Antimicrobial Effect of Partially Purified Bacteriocins on Pseudomonas aeruginosa

Abstract

Bacteriocins are commonly used in foods to inhibit the growth of spoilage and pathogenic bacteria, thus extending the shelf life of food products. Interest in bacteriocins is increasing because of the increasing tendency of consumers to use healthy, natural and additive-free products in foods. In this study, it was aimed to investigate the control of P. aeruginosa in milk by using partially purified bacteriocins produced from lactic acid bacteria (LAB) strains. Among the 13 reference LAB strains, four strains that showed the highest antimicrobial activity by the agar spot test were selected for bacteriocin production. The bacteriocins were partially purified with 40% ammonium sulfate. The antibacterial activity of bacteriocins on P. aeruginosa strains was determined in arbitrary unit by the well diffusion meth-od. Then, UHT milk samples inoculated with P. aeruginosa and bacteriocin cocktail were stored at +4°C for a week and bacterial counts were performed on the 1st, 3rd, 5th, and 7th days. The LAB strains that displayed the widest clear zones on P. aeruginosa were determined as L. plantarum, L. paraplantarum, L. fermentum and L. pentosus and the antimicro-bial activities of the partially purified bacteriocins of these strains were 640, 640, 160 and 80 AU/ml, respectively. Among tested P. aeruginosa strains, the highest antibacterial effect was observed against P. aeruginosa ATCC 15442 (>18mm). In the milk model, the bacteriocin cocktail caused a decrease of approximately 2 log cfu/ml in the number of bacteria for up to three days and the number remained constant until the end of the seventh day. However, the de-crease in the number of bacteria was not statistically significant (P>0.05). As a result, bacteriocins obtained from Lacto-bacillus strains showed antibacterial effect on P. aeruginosa on agar medium but could not achieve a significant de-crease on the milk. However, bacteriocins, which have generally been proven to efficient on Gram-positive bacteria, have been determined to be effective on P. aeruginosa, a Gram-negative bacterium. Moreover, this study emphasizes that in addition to in-vitro experiments, products to be used for biocontrol purposes in foods are also needed to comple-ment with food models.

Keywords

• Bacteriocin cocktail
• lactic acid bacteria
• milk
• P. aeruginosa
• storage

Kısmi Saflaştırılmış Bakteriyosinlerin Pseudomonas aeruginosa Üzerindeki Antimikrobiyal Etkisi

Abstract

Bakteriyosinler, patojen ve/veya gıdalarda bozulmaya neden olan bakterilerin büyümesini engellemek ve gıdaların raf ömrünü uzatmak amacıyla yaygın olarak kullanılmaktadır. Tüketicilerin gıdalarda sağlıklı, doğal ve katkı maddesi içermeyen ürünleri kullanma eğiliminin artması nedeniyle bakteriyosinlere olan ilgi de artmaktadır. Bu çalışmada laktik asit bakterisi (LAB) suşlarından üretilen kısmi saflaştırılmış bakteriyosinler kullanılarak sütte P. aeruginosa'nın kontrolü-nün araştırılması amaçlanmıştır. On üç adet referans LAB suşu arasında agar spot testiyle en yüksek antimikrobiyal aktiviteyi gösteren dört suş, amonyum sülfatla kısmi olarak saflaştırılmıştır. Bakteriyosinlerin P. aeruginosa suşları üze-rindeki antibakteriyel aktivitesi kuyu difüzyon yöntemiyle arbitrary unit (AU/ml) olarak belirlenmiştir. Ek olarak, P. aerugi-nosa ile kontamine edilen UHT süt örneklerine bakteriyosin kokteyli eklenmiş bir hafta boyunca +4°C'de inkübe edile-rek 1., 3., 5. ve 7. günlerde bakteri sayımları yapılmıştır. P. aeruginosa üzerinde en geniş zon sergileyen LAB suşları-nın L. plantarum, L. paraplantarum, L. fermentum ve L. pentosus olduğu ve bu suşlardan elde edilen kısmi saflaştırıl-mış bakteriyosinlerinin antimikrobiyal aktivitelerinin sırasıyla 640, 640, 160 ve 80 AU/ml olduğu belirlenmiştir. Test edi-len P. aeruginosa suşları arasında en yüksek antibakteriyel etki P. aeruginosa ATCC 15442'ye (>18mm) karşı gözlen-miştir. Süt modelinde ise, bakteriyosin kokteyli bakteri sayısında üç güne kadar yaklaşık 2 log kob/ml azalmaya neden olmuş ve sayı yedinci günün sonuna kadar sabit kalmıştır. Bununla birlikte, bakteri sayısındaki azalmanın istatistiksel olarak anlamlı olmadığı tespit edilmiştir (P>0.05). Sonuç olarak çalışmada elde edilen bakteriyosinlerin P. aerugino-sa'ya karşı in-vitro ortamda antibakteriyel etki gösterdiği tespit edilmesine karşın gıda modelinde anlamlı bir sonuç elde edilememiştir. Bu çalışmanın verileri gıdalarda biyokontrol amaçlı kullanılacak maddelerin in-vitro analizlerin yanı sıra gıda modelleri ile tamamlanmasının önemi vurgulamıştır.

Keywords

• Bakteriyosin kokteyli
• laktik asit bakterileri
• muhafaza
• P. aeruginosa
• süt

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