Lizozim ve Nisinin Gıda Kaynaklı Staphylococcus aureus Suşlarında Gelişim ve Biyofilm Oluşumu Üzerine Etkileri

Yıl 2013, Cilt: 39 Sayı: 2, 254 - 263, 10.03.2013

Mert Sudağıdan Ali Aydın

Öz

ÖZET

Staphylococcus aureus gıdalarda gelişebilen ve ürettiği toksinlerle gıda zehirlenmesine yol açan en önemli patojen bakterilerdendir. Bu çalışmada, besin maddelerinde sıklıkla biyokoruyucu olarak kullanılan lizozim ve nisinin, değişik gıdalardan izole edilen 14 adet S. aureus suşunun gelişme ve biyofilm oluşturması üzerindeki etkileri kantitatif mikroplaka yöntemi ile araştırılmıştır. Elde edilen veriler doğrultusunda 1, 2, 3, 4 ve 5 mg/ml konsantrasyonlardaki lizozimin, bakteri gelişmesini engelleme yönünde hiçbir etkisinin olmadığı ve tüm suşların lizozime karşı dirençli oldukları tespit edilmiştir. Diğer yandan, 12,5 μg/ml konsantrasyonda nisinin bazı suşlarda % 100 inhibisyon gösterdiği saptanmış, sadece bir suşta ise (S. aureus SE-22C) 37,5 μg/ml konsantrasyonda uzun inkübasyon sonrasında gelişme görülmüştür. Biyofilm testleri sonucunda, lizozimin artan konsantrasyonlarının dört suşta biyofilm yapımını aktive ettiği, bir suşta ise lizozim içermeyen besiyerine göre önce azalttığı, artan lizozim konsantrasyonlarında ise biyofilm oluşumunu arttırdığı tespit edilmiştir. Diğer yandan lizozimin artan konsantrasyonlarının iki suşta biyofilm oluşumunu azalttığı görülmüştür. Nisinin artan konsantrasyonlarının dört suşta bakterilerin biyofilm oluşumunu azalttığı, diğer suşlarda ise bir değişikliğe yol açmadığı bulunmuştur. Çalışmamız neticesinde lizozimin S. aureus suşlarına karşı etkisiz bir biyokoruyucu olduğu, fakat biyofilm yapımını aktive edebildiği; buna karşın nisinin yüksek konsantrasyonlarda bakteri gelişmesi üzerine inhibisyon etkisinin olduğu, ancak nisine dirençli S. aureus suşlarının bulunabileceği sonucuna varılmıştır.

Anahtar Kelimeler: Staphylococcus aureus, lizozim, nisin, biyofilm, gıda

ABSTRACT

EFFECTS OF LYSOZYME AND NISIN ON GROWTH AND BIOFILM FORMATION OF FOODBORNE STAPHYLOCOCCUS AUREUS STRAINS

Staphylococcus aureus is one of the most important pathogenic bacteria in food producing toxins and causing food poisoning. In this study, the effects of biopreservatives lysozyme and nisin on growth and biofilm formation of S. aureus strains (n=14) isolated from different foods were examined by quantitative microplate method. On the basis of obtained data, 1, 2, 3, 4, and 5 mg/ml of lysozyme concentrations did not inhibit the growth of strains and all strains were determined as resistant to lysozyme. On the other hand, nisin at 12,5 mg/ml concentration showed 100% inhibition on most of the strains, except S. aureus SE-22C strain at 37,5 mg nisin/ml concentration, the growth of strain was obtained after a long incubation time. As a result of biofilm tests, in four strains increasing concentrations of lysozyme were found to activate biofilm formation. In SE-22C strain, with compared to medium without lysozyme, firstly biofilm formation was decreased then with increasing lysozyme concentrations the increased biofilm formation was obtained. In addition, in two strains biofilm formation was decreased with increased lysozyme concentrations. Nisin reduced biofilm formation of four strains with increasing concentrations, but there was no effect of nisin on biofilm formation of the other strains. As a result of this study, lysozyme was determined as an ineffective biopreservative against S. aureus strains, but it can activate biofilm formation of the strains. Conversely, high concentrations of nisin had an inhibitory effect on bacterial growth, whereas the presence of nisin resistant S. aureus strains could exist.

