COMPARISON OF THE MICROTITER PLATE METHOD AND THE CONGO RED AGAR TECHNIQUE IN THE DETERMINATION OF STAPHYLOCOCCAL BIOFILM

Yıl 2024, Cilt: 10 Sayı: 1, 128 – 132, 30.06.2024

Sevda Er

https://doi.org/10.22531/muglajsci.1493167

Öz

Kolagülaz negatif stafiloloklar, insan florasında kommensal olarak yer alan fırsatçı patojenlerdir. Bu bakterilerin enfeksiyonlarının patogenezisinde bilinen en önemli virülans faktörlerinden biri biyofilm oluşumlarıdır. Biyofilm oluşumunun ortaya konmasında mikrotitre plaka yöntemi ve kongo kırmızısı agar tekniği yaygın bir şekilde kullanılmaktadır. Bu çalışmada, insan koagülaz negatif Staphylococcus spp. bakteri izolatlarının biyofilm oluşumlarının mikrotitre plaka yöntemi ve kongo kırmızısı agar tekniği ile karşılaştırılması amaçlanmıştır. Çalışmada, 41 insan koagülaz negatif stafilokok izolatından mikrotitre plaka yöntemine göre 35 tanesinin biyofilm oluşturmadığı, 6 izolatın zayıf derecede biyofilm oluşturduğu, kongo kırmızısı agar yüzeyinde ise hiçbir izolatın biyofilm oluşturmadığı sonucuna varılmıştır. Kongo kırmızısı agar tekniğinde sonucun yorumlanmasının güç ve gözleme dayalı olması sebebiyle, mikrotitre plaka yöntemi sonuçlarının daha güvenilir olduğu sonucuna varılmıştır. Literatürde koagülaz negatif stafilokokların biyofilm oluşumlarının mikrotitre plaka yöntemi ve kongo kırmızısı agar tekniğiyle karşılaştırıldığı neredeyse hiç çalışma olmaması sebebiyle, yapılan çalışma öncül çalışmalar arasında yer alıp, literatüre katkı sunacaktır.

Anahtar Kelimeler

Biyofilm, Koagülaz Negatif Stafilokok, Kongo Kırmızısı Agar Tekniği, Mikrotitre Plaka Yöntemi, Vajina

