11/1/2015
An Overview of Poultry Diseases:
Diagnosis and Control
Salama Shany
Lecturer of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University
Mobile: 01066970930
Email: s_abohamra@yahoo.com
Salama.shany@vet.bsu.edu.eg
Outline
 Introduction
 Classification
 Diagnosis
 Clinical diagnosis
 Laboratory diagnosis
 Prevention and control of poultry diseases
 Main lines for disease prevention
 Medication
 Vaccination
Health/Epidemiological triad
Infectious agent
(Virulence)
Health is a balance
Host
(Immunity)
Disease agents:
Resistance:
- Deficiencies
- Good feed
- Toxins
- Intestinal flora
- Viruses
- Immunity
- bacteria
- Parasites
Environment
* Local
* Systemic
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Introduction
Role of the environment in disease incidence
It is a fact that field case is a sum of interaction
between many factors including:
1.Host:
Species , Age , Breed , General health , Genetic and
Immunity.
2.Pathogen:
Type , Virulence , Dose , Route , Duration and
frequency of exposure .
3.Host parasite interaction:
• Favors livability of pathogens (cold, vectors, organic matter, wet
letter).
• Reduces host resistance (cold, hot, ammonia, gasses, dust).
• Facilitate spread of pathogens to a susceptible bird (wind, insects,
rodents, pets, reservoir, overcrowdings)
Poultry Diseases
Outline
 Introduction
 Classification
 Diagnosis
 Clinical diagnosis
 Laboratory diagnosis
 Prevention and control of poultry diseases
 Main lines for disease prevention
 Medication
 Vaccination
a. non- infectious
1.Nutrional deficiency
2.Environmental
3.Intoxication.
b. Infectious
1. Viral
2. Bacterial
3. Mycotic
4. Parasitic
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Classification according to clinical outcome
Classification according to etiology
Locomotor
Bact.
Viral






Septicemia




E.coli
M. synoviae
Pasteurrella
Salmonella
Staph.
Strept.
 Viral arthr.
E.coli
Pasteurrella
Salmonella
Spirochaet.
 AI
 ND
Kidney lesion
 Septicemia
 NE toxins
Respiratory
Egg production





Salmonella
Mycoplasma
Pasteurrella
Coryza
E.coli
 IBD
 IB
 Avian
nehritis
 Tumors
 IB
 EDS
 AI
 ND
 AE (transient)
 Ochratoxins
 Vit. A def.
 Gout
 Protein and
carbohydrates def.
Mycotic
Paras.
 Leg mange
Nutrit.
 Ca., Ph., vit. D,
biotin, niacin,
def.
Others
Immunosupp.






E.coli
Mycoplasma
Pasteurrella
Infectious coryza
Turkey coryza
Chlamydia
 Pasteurrella
 Spirochaetosis
 Paratyphoid in
pigeon
 Botulism
Viral






AI
ND
IB
ILT
QB
Pneumovirus




Mycotic
 Aspergillosis
 Aspergillosis
Parasitic
 Syngamus trachae
 Rabbit Ear mange
 Coccidiosis (indirect)
Nutritional
 Vit. A def.
 Vit. E , B1, B2 def.
 Mycotoxins
 Nutritional. def.
Others










AI
ND
Marek’s
AE
Marek’s
Leucosis
Reticuloend.
REO
IBD
Anemia
 Antimicrob.
Ammonia
Heat stress
Pox (indir.)
Trichomon. (indir.)
Diseases associated with age
0 day to
4 weeks
At 4 to
6 weeks
6 or 8 weeks
till point of lay
During
laying
variable
ages
Egg born
diseases
except….????
Omphalitis
IB
Fowl cholera
Avian leucosis.
Viral
AI.
ND.
IBD
Spirochetosis
Marek’s
disease.
Bacterial
Mycoplasmosis.
Salmonellosis.
E. Coli
IB
(Respiratory
form)
Pox
Infectious Coryza
Tuberculosis.
Others
Mycotoxicosis.
Nutritional
deficiency
IBD
Coccidiosis
ILT
EDS - IB (repr)
Quail
bronchitis
Paratyphoid
Marek's disease
Chronic
parasitism
Coccidiosis
Bumble foot
Nervous
Bacterial
Pox
DVH
Species
Egg born diseases
True egg born diseases
(ovarian transmission)
False
(shell)
Bacterial
Viral
Mycotic
E.COLI
Mycoplasma
Leucosis
Aspergellosis
Salmonella
(paratyphoid)
Salmonella
Reo
AE
CIA
ADENO(EDS)
Some disease problems affect particular species
•
•
•
•
IB
IBD
ILT affects only chickens.
Duck Virus Hepatitis affects only ducks.
Quail Bronchitis affects only quail…etc.
Turkey hemorrhagic enteritis
•
•
•
•
•
•
Paramyxoviruses
AI
Mycoplasmosis (but host specific)
Mycotoxicosis
Cholera
Pox
Some diseases have wide host range
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Morbidity and mortality
Mortality
Morbidity
Examples
High
High
Septicemic diseases
Viral(AI – ND)
Bacterial (Acute Spirochaetosis
Acute salmonellosis- Coli septicemia
Fowel cholera )
Acute ILT
Duck virus hepatitis.
