Geleneksel Yöntemlerle Yapılmış Peynirlerden Leuconostoc sp ve Pediococcus sp.’lerin İzolasyonu, Moleküler Tanımlanması ve Kısmi Karakterizasyonu

Year 2018, Volume: 21 Issue: 1, 44 - 50, 01.01.2018

Mehmet Sait Ekinci İsmail Akyol , Ahmed Karem Ramadan Ferit Can Yazdıç Emin Özköse

https://doi.org/10.18016/ksudobil.298876

Abstract

Laktik asit bakterileri heksoz şekerlerin laktik asite
fermentasyon prosesinde önemli rol oynayan ticari bakterilerdir ve “güvenli
mikroorganizmalar” olmaları nedeniyle süt endüstrisinde yaygın olarak
kullanılırlar. Laktik asit bakterilerinin izolasyonu amacıyla 40 adet fermente
gıda örneklenmiş ve bu örneklerden 160 bakteri izolatı saflaştırılarak
tanımlanmıştır. Morfolojik karakterlerine göre 62 izolat (38.75%) cocci olarak
sınıflandırılmış, bunların MRS agarda iyi yetişdiği ve tamamının
katalaz-negatif olduğu belirlenmiştir. Coccobacilli
ve bacilli formlar sırasıyla 66
(41.25%) ve 19 (11.875%) adet olarak gözlemlenmiş, 13 izolat ise morfolojik
özelliklerine göre “diğerleri” şeklinde tanımlanmış ve deneme dışı
bırakılmıştır. Pediococcus ve Leuconostoc suşlarının moleküler
tanımlaması sırasıyla 23S ve 16S rDNA’larının PZR yardımıyla amplifiye
edilmesiyle yapılmış ve 34 izolat Pediococcus
olarak tanımlanırken 4 izolatın Leuconostoc
suşu olduğu tespit edilmiştir. Yeni izole edilen bu suşların antimikrobiyal
aktiviteleri test edilmiş ve birden fazla izolatın S. paratyphi ve E. faecali’ye
karşı antimikrobiyal aktivitelerinin olduğu belirlenmiştir. Tüm test edilen
izolatların P. mirabilis, E. coli ve C. jejuni’a karşı göreceli olarak düşük antimikrobiyal etki
gösterirken P. aeruginosa’nın
izolatların sentezlediği tüm bakteriyosinlere karşı dirençlilik gösterdiği
belirlenmiştir. Sonuçlar test edilmiş izolatların (n=20) erythromycin,
chloramphenicol ve rifampine karşı dirençlilik gösterdiğini ortaya koymuştur.
Tüm izolatların plasmid içerikleri araştırılmış ve 10 izolatın en az 1 adet plasmid
vektör içerdiği bulunmuştur.

Keywords

Antibiyotik dirençlilik, Laktik asit bakterileri, Leuconostoc, Moleküler tanımlama, Pediococcus

Molecular Identification and Partial Characterization of Pediococcus sp. and Leuconostoc sp. Isolated from Traditionally Made Dairy Products

Abstract

Lactic acid bacteria (LAB) are group of commercially
important organisms that play an important role in the fermentation process of
hexose sugars into lactic acid and they are videly used in dairy industrial
applications since they were designated as “generally recognised as safe”
organisms. In order to isolate laktic acid bacteria (LAB), forty fermented food
samples were collected and 160 bacterial strains were purified and identified
from those samples. According to morphological characteristics, 62 isolates
(38.75%) were classified as cocci and all these cocci were found to be as
catalase negative and grown well on MRS agar plates. Coccobacilli and bacilli
forms were observed in 66 (41.25%) and 19 (11.875%) isolates respectively.
Total of 13 isolates (8.125%) were classified as “others” (according to
morphological characteristics) and excluded from rest of experimental process.
For molecular identification of Leuconostoc
sp. and Pediococcus sp., 16S and 23S
rDNAs (respectively) were amplified with the aid of PCR and 34 of them were
classified as Pediococcus whilst 4 of
them were designated as Leuconostoc.
These newly isolated strains were tested for their antimicrobial activity and
more than one strains exhibited antimicrobial activity against to S. paratyphi
and E. faecali. All tested strains
showed relatively lower (≤ 10mm in diameter) antimicrobial effect against P. mirabilis, E. coli and C. jejuni while
P. aeruginosa showed resistancy against all bacteriocins produced tested
isolates. Result of this study revealed that majority of the tested (n=20)
isolates were resistant to erythromycin, chloramphenicol and rifampin. All
isolates were investigated for the presence of plasmid vectors and 10 of them
found having at least 1 plasmid vector.

Keywords

Antibiotic resistance, molecular identification, Leuconostoc, Pediococcus

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