Kesim Hattının Farklı Aşamalarında Broiler Karkaslarında Arcobacter Türlerinin Varlığı ve Antimikrobiyel Dirençlerinin Belirlenmesi
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Dosyalar
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Broiler kesiminde en önemli bulaşma aşamalarından olan haşlama ve tüy
yolumu, iç organ çıkarma, soğutma ve paketleme sonrasında Arcobacter türlerinin
kontaminasyonunu araştırmak üzere üç farklı broiler kesimhanesinden değişik
zamanlarda her bir aşamadan 27 olmak üzere toplamda 108 numune alındı.
Numunelerden Arcobacter izolasyon ve identifikasyonu için kültürel ve
moleküler yöntemler kullanıldı. Numuneler steril torbalar içerisinde 400 ml
Maximun Recovery Diluent kullanılarak yıkandı. Yıkama suyundan 10 ml alınarak
90 ml CAT ilaveli ASB’a inoküle edildi. Mikroaerofilik ortamda 48 saat inkübe
edildi. Zenginleştirilmiş kültür Brucella Broth ile 1:10 oranında seyreltildi.
Seyreltiden 300 μl örnek %5 koyun kanlı Mueller Hinton Agar üzerine yerleştirilmiş
selüloz nitrat membran filtre vasıtasıyla agar üzerine yayıldıktan sonra üzerine 100 μl
Brucella Broth ilave edildi. 30°C'de 48 saat aerobik inkübasyondan sonra, şüpheli
koloniler Muller Hinton Agar’a ekildikten sonra 30°C'de 48 saat mikroaerofilik
olarak inkübe edildi. Mueller Hinton Agarda üreyen kolonilere gram boyama,
oksidaz, katalaz, nitrat redüksiyon, üreaz, hippurat testleri uygulandı. Arcobacter
spp. izolatlarının moleküler identifikasyonu için 16S ve 23S rRNA genlerini
hedefleyen multipleks PCR (mPCR) yöntemi uygulandı. Amplifiye DNA’lar
elektroforez tankında 90 volt, 70 miliamperde 60 dakika yürütme sonrası A. butzleri
401 bp’de, A. cryaerophilus 257 bp’de ve A. skirrowii 641 bp’de gözlemlendi.
İncelenen 108 numuneden kültürel yöntemlerle 104 tanesinde Arcobacter
spp. izole ve identifiye edildi. Elde edilen izolatların mPCR yöntemiyle incelenmesi
sonucunda, 51 numunede A. butzleri, A. cryaerophilus ve her iki Arcobacter türünün
karışık kontaminasyonu tespit edildi. 51 izolatın %52,9’u A. butzleri, %31,4’ü A.
cryaerophilus ve %15,7’si A. butzleri ve A. cryaerophilus karışık kontaminasyonu
olarak belirlendi. Araştırmada A. skirrowii tespit edilmedi. Haşlama ve tüy yolumu
sonrası numunelerin %25,9’u A. butzleri, %22,2’si A. cryaerophilus ve %11,1’i A.
buzleri ve A. cryaerophilus karışık kontaminasyonu olarak saptandı. İç organ
çıkarma sonrası numunelerin %26,1’i A. butzleri, %13’ü A. cryaerophilus ve %4,3’ü
A. buzleri ve A. cryaerophilus karışık kontaminasyonu olarak tespit edildi. Soğutma
sonrası numunelerin %22,2’si A. butzleri, %18,5’i A. cryaerophilus ve %3,7’si A.
buzleri ve A. cryaerophilus karışık kontaminasyonu olarak tespit edildi. Paketlenmiş
iv
ürünlerden toplanan numunelerin %29,6’sı A. butzleri, %7,4’ü A. cryaerophilus ve
%11,1’i A. buzleri ve A. cryaerophilus karışık kontaminasyonu olarak tespit edildi.
Tüm dünyada gıda kaynaklı bakteri olarak bilinen patojen Arcobacter
türlerinin incelenen broiler kesim aşamalarında oldukça yoğun bir kontaminasyona
sebep olduğu, broiler kesimindeki kritik kontrol noktalarını yönetme prensibine göre
her safhasının bir sonraki safhayı dikkate alarak geliştirilmesi ve son ürüne doğru
riskin en az düzeye indirilmesi göz önüne alındığında, araştırmamızda elde edilen
bulgular broiler kesimhanelerinde Kritik Kontrol Noktalarını (KKN) iyi bir şekilde
yönetilmediğini ve broiler etinin halk sağlığı açısından büyük bir risk teşkil
edebileceği sonucuna varıldı.
