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
BEFORE 1994
the bacterium was named
Pasteurella-like,
Kingella-like or
Pleomorphic Gram-Negative Rod (PGNR)
and also the name TAXON 28 was used.
1994
It was named Ornithobacterium rhinotracheale
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
ORT isolation from clinically infected flocks
% No. of +
flocks
Range of
mortality
Mean
age
(days)
No of
tested
chickens
No. of
examined
flocks
57.5% 23 0.2%-
16.8% 1 - 42
720 40
INCIDENCE OF ORT ISOLATION FROM
ORGANS
Organs No of
organs
No of
isolates
% isolation
Air sac 240 152 63%
Lung 160 95 59%
Joint 60 28 46.6%
Trachea 95 38 40%
Sinus 24 9 37.5%
Brain 40 12 30%
Yolk sac 150 40 26.6%
Reference
strains*
ORT positive
isolates Types of media
+ + -Sheep blood agar
+ + -Bovine blood agar
- - -MacCkonkey
+ + -Brain heart infusion agar
- - -PPLO broth
+ + -Tryptic soy agar
Different media used for Isolation of ORT
Biochemical identification
Reference
strains (18)
ORT isolates
(n=23) Reactions
Gram negative Gram negative (23) Gram stain
Negative (17) Negative (23) Catalase
Positive (17) Positive (23) Oxidase
Negative (18) Negative (23) Indole
Positive (18) Positive (13) Urease
Positive (17) Positive (17) Voges - Proskauer
Negative (18) Negative (23) Methyl red
Not tested Negative (23) Citrate utilization
Negative (18) Negative (23) Ornithine
decarboxylase
Positive (15)
Positive (17)
Positive (18)
(variable)
(variable)
Not tested
Positive (18)
(variable)
Positive (19)
(variable)
(variable)
(variable)
Oxidative/ferment
ation of
-glucose
-fructose
-lactose
-maltose
-galactose
-mannose
Non motile Non motile Motility test
The results of 18 reference strains puplished by Hinz, (1994).
BIOCHEMICAL CHARACTERISTICS OF ORT ISOLATED FROM
BROILER CHICKENS
ENZYMATIC REACTIONS OF ORT ISOLATES IN API 20 E
STRIP (24 HR.)
enzymatic activities of ORT isolates by Api ZYM
Reference ORT isolates Reactions
Positive Positive N-Acetyl-β-glucosaminidase
Positive Positive Phosphohydrolase
Positive Positive Acid phosphatase
Positive Positive Chymotrypsine
Positive Positive Alkaline phosphatase
Positive Positive Esterase
Positive Positive Esterase lipase
Positive Positive Leucine aminopeptidase
Positive Positive Valine aminipeptidase
Positive Positive Cystine aminopeptidase
Reference ORT isolates Reactions
Positive Positive Trypsine
Positive Positive -galactosidase
Positive Positive -glucosidase
Negative Negative -glucosidase
Negative Negative -glucoronidase
Negative Negative -fucosidase
Negative Negative -mannosidase
Negative Negative Lipase
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 10 out of 30 embryos (33.3%).
Application of ELISA kits in the diagnosis of
Ornithobacterium rhinotracheale (ORT)
infection in chickens
SAMPLES
511 serum samples from 24 clinically infected
broiler flocks at ages of 1-42 day
408 serum samples from 11 broiler-breeding
flocks at ages between 26-56 week, associated
with drop in egg production (1-4.5%).
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)
ORT
isolation
Disease
problem
Results No of
tested
samples
Flock Suspect Positive
% No % No
+* Respirat. 0 0 2.5 1 40 F1
+* Growth
retardat. 0 0 10.0 4 40 F2
+* Respirat. 33.3 15 20 9 45 F3
+* Arthritis
Respirat. 31.6 12 23.7 9 38 F4
- Respirat. 2.4 6 0 0 56 F5
- Growth
retardat. 0 0 0 0 20 F6
Results of antibody detection of ORT by ELISA test
in serum samples from broilers using BioChek kits
ORT
isolat
ion
Disease
problem
Results No of
samp
les
Flock Suspect Positive
% No % No
+* Nervous 5 2 85 34 40 F7
+ Arthritis
Respirat. 57.1 8 28.5 4 14 F8
+ Respirat. 18.7 3 81.2 13 16 F9
- Respirat. 8.3 1 0 0 12 F10
- Healthy 0 0 0 0 30 F11
- Healthy 41.5 5 0 0 12 F12
13.5 49 20.3 74 363 Total
Results of antibody detection of ORT by ELISA test
in serum samples from broilers using BioChek kits
ORT
Isolatio
n
results
Disease problem
ELISA
(positive) No of
tested
samples
Flock
% No
+ Respiratory. 100 20 20 F13
+ Growth retardation 100 10 10 F14
+ Respiratory 100 10 10 F15
+ Growth retardation. 100 20 20 F16
+ Growth retardation. 5.0 1 20 F17
+ Respirat., growth
retardation. 41.6 10 24 F18
Results of antibody detection of ORT by ELISA test
in serum samples from broilers using IDEXX kits
ORT
Isolation
results
Disease problem
ELISA
(positive) No of
tested
samples
Flock
% No
+ Respiratory 100 12 12 F19
+ Respiratory 100 10 10 F20
+ Respiratory 100 12 12 F21
+ Respirat. ,growth
retardation 0 0 10 F22
- Respiratory 100 10 10 F23
- Healthy 0 0 10 F24
77.7 115 148 Total
77.7 115 148 Total
Results of antibody detection of ORT by ELISA test
in serum samples from broilers using IDEXX kits
Detection of antibody titers by ELISA test in serum
samples of broiler breeders using BioChek kit.
ELISA results No. of
tested
sample
s
Age
weeks Flock* Mean
titer Suspect Positive
% No % No
6215 30 3 70 7 10 32 F1
4565 30 3 70 7 10 33 F2
2581 30 6 70 14 20 49 F3
7751 10 2 90 18 20 54 F4
7166 10 1 90 9 10 57 F5
----- 21.4 15 78.5 55 70 TOTAL
Detection of antibody titers by ELISA in serum
samples of broiler-breeders using IDEXX kit.
ELISA results No. of
tested
samples
Age
(weeks) Flock*
Mean titre Positive
% No
4619 95.0 38 40 26 F6
1880 76.2 65 90 31 F7
3653 75.6 28 37 37 F8
4495 75.5 37 49 50 F9
6526 95.1 59 62 56 F10
3896 98.3 59 60 57 F11
---- 84.6 286 338 TOTAL
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