High
Low
Per acute fowl cholera
Acute intoxication
Low
high
Infectious Coryza
Pox
Mild ILT
CRD
Mycoplasmosis
Low
Low
Chronic Long Standing Deplitating Diseases
Outline
 Introduction
 Classification
 Diagnosis
 Clinical diagnosis
 Laboratory diagnosis
 Prevention and control of poultry diseases
 Main lines for disease prevention
 Medication
 Vaccination
DIAGNOSES STEPS
a. Field/Clinical
1.History
2.Clinical exam.
3.Pathplogical exam.
4.Sampling
b. Laboratory
1. Virological
2. Bacteriological
3. Mycotic
4. Parasitological
Colibacillosis









Etiology:
G-ve, non spore forming, motile aerobic bacteria
More than 700 serotypes
Antigens: Somatic-O; Flagellar-H; Capsular-K; Fimbrial-F
Commensals or pathogenic (APEC) and mostly APEC are
extraintestinal.
Epidemiology
Occurs in all types and age groups of poultry and also mammals.
Infection is more frequent in young than mature birds specially with
– Low standard of sanitation, poor environmental conditions
– After a respiratory or immunosuppressive disease
– False egg transmission
False egg transmission is frequent
Colibacillosis ……cont.
Clinical forms
 Omphalitis and yolk sac infection
 Panothalmitis
 Colisepticemia
 Airsacculitis/CRD
 Cellulitis (Infectious process)
 Salpingitis
 Synovitis and osteoarthritis
 Coligranuloma (Hjärre’s disease)
 Swollen head syndrome (SHS)
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Nodules in intestine. Coli-Gra, Marek,
ALC., ATB, SGP,&LONG cestodes
Colibacillosis; Lab diagnosis.
Sample : Lesion + heart blood in case of septicemia
Isolation : MacConkey agar (pink colonies)
EMB media (green metallic sheen)
Serotyping: Biochemical, serological and molecular
Mycoplasmosis
 Etiology:
 Atypical bacteria with no cell wall (Pleomorphic in size and shape,
fragile, not affected by some antibiotics………?
 Very small, very fragile intracellular pathogens.
 Colonies are microscopic, fried egg appearance and require 21 days
to grow.
 Epidemiology
 Mycoplasmas tend to be quite host specific
 Major species are:
- M. gallisepticum: respiratory disease, embryo mortality, lameness, eye
lesions, ataxia.
- M. Synoviae: infectious tenosynovitis, respiratory disease, embryo
mortality.
- M. Meleagridis: respiratroy disease, leg weakness, embryo mortality.
- M. iowae: embryo mortality, leg abnormalities.
Mycoplasmosis ……cont.
Clinical outcome
 Airsac disease
 Infectious sinusitis
 Synovitis
 Drop in egg production, lower fertility and
hatchability
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Mycoplasmosis; Lab diagnosis
Sample : Lesion (Sinuses, Air sacs, Joints, ovary and Egg)
Isolation : PPLO
Frey’s media (Colonies are microscopic, fried egg
appearance)
Mycoplasmosis; vaccination
• Killed vaccines:
• Live vaccination: (F strain, Ts-11
and 6/85 vaccines).
 Displace field strains.
 reduce egg production drops in layers
and egg transmission in breeders.
 Two doses are recommended .
 One dose is enough.
 F strain is fully virulent for turkeys  Safe.
and of mild virulence for chickens.
Fowl cholera
 Etiology:
 Pathogenic Pasteurella species are:
o Pasteurella multocida type A
 Fowl Cholera in chicken.
o Pasteurella multocida type B
 Hemorrhagic septicemia in ruminant.
o Pasteurella haemolytica
 Pneumonic Pasteurellosis in cattle.
 Gram-negative, bipolar (bipolarity appears in stained blood film of septicemic
birds and in recent culture), capsulated aerobic bacilli.
 Epidemiology:
 Virulence among isolates is highly variable; encapsulated strains are usually
highly virulent.
 Insects play a role in transmission
 Frequently seen in mature birds
 Chronically infected birds and asymptomatic carriers are considered to be major
sources of infection
 More effective than killed
vaccines.
 Do not prevent infection (Not protect
against clinical resp. disease
 The birds remain infected for life.
 How to judge successful
vaccination ? (??????)
 Not proved to be effective in broilers.
Fowl cholera; serotyping
Capsular
antigens
5 serogroup •
A,B,D,E,F •
Somatic
antigens
16 serogroup •
1−16 (1,3,4 predominate) •
Capsular Ag (heat labile mucopolysaccharides)
5 serogroup (A, B, D, E and F)
(Most avian type is A)
(Rabbit type is D)
2- Somatic Ag (heat stable lipopolysaccharides
To 16 serotype (1, 2...16)
(1, 3, 4 are world wide)
(5, 8, 9 prevalent in Egypt)
Fowl cholera……cont.
Clinical forms
 Septicemic
 Localized/chronic
1-wattle form
2-sinustis form
3-foot bad form
4-Arthritic form
5-steranal bursitis
6-ovarian form
7-otitis media form (meningitis)
8-respiratoty tract form
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Ovary
Liver
Heart
Pneumonia: FC,
Sal.,ND &AI.
