IMMUNOLOGICAL AND BACTERIOLOGICAL STUDIES ON ORNITHOBACTERIUM RHINOTRACHEALIS

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1 IMMUNOLOGICAL AND BACTERIOLOGICAL STUDIES ON ORNITHOBACTERIUM RHINOTRACHEALIS PRESENTED BY RABAB AMIN KHALIFA UNDER THE SUPERVISION OF Prof. Dr. Mohamed Refai Prof. of Microbiology, Faculty of Vet. Med. Cairo Univ. and Prof. Dr. Abd El-Gelil El-Gohary Prof. of poultry diseases. Faculty of Vet. Med. Kafr El-Sheikh University

2 BEFORE 1994 the bacterium was named Pasteurella-like, Kingella-like or Pleomorphic Gram-Negative Rod (PGNR) and also the name TAXON 28 was used. It was named 1994 Ornithobacterium rhinotracheale

3 ORNITHOBACTERIUM RHINOTRACHEALE VANDAMME ET AL., 1994 Biota Domain Bacteria (Haeckel, 1894) Phylum Bacteroidetes Class "Flavobacteria" Order "Flavobacteriales" Family Flavobacteriaceae Reichenbach et al., 1992 Genus Ornithobacterium

4 ORT isolation from clinically infected flocks No. of examined flocks No of tested chickens Mean age (days) Range of mortality No. of flocks % %- 16.8% %

5 INCIDENCE OF ORT ISOLATION FROM ORGANS Organs No of organs No of isolates % isolation Air sac % Lung % Joint % Trachea % Sinus % Brain % Yolk sac %

6 Different media used for Isolation of ORT Types of media -Sheep blood agar -Bovine blood agar -MacCkonkey -Brain heart infusion agar -PPLO broth -Tryptic soy agar ORT positive isolates - - Reference strains* - -

7 PINPOINT COLONIES OF ORT ON TRYPTIC SOY AGAR

8 Biochemical identification Reactions Gram stain Catalase Oxidase Indole Urease Voges - Proskauer Methyl red ORT isolates (n=23) Gram negative (23) Negative (23) (23) Negative (23) (13) (17) Negative (23) Reference strains (18) Gram negative Negative (17) (17) Negative (18) (18) (17) Negative (18)

9 Citrate utilization Ornithine decarboxylase Oxidative/ferment ation of -glucose -fructose -lactose -maltose -galactose -mannose Negative (23) Negative (23) (18) (variable) (19) (variable) (variable) (variable) Not tested Negative (18) (15) (17) (18) (variable) (variable) Not tested Motility test Non motile Non motile The results of 18 reference strains puplished by Hinz, (1994).

10 BIOCHEMICAL CHARACTERISTICS OF ORT ISOLATED FROM BROILER CHICKENS ENZYMATIC REACTIONS OF ORT ISOLATES IN API 2 E STRIP (24 HR.)

11 enzymatic activities of ORT isolates by Api ZYM Reactions ORT isolates N-Acetyl-β-glucosaminidase Phosphohydrolase Acid phosphatase Chymotrypsine Alkaline phosphatase Esterase Esterase lipase Leucine aminopeptidase Valine aminipeptidase Cystine aminopeptidase Reference

12 Reactions Trypsine -galactosidase -glucosidase -glucosidase -glucoronidase -fucosidase -mannosidase Lipase ORT isolates Negative Negative Negative Negative Negative Reference Negative Negative Negative Negative Negative

13 PATHOGENICITY OF SOME ORT ISOLATES TO 11- DAY CHICKEN EMBRYOS. The inoculation of ORT into 11- day -old embryos in SPF eggs caused death of 1 out of 3 embryos (33.3%).

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15 Application of ELISA kits in the diagnosis of Ornithobacterium rhinotracheale (ORT) infection in chickens

16 SAMPLES 511 serum samples from 24 clinically infected broiler flocks at ages of 1-42 day 48 serum samples from 11 broiler-breeding flocks at ages between week, associated with drop in egg production (1-4.5%).

17 ELISA KITS: The ELISA Kits are available commercially and supplied by IDEXX-France BioChek-Holand Calculation and interpretation of the results using ELISA software (provided by kits suppliers)

18 Results of antibody detection of ORT by ELISA test in serum samples from broilers using BioChek kits Flock F1 F2 F3 F4 F5 F6 No of tested samples No Results % Suspect No % Disease problem Respirat. Growth retardat. Respirat. Arthritis Respirat. Respirat. Growth retardat. ORT isolation * * * * - -

19 Results of antibody detection of ORT by ELISA test in serum samples from broilers using BioChek kits Flock F7 F8 F9 F1 F11 F12 Total No of samp les No % Results Suspect No % Disease problem Nervous Arthritis Respirat. Respirat. Respirat. Healthy Healthy ORT isolat ion * - - -

20 Results of antibody detection of ORT by ELISA test in serum samples from broilers using IDEXX kits Flock No of tested samples ELISA (positive) No % Disease problem ORT Isolatio n results F Respiratory. F Growth retardation F Respiratory F Growth retardation. F Growth retardation. F Respirat., growth retardation.

21 Results of antibody detection of ORT by ELISA test in serum samples from broilers using IDEXX kits Flock No of tested samples No ELISA (positive) % Disease problem ORT Isolation results F Respiratory F Respiratory F Respiratory F22 1 Respirat.,growth retardation F Respiratory - F24 1 Healthy - Total Total

22 Detection of antibody titers by ELISA test in serum samples of broiler breeders using BioChek kit. Flock* Age weeks No. of tested sample s No % ELISA results Suspect No % Mean titer F F F F F TOTAL

23 Detection of antibody titers by ELISA in serum samples of broiler-breeders using IDEXX kit. Flock* Age (weeks) No. of tested samples ELISA results Mean titre No % F F F F F F11 TOTAL