SOME STUDIES ON ORNITHOBACTERIUM RHINOTRACHEALE INFECTION IN BROILER FLOCKS AT SHARKIA GOVERNORATE

Animal Health Research Institute, Zagazig Branch.

SOME STUDIES ON ORNITHOBACTERIUM RHINOTRACHEALE INFECTION IN BROILER FLOCKS AT SHARKIA GOVERNORATE

(With 5 Tables)

By

H.I.R. EL-Banna; E.A. Masoud and M.K. Rizk*

*Animal Health Research Institute, Mansoura Branch

(Received at 18/9/2008)

بعض الدراسات عن عدوىالاورنيثوباکتريم رينوتراکيالفيبداريالتسمين

بمحافظةالشرقية

حلمي إبراهيم رجب البنا ، السيد السعيد مسعود ، مدحت کمال رزق

تم أخذ عينات من 200 طائر تعاني من أمراض تنفسية من بعض المزارع في محافظة الشرقية، وتم إجراء الفحص الظاهري وعمل صفة تشريحية لهذه الطيور وتم أخذ عينات من الأحشاء الداخلية للزرع البکتيري. تم عزل 24 ميکروب من عترة ORT بنسبة (12%) من القصبة الهوائية، الرئتين، الأکياس الهوائية والجيوب الأنفية بنسب 10.5% و8% و5.5% و1% علي الترتيب، بينما لم يتم عزل ORT من الکبد. تم تصنيف العترات بيوکيميائياً وکذلک سيرولوجياً ووجد أنها جميعاً تنتمي للفصيلة A. وتم عمل اختبار حساسية لمعرفة مدى حساسية العترات المعزولة للمضادات الحيوية ووجد أن جميع العترات حساسة للأموکسيسيلين والأوکسي تتراسيکلين والأمبسيلين ووجد أنها غير حساسة للجنتاميسين والکولستين والدانو فلوکساسين. وتم عمل العدوى الصناعية بالعترات المعزولة بالرش في دجاج تسمين عمر 3 أسابيع ووجد أن الميکروب استطاع إحداث عدوى في الدجاج مع إحداث نفوق يصل إلي 5%. وکانت أهم الآفات التشريحية إلتهاب في الأکياس الهوائية والتهاب رئوي والتهاب القصبة الهوائية.

SUMMARY

The present study was carried out on 200 broiler chickens which had respiratory troubles on different farms of different districts at Sharkia Governorate, to throw a spot light on the clinical and microbiological investigations as well as prevalence rate of ORT infection in broiler flocks. Bacteriological examination revealed that 24 isolates (12%) of ORT were isolated from the examined birds. ORT was isolated from trachea, lungs, air sacs and sinuses in a percentage of 10.5%, 8%, 5.5% and 1% respectively. No ORT was isolated from liver. Serotyping of ORT using AGPT proved that all the isolates were belonged to serotype A. Antibiogram studies showed that all the isolates were highly sensitive to Amoxycillin, Ampicillin, Oxytetracyclin, and resistant to Gentamicin, Danofloxacin and colistin. Experimental infection with ORT evoked respiratory signs with mortality rate reached to 5%.  

Key words: Ornithobacterium rhinotracheale, broiler, respiratory troubles.

INTRODUCTION

Respiratory diseases due to infectious agents are an important problem in the broiler chicken industry (Turan and Akcadag, 2003). Ornithobacterium rhinotracheale (ORT) is a pathogenic agent causing respiratory diseases especially in chickens and turkeys(Hadimli and Erganis, 2004 and Kaukonen et al., 2004). Ornithobacterium rhinotracheale is a recent described species of bacterium associated with respiratory diseases, growth retardation, mortality and decreased egg production in chickens and turkeys. Pneumonia, pleuritis and air sacculitis characterize the infection (Canal et al., 2005 and Veen et al., 2005).

Ornithobacterium rhinotracheale (ORT) has been firstly isolated from broiler chickens by DuPreez (1991). Recently, Hafez (2001) isolated ORT from ducks, goose, ostrich, pheasants, pigeons, quails, rook and turkeys.  

In many countries of the worlds, ORT has been incriminated as a possible additional causative agent in respiratory disease complex in poultry. The organism cause substantial financial 1osses due to high rates of contamination up to 50% in slaughtered affected flocks (Charlton et al., 1993; Hinz et al., 1994; Dudouyt et al., 1995; Back      et al., 1997; Hafez and Sting, 1999 and Van Empel, 1999).

MATERIALS and METHODS

(1) Samples:

A total of 200 diseased and freshly dead birds suffering from respiratory troubles were collected from different farms of different districts at Sharkia Governorate. The examined birds were subjected to clinical signs and postmortem examination.

(2) Isolation and identification:

Loopfuls from trachea, lungs, air sacs, sinuses and liver were inoculated into defibrinated 10% sheep blood agar media containing 10ug gentamicin sulphate/ml blood agar (Back et al., 1996 and Rahimi and Banani, 2007) and incubated at 37^oC for 48 hours under 7.5 – 10% Co₂ tension by using gas bags (Oxoid) in candle jar according to Vandamme et al. (1994) and examined for suspected ORT colonies.

The obtained pure cultures of ORT were identified biochemically according to Hafez et al. (1993). The isolates were serotyped by using agar gel precipitation test (AGPT) according to Van Empel, (1997).

