Broiler Karkaslarından İzole Edilen Campylobacter jejuni İzolatlarının Makrolid, Kinolon ve Tetrasiklin Grubu Antibiyotiklere Karşı Direnç Durumu*

Year 2015, Volume: 12 Issue: 2, 81 - 92, 01.08.2015

Harun Hızlısoy Hüseyin Kılıç

Abstract

Çalışmada Haziran 2012 ve Ocak 2013 dönemlerinde, Kayseri şehir merkezinde bulunan farklı satış noktalarında tü-
ketime sunulan toplam 200 broiler karkasından Campylobacter jejuni izolasyonu ve izolatların makrolid, kinolon ve tetrasiklin
antibiyotiklerine karşı direnç durumlarının fenotipik ve genotipik yöntemlerle belirlenmesi amaçlanmıştır.
Çalışmada but, göğüs, kanat ve bütün tavuk olmak üzere toplam 200 örnek incelendi. Örneklerden elde edilen izolatların
identifikasyonu, fenotipik testler ve multipleks Polimeraz Zincir Reaksiyonu (mPZR) ile gerçekleştirildi. C. jejuni’lerin antibiyotiklere
direnç durumları disk difüzyon testi ile; MİK dağılımları ise E test ile saptandı. Aynı zamanda antibiyotiklere dirençlilikleri;
PZR ve sekans analizi gibi moleküler yöntemlerle test edildi.
Haziran döneminde toplanan 100 örneğin 81’i, Ocak döneminde ise 100 örneğin 77’si Campylobacter spp. yönünden pozitif
bulundu. mPZR testi sonucunda Haziran dönemindeki 124 izolatın 88’i; Ocak dönemindeki 109 izolatın 68’i C. jejuni olarak
tespit edildi. Disk difüzyon testinde en yüksek direnç oranları, Haziran ve Ocak dönemleri için sırasıyla nalidiksik asit (%78.4-
%92.6), siprofloksasin (%67.1-%86.7) ve tetrasikline (%55.6-%64.7) karşı, en düşük oranlar ise azitromisin (%1.1-%4.4), doksisiklin
(%2.2-%1.4) ve eritromisine (%11.3-%4.4) karşı bulundu. E test sonuçları ile disk difüzyon testi sonuçlarının uyumlu
olduğu belirlendi. Tetrasiklin direnci yönünden genomik tetO geni; Haziran ve Ocak döneminde sırasıyla %88 ve %85.7’inde
gösterilirken; plazmid aracılı tetO geni izolatların %93.7 ve %85.7’sinde tespit edildi. Makrolid direncinde A2074C mutasyonuna
izolatların hiçbirinde rastlanmazken; A2075G mutasyonuna Haziran ve Ocak dönemlerinde %18.4 ve %14.8 oranında
rastlandı. Genomik kinolon direncinde gyrA geni üzerindeki mutasyonlar, Haziran ve Ocak dönemlerinde, izolatların %83.5 ve
%81.8’inde tespit edilirken; plazmid aracılı kinolon direnci amacıyla incelenen qnr geni, izolatların hiçbirinde bulunamadı. PZR
ile saptanan kinolon ve makrolid direnci, sekans analizi ile doğrulandı. Sekans analizinde kinolon dirençli izolatlarda Tre-86-İle
mutasyonuna rastlanırken; duyarlı izolatlarda bu mutasyona rastlanmadı. Ayrıca, makrolid dirençli izolatlarda A2075G mutasyonu
tespit edildi. Ancak, hiçbir izolatta A2074C mutasyonuna rastlanmadı. Duyarlı izolatların hiçbirinde de bu iki mutasyon
görülmedi. Sonuç olarak; tavuk etindeki antibiyotiklere dirençli C. jejuni varlığı nedeniyle koruyucu tedbirlerin alınması, dar
spektrumlu antibiyotik kullanımının tercih edilmesi ve antibiyotiklerin organizmadan atılma süreleri tamamlanmadan hayvanların
kesilip tüketime sunulmaması, ayrıca tedavide ilk olarak tercih edilen eritromisine ilaveten azitromisinin ve doksisiklinin de
alternatif olarak kullanılabileceği kanısına varıldı.

Keywords

Broiler, Campylobacter jejuni, kinolon, makrolid, tetrasiklin

The Resistance State of Campylobacter jejuni Isolated from Broiler Carcasses Against to Macrolide, Quinolone and Tetracycline Groups of Antibiotics

Abstract

: In the study, in June 2012 and in January 2013, the isolation of Campylobacter jejuni from a total of 200 broiler carcasses
in different sales points in the city center of Kayseri and the determination of resistance states to macrolides, quinolone
and tetracycline groups of antibiotics by using phenotypical and genotypical methods were aimed.
Two hundered samples including chicken thigh, breast, wings and carcasses were examined in the study. The identification of
the isolates obtained from samples were performed by phenotypical tests and multiplex Polymerase Chain Reaction (mPCR) method. The antibiotic resistance and MIC values of C. jejuni isolates were detected with disk diffusion and E test method,
respectively. In addition, the antibiotic resistances were tested by using of PCR and sequencing.
Eighty-one of 100 samples collected in June and 77 of 100 samples in January were found to be positive in terms of Campylobacter
spp. As a result of mPCR, 88 of 124 and 68 of 109 Campylobacter spp. isolates were identified as C. jejuni from during
June and January terms, respectively. In disk diffusion test, while, the highest resistance rates were observed as nalidixic acid
(78.4% - 92.6%), ciprofloxacin (67.1% - 86.7%) and tetracycline (55.6% - 64.7%), the lowest resistance rates as azithromycin
(1.1% - 4.4%), doxycycline (2.2% - 1.4%) and erythromycin (11.3% - 4.4% ), for the period June and January, were respectively
found. It was determined that E test results were compatible with disk diffusion test. Whilst, chromosomal tetO gene was
shown in June and January isolates at 88 % and 85.7 % rates, plasmid mediated tetO gene was found at 93.7% and 85.7%
rates, respectively. In macrolide resistance, while A2075G mutation was detected in 23SrRNA at rates 18.4% and 14.8% of
June and January isolates, A2074C mutation was not observed at any of June or January isolates, respectively. When, in chromosomal
quinolone resistance, the mutations in gyrA gene were detected at rates of 83.5% and 81.8% in June and January
isolates, respectively, qnr gene examined for plasmid mediated quinolone resistance was found in none of C. jejuni isolates.
Quinolone and macrolide resistances detected by PCR were confirmed by DNA sequencing. While, Tre-86-Ile mutation was
found in quinolone resistant isolates with sequencing, but these mutations were not detected in susceptible isolates. Additionally,
A2075G mutation was detected in macrolide resistant isolates. However, A2074C mutation was observed in any isolates
by using sequencing. It was not observed that none of the macrolide susceptible isolates had any of these two mutations. In
conclusion, due to the presence of antibiotic resistant C. jejuni detected in chicken meat, protective measures should be taken
for, the using of narrow-spectrum antibiotics may be preferred, animals treated with antibiotics, before the completion of withdrawal
period of antibiotics from organisms, animals may not be slaughtered or supplied for consumption. Besides, it is considered
that in the treatment, azythromycin and doxycycline may be used as alternate to erythromycin in the drug of choice.

Keywords

Broiler carcass, Campylobacter jejuni, macrolide, quinolone, tetracycline

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