Ticari broylerlerde timokinonun bazı bakteriyel solunum sistemi hastalıklarında mukozal bağışıklık üzerine olan etkisinin araştırılması
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Dosyalar
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi / Sağlık Bilimleri Enstitüsü / Mikrobiyoloji (Veterinerlik) Ana Bilim Dalı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu tez çalışmasında ticari broylerlerde ilk defa, inhalasyon yolu ile uygulanan timokinonun, mukozal bağışıklık üzerine etkisi ve deneyesel Escherichia coli (E. coli) ve Ornithobacterium rhinotracheale (ORT) solunum sistemi enfeksiyonlarına karşı koruyucu etkinliğinin belirlenmesi amaçlanmıştır. Çalışmada, öncelikle challenge aşamasında kullanılacak, E. coli ve ORT suşlarının 10 günlük Ross 308 erkek broyler civcivler için LD50 dozları belirlendi. LD50 dozları, E. coli için; 1x109 CFU/ml, ORT için 1x1010 CFU/ml olarak hesaplandı. Deneysel aşamada; 1 kontrol ve 8 challenge olmak üzere 9 grup oluşturuldu. Her grupta 28 civciv olacak şekilde toplam 252 adet 1 günlük civciv üzerinde çalışıldı. Çalışma grupları; I-Kontrol, II- Concanavalin A (Con A), III- Timokinon (TQ), IV- E. coli, V- ORT, VI- E. coli + Con A, VII- E. coli + TQ, VIII- ORT + Con A, IX- ORT + TQ grubu olacak şekilde tasarlandı. Çalışmanın, 1, 3 ve 5. günlerinde, III., VII. ve IX. gruplarda bulunan her bir civcive 10 mg/kg TQ, II., VI. ve VIII. gruplarda bulunan her bir civcive 0,4 µg Con A nebulizer yardımıyla solunum yolundan uygulandı. Çalışmanın, 7. günü, IV, VI ve VII. gruplara LD50 dozunda E. coli, V., VIII. ve IX. gruplara ORT ile intratrakeal yolla challenge yapıldı. Deney süresince 1, 5, 15, 25, 35 ve 45. günlerde kan ve trakeal yıkantı örnekleri toplandı, ayrıca canlı ağırlıklar ve ölümler kaydedildi. Çalışma 45. gün sonlandırıldı. Çalışma sonunda tavukların nekropsisi ve iç organların makroskobik değerlendirmeleri yapıldı. Akciğer dokularından kültür yapılarak bakteri geri izolasyon oranları hesaplandı. Alınan kan örneklerinden ticari ELISA ile TNF-α, IFN-γ ve IL-4 ölçümleri yapıldı. Trakeal yıkantı örneklerinde ticari ELISA ile IgA ve qRT-PCR ile AvBD 1 ve AvBD 3 β-defensin gen ekspresyon seviyeleri ölçüldü. Timokinon uygulanan gruplarda, uygulanmayan gruplara göre canlı ağırlık artışının ve kesim ağırlığının yüksek olduğu görüldü. E. coli ve ORT challenge gruplarında tipik enfeksiyon bulguları görülürken, TQ uygulamasının enfeksiyondan kaynaklı patolojik bulguları azalttığı görüldü. TQ uygulamasını takiben, akciğerlerden bakteri geri izolasyon oranları E. coli + TQ grubu için %25, ORT + TQ grubu için %17,86 olarak hesaplandı. TQ uygulanan grupta, kontrol grubuna göre TNF-α konsantrasyonunda önemli bir fark bulunmazken (p<0,05), ORT + TQ grubunda yüksek olduğu görüldü. IL-4 seviyelerinde gruplar arasında farklılık görülmedi (p<0,05). IgA konsantrasyonun TQ uygulanan grupta kontrol grubuna göre 25. günde yükseldiği, en fazla artışın E. coli + TQ grubunda olduğu görüldü. AvBD 1 gen ekspresyon seviyesindeki en fazla artış 15. günde ORT + TQ grubunda görülürken, AvBD 3 gen ekspresyon seviyesindeki en fazla artış 15. günde E. coli + TQ ve ORT + TQ gruplarında görüldü. İnhalasyon yoluyla TQ uygulanmasının solunum sisteminde ifade edilen antimikrobiyal peptitleri (AvBD 1 ve AvBD 3), mukozal immünglobulin (IgA), Th1 hücre yanıtı üzerinde immünmodülatör etkilere sahip olduğu belirlendi. Ayrıca, enfeksiyonlardan kaynaklanan canlı ağırlık azalışı ve verim kaybını azalttığı, bakteri eliminasyonunu artırarak hastalığın klinik seyrini ve enfeksiyon prognozunu iyileştirdiği görüldü. Sonuç olarak, ticari broylerlerde TQ uygulamasının bakteriyel solunum sistemi enfeksiyonlarında mukozal bağışıklığı artırarak hastalıkların önlenmesinde yararlı olabileceği belirlendi. Antibakteriyellerin alternatifi olarak kullanılabileceği ancak etkinliğin doza ve kullanım dönemine bağlı olduğu görüldü. Çalışmada elde edilen veriler ışığında, broylerlerde 42-45 günlük yetiştirme döneminde enfeksiyonlardan korunmak amacıyla TQ'nun birden fazla uygulanması gerektiği kanaatine varıldı.