Proje Numarası

1710S548

Kaynakça

• Gonçalves, T.G., Timm, C.D., “Biofilm production by coagulase-negative Staphylococcus: a review”, Arq. Inst. Biol., 87, 1-9, 2020.
• Tenke, P., Koves, B., Nagy, K., Uehara, S., Kumon, H., Hultgren, S.J., et al., “Biofilm and Urogenital Infections”, In Clinical Management of Complicated Urinary Tract Infection, IntechOpen, 2011.
• Hardy, L., Cerca, N., Jespers, V., Vaneechoutte, M., Crucitti, T., “Bacterial biofilms in the vagina”, Research in microbiology, 168(9-10), 865-874, 2017.
• Qu, Y., Daley, A.J., Istivan, T.S., Garland, S.M., Deighton, M.A., “Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation”, Annals of clinical microbiology and antimicrobials, 9(1), 16, 2010.
• Arciola, C.R., Campoccia, D., Ravaioli, S., Montanaro, L., “Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects”, Frontiers in cellular and infection microbiology, 5, 7, 2015.
• Jain, A., Agarwal, A., “Biofilm production, a marker of pathogenic potential of colonizing and commensal staphylococci”, Journal of microbiological methods, 76(1), 88-92, 2009.
• Harris, L.G., Murray, S., Pascoe, B., Bray, J., Meric, G., Magerios, L., et al., “Biofilm morphotypes and population structure among Staphylococcus epidermidis from commensal and clinical samples”, PLoS One, 11(3), 1-15, 2016.
• Peters, J., Price, J., Liewelyn, M., “Staphylococcal and streptococcal infections”, Medicine, 45(12), 727-734, 2017.
• Otto, M., “Bacterial biofilms: Staphylococcal biofilms”, Ed: Tony Romeo. Springer, 227-228, 2008.
• Soumya, K.R., Jishma, P., Sugathan, S., Mathew, J., Radhakrishnan, E.K., “Biofilm Changes of Clinically Isolated Coagulase Negative Staphylococci, Proceedings of the National Academy of Sciences, India Section B: Biological Sciences, 90(1), 199-206, 2020.
• Melo, P.D.C., Ferreira, L.M., Nader Filho, A., Zafalon, L.F., Vicente, H.I.G., Souza, V.D., “Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis”, Brazilian Journal of Microbiology, 44, 119-124, 2013.
• Stepanovic, S., Vukovic, D., Dakic, I., Savic, B., Svabic-Vlahovic, M., “A modified microtiter-plate test for quantification of staphylococcal biofilm formation”, J. Microbiol. Methods., 40, 175–179, 2000.
• Kaiser, T.D.L., Pereira, E.M., Dos Santos, K.R.N., Maciel, E.L.N., Schuenck, R.P., Nunes, A.P.F., “Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis”, Diagnostic Microbiol Infect Dis, 75(3), 235–9, 2013.
• Büttner, H., Mack, D., Rohde, H., “Structural basis of Staphylococcus epidermidis biofilm formation: mechanisms and molecular interactions,” Frontiers in cellular and infection microbiology, 5: 14, 2015.
• Boynukara, B., Gulhan, T., Gurturk, K., Alisarli, M., Ogun, E., “Evolution of slime production by coagulase-negative staphylococci and enterotoxigenic characteristics of Staphylococcus aureus strains isolated from various human clinical specimens”, Journal of medical microbiology, 56(10), 1296-1300, 2007.
• Raimundo, O., Heussler, H., Bruhn, J.B., Suntrarachun, S., Kelly, N., Deighton, M.A., Garland, S.M., “Molecular epidemiology of coagulase-negative staphylococcal bacteraemia in a newborn intensive care unit”, Journal of Hospital Infection, 51(1), 33-42, 2002.
• Koksal, F., Yaşar, H., Samasti, M., “Antibiotic resistance patterns of coagulase-negative staphylococcus strains isolated from blood cultures of septicemic patients in Turkey”, Microbiological research, 164(4), 404-410, 2009.
• Özgüneş, İ., Yıldırım, D., Çolak, H., Durmaz, G., Usluer, G., Akgün, Y., “Koagülaz Negatif Stafilokokların Patojenitesi ve Antibiyotik Duyarlılığı ile Slime Pozitifliği Arasındaki İlişki”, Hastane İnfeksiyonları Dergisi, 4, 106-111, 2000.
• Garza-Gonzalez, E., Morfin-Otero, R., Llaca-Diaz, J.M., Rodriguez-Noriega, E., “Staphylococcal cassette chromosome mec (SCCmec) in methicillin-resistant coagulase-negative staphylococci. A review and the experience in a tertiary-care setting”, Epidemiology & Infection, 138(5): 645-654, 2010.
• Shakya, P., Nayak, A., Sharma, R.K., Singh, A.P., Singh, R.V., Jogi, J., et al., “Phenotypic detection and comparison of biofilm production in methicillin resistant Staphylococcus aureus”, The Pharma Innovation Journal, 11(3), 1352-1357, 2002.
• Al-Jubory, A., Essa, M. ., “Comparison of Three Biofilm Detection Methods in Coagulase Negative Staphylococci Species”, Rafidain Journal of Science, 30(2), 1-15, 2021.
• Reddy, P. S., “Biofilm detection and Clinical significance of Coagulase negative Staphylococci isolates in a tertiary care centre”, IOSR Journal of Dental and Medical Sciences, 20, 58-65, 2021.
• AL-Mojamaee, N. A. H., ALtaii, H. A. J., “Comparison of two methods for the detection of Pseudomonas aeruginosa biofilm formation isolated from different clinical samples”, Iragi Journal of Humanitarian, Social and Scientific Research, 11, 651-668, 2023.

