Biofilm Formation and Extended Spectrum Beta Lactamase (ESBL) Production of Bacterial Strains Isolated From Sewage Water

Abstract

The production extended spectrum beta lactamase (ESBL) and metallo-beta lactamase of strains grown biofilm that were isolated from Kilis State Hospital sewage water were investigated in this study. Congo red agar, tube adherence, tissue culture plate tests and SEM analysis were performed for the detection of biofilm production. The extended spectrum beta lactamase and metallo-beta lactamase expression of biofilm producing bacteria was tested by using Kirby-Bauer disc diffusion method according to CLSI guidelines. 5 clinical isolates were revealed to be biofilm producer. From of these isolates, Enterococcus spp. (C2) and E. coli strains showed strong biofilm producing. One of Enterococcus isolates, SEM micrograph of C1 was viewed the weak adherence on glass surface. Only one from 5 isolates, Enterococcus spp. (C2) strain was determined to synthesis ESBL enzyme. None of strains was observed to not produce metallo-beta lactamase. ESBL and biofilm production provide an important pathogenic character allowing protection against antibiotic treatments to Enterococcus spp. (C2).

Keywords

• Biofilm,ESBL,metallo-beta lactamase,SEM

Atık Sudan İzole Edilen Bakteri Suşlarının Biyofilm Oluşumu ve Genişletilmiş Spektrum Beta Laktamaz (ESBL) Üretimi

Öz

Bu çalışmada Kilis Devlet Hastanesi kanalizasyon suyundan izole edilen biyofilm oluşturan suşların genişlemiş spektrumlu beta laktamaz ve metallo-beta laktamaz üretimi incelenmiştir. Biyofilm üretiminin belirlenmesi için kongo kırmızısı agar, tüp adherens, doku kültürü plak testleri ve SEM analizi uygulanmıştır. Biyofilm üreten suşların genişlemiş spektrumlu beta laktamaz ve metallo-beta laktamaz ekspresyonu CLSI yönetmeliğine göre Kirby-Bauer disk difüzyon metodu kullanılarak test edilmiştir. 5 klinik izolatın biyofilm üreticisi olduğu belirlenmiştir. Bu izolatlardan Enterococcus spp. (C2) ve E. coli suşları güçlü biyofilm üretmiştir. Enterococcus izolatlarından C1’in SEM mikrografında cam yüzeye zayıf tutunma gözlenmiştir. 5 izolattan yalnızca biri, Enterococcus spp.’nin (C2) GSBL enzimi sentezlediği ortaya konmuştur. Suşlardan hiçbirinin metallo-beta laktamaz üretmediği belirlenmiştir. GSBL ve biyofilm üretimi Enterococcus spp. (C2) suşuna antibiyotik tedavilerine karşı koruyarak önemli bir patojenik karakter sağlamaktadır.

Anahtar Kelimeler

• Biyofilm,GSBL,metallo-beta laktamaz,SEM

Kaynakça

1. Referans1 Adamus-Białek, W., Kubiak, A., & Czerwonka, G. (2015). Analysis of uropathogenic Escherichia coli biofilm formation under different growth conditions. Acta Biochimica Polonica, 62 (4), 765-771. https://doi.org/10.18388/abp.2015_1127.
2. Referans2 Akyıldız, S. (2015). Investigation of Biofilm Formation in the Food-Borne Salmonella Isolates. Dissertation, Gazi University.
3. Referans3 Asati, S., & Chaudhary, U. (2018). Evaluation of Two Methods of Biofilm Detection in Members of Family Enterobacteriaceae. Journal of Clinical and Diagnostic Research, 12(7), 20-23. https://doi.org/10.7860/JCDR/2018/34795.11827.
4. Referans4 Barnhart, M.M., & Chapman, M.R. (2006). Curli Biogenesis and Function. Annual Review of Microbiology, 60, 131–147. https://doi.org/10.1146/annurev.micro.60.080805.142106.
5. Referans5 Bogino, P.C., Oliva Mde, L., Sorroche, F.G., & Giordano, W. (2013). The Role of Bacterial Biofilms and Surface Components in Plant-Bacterial Associations. International Journal of Molecular Sciences, 14(8), 15838-15859. https://doi.org/10.3390/ijms140815838.
6. Referans6 Christensen, G.D., Simpson, W.A., Bisno, A.L., & Beachey, E.H. (1982). Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces. Infection and Immunity, 37(1), 318-26.
7. Referans7 Lima, J.L.D.C, Alves, L.R., Paz, J.N.P.D, Rabelo, M.A., Maciel, M.A.V., & Morais, M.M.C. (2017). Analysis of biofilm production by clinical isolates of Pseudomonas aeruginosa from patients with ventilator-associated pneumonia. Revista Brasileira de Terapia Intensiva, 29(3), 310-316. https://doi.org/ 10.5935/0103-507X.20170039.
8. Referans8 Darwish, S.F., & Asfour, H.A.E. (2013). Investigation of Biofilm Forming Ability in Staphylococci Causing Bovine Mastitis Using Phenotypic and Genotypic Assays. The Scientific World Journal 13, 1-9. http://dx.doi.org/10.1155/2013/378492.