Key Words: Staphylococcus aureus, lysozyme, nisin, biofilm, food

Anahtar Kelimeler

Staphylococcus aureus, lizozim, nişin, biyofilm, gıda

Kaynakça

• Akkoç, N., Şanlıbaba, P., Akçelik, M., 2009. Bakteriyosinler: Alternatif gıda koruyucuları. Erciyes Üniversitesi Fen Bilimleri Enstitüsü Dergisi 25, 59-70.
• Aydin, A., Muratoglu, K . , Sudagidan, M., Bostan, K . , Okuldu, B., Harsa, S., 2011. Prevalence 262 Mert Sudağıdan, Ali Aydın and antibiotic resistance o f foodborne Staphylococcus aureus isolates in Turkey. Foodborne Pathogens and Disease 8, 63-69.
• Bera, A., Herbert, S., Jakob, A., Vollmer, W., Götz, F . , 2005. Why are pathogenic staphylococci so lysozyme resistant? The peptidoglycan Oacetyltransferase OatA is the major determinant for lysozyme resistance of Staphylococcus aureus. Molecular Microbiology 55, 778-787.
• Blake, K . L . , Randall, C P . , O'Neill, A.J., 2011. In vitro studies indicate a high resistance potential for the lantibiotic nisin in Staphylococcus aureus and define a genetic basis for nisin resistance. Antimicrobial Agents and Chemotherapy 55, 2362-2368.
• Carlson, S., Bauer, H.M., 1957. Nisin, ein antibakterieller Wirkstoff aus Streptococcus lactis unter berucksichtigung des resistenz problems. Archiv für Hygiene und Bakteriologie 141,445^59.
• Chen, H . , Hoover, D.G., 2003. Bacteriocins and their food applications. Comprehensive Reviews in Food Science and Food Safety 2, 82-100.
• Cotter, P.D., Hill, C , Ross, R.P., 2005. Bacteriocins: developing innate immunity for food. Nature Reviews Microbiology 3, 777-788.
• Davidson, P.M., Harrison, M.A., 2002. Resistance and application to food antimicrobials, sanitizers, and other process controls. Food Technology 56, 69-78.
• Davidson, P.M., Juneja, V . K . , Branen, J . K . , 2002. Antimicrobial Agents. In: Branen, A . L . , Davidson, P.M., Salminen, S. and Thorngate J.H. (Eds). Food Additives. Marcel Dekker, New York.
• Donlan, R.M., 2001. Biofilms and device associated infections. Emerging Infectious Disease 7, 277¬ 2
• Fedtke, I . , Götz, F . , Peschel, A., 2004. Bacterial evasion o f innate host defenses-the Staphylococcus aureus lesson. International Journal of Medical Microbiology 294, 189-194.
• Gill, A.O., HoUey, R.A., 2000. Inhibition of bacterial growth on ham and bologna by lysozyme, nisin and EDTA. Food Research International 33, 83¬
• Grade, S., Avila, M., Nunez, M., 2004. Fast induction of nisin resistance in Streptococcus thermophilus INIA 463 during growth in milk. International Journal of Food Microbiology 96, 165-1
• Harris, L . J . , Fleming, H.P., Klaenhammer, T . R , 19 Sensitivity and resistance of Listeria monocytogenes ATCC 19115, Scott A and UAL500 to nisin. Journal of Food Protection 54, 836-840.
• Herbert, S., Bera, A., Nerz, C , Graus, D., Paschel, A., Goerke, C , Meehl, M., Cheung, A., Götz, F . , 2007. Molecular basis of resistance to muramidase and cationic antimicrobial peptide activity of lysozyme in Staphylococci. PLoS Pathogens 3, 981-994.
• Jarvis, B . 1967. Resistance to nisin and production of nisin-inactivating enzymes by several Bacillus species. Journal of General Microbiology 47, 33Koponen, O., 2004. Studies of producer self-protection and nisin biosynthesis of Lactococcus lactis.
• Institute of Biotechnology and Department o f Applied Chemistry and Microbiology, Helsinki, Finland. Martinez, B., Obeso, J . M . , Rodriguez, A., Garcia, P., 2008. Nisin-bacteriophage cross resistance in Staphylococcus aureus. International Journal of Food Microbiology 122,253-258.
• Ming, X., Daeschel, M.A., 1993. Nisin resistance of foodborne bacteria and the specific resistance responses of Listeria monocytogenes Scott A .
• Journal of Food Protection 56,944-948. Moretro, T., Hermansen, L . , Hoick, A . L . , Sidhu, M.S., Rudi, K . , Langsrud, S., 2003. Biofilm formation and the presence of the intercellular adhesion locus ica among Staphylococci from food and food processing environments.
• Applied and Environmental Microbiology 69, 5648-5655.
• Naidu, A.S., 2000. Overview. In: Naidu, A.S., (Ed).
• Natural Food Antimicrobial Systems, Chapter CRC Press, Boca Raton. Parente, E . , Riciardi, A. 1999. Production, recovery and purification of bacteriocins from lactic acid bacteria. Applied Microbiology and Biotechnology 52, 628-638.
• Peschel, A., Otto, M., Jack, R.W., Kalbacheri, H . , Jung, G., Götz, F . , 1999. Inactivation of the dit operon in Staphylococcus aureus confers sensitivity to defensins, protegrins and other antimicrobial peptides. The Journal of
• Biological Chemistry 274, 8405-8410.
• Pinto, M.S., de Carvalho, A.F., Pires, A.C.S., Souza, A.A.C., da Silva, P.H.F., SobraL D., 2011.
• The effects o f nisin on Staphylococcus aureus count and the physicochemical properties of traditional minas serro cheese. International Dairy Journal 21,90-96. Ross, R.P., Morgan, S., Hill, C , 2002. Preservation and fermentation: Past, present and future. International Journal o f Food Microbiology 79, 3Samelis, J . , Kakouri, A., Rogga, K . J . , Sawaidis, I.N., Kontominas, M.G., 2003. Nisin treatments to control Listeria monocytogenes post-processing contamination on Anthotyros, a traditional Greek whey cheese, stored at 4°C in vacuum packages. Food Microbiology 20, 661¬ 6
• Sudagidan, M., Yemenicioglu, A., 2012. Effects o f nisin and lysozyme on growth inhibition and biofilm formation capacity o f Staphylococcus aureus strains isolated from raw milk and cheese samples. Journal o f Food Protection 75, 1627-1633.
• Ünlütürk, A., 2003. Mikrobiyal Gelişmenin Inhibisyonu. In: Ünlütürk, A., Turantaş, F.
• (Eds), Gıda Mikrobiyolojisi, 3. baskı. Meta basım Matbaacılık hizmetleri, Izmir, pp. 188. Üstünkol, N. 2006. Farklı ortam koşullarında nisin, lizozim ve bazı bitkisel kaynakların küf gelişiminin kontrol altına alınması üzerine etkileri. Yüksek Lisans Tezi, İstanbul Teknik
• Üniversitesi. Fen Bilimleri Enstitüsü. İstanbul.