Bazı Bakteriyel Balık Patojenlerinde Biyofilm Oluşumunun Farklı İn Vitro Metodlarla Tespiti

Yıl 2019, Cilt: 15 Sayı: 3, 378 - 390, 15.09.2019

Fethi Filik , Ayşegül Kubilay

https://doi.org/10.22392/actaquatr.537796

Cited By: 5

Öz

Bu
çalışmada, fenomen bir virülens faktörü olan biyofilmin bazı bakteriyel balık
patojenlerinden Staphylococcus warneri,
Aeromonas sobria, Yersinia ruckeri, Flavobacterium psychrophilum, Vibrio
anguillarum bakterileri tarafından üretilmesi tüp, mikroplak ve kongo red
agar yöntemleriyle araştırılmıştır.

Tüp yöntemi ile biyofilm oluşum
dereceleri tüpün iç yüzeyinde oluşan film tabakasının kalınlığına göre sırayla F. psychrophilum’da (++), A. sobria’da
(++), S. warneri’de (+), Y. ruckeri’de (-), V. anguillarum’da (-) olarak tespit
edilmiştir. Mikroplaklarla yapılan biyofilm testinde plaklarda oluşan biyofilm
negatif kontrolde 0,048, S. warneri’de
0,121, A. sobria’da 0,158, Y. ruckeri’de 0,071, F. psychrophilum’da 0,172, V.
anguillarum’da 0,212 absorbans değeri tespit edilmiştir. Mikroplak testinde
biyofilm oluşumu sırayla V. anguillarum,
F. psychrophilum, A. sobria, S. warneri, Y. ruckeri olarak
belirlenmiştir. Kongo red agar testinde biyofilm (slime) oluşturan bakterilerin
koloni morfolojisinin renk değiştirmesine göre biyofilm oluşumu sırayla V. anguillarum, A. sobria, S. warneri,
F. psychrophilum, Y. ruckeri olarak belirlenmiştir. Kongo red agarda kuru
kristalize siyah koloni üreterek en güçlü biyofilm oluşumu V. anguillarum, A. sobria, S. warneri’de bulunmuştur. Pembe
renkli koloni üreten F. psychrophilum ve
Y. ruckeri bakterileri ise bu teste
biyofilm oluşumu göstermemiştir.

     Sonuç
olarak; biyofilm kaynaklı enfeksiyonlar balıklar üzerinde ölümcül etki
göstermektedir. Bu etkilerin araştırılması ve durdurulması balık ölümlerinin
azaltılmasında önemlidir. Bakteriyel balık patojenlerinden olan S. warneri, A. sobria, V.
anguillarum, F. psychrophilum, Y.
ruckeri’nin araştırmamızda oldukça güçlü biyofilm oluşturdukları farklı
yöntemlerle tespit edilmiştir. 

Anahtar Kelimeler

Bakteriyel balık patojenleri, biyofilm, tüp yöntemi, mikroplak yöntemi, kongo red agar yöntem

