Pseudomonas fluorescens BALIK PATOJENİNDE ÇEVREYİ ALGILAMA SİSTEMİ VE VİRÜLENS FAKTÖRLERİ

Year 2016, Volume: 31 Issue: 2, 68 - 78, 03.12.2016

Nurdan Nurcan Ayşegül Kubilay

Abstract

Araştırmada Gram negatif bakteriyel balık
patojenlerinden Pseudomonas fluorescens’in üç adet suşu
kullanılmıştır. Suşlar da öncelikle N-butanoyl-L-homoserin lakton (BHL)
ve N-(3-okzododekanoyl)-L-homoserin lakton (OdDHL) sinyal moleküllerinin
üretimi, Chromobacterium violaceum CV026 ve Agrobacterium tumafeciens
NT1 biyosensör suşları kullanılarak test edilmiştir. Pseudomonas aeruginosa
PAO1 suşu pozitif kontrol olarak kullanılmıştır. Sonuç olarak, P. fluorescens suşları, C.
violaceum CV026 suşu kullanıldığında BHL sinyal molekülünü üretmediği,
bununla birlikte, A. tumafeciens NT1 kullanıldığında, P. fluorescens
suşlarının OdDHL sinyal molekülünü ürettiği tespit edilmiştir. P. fluorescens
suşları; biyofilm, pigment, ramnolipid, elastaz, amilaz, hemoliz ve proteaz
aktiviteleri gibi virülens faktörlerini üretmiştir. 

Keywords

P. fluorescens, Çevreyi algılama sistemi, Virülens faktörleri, Biyofilm, Pigment, Ramnolipid

QUORUM SENSING SYSTEM AND VIRULENCE FACTORS IN Pseudomonas fluorescens FISH PATHOGEN

Abstract

In this study three Pseudomonas fluorescens strains, Gram-negative fish
pathogen were used.  Previously N-butanoyl-L-homoserine
lacton (BHL) and N-(3
okzododekanoyl)-L-homoserine lacton (OdDHL) signal molecules of production of strains were
examined using biosensor strains, Choromobacterium violaceum CV026
strain and Agrobacterium tumefaciens NT1. Pseudomonas
aeruginosa PAO1 strain used to as positive control. As a results of this study; P. fluorescens strains did not produce BHL signal molecules using C.
violaceum CV026.  However, the
strains produced OdDHL signal molecules using A. tumafeciens NT1. It was
determined production
of biofilm, pigment, rhamnolipid, elastase,
protease, amilase and hemolysis activity
as phenotype tested as virulence factors
in the strains.  

