1. ISOLATION AND IDENTIFICATION ORNITHOBACTERIUM RHINOTRACHEALE FROM CHICKEN IN VIETNAM
Nguyen Thi Bich Lien1, Vo Thi Tra An1, Tran Thi Ngoc Han1, Ho Hoang Dung1 and Niwat Chansiripornchai2
Nong Lam University, Vietnam
Chulalongkorn University, Thailand

2. Content ORT infection ORT - Isolation ORT - Identification
Further study

3. Introduction

4. Ornithobacterium infection
Ornithobacterium rhinotracheale (ORT)
Symptoms: sneezing, moist eyes, swollen heads, difficult breathing, mortality (2-10%)
Lesions: airsacculitis, tracheitis, pneumonia
Risk factors: virus (ND, AI, IBD, IB), ventilation (NH3, CO2)
Transmission: horizontal (aerosols, direct contact, drinkers), vertical (egg shell)

5. Ornithobacterium rhinotracheale
Gram-negative, rod shaped bacterium, pleomorphic
Slow growing, easily overgrown by less fastidious growing bacteria (gentamicin supp.)
For optimal growth
5–10% sheep blood agar
For 48 hours at 37°C
Microaerophilic conditions (5-10% CO2)
Small circular grey to grey-white colonies

6. Antibiotic susceptibility Pathogenic characteristics
1981
Turkey
1987
duck
1991
chicken
1993
Characterized
Isolation
Antibiotic susceptibility
Pathogenic characteristics
Vietnam: no publication on isolation ORT

7. The study

8. swab (30), trachea/ lungs (75)
Chicken: difficult breathing , sneezing , facial edema
Samples
swab (30), trachea/ lungs (75)
DNA extraction
Columbia +
5% sheep blood
+ gentamicin 5 µg/ml
PCR amplification
pinpoint small-colony, grey, greydish white, non-heamolysis
16S rRNA ORT
Gram stain (-), mobility (-), oxidase (+), catalase (-), MacConkey (-)
Nucleotide sequencing

9. Results Generally, positive 4.7% No swab samples found carrying ORT
5 isolates cultured from lung/ tracheal samples (6.6%)
7 samples were positive by PCR assay (6.7%) (9.3%)
Asadpour et al (2008), 2.3%
Turan và Ak (2002), 11,46%

10. RESULT: isolation Samples from No (swab No.) Pos. Rate (%)
Breeding hens
5 (3) 1 20
Layer
8 (5)
Broiler
21 (2) 2
9,5
Native chicken
40 (17)
Local breed
18 (3)
5,5
Black-Bone Silky Fowl 
13 (0)
7,7
Total
105 (30) 5
4,7
Seifi, 2002 (2,6%); Turkyilmaz, 2005 (1,2%), Ghanbarpour and Salehi, 2009 (3,5%)

11. RESULT: PCR
Positive control (ORT, N. Chansiripornchai, 2007)
Negative control (E.coli)
- 6: positive samples
negative sample
. PCR product was nucleotide-sequenced and compared with reference gene in GenBank (KC454288). . The homology of 100% found (several ORT sequences)

12. RESULT: PCR (direct from samples)
Samples from No
Pos.
Rate (%)
Breeding hens
5 (3) 1 20
Layer
8 (5)
Broiler
21 (2) 2
9,5
Native chicken
40 (17)
Local breed
18 (3)
5,5
Black-Bone Silky Fowl 
13 (0)
7,7
Total
105 (30) 7
4,7
Asadpour et al (2008): ELISA, 62,8%; PCR, 3,1%;
Ozbey et al (2004): ELISA 10,2%; PCR, 2,4%

13. Conclusion
First study on isolation and identification of ORT in Vietnam.
Further study on antimicrobial susceptibility, pathogenic characteristics

14. Thank you for your attention!