x. View

Speziale P, Pietrocola G, Foster T, Geoghegan J . Protein-based biofilm matrices in Staphylococci. Front Cell Infect Microbiol. 2014; 4:171. PMC: 4261907. DOI: 10.3389/fcimb.2014.00171. View

Ridley R, Douglas I, Whawell S . Differential adhesion and invasion by Staphylococcus aureus of epithelial cells derived from different anatomical sites. J Med Microbiol. 2012; 61(Pt 12):1654-1661. DOI: 10.1099/jmm.0.049650-0. View

10.

Figueiredo A, Ferreira F, Beltrame C, Cortes M . The role of biofilms in persistent infections and factors involved in ica-independent biofilm development and gene regulation in Staphylococcus aureus. Crit Rev Microbiol. 2017; 43(5):602-620. DOI: 10.1080/1040841X.2017.1282941. View

11.

Rohde H, Burandt E, Siemssen N, Frommelt L, Burdelski C, Wurster S . Polysaccharide intercellular adhesin or protein factors in biofilm accumulation of Staphylococcus epidermidis and Staphylococcus aureus isolated from prosthetic hip and knee joint infections. Biomaterials. 2006; 28(9):1711-20. DOI: 10.1016/j.biomaterials.2006.11.046. View

12.

Christensen G, Simpson W, Younger J, Baddour L, Barrett F, Melton D . Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985; 22(6):996-1006. PMC: 271866. DOI: 10.1128/jcm.22.6.996-1006.1985. View

13.

Piechota M, Kot B, Frankowska-Maciejewska A, Gruzewska A, Wozniak-Kosek A . Biofilm Formation by Methicillin-Resistant and Methicillin-Sensitive Strains from Hospitalized Patients in Poland. Biomed Res Int. 2019; 2018:4657396. PMC: 6327255. DOI: 10.1155/2018/4657396. View

14.

Sharifinejad M, Shokatpour N, Farnaghi F, Abedinyfar Z, Amoli F, Doustdar F . Different Genotypes of Adhesion Operon Genes in Staphylococcus epidermidis Isolates From Various Ocular Infections. Eye Contact Lens. 2017; 44 Suppl 2:S277-S280. DOI: 10.1097/ICL.0000000000000464. View

15.

Smith K, Perez A, Ramage G, Lappin D, Gemmell C, Lang S . Biofilm formation by Scottish clinical isolates of Staphylococcus aureus. J Med Microbiol. 2008; 57(Pt 8):1018-1023. DOI: 10.1099/jmm.0.2008/000968-0. View

16.

Bur S, Preissner K, Herrmann M, Bischoff M . The Staphylococcus aureus extracellular adherence protein promotes bacterial internalization by keratinocytes independent of fibronectin-binding proteins. J Invest Dermatol. 2013; 133(8):2004-12. DOI: 10.1038/jid.2013.87. View

17.

Zaidi T, Zaidi T, Yoong P, Pier G . Staphylococcus aureus corneal infections: effect of the Panton-Valentine leukocidin (PVL) and antibody to PVL on virulence and pathology. Invest Ophthalmol Vis Sci. 2013; 54(7):4430-8. PMC: 3700385. DOI: 10.1167/iovs.13-11701. View

18.

Krut O, Utermohlen O, Schlossherr X, Kronke M . Strain-specific association of cytotoxic activity and virulence of clinical Staphylococcus aureus isolates. Infect Immun. 2003; 71(5):2716-23. PMC: 153241. DOI: 10.1128/IAI.71.5.2716-2723.2003. View

19.

Flores-Paez L, Zenteno J, Alcantar-Curiel M, Vargas-Mendoza C, Rodriguez-Martinez S, Cancino-Diaz M . Molecular and Phenotypic Characterization of Staphylococcus epidermidis Isolates from Healthy Conjunctiva and a Comparative Analysis with Isolates from Ocular Infection. PLoS One. 2015; 10(8):e0135964. PMC: 4537226. DOI: 10.1371/journal.pone.0135964. View

20.

Elkhashab T, Adel L, Nour M, Mahran M, Elkaffas M . Association of intercellular adhesion gene A with biofilm formation in staphylococci isolates from patients with conjunctivitis. J Lab Physicians. 2018; 10(3):309-315. PMC: 6052808. DOI: 10.4103/JLP.JLP_122_17. View