Keywords

P. fluorescens, Quorum sensing system, Virulence factors, Biofilm, Pigment, Rhamnolipid

References

• Ağalar, C., (2011). Yabancı Cisimlerde Biyofilm Gelişimi Engellenebilir Mi? Bir Multidisipliner Araştırma Grubu Modeli, ANKEM Derg 2011;25(2):125-129 26. ANKEM Antibiyotik ve Kemoterapi Kongresi Kahvaltılı Oturum-4 sunumundan (18-22 Mayıs 2011, Kızılağaç/Manavgat) 125 doi: 10.5222/ankem.2011.125
• Aleksandra, D., (2010). The Dawn of the Living, Talking Robots. The Journal of Young Investigators: An Undergraduate, Peer-Reviewed Science Journal, 19, 22, http://www.jyi.org. http://www.jyi.org/features/ft.php?id=285. Erişim Tarihi: 03.05.2010.
• Allison, D.G., Ruiz, B., SanJose, C., Jaspe, A., & Gilbert, P. (1998). Extracellular products as mediators of the formation and detachment of Pseudomonas fluorescens biofilms. FEMS Microbiology Letters, 167, 179-184.
• Akan, E., & Kınık, A. (2014). Biyofilm Oluşum Mekanizması ve Biyofilmlerin Gıda Güvenliğine Etkisi, Gıda ve Yem Bilimi - Teknolojisi Dergisi / Journal of Food and Feed Science – Technology, 14, 42-51.
• Arda, M., Seçer, S., & Sarıeyyüpoğulları, M. (2002). Balık Hastalıkları. Medisan Yayın Serisi 56. Ankara.
• Arda, M. (1997). Temel Mikrobiyoloji. Medisan Yayınevi, Medisan Yayın Serisi No:25, 490. Ankara.
• Ausubel, F., Brent, R., Kingston, R., Moor, D., Seidman, J., Smith, J., & Stauhle, K. (1988). Current Protocols in Molecular Biology. New York: Wiley Intersciences.
• Bruhn, J.B, Dalsgaard, I., Nielsen, K.F., Buchholtz, C., Larsen, J.L., & Gram L. (2005). Quorum sensing signal molecules (acylated homoserine lactones) in Gram-negative fish pathogenic bacteria. Diseases of Aquatic Organisms, (65), 43-52.
• Camara, M. (2007). Quorum sensing: A cell-cell signalling mechanism used to coordinate behavioural changes in bacterial populations. http://wmc7.liacs.nl/proceedings/WMC7Camara.pdf
• Cha, C., Gao, P., Chen, Y.C., Shaw, P.D., & Farrand, S.K. (1998). Production of acylhomoserine lactone signals by gram-negative plant-associated bacteria. Molecular Plant-Microbe Interactions, 11, 1119-1129.
• Chu, T., Ni, C., Zhang, L., Wang Q., Xiao J., Zhang, Y., & Liu, Q. (2015). A quorum sensing-based in vivo expression system and its application in multivalent bacterial vaccine. Microbial Cell Factories, 14, 37.
• Croxatto, A., Chalker, V.J., Lauritz, J., Jass, J., Hardman, A., Williams, P., Camara, M., & Milton D.L. (2002). VanT, a Homologue of Vibrio harveyi LuxR, Regulates Serine, Metalloprotease, Pigment and Biofilm Production in Vibrio anguillarum. Journal of Bacteriology, 184(6), 1617-1629.
• Deepa, S., Venkateswaran, P., Vinithkumar N. V., & Kirubagaran R. (2014). Prevention of Acyle Homoserine Lactone (AHL) mediated biofilm Formation by selected flora of Andaman & Nicobar Island. Indian Journal of Geo-Marine Sciences, 43, 11.
• Defoirdt, T., Boon, N., Bossier, P., & Verstraete, W. (2004). Disruption of bacterial quorum sensing: an unexplored strategy to fight infections in aquaculture. Aquaculture, 240, 69-88.
• Delille, A., Quile`s, F., & Humbert, F. (2007). In Situ Monitoring of the Nascent Pseudomonas fluorescens Biofilm Response to Variations in the Dissolved Organic Carbon Level in Low-Nutrient Water by Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy, Applied and Environmental Microbiology, 5782-5788.
• Dong, Y., Zhang, X., Soo, H.L., Greenberg, P., & Zhang, L. (2005). The two-component response regulator PprB modulates quorum-sensing signal production and global gene expression in P. aeruginosa, Molecular Microbiology, 56, 1287-1301.
• Ersoy, E., Karaboz, İ., Sukatar, A., Aldağ, D.S., Sinecan, E., & Güner, G. (2005). Biyolüminesent Photobacterium leiognathi TEM04S2 Straini ile Bakterilerde Quorum Sensing 'in Kanıtlanması, Orlab On-Line Mikrobiyoloji Dergisi, 3(2), 1-6.
• Federle, M.J., & Bassler, B. (2003). Interspecies communication in bacteria. The Journal of Clinical Investigation, 112(9), 1291-1299.
• García-Aljaro, C., Melado-Rovira, S., Milton D.L., & Blanch A.R. (2012). Quorum-sensing regulates biofilm formation in Vibrio scophthalmi, BMC Microbiology, 12: 287.
• Gatesoupe, F.J. (1999). The use of probiotics in aquaculture. Aquaculture, 180, 147-165.
• Göze, E.Y. (2009). Çeşitli Bitki Ekstraklarının Pseudomonas aeruginosa’da Çevreyi Algılama Sistemi Üzerine Etkilerinin İncelenmesi. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, 64s, Isparta.