Fowl cholera; Lab diagnosis
Sample : Lesion + heart blood in case of septicemia
Blood film stained with Giemsa (Bipolar M.O.)
Isolation : Grows readily on blood agar or dextrose starch agar with 5
% avian serum. In recent culture; colonies appear smooth,
circular, glisthing & iridescent while in old cultures colonies are
blue, rough & opaque (also in case of low virulent strains
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Fowl cholera; vaccination
Riemerella anatipestifer
Live vaccines
1. CU (Clemson University), a strain of low virulence
2. M-9, a mutant of CU with very low virulence
3. PM-1, a mutant of CU intermediate in virulence between
CU and M-9.
Etiology:
Pasteurella Heamolytica:
 G-ve, non motile, capsulated with bipolarity
 Cultivated on blood agar+(0.05% yeast extract, 5% calf serum ) in
anaerobic condition (CO2)
 Stained by Indian ink to see capsules
 20 serotypes
Bacterins
 They usually contain whole cells of serotypes 1, 3, and 4
emulsified in an oil adjuvant.
 Autogenous vaccines are preferred
Epidemiology:
 Duckling of 1-8 weeks of age highly susceptible.
 Duckling under 5 weeks of age of age usually die 2-12 days after
signs appear. Older birds may survive longer.
 The disease is rare in breeder ducks.
 Naturally occurring outbreaks of RA have been reported in turkeys.
Riemerella anatipestifer ……… cont.
Signs:
 Listlessness
 Ocular and nasal discharge
 Mild coughing and sneezing
 Greenish diarrhea
 Ataxia, tremor of head and neck and coma.
 Affected duckling show inability to move with the brood. Surviving
ducks may be stunted.
Lesions:
 The most obvious gross lesion in ducks is fibrinous exudates, which
involves serosal surface in general. Fibrinous air sacculitis is
common.
 Spleen maybe enlarged and mottled.
 Mucopurelent exudates in nasal sinuses
 caseous exudates in oviducts have been observed in RA infection.
Riemerella anatipestifer; vaccine
Infectious coryza
 Avibacterium paragallinarum (Haemophilus paragallinarum)
Live RA vaccine
 Developed against serotypes 1, 2 and 5
 Administered to 1-day-old duckling by aerosol or in drinking water.
 Single vaccination provided protection for least 42 days.
Inactivated bacterins
 Duckling is vaccinated at 2 and 3 weeks of age to provide adequate
protection up to market age.
– Gram-negative bipolar staining, non motile-non-spore-forming and
capsulated rod-shaped bacterium
– Requires special media to be isolated
– There are A, B, C serovars (serovar B a pathogenic).
– Virulence factors: capsule & haemagglutination antigen.
 Upper resp. tract infection and drop in egg production
 High morbidity, low mortality
 Bacterins provide sero-group-specific immunity (no cross protection)
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Infectious coryza
Infectious coryza ….. Lab diagnosis
Sample : Nasal sinuses
Isolation: Casman blood agar containing NAD - NaCl Chicken serum 1%. Colonies are 1-2 mm in size (dew like
drop- iridescent) within 3 wks.
Turkey coryza
 Colibacillosis
 Mycoplasmosis
 Pasteurrellosis
 Infectious coryza
 Turkey coryza
 Chlamydiosis
 Salmonellosis
 Spirochaetosis
 Clostridial diseases
•
•
•
•
•
•
•
•
Highly contagious disease (Morbidity of 80–100% in young turkeys)
Turkey poults and broilers and turkeys breeders are susceptible
More frequent in 2-6 week old turkeys
The causative agent is Bordetella avium
B. avium is pathogenic for turkeys and opportunistic in chickens
B. avium survives in litter for 1-6 months.
G-ve, non fermentative, motile, aerobic bacillus.
Grow on MacConkey agar, blood agar, and brain-heart infusion
broth.
• Colonies: small, compact, translucent, glistening pearl-like colonies
with smooth edges.
Turkey coryza ; lab diagnosis
• Isolation and identification of B. avium spp
• Pathogenicity test for susceptible poults
• Serology
A) Macroagglutination (detects IgM ~1 wk after infection)
B) ELISA (detects IgG >2 wk after infection).
C) Indirect immunofluorescent tests
• PCR
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Turkey coryza ; lab diagnosis
 Colibacillosis
 Mycoplasmosis
 Pasteurrellosis
 Infectious coryza
 Turkey coryza
 Chlamydiosis
 Salmonellosis
 Spirochaetosis
 Clostridial diseases
Avían Chlamydiosis
Avían Chlamydiosis
Thickened abdominal
airsac totally covered
with fibrin plaques .
Nasal discharge,conj. and lacremation
Serous fluid
and fibrin in the
pericardial sac.
Severe hepatomegaly.
Difficult breathing
Avían Chlamydiosis
Avían Chlamydiosis; Lab diagnosis
Sample : heart blood and lesions
Isolation : ECE
Impression smear from lesion stained with Giemsa
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Salmonellosis
Salmonellosis
Gram negative, non-sporulating rods.produce acid and gas form
glucose and mannitol, utilize citrate but do not ferment lactose or
form indole
Salmonella gallinrum-pullorum infection (non motile salmonellae)
It is also known Pullorum disease, fetal septicemia of young chicks,
bacillary white diarrhea (B. W. D.) or fowl Typhoid.