Kaynakça

• Abdallah, F.B., Chaieb, K., Zmantar, T., Kallel, H. & Bakhrouf, A. (2009). Adherence assays ve slime production of Vibrio alginolyticus ve Vibrio parahaemolyticus. Brazilian Journal of Microbiology, 40(2), 394-398.
• Arciola, C.R., Campoccia, D., Ravaioli, S. & Montanaro, L. (2015). Polysaccharide intercellular adhesin in biofilm: structural ve regulatory aspects. Frontiers in Cellular and Infection Microbiology, 5,7, doi: 10.3389/fcimb.2015.00007.
• Arciola, C.R., Campoccia, D., Gamberini, S., Cervellati, M., Donati, E. & Montanaro, L. (2002). Detection of Slime Production by Means of an Optimised Congo Red Agar Plate Test Based on a Colourimetric Scale in Staphylococcus epidermidis. Clinical İsolates Genotyped for İca Locus, Biomaterials, 23, 4233-4239.
• Aslan, H. (2015). Cveida Türlerinde Biyofilm Oluşumu ve Antifungal. T.C. Hacettepe Üniversitesi Sağlık Bilimleri Enstitüsü Mikrobiyoloji Programı, Yüksek Lisans Tezi, Ankara.
• Bjarnsholt, T. (2013). The role of bacterial biofilms in chronic infections. APMIS Supplement Acta Pathologica, Microbiologica Et Immunologica Scandinavica, 136, 1-51.
• Bryers, J.D. (2008). Medical biofilms. Biotechnology ve bioengineering. 100(1), 1-18, PubMed PMID: 18366134.
• Černohorská, L. & Votava M. (2004). Determination of Minimal Regrowth Concentration (MRC) in Clinical Isolates of Various Biofilm-Forming Bacteria. Folia Microbiologica, 49 (1), 75-78.
• Christensen, G.D., Simpson, W.A., Younger, J.A., Baddour, L.M., Barrett, F.F. & Melton, D.M. vd. (1985). Adherence of coagulase negative staphylococci to plastic tissue cultures: A quantitative model for the adherence of staphylococci to medical devices. Journal of Clinical Microbiology, 22 (6), 996-06.
• Coquet, L., Cosette, P., Junter, G. A., Beucher, E., Saiter, J. M. & Jouenne, T. (2002). Adhesion of Yersinia ruckeri to fish farm materials: influence of cell ve material surface properties. Colloids and surfaces B: Biointerfaces, 26 (4), 373-378.
• Coquet, L., Cosette, P., Quillet, L., Petit, F., Junter, G.A. & Jouenne, T. (2002). Occurrence ve Phenotypic Characterization of Yersinia ruckeri Strains with Biofilm-Forming Capacity in a Rainbow Trout Farm. Applied and Environmental Microbiology, 68(2), 470-475.
• Çiftçi, Z. (2005). Kronik Tonsillitte Biofilmin Rolü. T.C. Taksim Eğitim ve Araştırma Hastanesi KBB Kliniği, Uzmanlık Tezi, 69s, İstanbul.
• Davies, D. (2003). Understveing biofilm resistance to antibacterial agents. Nature Reviews Drug Discovery, 2(2), 114-22.
• Deighton, M.A., Capstick, J., Domalewski, E. & Van Nguyen, T. (2001). Methods for studying biofilms produced by Staphylococcus epidermidis Methods in Enzymology, 336, 177-95.
• Delshad, S.T., Soltanian, S., Sharifiyazdi, H. & Bossier, P. (2018). Effect of quorum quenching bacteria on growth, virulence factors and biofilm formation of Yersinia ruckeri in vitro and an in vivo evaluation of their probiotic effect in rainbow trout. Journal of Fish Diseases, https://doi.org/10.1111/jfd.12840.
• Demir, C. & İnanç, B.B. (2015). Investigate Nasal Colonize Staphylococcus Species Biofilm Produced (Nazal Kolonize Stafilokok Türlerinde Biyofilm Üretiminin Araştırılması). Journal of Clinical and Analytical Medicine, 414-418.
• Donlan, R.M. & Costerton, W.J. (2002). Biofilms: Survival mechanisms of clinically relevant microorganisms. Clinical Microbiology Reviews, 15(2), 167-93.
• Donlan, R.M. (2002). Biofilms: microbial life on surfaces. Emerg Infect Dis, 8(9), 881-890.
• Freeman, D.J., Falkiner, F.R. & Keane, C.T. (1989). New method for detecting slime production by coagulase negative staphylococci. Journal of Clinical Pathology 1989, 42(1), 872-4.
• Harraghy, N., Seiler, S., Jacobs, K., Hannig, M., Menger, M.D. & Herrmann, M. (2006). Advances in in vitro ve in vivo models for studying the staphylococcal factors involved in implant infections. The International Journal Artificial Organs. 29, 368-78.
• Högfors‐Rönnholm, E., Norrgård, J. & Wiklund, T. (2015). Adhesion of Smoothan Drough Pheno types of Flavobacterium psychrophilum to Poly Styrene Surfaces, Journal of Fish Diseases, 38(5), 429-437.
• Jahid, I.K., Lee, N.Y., Kim, A. & Ha, S.D. (2013). Influence of glucose concentrations on biofilm formation, motility, exoprotease production, ve quorum sensing in Aeromonas hydrophila. Journal of Food Protection, 76(2), 239-247.
• Jurcisek, J.A. & Bakaletz, L.O. (2007). Biofilms formed by nontypeable Haemophilus influenzae In vivo contain both double-strveed DNA ve type IV pilin protein. Joutnal of Bacteriology, 189(10), 3868-75.
