Mersin ilinde çiğ sütlerden Mycobacterium bovis ve tüberküloz dışı mikobakterilerin izolasyonu ve tanımlanması
Küçük Resim Yok
Dosyalar
Tarih
2012
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Öz: Çalışmamızda, sığır çiğ süt örneklerinde Mycobacterium bovis ve tüberküloz dışı mikobakteri (non-tu- berculous mycobacteria; NTM) türlerinin izolasyonu, tanımlanması ve kullanılan yöntemlerin tanısal per- formanslarının karşılaştırılması amaçlanmıştır. Çalışmada, Nisan 2008-Haziran 2008 döneminde Mersin ilindeki beş köyden toplanan 145 sığır çiğ süt örneği incelenmiş; bu örneklerde mikobakteri varlığı Ehr- lich-Ziehl-Neelsen (EZN) boyama yöntemi, Löwenstein-Jensen (LJ) besiyerinde kültür ve polimeraz zincir reaksiyonu (PCR) ile araştırılmıştır. Toplam 145 çiğ süt örneğinden hazırlanan yaymaların EZN boyama sonucunda sadece 1 (%0.7) örnekte aside dirençli basil (ARB) görülmüş, 11 (%7.6) örneğin LJ besiyerin- de kültürü pozitif olarak belirlenmiş ve 6 (%4.1) örnekte PCR ile mikobakteri DNAsı tespit edilmiştir. Kül- tür pozitif bir örneğin, hem kaymak hem de pellet tabakasından mikobakteri izolasyonu yapılmıştır. İzo- latların tanımlanması, geleneksel biyokimyasal testler, PCR-RFLP (PCR-Restriction Fragment Length Poly- morphisms) ve spoligotiplendirme (spacer oligonucleotide typing) yöntemleriyle gerçekleştirilmiştir. Kül- türde izole edilen toplam 12 izolattan biri Mycobacterium tuberculosis kompleks (MTC), 11i de NTM ola- rak tanımlanmıştır. PCR-RFLP sonuçlarına göre 11 NTM izolatının 6 (%54.5)sı Mycobacterium genavense , 2 (%18.2)si Mycobacterium simiae , 2 (%18.2)si Mycobacterium szulgai ve 1 (%9.1)i Mycobacterium for- tuitum olarak belirlenmiştir. MTC izolatı spoligotiplendirme yöntemiyle M.bovis olarak tanımlanmıştır. Ça- lışmamızın sonuçlarına göre, süt örneklerindeki izolasyon şansını artırmak için; hem pellet hem de kay- mak tabakalarından selektif LJ besiyerine (gliserolsüz, %0.4 sodyum pirüvatlı) ekim yapılmalı ve kültürle- rin mutlaka sekiz haftaya kadar inkübe edilmesi gerekmektedir. Sonuç olarak bölgemizde sığır çiğ süt ör- neklerinde NTM %6.9 ve M.bovis %0.7 oranında izole edilmiş; insanların bu mikroorganizmalara doğru- dan temas yoluyla veya pastörize edilmemiş süt ürünlerinin tüketilmesiyle maruz kalabilecekleri bir kez daha vurgulanmıştır.
Abstract: This study was aimed to isolate and identify Mycobacterium bovis and non-tuberculous mycobacte- ria (NTM) species in raw milk samples from cattles and to compare the diagnostic performance of the methods used for that purpose. A total of 145 raw milk samples from cattles were collected from five vil- lages in Mersin province (located on Mediterrenean region of Turkey) between April and June 2008. Pre- sence of mycobacteria was investigated by Ehrlich Ziehl Neelsen (EZN) staining method, culture in Lö- wenstein-Jensen (LJ) medium and polymerase chain reaction (PCR). Only 1 (0.7%, 1/145) raw milk sample was found to be acid fast bacilli (AFB) positive with EZN staining. Eleven (7.6%) samples were positive by culture and mycobacterial DNA was detected in 6 (4.1%) samples by PCR. Mycobacterium was isolated from both creamy and pellet layer of a culture positive sample. Identification was carried out with conventional biochemical tests, PCR-Restriction Fragment Length Polymorphisms (PCR-RFLP) and spoligotyping (spacer oligonucleotide typing) methods. One isolate was identified as Mycobacteri- um tuberculosis complex (MTC) and 11 isolates were identified as NTM out of 12 isolates those were iso- lated from culture. According to PCR-RFLP analysis of these 11 NTM isolates, 6 (54.5%) were Mycobac- terium genavense , 2 (18.2%) were Mycobacterium simiae , 2 (18.2%) were Mycobacterium szulgai and 1 (9.1%) was Mycobacterium fortuitum . MTC isolate was identified as M.bovis by spoligotyping. According to the results of our study, both pellet and creamy layers from raw milk samples should be cultured to selective LJ medium (without glycerol, with 0.4% sodium pyruvate) to improve the chance of isolation and must be incubated for up to eight weeks. In our region, NTM were isolated in 6.9% and M.bovis in 0.7% of the raw milk samples from cattles and this emphasized the risk of transmission of mycobacteria to man via direct contact or ingestion of unpasteurized milk products.
Abstract: This study was aimed to isolate and identify Mycobacterium bovis and non-tuberculous mycobacte- ria (NTM) species in raw milk samples from cattles and to compare the diagnostic performance of the methods used for that purpose. A total of 145 raw milk samples from cattles were collected from five vil- lages in Mersin province (located on Mediterrenean region of Turkey) between April and June 2008. Pre- sence of mycobacteria was investigated by Ehrlich Ziehl Neelsen (EZN) staining method, culture in Lö- wenstein-Jensen (LJ) medium and polymerase chain reaction (PCR). Only 1 (0.7%, 1/145) raw milk sample was found to be acid fast bacilli (AFB) positive with EZN staining. Eleven (7.6%) samples were positive by culture and mycobacterial DNA was detected in 6 (4.1%) samples by PCR. Mycobacterium was isolated from both creamy and pellet layer of a culture positive sample. Identification was carried out with conventional biochemical tests, PCR-Restriction Fragment Length Polymorphisms (PCR-RFLP) and spoligotyping (spacer oligonucleotide typing) methods. One isolate was identified as Mycobacteri- um tuberculosis complex (MTC) and 11 isolates were identified as NTM out of 12 isolates those were iso- lated from culture. According to PCR-RFLP analysis of these 11 NTM isolates, 6 (54.5%) were Mycobac- terium genavense , 2 (18.2%) were Mycobacterium simiae , 2 (18.2%) were Mycobacterium szulgai and 1 (9.1%) was Mycobacterium fortuitum . MTC isolate was identified as M.bovis by spoligotyping. According to the results of our study, both pellet and creamy layers from raw milk samples should be cultured to selective LJ medium (without glycerol, with 0.4% sodium pyruvate) to improve the chance of isolation and must be incubated for up to eight weeks. In our region, NTM were isolated in 6.9% and M.bovis in 0.7% of the raw milk samples from cattles and this emphasized the risk of transmission of mycobacteria to man via direct contact or ingestion of unpasteurized milk products.
Açıklama
Anahtar Kelimeler
Kaynak
Mikrobiyoloji Bülteni
WoS Q Değeri
Scopus Q Değeri
Cilt
46
Sayı
2
Künye
AYDIN F. E,ÜLGER M,EMEKDAŞ G,ASLAN G,GÜNAL S (2012). Mersin ilinde çiğ sütlerden Mycobacterium bovis ve tüberküloz dışı mikobakterilerin izolasyonu ve tanımlanması. Mikrobiyoloji Bülteni, 46(2), 283 - 289.