The most important contamination stages in broiler slaughter are scalding and feather plucking, evisceration, cooling and packaging. Then, to investigate the contamination of Arcobacter species, a total of 108 samples, 27 from each stage, were taken from three different broiler slaughterhouses at different times. Cultural and molecular methods were used for the isolation and identification of Arcobacter from the samples. Samples were washed using 400 ml Maximun Recovery Diluent in sterile bags. 10 ml of wash water was inoculated into ASB with 90 ml of CAT. It was incubated microaerophilically for 48 hours. The enriched culture was diluted 1:10 with Brucella Broth. After 300 μl of the dilution was spread on the agar through a cellulose nitrate membrane filter placed on Mueller Hinton Agar with 5% sheep blood, 100 μl of Brucella Broth was added. After 48 hours of aerobic incubation at 30°C, suspicious colonies were incubated microaerophilically at 30°C for 48 hours after planting on Mueller Hinton Agar. Colonies grown on Mueller Hinton Agar were examined by gram staining, oxidase, catalase, nitrate reduction, urease and hippurate tests. Arcobacter spp. multiplex PCR (mPCR) method targeting 16S and 23S rRNA genes was used for molecular identification of isolates. Amplified DNAs were observed in A. butzleri at 401 bp, A. cryaerophilus at 257 bp and A. skirrowii at 641 bp after 60 minutes of running at 90 volts, 70 milliamperes in the electrophoresis tank. Arcobacter spp. isolated and identified. As a result of the examination of isolates obtained from 104 samples by mPCR method, mixed contamination of A. butzleri, A. cryaerophilus and both Arcobacter species was detected in 51 samples. 52.9% of the 51 isolates were determined as A. butzleri, 31.4% as A. cryaerophilus and 15.7% as A. butzleri and A. cryaerophilus mixed contamination. A. skirrowii was not detected in the study. After boiling and feather plucking, 25.9% of the samples were found to be A. butzleri, 22.2% to A. cryaerophilus, and 11.1% to A. butzleri and A. cryaerophilus mixed contamination. After evisceration, 26.1% of the samples were detected as A. butzleri, 13% as A. cryaerophilus and 4.3% as A. butzleri and A. cryaerophilus mixed contamination. After cooling, 22.2% of the samples were detected as A. butzleri, 18.5% as A. cryaerophilus and 3.7% as A. butzleri and A. cryaerophilus mixed contamination. Of the samples collected from the packaged products, 29.6% were detected as A. vi butzleri, 7.4% as A. cryaerophilus and 11.1% as A. butzleri and A. cryaerophilus mixed contamination. Considering that the pathogenic Arcobacter species, known as foodborne bacteria all over the world, cause very intense contamination in the broiler slaughter stages, each stage is developed according to the principle of managing critical control points in the broiler slaughter, taking into account the next stage and minimizing the risk towards the final product, the findings of our study concluded that Critical Control Point (CCP) is not well managed in broiler slaughterhouses and broiler meat may pose a great risk for public health.
The most important contamination stages in broiler slaughter are scalding and feather plucking, evisceration, cooling and packaging. Then, to investigate the contamination of Arcobacter species, a total of 108 samples, 27 from each stage, were taken from three different broiler slaughterhouses at different times. Cultural and molecular methods were used for the isolation and identification of Arcobacter from the samples. Samples were washed using 400 ml Maximun Recovery Diluent in sterile bags. 10 ml of wash water was inoculated into ASB with 90 ml of CAT. It was incubated microaerophilically for 48 hours. The enriched culture was diluted 1:10 with Brucella Broth. After 300 μl of the dilution was spread on the agar through a cellulose nitrate membrane filter placed on Mueller Hinton Agar with 5% sheep blood, 100 μl of Brucella Broth was added. After 48 hours of aerobic incubation at 30°C, suspicious colonies were incubated microaerophilically at 30°C for 48 hours after planting on Mueller Hinton Agar. Colonies grown on Mueller Hinton Agar were examined by gram staining, oxidase, catalase, nitrate reduction, urease and hippurate tests. Arcobacter spp. multiplex PCR (mPCR) method targeting 16S and 23S rRNA genes was used for molecular identification of isolates. Amplified DNAs were observed in A. butzleri at 401 bp, A. cryaerophilus at 257 bp and A. skirrowii at 641 bp after 60 minutes of running at 90 volts, 70 milliamperes in the electrophoresis tank. Arcobacter spp. isolated and identified. As a result of the examination of isolates obtained from 104 samples by mPCR method, mixed contamination of A. butzleri, A. cryaerophilus and both Arcobacter species was detected in 51 samples. 52.9% of the 51 isolates were determined as A. butzleri, 31.4% as A. cryaerophilus and 15.7% as A. butzleri and A. cryaerophilus mixed contamination. A. skirrowii was not detected in the study. After boiling and feather plucking, 25.9% of the samples were found to be A. butzleri, 22.2% to A. cryaerophilus, and 11.1% to A. butzleri and A. cryaerophilus mixed contamination. After evisceration, 26.1% of the samples were detected as A. butzleri, 13% as A. cryaerophilus and 4.3% as A. butzleri and A. cryaerophilus mixed contamination. After cooling, 22.2% of the samples were detected as A. butzleri, 18.5% as A. cryaerophilus and 3.7% as A. butzleri and A. cryaerophilus mixed contamination. Of the samples collected from the packaged products, 29.6% were detected as A. vi butzleri, 7.4% as A. cryaerophilus and 11.1% as A. butzleri and A. cryaerophilus mixed contamination. Considering that the pathogenic Arcobacter species, known as foodborne bacteria all over the world, cause very intense contamination in the broiler slaughter stages, each stage is developed according to the principle of managing critical control points in the broiler slaughter, taking into account the next stage and minimizing the risk towards the final product, the findings of our study concluded that Critical Control Point (CCP) is not well managed in broiler slaughterhouses and broiler meat may pose a great risk for public health.
Açıklama
Anahtar Kelimeler
Arcobacter spp., broiler, kesimhane, gıda güvenliği, broiler, slaughterhouse, food safety
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Akkemik, Y., (2022). Kesim Hattının Farklı Aşamalarında Broiler Karkaslarında Arcobacter Türlerinin Varlığı ve Antimikrobiyel Dirençlerinin Belirlenmesi. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