Lungs with congestion
Lung Brown discolor Sal.
Paratyphoid infections
Salmonella gallinrum-pullorum infection (non motile salmonellae)
Denote generally the diseases caused by any of the microorganisms
of the salmonella group other than Salmonella gallinrum-pullorum
Salmonellosis
Grayish white necrotic foci in
lung
Salmonellosis
Swollen hock joint containing
yellow viscous fluid.
Heart: Thickened yellowish
pericarditius
White nodules in the heart;
Bronzy liver and congested spleen
Salmonellosis
Salmonellosis; Lab diagnosis
Sample : heart blood and lesions
Isolation : pre-enrichement (Selenite-F-broth) then selective
media
Serology : Agglutination test
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Spirochaetosis
 Etiology
Borrelia anserina
 Aerobic, motile spiral bacterium, Gram negative
 Found between RBCs in acute feverish stage
 Cultivated on E.C.E via yolk sac
 Stained blood film from acute feverish stage by Gemsia stain or
Wright’s stain or Leishman’s stain-----spiral M.O. between RBCs
 Summer disease
 Soft tick is the main method of transmission
 Characterized by paresis or paralysis, thrist, mortality, mottled
enlarged spleen,enlarged liver with small hemorrhagic foci,
parboiled appearance of myocardium with fibrinous preicarditis
 Dark field microscopy or stained blood film
Clostridial diseases
 Colibacillosis
 Mycoplasmosis
 Pasteurrellosis
 Infectious coryza
 Turkey coryza
 Chlamydiosis
 Salmonellosis
 Spirochaetosis
 Clostridial diseases
Clostridial diseases
Liver necrosis Kidney swollen with cholecystitis , yellow or
green fibrinonecrotic pseudomembrane
C. colinum
C. perferingens type C
Clostridial diseases
Discolored muscle and under serosanguinous fluid.
Clostridial diseases
GD in wing
C. speticum, C, perfingins, C. novyi, and C. sporogenes
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Avian Tuberculosis
Liver Tuberculosis
Osteomyelitis of proximal
tibiotarsus.
TB
Swollen hock with
inflammatory exudate
along tendon sheaths
Multiple foci of necrosis in liver
in septicemic infection
Liver, Spleen ATB
Green liver in turkeys with
osteomyelitis.
Bilateral osteomyelitis of femoral head
and the joint into the body cavity.
Introduction
•
•
•
•
•
•
•
Nature of viruses
Maintenance of virus
Enveloped/non enveloped viruses
Immunosuppressive viruses
Egg transmitted viruses
Speticemic viruses
Immunity against different viruses
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18 H+ 11 N
Swabs from trachea ,cloaca, sinus& organs
Al. SAC-death at 24h +hemorrhages HA
IDENTIFICATION: HI monoclonal ,ELISA & PCR.
Resp. , Renal , reproductive.
VIRUS: sRNA, enveloped.
Corona ,RNA ,All Sac 3-5 pass curling and
dwarfing. AGP, HI,ELISA.
SAMPLES: Trachea, liver , spleen and brain in AL SAC -death
hemorrhagic embryo 2-6 days with HA +ve Fluid.
Trachea ’lung ,oviduct & kidney
IDENTIFICATION: HI , ELISA & PCR.
Comparative clinical signs
Signs
Coughing
Sneezing
Rales
Gasping
Shaking head
Ocular-nasal discharges
Swelling of face & wattles
Herpes ,grow on CAM with pocks and INIB.
Bluish-purple discoloration of face
Death 2-8 ds .AGP ,NT,
Red/white spots on legs and comb
Twisting of head and neck
Tracheal and eye mucosa
Green, watery diarrhea
HPAI
+
+
+
+
+
+
+
+/-
LPAI
+/+/+
+
+
-
NDV
+
+
+
+
+
+
+
+
+
+
IB
+
+
+
-
ILT
+
+
+
+
+
+
+
-
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Comparative PM lesions
Signs
Hemorrhagic tracheitis
General septicemia
Hemorrhage in proventriculus
Hemorrhage on cecal tonsils
Tracheal caseation
Nephritis and gout
Various methods of diagnosis
HPAI
LPAI
NDV
+
+
+
-
+/-
+
+
+
+
-
IB
ILT
- +
- - - ++ ++ -
Virus
Diagnostic technique
NDV
• Flock history, signs and necropsy, typical course of disease & mortality
• Virus isolation in 9-11 day old SPF eggs
• Serology: HA/HAI, FAT, ELISA test.
AI
• Clinical sings and lesions (HPAI).
• Isolation and identification of the virus in 9-10 day old SPF eggs
• Serology HA/HAI, AGP, VN, Neuramindase inhibition test and ELISA.
