Bursa yöresinde tavuk, güvercin ve kumrulardan Newcastle Hastalığı Virusu'nun (NHV) izolasyonu ve patotiplendirilmesi
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Date
1995
Authors
Öncel, Taner
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Publisher
Uludağ Üniversitesi
Abstract
Bu çalışmada, Bursa ili ve çevresinde tavuk, güvercin ve kumrularda Newcastle hastalığının (NH) varlığı incelenmiş ve izole edilen Newcastle hastalığı viruslarının (NHV) patotiplendirilmeleri yapılmıştır. Aralık 1992 ve Eylül 1993 tarihleri arasında toplam 19 hastalık olgusu incelenerek 1 no'lu izolat Geçit'teki broiler işletmesinden, 2 no'lu izolat Edincik'teki ticari yumurtacı işletmeden, 3, 4 no'lu izolatlar Bursa merkezdeki ve 8 no'lu izolat İznik'teki köy tavuklarından, 5, 7 ve 9 no'lu izolatlar Mustafakemalpaşa' daki güvercinlerden, 6, 10vq 11 no'lu izolatlar Bursa merkezdeki güvercinlerden izole edilmiştir. Virus izolasyonu için 10 günlük spesifik patogen free (SPF) embriyolu yumurtalar kullanılmıştır. İdentifikasyon, Hemaglütinasyon-İnhibisyon (HI) testi ile hem tavşanda Lasota susuna karşı hazırlanan NHV antiserumu, hemde hasta güvercinlerden elde edilen >Log2 4 HI antikor titresine sahip serumlar kullanılarak yapılmıştır. İzolatlarm patotipleri, her izolat için embriyolu SPF yumurtaları ortalama öldürme zamanı (OÖZ), maternal olarak ortalama Log2 8 HI antikor titresine sahip günlük civcivlerde Intracerebral Patojenite İndeksi (İCPİ) ve civciv embriyo fibroblast (CEF) hücre kültüründe plak oluşturma yeteneği saptanarak belirlenmiştir. Patojenite testlerinde tavuk izolatları 1, 2, 3, 4 ve 8 sırasıyla 1,55, 1,61, 1,62, 1,60 ve 1,47 ortalama 1,57 İCPİ ve 49,8, 54,4, 49,6, 48 ve 51,2 saat ortalama 50,6 saat OÖZ değerlerine, güvercin izolatları 5, 6, 7, 9, 10 ve 11 ise sırasıyla 1,45, 1,35, 1,33, 1,40, 1,38 ve 1,41 ortalama 1,38 İCPİ ve 57,6, 56, 56, 56, 57,6 ve 54,4 saat ortalama 56,2 saat OÖZ değerlerine sahip olarak bulunmuşlardır. Tüm izolatlar CEFte plak oluşturmuşlardır. OÖZ ve CEF'te plak oluşturma sonuçlarına göre güvercin ve tavuk izolatları Velojenik patotipte bulunurken, İCPİ sonuçlarına göre tavuk izolatları Velojenik, güvercin izolatları ise Mezojenik patotipte bulunmuşlardır. İzolatlar biyolojik özellikleri yönünden Hemaglutinin termostabiliteleri ve Hemaglutinasyon-elusyon tipleri belirlenerek karakterize edilmişlerdir. Tüm izolatlarm Hemaglutinin termostabiliteleri dar bir sınır içerisinde 15-30 dakika arasında ve Hemaglutinasyon-elusyon tipleri 2 no'lu izolat haricinde, yavaş olarak tespit edilmiştir. İncelenen NH olgularının benzer özelliklere sahip viruslar tarafından oluşturulduğu görülmüştür. İzolatlardan bazılarının^, 3, 5, 6, 7, 9 ve 10) Dr. DJ. Alexander tarafından mAb'ler kullanılarak incelenmesi, bunların klasik ve variant yapılı viruslardan farklı, aynı gruba ait vi ruslar olduğunu göstermiştir. Sonuç olarak, Bursa ve çevresinde halen önemli bir problem olan NH'na karşı aşıla maların köy tavukları ve güvercinleride içermesi gerektiği düşünülmektedir.