• Greenberg, E.P. (2003). Bacterial communication and group behavior. The Journal of Clinical Investigation, 112(9), 1288-1290.
• Gudding, R., Lillehaug, A., & Evensen, Ø., (1999). Recent developments in fish vaccinology. Veterinary Immunology and Immunopathology, 72, 203-212.
• Gün, İ., & Ekinci, F.Y. (2009). Biyofilmler: Yüzeylerdeki Mikrobiyal Yaşam. Gıda, Derleme/Review, 34(3), 165-173.
• Heppell, J., & Davis, H.L. (2000). Application of DNAvaccine technology to aquaculture. Advanced Drug Delivery Reviews, 43, 29-43.
• Karaboz, İ., & Sukatar, A. (2004). Bakterilerde Sosyal Davranışlar (Bakterilerde İletişim Mekanizmaları) Orlab On-Line Mikrobiyoloji Dergisi, 2(5), 23-32.
• Kjelleberg, S., Steinberg, P., Givskov, M., Gram, L., Manefield, M., & De Nys, R. (1997). Do marine natural products interfere with prokaryotic AHL regulatory systems?. Aquatic Microbial Ecology, 13, 85-93.
• McClean, K.H., Winson, M.K., Fish, L., Taylor, A., Chhabra, S.R., Camara, M., Daykin, M., Lamb, J.H., Swift, S., Bycroft, B.W., Stewart, G.S.A.B., & Williams, P. (1997). Quorum sensing and Chromobacterium violeceum: exploitation of violacein production for the detection of N-acylhomoserine lactones. Microbiology, 143, 3703-3711.
• Mıdık, F., Tokatlı, M., & Özçelik, F. (2011). Laktik asit bakterileri tarafından üretilen ekzopolisakkaritler ve üretimini etkileyen faktörler. Syf: 106.
• Moghaddam, M.M., Khodi, S., & Mirhosseini, A. (2014). Quorum Sensing in Bacteria and a Glance on Pseudomonas aeruginosa. Clinical Microbiology: Open Access, 3, 4, ISSN: 2327-5073.
• Molobela, I.P., Cloete, T.E., & Beukes M. (2010). Protease and amylase enzymes for biofilm removal and degradation of extracellular polymeric substances (EPS) produced by Pseudomonas fluorescens bacteria., African Journal of Microbiology Research Vol. 4(14), 1515-1524.
• Myszka, K. & Czaczyk, K. (2012). N-Acylhomoserine Lactones (AHLs) as Phenotype Control Factors Produced by Gram – Negative Bacteria in Natural Ecosystems. Polish Journal of Environmental Studies, 21, 1, 15-21.
• Newman, S.G. (1993). Bacterial vaccines for fish. Annual Review of Fish Diseases, 3, 145-185.
• Ohman, D.E., Cryz, S.J., & Iglewski, B.H. (1980). Isolation and characterization of a P. aeruginosa PAO mutant that produces altered elastase. Journal of Bacteriology, 142, 836-842.
• O’Toole, G.A. & Kolter R. (1998). Initiation of biofilm formation in Pseudomonas fluorescensWCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Molecular Microbiology, 28(3), 449-461.
• Ravn, L., Christensen, A.B., Molin, S., Givskov, M., & Gram, L. (2001). Methods for acylated homoserine lactones produced by Gram-negative bacteria and their application in studies of AHL-production kinetics. Journal of Microbiological Methods, 44, 239-251.
• Sakai, M. (1999). Current status of fish immunostimulants. Aquaculture, 172, 63-92.
• Shaw, P.D., Ping, G., Daly, S.L., Cha, C., Cronan, J.E., JR., Rınehart, K.L.,& Farrand, S.K. (1997). Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography, Proceedings of the National Academy of Sciences USA,94, 6036-6041.
• Swift, S., Lynch, M.J., Fish, L., Kirke, D.F., Tomas, J.M., Stewart, G.S.A.B., & Williams, P. (1999). Quorum Sensing-Dependent Regulation and Blockade of Exoprotease Production in Aeromonas hydrophila. Infection and Immunity, 67(10), 5192-5199.
• Timur, G., & Timur, M. (2003). Balık Hastalıkları. İstanbul Üniversitesi Su Ürünleri Fakültesi, Yayın No:5. 538 s. ISBN.975-404699-5.
• Ulusoy, S. (2007). Yoğun Bakım Ünitelerinden İzole Edilen Pseudomonas aeruginosa Suşlarında N-Açil Homoserin Lakton Üretiminin Araştırılması. Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü, Doktora Tezi, 100s, Isparta.
• Verschuere, L., Rombaut, G., Sorgeloos, P., & Verstraete, W. (2000). Probiotic bacteria as biological control agents in aquaculture. Microbiology and Molecular Biology Reviews, 64, 655-671.
• Wei, H.L., & Zhang, L.Q., (2006). Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24, Antonie van Leeuwenhoek, 89, 267-280.
• Williams, P. (2007). Quorum sensing, communication and cross-kingdom signalling in the bacterial world. Microbiology, 153, 3923-3938.
• 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.
• Yılmaz-Yıldıran, H., Karahan, A.G., & Başyiğit-Kılıç, G. (2015). Laktik asit bakterilerinde çoğunluğu algılama mekanizması, Türk Hijyen ve Deneysel Biyoloji Dergisi, 72(1), 79-90.