IB
• History, symptoms (gasping, and caseous plug at tracheal bifurcation)
• Isolation in 9-11 days old SPF chicken eggs
• Serology: AGP, ELISA and HA/HI
ILT
• History (acute), disease trend (rapid spreading), symptoms (typical coughing,
expulsion of blood and high mortality)
• Isolation in 11-12 days old SPF chicken eggs
• Serology: VN, AGP, ELISA and direct FAT
What is the best samples to submit
Virus
Best samples
NDV
• Trachea, Cloacal swabs
• Spleen, brain and liver kidney
AI
• Trachea, sinus exudates
Main lesions in ECE
Virus
Best samples
Confirmation
NDV
• Hemorrhage embryo
• HI using NDV antisera
• Embryonic deaths
• PCR
AI
• Positive HA allantoic fluids
• HI using AIV antisera
IB
• Stunted, curled and dwarfed embryos (after
• Tracheal and cloacal swabs
• Spleen and liver.
IB
• PCR
• Trachea, tracheal swabs
• Oviducts
• Green liver
• Kidneys.
ILT
• Larynx, tracheal exudate
• Tracheal and conjunctiva! swab and lungs
• AGPT using IBV antisera
• PCR
passaging)
• Kidney urates
ILT
• CAM lesions
• Chicken infection cause
typical c.signs
• PCR
Adenoviruses
Resp & mortality in quails.
Resp & egg prod. Change in chickens.
Adeno type 1 ( CELO ) , Agglut human O ,
All. Sac Curling and dwarfing.
Tracheal and lungs and air sacs.
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TURKEY HEMORRAGIC ENTERITIS
Drop in egg with change in
shell color and quality at
the beak in brawn egg
layers.
Spleenomegally
salpengitits in shell gland.
Rabid progress in signs within 24 hs , depression ,bloody dropping ,and death
massive hemorrhages. Small hemorrhages on heart
,muscles liver and proventriculus. Intestine filed with blood
Spleen enlarged mottled
IN IBs can be seen in liver, spleen, bone marrow, pancreas,
lug intestine and blood lymphocytes.
Adenovirus-3 , Ha + ve in
All sac duck embryo -> IN
IB. HI , ELISA&ELISA.
Shell gland ,Buffy coat egg
albumin.
Adeno type-2 , Turkey embryos.
Organs , Blood , Intestinal content.
Nervous in baby chicks.
Prod. Problems in breeder.
No gross lesion only
microscopic.
Entero virus, Yolk sac
muscular dystrophy and
leg paralyses after passage.
Brain. IF,AGP,NT
,ELISA,PCR and egg test
POX VIRUS , CAM 
Pocks + IC IB..
LESIONS
Mortality ,diarrhea,
dehydration
immunosup
Anemia , hemorr. ,
immunosupp. And
low PCV < 27 %
Birnavirus non enveloped
Yolk sac stunted ,liver
CIRCO VIRUS.
Whole blood ,Buffy coat , Semen ,oviduct sec. Eggs.
Injection of 1 day chicks in foot bad ,EM,PCR or
ELISA.
Necroses , hemor. in feather
AGP , NT , ELISA.
Bursa and spleen
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Swollen joint , stunted , lameness ,
DITHS,LOW
PROD&IMM
-UNOSUPP..
Low fertility immunosuppresion.
Joint , synovial mm. articular
surface and tendons
Reovirus, yolk sac or CAM 
stunted liver necroses and pocks.
AGP, NT , ELISA. Foot bad
inoculation of 1 day chicks.
Herpes DND, 3 serotypes, Yolk sac  pocks on CAM.
Synovial content ,
tracheal swab .
Detection of virus, provirus or transcriptase enzyme.
cloacal or
Buffy coat and ,Feather follicle
Differential diagnosis between lymphoid leucosis and Marek's
disease
Chronic disease ch. by anemia emaciation ,low productivity ,variable mortality and immunosup..
Feature
Lymphoid leucosis
Age of onset
16weeks
6– 4weeks or older
Symptoms
Absent
Frequently paralysis or paresis
Incidence
Seldom above 5%
Usually above 5%
Peripheral nerve enlargement
Absent
Usually present
Bursa of Fabricius
Nodular tumours
Diffuse enlargement or atrophy
Skin, muscle or proventriculus tumours
Usually absent
May be present
Peripheral nerve infiltration
Absent
Present
Cuffing in white matter ofcerebellum
Absent
Present
Tumour in the liver
Focal or diffuse
Frequently perivascular
Retrovirus carry its transcreptase enzyme .There are 6 groups .
Bursa of Fabricius
Intra-follicular tumour
Inter-follicular tumours or atrophy
Both exogenous and endogenous type. In ECE all sac  tumors at 200 days after
hatch . In TC no CPE but interfere RSV of the same group.
Follicular patterns of lymphoid cells
infiltration in the skin
Absent
Present
Cytology
Uniform lymphoblasts
Pleomorphic mature and immature cells
including lymphoblast, small medium and large
lymphocytes and reticulum cells.
Disease forms:The most important are:
1.Lymphoid .( BLD ). 2.Erythroid .
3.Myloblast
. 4. Myloid
5.Ostiopetrosis.
Marek's diseases
Gross Lesions
Microscopic Lesions
PCR,ELISA,COFAL and RIF tests used .Detection of virus or enzyme.
From blood, serum, eggs and lesions.
1-Acute reticulum cell neoplacia.
2.Runting disease syndrome.
3.Upnormal feathering (Nukanake ).
Sudden high mortality in 1-3
week old duckling with nervous
paddling.
