nong lam university, ho chi minh city faculty of animal science and veterinary medicine isolation...
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Nong lam University, Ho Chi Minh City
Faculty of Animal Science and Veterinary Medicine
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, VietnamChulalongkorn University, Thailand
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
Content
• ORT infection• ORT - Isolation• ORT - Identification• Further study
INTRODUCTION
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
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)
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
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
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
1981Turkey
1987duck
1991chicken
1993Characterized
• Isolation• Antibiotic susceptibility• Pathogenic characteristics• Vietnam: no publication on isolation ORT
THE STUDY
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
Samplesswab (30), trachea/ lungs (75)
Chicken: difficult breathing , sneezing , facial edema
DNA extraction Columbia + 5% sheep blood+ gentamicin 5 µg/ml
Gram stain (-), mobility (-), oxidase (+), catalase (-), MacConkey (-)
PCR amplification pinpoint small-colony, grey, greydish white, non-heamolysis16S rRNA ORT
Nucleotide sequencing
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
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%)
30
75
Samples
SwabLung/ trachea
Asadpour et al (2008), 2.3%Turan và Ak (2002), 11,46%
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
RESULT: isolationSamples from No (swab No.) Pos. Rate (%)
Breeding hens 5 (3) 1 20Layer 8 (5) 0 0
Broiler 21 (2) 2 9,5Native chicken 40 (17) 0 0
Local breed 18 (3) 1 5,5Black-Bone Silky Fowl 13 (0) 1 7,7
Total 105 (30) 5 4,7
Seifi, 2002 (2,6%); Turkyilmaz, 2005 (1,2%), Ghanbarpour and Salehi, 2009 (3,5%)
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
RESULT: PCR
. PCR product was nucleotide-sequenced and compared with
reference gene in GenBank (KC454288).
. The homology of 100% found (several ORT sequences)
1. Positive control (ORT, N. Chansiripornchai, 2007)
2. Negative control (E.coli)3. - 6: positive samples7. negative sample
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
RESULT: PCR (direct from samples)Samples from No Pos. Rate (%)
Breeding hens 5 (3) 1 20Layer 8 (5) 0 0
Broiler 21 (2) 2 9,5Native chicken 40 (17) 1 0
Local breed 18 (3) 2 5,5Black-Bone Silky Fowl 13 (0) 1 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%
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
Conclusion
• First study on isolation and identification of ORT in Vietnam.
• Further study on antimicrobial susceptibility, pathogenic characteristics
Faculty of Animal Science and Veterinary Medicine – NLU - HCMC
Thank you for your attention!