(3) Sensitivity test:

It was carried out according to the technique described by Back et al. (1997).

(4) Pathogenicity test:

This experiment was done to study the effect of ORT isolates on threeweeks old broiler chickens.

One hundred and fifty, one day old broilers obtained from       Al-Brmawy Company. Ten of them sacrificed and were subjected to postmortem and bacteriological examination. These birds were free from infection with ORT. The remaining 140 chicks were grouped into four equal groups (A, B, C and D).

All birds were vaccinated with LasotaND vaccine. First group (A) remained as control not infected and not treated while groups (B, C and D) infected with ORT. The Group (B) was remained not treated but group (C) was treated with Amoxycillin 10mg/kg b.wt. for 3 days. Group (D) was treated with Amoxycillin 10mg/kg b.wt. for 5 successive days. The dose of ORT was 100ml of cultured brain heart infusion broth (BHI), each ml of culture contain x 10⁹ c.f.u. The ORT was inoculated by coarse spray (Van Empel et al., 1996).

Birds were kept under observation for 4 weeks post infection. Clinical signs, postmortem lesions and reisolation trials and mortality were recorded.

RESULTS

Table 1: Results of isolation of ORT from naturally infected chicken in different farms at Sharkia Governorate.

Table 2: Biochemical character for isolated ORT.

Table 3: Isolation of ORT from internal organs of naturally infected birds.

Table 4: Result of in vitro sensitivity test of ORT isolates to different antimicrobial agents.

Table 5: Experimental infection of 3 weeks-old broiler chickens with ORT isolates

DISCUSSION

Natural infection of ORT in chickens at 4 weeks which was suffering from relatively mild respiratory signs (nasal exudates, sneezing, wetting of eyes, swelling of infraorbital sinus, retarded growth) accompanied by increased mortalities. The course of the disease was about 10- 14 days.

These finding were concurrent with findings of Van Beek et al. (1994); Sakai et al. (2000) and Heyla et al. (2005).

The main postmortem lesions in examined birds were congestion of respiratory passage and trachea, pneumonia, exudate in the air sacs, predominantly in the abdominal air sacs were the most striking feature. Similar findings were observed by Hafez et al. (1993) and Abd El-Ghany Wafaa, (2000).

The isolation of 24 (12%) isolates of ORT from 200 diseased and freshly dead chickens were collected from different farms of different districts at Sharkia Governorate were reflected their wide distribution through out these province (Table 1).

The distribution of ORT in internal organs showed that the highest incidence were observed in trachea (10.5%), lungs (8%), air sac (5.5%), sinuses (1%) and liver (0%) (Table 3). These findings were supported with the results reported by Chin (1996); Cauwerls et al. (2002) and El-Gohary and Sultan (2002).

Morphologically, the ORT colonies were observed on 10% sheep blood agar as non-haemolytic, non-pigmented grayish white colonies.

Microscopical examination of ORT colonies, stained with Gram stain revealed highly pleomorphic Gram negative rods, non-motile, non-spore former and non-capsulated. These findings inconformity with those obtained by Hafez et al. (1993); Van Empel et al. (1996) and Hanan et al. (2008).

The biochemical reaction of the ORT isolates in Table (2). These findings agree with those described by Charlton et al. (1993); Hinz,       et al. (1994); De Rosa et al. (1996) and El-Gohary et al. (1998).

Serotyping of 24 isolates of ORT using agar gel precipitation test with specific antisera against 18 ORT serotypes (A-R) revealed that all tested isolates were belonged to serotype A. These findings were similar to the previous results recorded by Hafez, (1996); El-Gohary and Awaad, (1998) and Abd El-Ghany, Wafaa (2000).

The result of sensitivity test for ORT isolates against antimicrobial agents, by using disc diffusion technique (Table 4) revealed that the isolates were sensitive to Amoxycillin, Oxytetracycline and Ampicillin while were variably to Ciprofloxacine, Streptomycin and Enrofloxacine and resistant to Neomycin, Spectinomycin, Gentamicin, Colistin and Danofloxacin. These result agreed partially with those described by Hinz et al. (1994).

Experimental infection with ORT are given in Table (5). Birds in group (B) "infected with ORT and not treated", the ORT was able to reproduce the respiratory disease as the same characteristics as seen in the natural outbreak and the mortality was 5% and morbidity rat was 57%. The birds in group (C) "infected with ORT and treated with Amoxycillin for 3 days", the ORT was cause light respiratory disease and mortality rate was 1% and morbidity rate was 22%. While the birds in group (D), the mortality rate was 0.0% and morbidity rate was 10%. Similar results observed by Hinz et al. (1994).

The result of treatment of experimentally infected broiler revealed that the treatment with Amoxycillin 10mg/kg b.wt. for 5 days decreased the mortality and morbidity rates than those treated with Amoxycillin 10mg/kg b. wt. for 3 days. Reisolation rate of ORT from experimentally infected and not treated broilers was 80% but 20% reisolation rate was recorded in infected and treated birds. These results were in agreement with those obtained by Hafez et al. (1993)andAwaad et al. (2002).

It is concluded that the ORT infection are incorporated in complicated respiratory infection and the utmost need for use of sensitive antibiotic for control measures considered an important direction to avoid loss of body weight, respiratory signs and economic losses.

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