In this study existence of Newcastle Disease (ND) in chicken, pigeon and doves were investigated in Bursa region and pathotyping of isolated Newcastle Disease virus (ND V) were made. Between December of İ992 to September of 1993 a total of 19 cases was examined, and as a result, following isolations obtained; isolate no. I from a broiler flock in Geçit, Bursa, isolate no.2 from a commercial layer flock in Edincik, Balıkesir, isolates nos. 3,4 and 8 from village chickens in Bursa and İznik respectively, isolates nos. 5,7 and 9 from pigeons in Mustafakemalpaşa, Bursa, and isolates nos. 6,10 and 11 from pigeons in Bursa. For isolation 10 day-old specific pathogen free (SPF) embryonated eggs were used. Identification was performed via Hemagglutination-Inhibition (HI) test using both NDV positive sera produced against Lasota strain in rabbit and pigeon positive sera with >Log2 4 HI antibody titre obtained from affected pigeons. Pathotypes of each isolate was evaluated using three different tests. The Mean Death Time (MDT) in 10 day-old SPF embryonated chicken eggs, the Intracerebral Pathogenicity Index (ICPI) in day-old maternally immune chicks with Log2 8 FH antibody titre, and the plaque formation ability in chicken embryo fibroblast (CEF). In pathogenicity tests, chicken isolates 1,2,3,4 and 8 have been found to have ICPIs of 1,55, 1,61, 1,62, 1,60 and 1,47 (average 1,57) and MDTs of 49,8, 54,4, 48, 48 and 51,2 hours (average 50,6 hours) respectively and pigeon isolates 5,6,7,9,10 and 11 have been found to have ICPIs of 1,45, 1,35, 1,33, 1,40, 1,38 and 1,41 (average 1,38) and MDTs of 57,6 56, 56, 56, 57,6 and 54,4 hours (average 55,9 hours) respectively. All isolates formed plaques on CEF. While all the isolates were of Velogenic pathotype on the basis of their MDT and plaque formation results, the chicken isolates were of Velogenic pathotype and pigeon isolates were of Mesogenic pathotype according to their ICPI results. Characterisation of the isolates, according to their biological properties was made by using Hemagglutination thermostability and Hemagglutination-elution tests. All of the isolates produced close grouping values ranged from 15 to 30 minutes in the Hemagglutinin thermostability test. In the Hemagglutination-elution test all of the isolates were in slow type, except isolate no.2. Results showed that examined ND cases caused by the viruses with similar properties. Monoclonal antibody typing of some of the isolates ( nos. 2, 3, 5, 6, 7, 9 and 10) was carried out in Central Veterinary Laboratory, Weybridge by Dr. D.J. Alexander and showed that isolates tested were indistinguishable each other, but unlike other NDVs so far tested.In conclusion, it was thought that ND still seems an important problem in Bursa region and vaccination against ND should be included village chickens and pigeon
In this study existence of Newcastle Disease (ND) in chicken, pigeon and doves were investigated in Bursa region and pathotyping of isolated Newcastle Disease virus (ND V) were made. Between December of İ992 to September of 1993 a total of 19 cases was examined, and as a result, following isolations obtained; isolate no. I from a broiler flock in Geçit, Bursa, isolate no.2 from a commercial layer flock in Edincik, Balıkesir, isolates nos. 3,4 and 8 from village chickens in Bursa and İznik respectively, isolates nos. 5,7 and 9 from pigeons in Mustafakemalpaşa, Bursa, and isolates nos. 6,10 and 11 from pigeons in Bursa. For isolation 10 day-old specific pathogen free (SPF) embryonated eggs were used. Identification was performed via Hemagglutination-Inhibition (HI) test using both NDV positive sera produced against Lasota strain in rabbit and pigeon positive sera with >Log2 4 HI antibody titre obtained from affected pigeons. Pathotypes of each isolate was evaluated using three different tests. The Mean Death Time (MDT) in 10 day-old SPF embryonated chicken eggs, the Intracerebral Pathogenicity Index (ICPI) in day-old maternally immune chicks with Log2 8 FH antibody titre, and the plaque formation ability in chicken embryo fibroblast (CEF). In pathogenicity tests, chicken isolates 1,2,3,4 and 8 have been found to have ICPIs of 1,55, 1,61, 1,62, 1,60 and 1,47 (average 1,57) and MDTs of 49,8, 54,4, 48, 48 and 51,2 hours (average 50,6 hours) respectively and pigeon isolates 5,6,7,9,10 and 11 have been found to have ICPIs of 1,45, 1,35, 1,33, 1,40, 1,38 and 1,41 (average 1,38) and MDTs of 57,6 56, 56, 56, 57,6 and 54,4 hours (average 55,9 hours) respectively. All isolates formed plaques on CEF. While all the isolates were of Velogenic pathotype on the basis of their MDT and plaque formation results, the chicken isolates were of Velogenic pathotype and pigeon isolates were of Mesogenic pathotype according to their ICPI results. Characterisation of the isolates, according to their biological properties was made by using Hemagglutination thermostability and Hemagglutination-elution tests. All of the isolates produced close grouping values ranged from 15 to 30 minutes in the Hemagglutinin thermostability test. In the Hemagglutination-elution test all of the isolates were in slow type, except isolate no.2. Results showed that examined ND cases caused by the viruses with similar properties. Monoclonal antibody typing of some of the isolates ( nos. 2, 3, 5, 6, 7, 9 and 10) was carried out in Central Veterinary Laboratory, Weybridge by Dr. D.J. Alexander and showed that isolates tested were indistinguishable each other, but unlike other NDVs so far tested.In conclusion, it was thought that ND still seems an important problem in Bursa region and vaccination against ND should be included village chickens and pigeon
Description
Keywords
Newcastle hastalığı, İzolasyon, Patotiplendirme, Newcastle disease, Isolation, Pathotyping
Citation
Öncel, T. (1995). Bursa yöresinde tavuk, güvercin ve kumrulardan Newcastle Hastalığı Virusu'nun (NHV) izolasyonu ve patotiplendirilmesi. Yayınlanmamış doktora tezi. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.