4.Preferal nerve enlargement.
5.Chronic ulcerative proventriculitis.
Liver hemorrhages
6.Others.
Focal or defuse tumors in liver spleen ,
bursa, gonads, heart and kidney.
DVH-1 ,-2, and-3 ,yolk or all
sac stinted , liver lesion. AGP
,NT, ELISA & PCR.
Retrovirus + reverstransccriptase enzyme.
In embryo  specific lesions.
Whole blood, plasma, tumer for detection of
virus antigen or enzyme.
Liver, blood and feces.
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11/1/2015
Supclinical infection of turkey.
Acute highly fatal disease of
MUSKOVEY ducks and geese
Liver enlarged with necrosis of
several mm . Pancreas is roughly
circular ,gray-pink may and may
extend across a lobe.
Casinos precarditis and
prehepatitis
Picornalike ,in yolk sac death
4-11 dpi with coetaneous
congestion ,dwarfed and edema.
Parvovirus
Liver, pancreases, spleen ,kidney.
And feces. Histopathology and
EM..
Liver ,spleen.
DIAGNOSES STEPS
a. Field/Clinical
E. Tenella
Eimeria
Necatrix
Histom
onasis
1.History
2.Clinical exam.
3.Pathplogical exam.
4.Sampling
UE
b. Laboratory
1. Virological
2. Bacteriological
3. Mycotic
4. Parasitological
NE
Summary of laboratory diagnosis of poultry diseases
I.
II.
III.
IV.
V.
Field diagnosis
Sampling
Direct detection of pathogens, antigens and
parasitic eggs
Pathological examination
Isolation and identification
organ
swab
sample
Whole
blood
serum
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11/1/2015
Summary of laboratory diagnosis of poultry diseases
I.
II.
III.
IV.
V.
Field diagnosis
Sampling
Direct detection of pathogens, antigens and
parasitic eggs
Pathological examination
Isolation and identification
III- Direct pathogen detection
Blood film
• Impression smears and wet preparation
• Droplet examination
• Swabs
Blood film stained with giemsa
Fowl cholera
Spirochaetosis
Chlamydiosis
Blood parasites
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11/1/2015
III- Direct pathogen detection… cont
• Blood film
Impression smears and wet preparation
• Droplet examination
• Swabs
Impression smears and wet preparation
Fungal hyphae (PAS)
Chlamydia elementary bodies (Giemsa)
Trichomoniasis (Hanging drop technique)
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11/1/2015
III- Direct pathogen detection… cont
• Blood film
• Impression smears and wet preparation
Droplet examination
• Swabs
Droplet examination
• Uses: Parasitic egg and Eimeria Oocyst
• Methods:
A- Direct smear
B- Concentration technique:
a- Saturated salt solution technique.
b- Centrifugation concentration technique.
Cestodes and nematodes eggs
Parasites Egg
1- Cestodes.
Small, thin walled and contain embryonic
mass (embryo).
2- Ascaridia.
Large, double walled and contain embryonic
mass.
3- Hetrakis.
Smaller than Ascaridia but thicker.
4-Capillaria.
Lemon shape with embryonic masses in its edges
5- S. trachea.
Elliptical and operculated.
Eimeria Oocycst
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III- Direct pathogen detection… cont
• Blood film
• Impression smears and wet preparation
• Droplet examination
Swabs
Swabs
Viral antigen detection
(many viruses)
Summary of laboratory diagnosis of poultry diseases
I.
II.
III.
Isolation
IV. Pathological examination
INIB
ICIB
Tumor differentiation
IV.
V.
Field diagnosis
Sampling
Direct detection of pathogens, antigens and
parasitic eggs
Pathological examination
Isolation and identification
Summary of laboratory diagnosis of poultry diseases
I.
II.
III.
IV.
V.
Field diagnosis
Sampling
Direct detection of pathogens, antigens and
parasitic eggs
Pathological examination
Isolation and identification
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11/1/2015
Virus isolation
V. Isolation and identification of avian pathogens
Virus
isolation
• ECE
• T culture
Viral
• Bacterial
• Fungal
Vaccine
preparation
Uses
Virus
titertion
Virus
identification
Routes of egg inoculation
Yolk sac
5-7day
Amniotic sac
8-10 day
Allantoic sac
9-11 day
IBD
AI
AI
Chorio
allantoic
11-13 day
ILT
Mareks
DVH type3
ND
POX
AE
IB
IBD
DVH type 2
DVE
Mycoplasma
DVH type1
chlamydia
EDS
Virus identification
1-Pathology on ECE
2-Laboratory identification
A- Detect virus (Ag)
B- Virion
C-Viral nucleic acid
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11/1/2015
Pathology on ECE
1-Embryo death.
2- CAM
a- Plaque formation (Pock lesions).
b- Oedema.
3- Embryos
a-Stunting.
b- Cutaneous haemorrhage.
c- Abnormal development of muscles and feathers.
4-Abnormalities in visceral organs as
a- Enlargement of liver and spleen.
b- Greenish discoloration of liver.
c-Necrotic foci on heart and liver.
d-Formation of ureates deposits on mesonephrons.