• Kirov, S.M., Castrisios, A. & Shaw, J.G. (2003). Aeromonas Flegella (Polar ve Lateral) are Enterocyte Adhesins That Contribute to Biofilm Formation on Surfaces. American Society for Micrbiology, 72, 1939-45.
• Levipan, H.A. & Avendaño-Herrera, R. (2017). Different phenotypes of mature biofilm in Flavobacterium psychrophilum share a potential for virulence that differs from planktonic state. Frontiers in Cellular and Infection Microbiology, www.frontiersin.org, 7(76), doi: 10.3389/fcimb.2017.00076.
• Madetoja, J. (2002). Flavobacterium psychrophilum: Characterisation, Experimental Transmission and Ocuurence in Fish and Fish Farming Enviroments. Academic Dissertation, Finland, 1-37.
• 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 Medical Microbiology, 24, 25-9.
• Molobela, I. P., Cloete, T.E. & Beukes, M. (2010). Protease ve amylaseenzymes for biofilm removal vedegradation of extracellular polymericsubstances (EPS) produced by Pseudomonas fluorescens bacteria. African Journal of Microbiology Research, 4(14), 1515-1524.
• Monroe, D. (2007). Looking for Chinks in the Armor of Bacterial Biofilms. PLoS Biology, 5,11. Erişim tarihi: 14.10.2018. http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.0050307
• Mortensen, B.K. (2014). Problems caused by biofilms. Bactoforce International A/S. http://www.bactoforce.se/wp-content/uploads/2014/09/Problems-caused-by biofilms.pdf.
• Nurcan, N. (2010). Bazı Gram-Negatif Bakteriyel Balık Patojenlerinde Çevreyi Algılama Sisteminin İncelenmesi. Süleyman Demirel Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi.
• Ormancı, S. (2010). Hayvansal ve Çevresel Kaynaklardan İzole Edilen Hareketli Aeromonas Türlerinde Biyofilm Oluşumunun İncelenmesi. Gazi Üniversitesi, Yüksek Lisans Tezi.
• O’Toole, G.A. & Kolter, R. (1998). Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Molecular Microbiology, 28(3), 449-461.
• Post, J.C., Stoodley, P., Hall-Stoodley, L. & Ehrlich, G.D. (2004). The role of biofilms in otolaryngologic infections. Current Opinion Otolaryngology & Head Neck Surgery, Review, 12(3), 185-90.
• Rahman, M., Simm, R., Kader, A., Basseres, E., Römling, U. & Möllby, R. (2007). The role of c-di-GMP signalining an Aeromonas veronii biovar sobria strain. FEMS Microbiology Letters, 273, 172-179.
• Rendueles, O. & Ghigo, J.M. (2012). Multi-species biofilms: how to avoid unfriendly neighbors. FEMS Microbiology Reviews, 1-18, doi: 10.1111/j.1574-6976.2012.00328.x.
• Richards, J.J. & Melveer, C. (2009). Controlling bacterial biofilms. Chembiochem, 10(14), 2287-94.
• Stepanovic, S., Vukovic, D., Hola, V., Ventura, G.B., Djukic, S., Irkovic, I.C. & Ruzicka, F. (2007). Quantification of biofilm in microtiter plates: overview of testing conditions ve practical recommendations for assessment of biofilm production by staphylococci. Apmis ResearchGate, 115, 891-9.
• Stepanovic, S., Vukovic, D., Dakic I., Savic, B. & Svabic-Vlahovic, M. (2000). A modified microtiter-plate test for quantification of staphylococcal biofilm formation. Journal of Microbiology Methods, 40, 175-9.
• Sundell, K. & Wiklund, T. (2011). Effect of biofilm formation on antimicrobial tolerance of Flavobacterium psychrophilum. Journal of Fish Diseases, 34, 373-383 doi:10.1111/j.1365-2761.2011.01250.x.
• Szczuka, E. & Kaznowski, A. (2014). Antimicrobial activity of tigecycline alone or in combination with rifampin against Staphylococcus epidermidis in biofilm. Folia Microbiology (Praha), 59, 283-8, http://dx.doi.org/10.1007/s12223-013-0296-9.
• Wiklund, T., Kaas, K., Lönnström, L. & Dalsgaard, I. (1994). Isolation of Cytophaga psychrophila (Flexibacter psychrophilus) from wild and farmed rainbow trout (Oncorhynchus mykiss) in Finland. Bulletin European Associate Fish Pathology, 14, 44-46.
• Wu, H., Moser, C., Wang, H.Z., Hoiby, N. & Song, Z.J. (2015). Strategies for combating bacterial biofilm infections. International Journal of Oral Science, 7(1), 1-7.
• Yasuda, H., Ajiki, Y., Aoyama, J. & Yolota, I. (1994). Interaction between human polymorphonuclear leucocytes ve bacteria relased from In vitro bacteriol biofilms models. Journal of Medical Microbiology, 41, 359-67.
• Ye, J., Ma, Y., Liu, Q., Zhao, D.L., Wang, Q.Y. & Zhang, Y.X. (2008). Regulation of Vibrio alginolyticus virulence by the LuxS quorum-sensing system. Journal of Fish Diseases, 31, 161-169.
• Yildiz, F.H. & Visick, K.L. (2009). Vibrio biofilms: so much the same yet so different. Trends in Microbiology, 17(3), 109-118.