COMPARISON OF THE MICROTITER PLATE METHOD AND THE CONGO RED AGAR TECHNIQUE IN THE DETERMINATION OF STAPHYLOCOCCAL BIOFILM

Yıl 2024, Cilt: 10 Sayı: 1, 128 – 132, 30.06.2024

Sevda Er

https://doi.org/10.22531/muglajsci.1493167

Öz

Coagulase-Negative Staphylococci are opportunistic pathogens that are commensal in human flora. One of the most important virulence factors known in the pathogenesis of infections of these bacteria is biofilm formation. The Microtiter Plate Method and The Congo Red Agar Technique are widely used to reveal biofilm formation. This study aims to compare human coagulase negative Staphylococcus spp. bacterial isolates, biofilm formations with the Microtiter Plate Method and Congo Red Agar Technique. In the study, it was concluded that 35 of 41 human coagulase negative staphylococcal isolates did not form biofilms according to the microtiter plate method, 6 isolates formed a weak biofilm, and none of the isolates formed a biofilm on the Congo Red Agar surface. It has been concluded that the results of the Microtiter Plate Method are more reliable, since the interpretation of the result in the Congo Red Agar Technique is difficult and subjective, based on observation. Since there are very few studies in the literature comparing the biofilm formation of coagulase negative staphylococci with the Microtiter Plate Method and the Congo Red Agar Technique, this study will be among the preliminary studies and will contribute to the literature.

Anahtar Kelimeler

Biofilm, Coagulase-Negative Staphylococci, Congo Red Agar Technique, Microtiter Plate Method, Vagina

Etik Beyan

Not applicable.

Destekleyen Kurum

Anadolu University

Proje Numarası

1710S548

Teşekkür

This study was supported within the scope of the project numbered 1710S548 accepted by Anadolu University Scientific Research Projects Commission. The author(s) would like to thank Prof. Dr. Merih Kıvanç for her endless support.