In this thesis, it was aimed to determine the effect of thymoquinone administered by inhalation in commercial broilers for the first time on mucosal immunity and its protective efficacy against experimental Escherichia coli (E. coli) and Ornithobacterium rhinotracheale (ORT) respiratory system infections. In the study, LD50 doses of E. coli and ORT strains which was used in the challenge phase were determined for 10-day old Ross 308 male broiler chicks firstly. LD50 doses were calculated as 1x109 CFU/ml for E. coli and 1x1010 CFU/ml for ORT. In the experimental stage; 9 groups, including 1 control and 8 challenge, were created. A total of 252 pieces, 1-day-old chicks were studied, with 28 chicks in each group. Working groups was designed as I-Control, II- Concanavalin A (Con A), III- Thymoquinone (TQ), IV- E. coli, V- ORT, VI- E. coli + Con A, VII- E. coli + TQ, VIII- ORT + Con A, IX- ORT + TQ group. On the 1st, 3rd and 5th days of the study, 10 mg/kg TQ was administered to each chick in the groups of III, VII, IX and 0.4 µg Con A administered to groups of II, VI and VIII through the respiratory tract with the help of a nebulizer. On the 7th day of the study, the groups of IV, VI and VII were challenged with LD50 dose of E. coli and the groups of V, VIII and IX challenged with ORT by intratracheal route. During the experiment, blood and tracheal wash samples were collected, also body weights and deaths were recorded on days 1, 5, 15, 25, 35 and 45. The study was terminated on the 45th day. At the end of the study, the chickens were necropsy and the internal organs were evaluated macroscopically. Bacterial re-isolation rates were calculated by culturing lung tissues. TNF-α, IFN-γ and IL-4 levels were deteced with commercial ELISA from blood samples. In tracheal wash samples, IgA levels were measured with commercial ELISA and AvBD 1 and AvBD 3 β-defensin gene expression levels were determined by qRT-PCR. It was observed that the body weight gain and slaughter weight were higher in the thymoquinone applied groups compared to the non-administered groups. While typical infection findings were observed in E. coli and ORT challenge groups, it was observed that TQ application reduced pathological findings caused by infection. After TQ application, bacterial re-isolation rates from lungs were calculated as 25% for E. coli + TQ group and 17.86% for ORT + TQ group. While there was no significant difference in TNF-α concentration in the TQ-treated group compared to the control group (p<0.05), an increase was observed in the ORT + TQ group. There was no difference between the groups in IL-4 levels (p<0.05). It was observed that the IgA concentration increased on the 25th day in the TQ-treated group compared to the control group, and the highest increase was observed in the E. coli + TQ group. While the highest increase in AvBD 1 gene expression level was seen in the ORT + TQ group on day 15, the highest increase in AvBD 3 gene expression level was observed in E. coli + TQ and ORT + TQ groups on day 15. It was determined that administration of TQ by inhalation had immunomodulatory effects on antimicrobial peptides (AvBD 1 and AvBD 3) and mucosal immunoglobulin (IgA) in the respiratory system, and Th1 cell response. In addition, it was observed that it reduced the weight loss and yield loss caused by infections, and improved the clinical course of the disease and the prognosis of infection by increasing bacterial elimination. As a result, it was determined that TQ application in commercial broilers may be beneficial in preventing diseases by increasing mucosal immunity in bacterial respiratory system infections. It was observed that it can be used as an alternative to antibacterials, but the effectiveness depends on the dose and the period of use. In the light of the data obtained in the study, it was concluded that TQ should be applied more than once in order to prevent infection in the 42-45 day rearing period in broilers.
