Evaluation of biofilm formation and associated slime encoding determinants in Staphylococcus aureus isolated from clinically diseased pets

Sohier M. Syame; Mona Elshabrawy; Hussien A. Abouelhag; Randa Mohamed Ismail; Engy Farahat; E. S. Gaber; Hanan M. El-Hewairy; Ashraf Hakim

doi:10.53730/ijhs.v6nS9.12934

Authors

Sohier M. Syame
sohiersyame@yahoo.com
Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Mona Elshabrawy Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Hussien A. Abouelhag Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Randa Mohamed Ismail Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Engy Farahat Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Gaber, E. S. Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622
Hanan M. El-Hewairy Immunology department, Animal Health Research Institute, Agriculture Research Center, Dokki, Giza, Postal code 12611
Ashraf Hakim Department of Microbiology and Immunology, National Research Centre, 33Bohouth St., Dokki, Cairo, Egypt, Postal code 12622

Keywords:

biofilm, ica genes, dog and cat, Staphylococcus aureus, fluorescence, SEM

Abstract

Staphylococcus aureus (S.aureus) is an important zoonotic pathogen implemented in various hospital, community as well as livestock infections. Pets as dogs and cats have increased their close social relation with human leading to significant elevation in transmission of zoonoticmultidrug resistant virulent pathogens.Diplex polymerase chain reaction was applied on ten clinical Staphylococcus aureus isolates obtained from wound and nasal samples from dogs and cats for detection of twoslime formation encoding genes;icaA andicaD. The positive genes carrying isolates were encouraged to produce biofilm and evaluated phenotypically by cultivation onto Congo red medium. Quantitative assessment was done using a microtiter plate assay, also the formed biofilm examined by fluorescent and scanning electron microscopy. Four out of tested tenStaphylococcus aureus isolates were found to harbor the two genes separately.The four isolates displayed positive biofilm production onto Congo red medium while only three isolates produced biofilmin sterile polystyrene 96-well microtiter plate. One isolate associated dog infection developed strong biofilm formation which examined byboth fluorescent and scanning electron microscopy.Staphylococcus aureus isolates obtained from diseased dogs and cats can produce biofilm that increasing their virulence and pathogenicity as well as public health concern.

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References

Arciola CR, Baldassarri L, Montanaro L. Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. J ClinMicrobiol.2001; 39:2151–6.

Avila-Novoa M, Iñiguez-Moreno M, Solís-Velázquez O, González-Gómez J.P. Biofilm formation by Staphylococcus aureus isolate from food contact surfaces in the dairy industry of Jalisco, Mexico. J. Food Qual. 2018;2018:1746139.

Bruce S.A., Smith J.T., Mydosh J.L., Ball J., Needle D.B., Gibson R., Andam C.P. Shared antibiotic resistance and virulence genes in Staphylococcus aureus from diverse animal hosts. Sci. Rep. 2022;12:4413.

Chen L, Tang ZY, Cui SY, Ma ZB, Deng H, Kong WL, Yang LW, Lin C, Xiong WG, Zeng ZL. Biofilm Production Ability, Virulence and Antimicrobial Resistance Genes in Staphylococcus aureus from Various Veterinary Hospitals.Pathogens. 2020 Apr 4;9(4):264.

Cheville NF, Stasko J. Techniques in electron microscopy of animal tissue. Vet Pathol. 2014;51(1):28–41.

Cramton SE, Gerke C, Schnell NF, Nichols WW, G¨otz F (1999)The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun 67:5427–5433

Dai J, Wu S, Huang J, Wu Q, Zhang F, Zhang J, Wang J, Ding Y, Zhang S, Yang X, Lei T, Xue L, Wu H. Prevalence and Characterization of Staphylococcus aureus Isolated From Pasteurized Milk in China. Front Microbiol. 2019 2(10)641.

Elshabrawy, M.A., Abouelhag, H.A., Khairy, E.A., Marie, H.S., Hakim, A.S. (2020): Molecular divergence of Staphylococcus aureus isolated from dogs and cats. Jordan Journal of Biological Sciences, 13(2): 139–144.

Ghasemian A, NajarPeerayeh S, Bakhshi B, Mirzaee M. Comparison of Biofilm Formation between Methicillin-Resistant and Methicillin-Susceptible Isolates of Staphylococcus aureus. Iran Biomed J. 2016; 20(3):175–81.