9. Referans10 Dias, C., Borges, A., Oliveira, D., Martinez-Murcia, A., Saavedra, M.J., & Simões, M. (2018). Biofilms and antibiotic susceptibility of multidrug-resistant bacteria from wild animals. Peer J, 6:e4974; https://doi.org/10.7717/peerj.4974.
10. Referans11 Dufour, D., Leung, V., & Lévesque, C.M. (2012). Bacterial biofilm: structure, function, and antimicrobial resistance. Endodontic Topics, 22, 2–16. https://doi.org/10.1111/j.1601-1546.2012.00277.x
11. Referans12 Dumaru, R., Baral, R., & Shrestha, L.B. (2019). Study of biofilm formation and antibiotic resistance pattern of gram‑negative Bacilli among the clinical isolates at BPKIHS, Dharan. BMC Research Notes, 12(1): 38. http://dx.doi.org/10.1186/s13104-019-4084-8.
12. Referans13 Hassan, A., Usman, J., Kaleem, F., Omair, M., Khalid, A., & Iqbal, M. (2011). Evaluation of different detection methods of biofilm formation in the clinical isolates. Brazilian Journal of Infectious Diseases, 15(4), 305-311.
13. Referans14 Ivana, C., Marija, S., Jovanka, L. Bojana, K., Nevena, B., Dubravka, M., & Ljiljana, S. (2015). Biofilm Forming Ability of Salmonella enteritidis in vitro. Acta Veterinaria-Beograd, 65(3), 1-19. https://doi.org/10.1515/acve-2015-0031.
14. Referans15 Jamal, M., Tasneem, U., Hussain, T., & Andleeb, S. (2015). Bacterial Biofilm: Its Composition, Formation and Role in Human Infections. Research & Reviews: Journal of Microbiology and Biotechnology, 4(3), 1-14.
15. Referans16 Kaiser, T.D., Pereira, M.E., dos Santos, K.R., Maciel. E.L., Schuenck, P.R., & Nunes, A.P. (2013). Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis. Diagnostic Microbiology and Infectious Disease, 75(3), 235-239. https://doi.org/10.1016/j.diagmicrobio.2012.11.014.
16. Referans17 Karaca, B. (2011). Identification Of Biofilm Forming Abilities Of Salmonella Strains Isolated From Turkey. Dissertation, Ankara University.
17. Referans18 Kırmusaoglu, S. (2017). The comparison of methods used for the detection of biofilm formation that cause antibiotic resistance of Staphylococcus epidermidis and Staphylococcus aureus. Ortadogu Medical Journal, 9 (1): 28-33. https://doi.org/10.21601/ortadogutipdergisi.299940.
18. Referans19 Marić, S., & Vraneš, J. (2007). Characteristics and significance of microbial biofilm formation. Periodicum Biologorum, 109(2), 115-121.
19. Referans20 Mathur, T., Singhal, S., Khan, S., Upadhyay, D.J., Fatma, T., & Rattan, A. (2006). Detection of Biofilm Formation Among the Clinical Isolates of Staphylococci: An Evaluation of Three Different Screening Methods. Indian Journal of Medical Microbiology, 24(1), 25-29.
20. Referans21 Mohamed, A., Rajaa, A.M., Khalid, Z., Fouad, M., & Naima, R. (2016). Comparison of Three Methods for the Detection of Biofilm Formation by Clinical Isolates of Staphylococcus aureus Isolated in Casablanca. International Journal of Science and Research, 5(10), 1156-1159. https://doi.org/1159. 10.21275/ART20162319.
21. Referans22 Nachammai, S.M., Jayakumar, K., & Aravazhi, A.N. (2016). The effectiveness of antibiotics against a major uropathogen- Escherichia coli and its biofilm assay by phenotypic methods. International Journal of Research in Medical Sciences, 4(11), 4820-4828. http://dx.doi.org/10.18203/2320-6012.ijrms20163773.
22. Referans23 Ozdemir, F., & Arslan, S. (2018). Biofilm Production and Antimicrobial Susceptibility Profiles of Bacillus spp. from Meats. Sakarya University Journal of Science, 22(6), 1674-1682. https://doi.org/10.16984/saufenbilder.395016.
23. Referans24 Sala, L., Morar, A., Colibar, O., & Morvay, A.A. (2012). Antibiotic resistance of gram negative bacteria isolated from meat surface biofilm. Romanian Biotechnology Letters, 17(4), 7483-7492.
24. Referans25 Saxena, N., Maheshwari, D., Dadhich, D., & Singh, S. (2014). Evaluation of Congo Red Agar For Detection of Biofilm Production by Various Clinical Candida Isolates. Journal of Evolution of Medical and Dental Sciences, 3(59), 13234-13238. https://doi.org/10.14260/jemds/2014/3761.
25. Referans26 Sharma, G., Sharma, S., Sharma, P., Chandola, D., Dang, S., Gupta, S., & Gabrani, R. (2016). Escherichia coli biofilm: development and therapeutic strategies. Journal of Applied Microbiology, 121(2), 309-319. https://doi.org/10.1111/jam.13078.
26. Referans27 Temel, A., & Erac, B. (2018). Bacterial Biofilms: Detection Methods and Role in Antibiotic Resistance. Turk Mikrobiyoloji Cemiyeti Dergisi, 48(1):1-13. https://doi.org/10.5222/TMCD.2018.00.
27. Referans28 Zheng, J.X., Bai, B., Lin, Z.W., Pu, Z.Y., Yao, W.M., Chen, Z., Li, D.Y., Deng, X.B., Deng, Q.W., & Yu, Z.J. (2018). Characterization of biofilm formation by Enterococcus faecalis isolates derived from urinary tract infections in China. Journal of Medical Microbiology, 67, 60-67. https://doi.org/10.1099/jmm.0.000647.