CPE following yolk sac inoculation
5-7days
Virus
CPE
AE
Muscle atrophy
Curling, dwarfing
Living embryo shows nervous manifestations
Mareks
POCKS like lesion on CAM
- INIB
IBD
Haemorrhage along feather tract of embryo – greenish necrosed liver
DVH type 2
Liver necrosis
Allantoic sac inoculation
9-11days
Virus
CPE
AI –ND
High mortality-diffuse haemorrhages
IB
Curling –dwarfing (stunting) of embryo after serial blind passage (up
to 7 passages)
EDS
No specific lesion
DVH type 1
Liver necrosis
DVE
Embryo death- INIB
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CAM inoculation
11-13 days
Virus
CPE
POX
POCKS like lesion , oedema ,thickening , peteceal haemorrhage and necrosis
ICIB
IBD
As previous
ILT
POCKS like lesion yellowish , necrosis (button)
INIB in acute stage (4dayes)
After that masked by cell lysis
(7 days)
So trypsinized membrane test to detect INIB
Virus Identification… cont
1-CPE on ECE
2-Laboratory identification
A- Detect virus antigen
–
–
–
–
–
–
FAT
IP
ELISA
AGPT
Dot ELISA
HA
B- Detect virion
– Electron microscopy
C- Neutralize virus activity
– SNT
– HI test
D-Detect viral nucleic acid
– PCR and RT-PCR
II. Hemagglutination test
Ⅰ. Rapid plate agglutination test
 Principle: Some HA-bearing viruses agglutinate RBCs
in vitro.
 Procedures:
discard 0.25ml
Tubes
1
2
3
4
5
6
7
NS
0.25
0.25
0.25
0.25
0.25
0.25
0.25
virus
0.25
0.25
0.25
0.25
0.25
0.25
(1:4)
(1:8)
(1:16)
(1:32)
(1:64)
(1:128) (1:256)
0.5%RBC 0.25
0.25
0.25
0.25
0.25
0.25
0.25
(negative control)
mix, RT, 15-30min，observation
25
11/1/2015
HI test
II. Hemagglutination test
 Measure 4 or 8 HA units of antigen
 Interpretation
Hemagglutination titer: is determined as the highest dilution
 Example:
of virus that can cause an obvious ( ++) hemaglutination.
1
2
(1:8) 7(1:16)
3
4
5
 HA of 1:64, divided by 16 = 4 (dilute the isolates in
6
PBS 1:4 to obtain 8 HAU/25 ul)
(1:32) (1:64) (1:128) (1:256)
++++ +++
++
++
+
- control
III. Hemagglutination inhibition
test
III. Hemagglutination inhibition
test
 Principle:

Procedures:
Discard 0.25ml
If a person is infected by HA-bearing virus, anti-HA Ab may
appear
in
his
serum.
These
antibodies
can
block
Tube
1
hemagglutination and lead to the hemagglutination inhibition
NS(ml) 0.25
phenomenon.
serum
2
0.25
0.25
0.25
(1:2)
(1:4)
3
4
0.25
0.25
0.25
(1:8)
5
6
7
0.25
0.25
0.25
0.25
0.25
0.25
--
0.25
0.25
(1:16)
(1:32)
8
(1:64) (1:128)
Virus(4U) 0.25
0.25
0.25
0.25
0.25
0.25
0.5% RBC 0.5
0.5
0.5
0.5
0.5
0.5
0.25
0.5
0.5
Mix, RT, 30min, observation
III. Hemagglutination inhibition
test
III. Hemagglutination inhibition test
 Interpretation:
Hemagglutination inhibition titer is determined as
the highest dilution of serum that can completely
inhibit hemagglutination.
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Molecular Techniques
Temperature
100
PCR
 principle:
the PCR is an artificial way of doing DNA replication
 Advantages:
Melting
94 oC
Annealing
Primers
50 oC
50
0
o Less amount of pathogens
o Safe for high contagious and zoonotic
30x
Melting
o
Extension 94 C
72 oC
T i m e
3’
3’
5’
5’
3’
pathogens
3’
5’
5’
3’
5’
5’
3’
5’
5’
5’
5’
5’
3’
5’
5’
5’
3’
5’
3’
5’
3’
V. Isolation and identification of avian pathogens
PCR product stained with PCR product stained with ethidium ethidium
bromide bromide
• Viral
Bacterial
• Fungal
Lane 1: 100 bp DNA ladder,
Next lanes samples of 190bp
Organism
Salmonellae.
Media
Pre enrichment media
Shape of colony
Clear supernatant
Granular ,boundary deposit
a-Selenite-F-broth
b-Tetrathionate broth
Selective media:
a-Salmonella shigella agar
Fine, small, transparent
b-Brilliant green agar
Differential media
Colony morphology
MaCconky agar
non lactose fermenter.(colorless)
Proteus.
MaCconky agar
Large opaque, swarmy and non lactose
E.coli.
MaCconky agar
Smooth, convex and pink colonies
EMB media
Green metallic shine
fermenter .(motility (swarmy))
Mycoplasma
Pleuropneumonia like organism media Circular and transparent colonies with dense
(PPLO)
raised center (fried egg appearance)
Frey's media
Pasteurella
multocida
Blood agar
Glestining, very transparent and iridescent.