Gorokhova E, Mattsson L, and Sundstrom AM.A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy,” Journal of Microbioligical Methods, 2012: (89)3: 216–221.

Gowrishankar S, Kamaladevi A, Balamurugan K, Pandian S.K. In vitro and in vivo biofilm characterization of methicillin-resistant Staphylococcus aureus from patients associated with pharyngitis infection. Biomed Res. Int. 2016; 2016:1289157.

Hamel Mohaisen S, Hussain Ali M, HashemShehab Z, Al-Mayyahi AW, Abdulhassan AA. Effect of Ultraviolet Light on the Expression of icaD Gene in Staphylococcus aureus Local Isolates in Iraq. Arch Razi Inst. 2021 Nov 30;76(5):1221-1227.

Kern Z.T., Jacob M.E., Gilbertie J.M., Vaden S.L., Lyle S.K. Characteristics of Dogs with Biofilm-Forming Escherichia Coli Urinary Tract Infections. J. Vet. Intern. Med. 2018;32:1645–1651.

Kranjec C, Morales Angeles D, TorrissenMårli M, Fernández L, García P, Kjos M, Diep DB. Staphylococcal Biofilms: Challenges and Novel Therapeutic Perspectives. Antibiotics (Basel). 2021 Jan 29;10(2):131.

Marques VF, Motta CC, Soares BD, Melo DA, Coelho SM, Coelho ID, Barbosa HS, Souza MM. Biofilm production and beta-lactamic resistance in Brazilian Staphylococcus aureus isolates from bovine mastitis. Braz J Microbiol. 2017 Jan-Mar;48(1):118-124.

Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A(2006) Detection of biofilm formation among the clinical isolates ofstaphylococci: an evaluation of three different screening methods.Indian J Med Microbiol 24:25–29.

Mlynek KD, Bulock LL, Stone CJ, Curran LJ, Sadykov MR, Bayles KW, Brinsmade SR. Genetic and Biochemical Analysis of CodY-Mediated Cell Aggregation in Staphylococcus aureus Reveals an Interaction between Extracellular DNA and Polysaccharide in the Extracellular Matrix. J Bacteriol. 2020 Mar 26;202(8):e00593-19.

MohammadiMollaahmadi C, Anzabi Y, Shayegh J. Comparison of the Frequency of Biofilm-Forming Genes (icaABCD) in Methicillin-Resistant S. aureus Strains Isolated from Human and Livestock. Arch Razi Inst. 2021 Dec 30;76(6):1655-1663.

Namvar A.E, Asghari B, Ezzatifar F, Azizi G, Lari A.R. Detection of the intercellular adhesion gene cluster (Ica) in clinical Staphylococcus aureus isolates. GMS Hyg. Infect. Control. 2013;8(1):Doc03.

Olaniyi R., Pozzi C., Grimaldi L., Bagnoli F. Staphylococcus aureus. Springer; Berlin/Heidelberg, Germany: 2016. Staphylococcus aureus-associated skin and soft tissue infections: Anatomical localization, epidemiology, therapy and potential prophylaxis; pp. 199–227.

Overgaauw PAM, Vinke CM, Hagen MAEV, Lipman LJA.A One Health Perspective on the Human-Companion Animal Relationship with Emphasis on Zoonotic Aspects. Int J Environ Res Public Health. 2020;17(11):3789.

Pedersen R.R., Krömker V., Bjarnsholt T., Dahl-Pedersen K., Buhl R., Jørgensen E. Biofilm Research in Bovine Mastitis. Front. Vet. Sci. 2021;8:449.

Peeters E., Nelis H.J., Coenye T. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J. Microbiol. Methods. 2008;72:157–165.

Schmidt P.L. Companion animals as sentinels for public health. Vet. Clin.N. Am. Small Anim. Pract. 2009;39:241–250.

Silva V, Correia E, Pereira JE, González-Machado C, Capita R, Alonso-Calleja C, Igrejas G, Poeta P. Biofilm Formation of Staphylococcus aureus from Pets, Livestock, and Wild Animals: Relationship with Clonal Lineages and Antimicrobial Resistance. Antibiotics (Basel). 2022 Jun 4; 11(6):772.

Szczuka E, Jabłońska L, Kaznowski A. Coagulase-negative staphylococci: pathogenesis, occurrence of antibiotic resistance genes and in vitro effects of antimicrobial agents on biofilm-growing bacteria. J Med Microbiol.2016; 65:1405–13.