In this thesis, it was aimed to determine the effect of thymoquinone administered by inhalation in commercial broilers for the first time on mucosal immunity and its protective efficacy against experimental Escherichia coli (E. coli) and Ornithobacterium rhinotracheale (ORT) respiratory system infections. In the study, LD50 doses of E. coli and ORT strains which was used in the challenge phase were determined for 10-day old Ross 308 male broiler chicks firstly. LD50 doses were calculated as 1x109 CFU/ml for E. coli and 1x1010 CFU/ml for ORT. In the experimental stage; 9 groups, including 1 control and 8 challenge, were created. A total of 252 pieces, 1-day-old chicks were studied, with 28 chicks in each group. Working groups was designed as I-Control, II- Concanavalin A (Con A), III- Thymoquinone (TQ), IV- E. coli, V- ORT, VI- E. coli + Con A, VII- E. coli + TQ, VIII- ORT + Con A, IX- ORT + TQ group. On the 1st, 3rd and 5th days of the study, 10 mg/kg TQ was administered to each chick in the groups of III, VII, IX and 0.4 µg Con A administered to groups of II, VI and VIII through the respiratory tract with the help of a nebulizer. On the 7th day of the study, the groups of IV, VI and VII were challenged with LD50 dose of E. coli and the groups of V, VIII and IX challenged with ORT by intratracheal route. During the experiment, blood and tracheal wash samples were collected, also body weights and deaths were recorded on days 1, 5, 15, 25, 35 and 45. The study was terminated on the 45th day. At the end of the study, the chickens were necropsy and the internal organs were evaluated macroscopically. Bacterial re-isolation rates were calculated by culturing lung tissues. TNF-α, IFN-γ and IL-4 levels were deteced with commercial ELISA from blood samples. In tracheal wash samples, IgA levels were measured with commercial ELISA and AvBD 1 and AvBD 3 β-defensin gene expression levels were determined by qRT-PCR. It was observed that the body weight gain and slaughter weight were higher in the thymoquinone applied groups compared to the non-administered groups. While typical infection findings were observed in E. coli and ORT challenge groups, it was observed that TQ application reduced pathological findings caused by infection. After TQ application, bacterial re-isolation rates from lungs were calculated as 25% for E. coli + TQ group and 17.86% for ORT + TQ group. While there was no significant difference in TNF-α concentration in the TQ-treated group compared to the control group (p<0.05), an increase was observed in the ORT + TQ group. There was no difference between the groups in IL-4 levels (p<0.05). It was observed that the IgA concentration increased on the 25th day in the TQ-treated group compared to the control group, and the highest increase was observed in the E. coli + TQ group. While the highest increase in AvBD 1 gene expression level was seen in the ORT + TQ group on day 15, the highest increase in AvBD 3 gene expression level was observed in E. coli + TQ and ORT + TQ groups on day 15. It was determined that administration of TQ by inhalation had immunomodulatory effects on antimicrobial peptides (AvBD 1 and AvBD 3) and mucosal immunoglobulin (IgA) in the respiratory system, and Th1 cell response. In addition, it was observed that it reduced the weight loss and yield loss caused by infections, and improved the clinical course of the disease and the prognosis of infection by increasing bacterial elimination. As a result, it was determined that TQ application in commercial broilers may be beneficial in preventing diseases by increasing mucosal immunity in bacterial respiratory system infections. It was observed that it can be used as an alternative to antibacterials, but the effectiveness depends on the dose and the period of use. In the light of the data obtained in the study, it was concluded that TQ should be applied more than once in order to prevent infection in the 42-45 day rearing period in broilers.
Açıklama
Anahtar Kelimeler
Mikrobiyoloji = Microbiology ; Veteriner Hekimliği = Veterinary Medicine