Kaynakça

• Gonçalves, T.G., Timm, C.D., “Biofilm production by coagulase-negative Staphylococcus: a review”, Arq. Inst. Biol., 87, 1-9, 2020.
• Tenke, P., Koves, B., Nagy, K., Uehara, S., Kumon, H., Hultgren, S.J., et al., “Biofilm and Urogenital Infections”, In Clinical Management of Complicated Urinary Tract Infection, IntechOpen, 2011.
• Hardy, L., Cerca, N., Jespers, V., Vaneechoutte, M., Crucitti, T., “Bacterial biofilms in the vagina”, Research in microbiology, 168(9-10), 865-874, 2017.
• Qu, Y., Daley, A.J., Istivan, T.S., Garland, S.M., Deighton, M.A., “Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation”, Annals of clinical microbiology and antimicrobials, 9(1), 16, 2010.
• Arciola, C.R., Campoccia, D., Ravaioli, S., Montanaro, L., “Polysaccharide intercellular adhesin in biofilm: structural and regulatory aspects”, Frontiers in cellular and infection microbiology, 5, 7, 2015.
• Jain, A., Agarwal, A., “Biofilm production, a marker of pathogenic potential of colonizing and commensal staphylococci”, Journal of microbiological methods, 76(1), 88-92, 2009.
• Harris, L.G., Murray, S., Pascoe, B., Bray, J., Meric, G., Magerios, L., et al., “Biofilm morphotypes and population structure among Staphylococcus epidermidis from commensal and clinical samples”, PLoS One, 11(3), 1-15, 2016.
• Peters, J., Price, J., Liewelyn, M., “Staphylococcal and streptococcal infections”, Medicine, 45(12), 727-734, 2017.
• Otto, M., “Bacterial biofilms: Staphylococcal biofilms”, Ed: Tony Romeo. Springer, 227-228, 2008.
• Soumya, K.R., Jishma, P., Sugathan, S., Mathew, J., Radhakrishnan, E.K., “Biofilm Changes of Clinically Isolated Coagulase Negative Staphylococci, Proceedings of the National Academy of Sciences, India Section B: Biological Sciences, 90(1), 199-206, 2020.
• Melo, P.D.C., Ferreira, L.M., Nader Filho, A., Zafalon, L.F., Vicente, H.I.G., Souza, V.D., “Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis”, Brazilian Journal of Microbiology, 44, 119-124, 2013.
• Stepanovic, S., Vukovic, D., Dakic, I., Savic, B., Svabic-Vlahovic, M., “A modified microtiter-plate test for quantification of staphylococcal biofilm formation”, J. Microbiol. Methods., 40, 175–179, 2000.
• Kaiser, T.D.L., Pereira, E.M., Dos Santos, K.R.N., Maciel, E.L.N., Schuenck, R.P., Nunes, A.P.F., “Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis”, Diagnostic Microbiol Infect Dis, 75(3), 235–9, 2013.
• Büttner, H., Mack, D., Rohde, H., “Structural basis of Staphylococcus epidermidis biofilm formation: mechanisms and molecular interactions,” Frontiers in cellular and infection microbiology, 5: 14, 2015.
• Boynukara, B., Gulhan, T., Gurturk, K., Alisarli, M., Ogun, E., “Evolution of slime production by coagulase-negative staphylococci and enterotoxigenic characteristics of Staphylococcus aureus strains isolated from various human clinical specimens”, Journal of medical microbiology, 56(10), 1296-1300, 2007.
• Raimundo, O., Heussler, H., Bruhn, J.B., Suntrarachun, S., Kelly, N., Deighton, M.A., Garland, S.M., “Molecular epidemiology of coagulase-negative staphylococcal bacteraemia in a newborn intensive care unit”, Journal of Hospital Infection, 51(1), 33-42, 2002.
• Koksal, F., Yaşar, H., Samasti, M., “Antibiotic resistance patterns of coagulase-negative staphylococcus strains isolated from blood cultures of septicemic patients in Turkey”, Microbiological research, 164(4), 404-410, 2009.
• Özgüneş, İ., Yıldırım, D., Çolak, H., Durmaz, G., Usluer, G., Akgün, Y., “Koagülaz Negatif Stafilokokların Patojenitesi ve Antibiyotik Duyarlılığı ile Slime Pozitifliği Arasındaki İlişki”, Hastane İnfeksiyonları Dergisi, 4, 106-111, 2000.
• Garza-Gonzalez, E., Morfin-Otero, R., Llaca-Diaz, J.M., Rodriguez-Noriega, E., “Staphylococcal cassette chromosome mec (SCCmec) in methicillin-resistant coagulase-negative staphylococci. A review and the experience in a tertiary-care setting”, Epidemiology & Infection, 138(5): 645-654, 2010.
• Shakya, P., Nayak, A., Sharma, R.K., Singh, A.P., Singh, R.V., Jogi, J., et al., “Phenotypic detection and comparison of biofilm production in methicillin resistant Staphylococcus aureus”, The Pharma Innovation Journal, 11(3), 1352-1357, 2002.
• Al-Jubory, A., Essa, M. ., “Comparison of Three Biofilm Detection Methods in Coagulase Negative Staphylococci Species”, Rafidain Journal of Science, 30(2), 1-15, 2021.
• Reddy, P. S., “Biofilm detection and Clinical significance of Coagulase negative Staphylococci isolates in a tertiary care centre”, IOSR Journal of Dental and Medical Sciences, 20, 58-65, 2021.
• AL-Mojamaee, N. A. H., ALtaii, H. A. J., “Comparison of two methods for the detection of Pseudomonas aeruginosa biofilm formation isolated from different clinical samples”, Iragi Journal of Humanitarian, Social and Scientific Research, 11, 651-668, 2023.

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