Dextrose starch agar + 5% avian serum
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V. Isolation and identification of avian pathogens
Organism
Strept. pyogens
Media
Blood agar for primary isolation
Shape of colony
Dew drop, non pigmented and surrounded
by zone of B-haemolysis.
Staph. aureus
Haemophilus
paragallinarum
(IC)
Blood agar
Circular coloniues surrounded by zone of
Nutrient agar
haemolysis (white yellow to orange)
BHIA + reduced NAD + Salt
Dew drop, smooth and iridescent
Blood agar + reduced NAD + Salt
Staph. may be used as NAD source
Campylo bacter
Campy cefex agar
Convex,water drop
clostridium
Blood agar
Zone of hemolysis
• Viral
• Bacterial
Fungal
V. Isolation of avian pathogens
• Viral
• Bacterial
Fungal
28
11/1/2015
Problems facing egg production
Stages of egg formation
part
infundibulum
Time
function
15-30 min
Catch ova
Outer vitelline memberan formation
Chalaza formation
neck
magnum
2-3 hr
isthmus
1-1.25 hrs
Isthmo uterine
junction
Uterus
(shell gland)
vagina
Drop in egg production
Albumen formation
(fertilization site)
Shell memberan formation
Mammary site of mineralisation
18-20 hr
less than 1
minute
Shell formation
Cutical formation
Quality of egg
Drastic drop
Gradual
Decrease
Transient drop
AI
Salmonella
AE
ND
E.COLI
EDS
pox
IB
IC
Shell
quality
Internal
quality
Air
cell
Yolk
Albumen
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11/1/2015
Quality of egg
1-External quality
Shape
I- External quality of egg (Shell quality )
Colour
Size
1-Thin shell
2-Rough shell
3-Miss
shaped
4-Cracked egg
Small
1-Loss colour
2-Yellow shell
3-Mottled
- Thin or soft shell :
Calcium def.
IB
- Rough shell :
High Ca
IB
- Miss-shaped egg :
EDS
- Cracked shell :
High stocking density
- Loss of color :
IB
EDs
ND
Old age
Genetic
Old age
Genetic
Improper collection
IB
ND
- Mottled shell :
High RH
- Yellow shell :
High doses of Oxytetracycline
Genetic
- Small egg size:
Low nutrient and protein intake
young age
Cracked egg
Miss shaped
Thin shell
Stained egg
Rough egg
II- Internal quality of egg
color
1- Air cell
Loss of air cell
•Watery Albumen
•Storage with Flat end downward
•Rough handling
Egg yolk
Blood or meat spots
•Cold environment.
•Vit K deficiency.
•AE.
•Mycotoxins.
•Genetic causes.
odour
shape
Abnormal
color
Mottled yolk
Taint egg
Flat yolk
Cheesy
yolk
Cotton seed meal
High cotton seed
meal
Robenedine
IB
Chilling
Increased oil in
ration
Peprazine
Detergents
Watery
albumen
Some bacterial
diseases
Phenothiazine
Moulds
Storage with
Flat end
downward
Sulphonamides
Increased
ammonia
Storage near a
strong odour
Nicarbazine
(anticoccidial)
Some antibiotics
(tetra.)
Mould growth
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11/1/2015
3-Albumen
Watery white
Mottled yolk
Abnormal colour
IB
Low protein in ration
Prolonged storage
Old age
Genetic causes
ND
Pink white
Bacterial infection
High iron level in
ration.
Fertility
-Cock
• Arthritis
• semen quality (some M.O transmitted in semen)
• nutrition deficiency
• System of housing
-Hen
(age-genetic-nutrition)
Hatchability of the egg
Vaccination
Early embryonic
Mid embryonic death Late embryonic death
death
(2 week)
(3 week)
(1 week)
Improper egg
Nutritional deficiency Improper incubation
sanitization
of egg
Nutritional deficiency Improper incubation
IB
of egg( temp and
humidity)
Ovi duct tumour
AE
V ND
M ND
L ND
HPAI
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11/1/2015
How could you protect your flock?
Control in an applied term
Lab. Monitoring
Vaccination and medication
Cleaning/Disinfection
1
2
3
Biosecurity
Diagnostics
&
surveillance
5
4
Feed and water quality
All-in All-out
Education
Control of rodents & insects
Elimination
of infected
Poultry
Decreasing
Host
Susceptibility
5
Flock management
Defense System of Chickens against Infections
•
• Peripheral lymphoid
tissue
Primary Organs
– Thymus gland
• T-cell system
 cell-mediated immunity
– Bursa of Fabricius
• B-cell system
– Harderian gland
– Caecal tonsilles
 humoral immunity
– Bone marrow
• Precursor blood cells
– Spleen
– Yolk sac
• Maternal immunity
– GALT
Introduction
Introduction
Immunizing agents
Immunity
Active immunity
Following clinical infection
Passive immunity
natural
Immunizing agents
Transfer of maternal
Antibodies through yolk
Following subclinical infection
Antigen
acquired
Following vaccination
Immunoglobulin
Following administration of
Immunoglobulin or antiserum
32
11/1/2015
Maternally derived immunity
In chickens, Ig are the principal mode of transfer of immunity. Little evidence shows that the
mother’s immune cells are passed on to the embryo.
Ig from hen’s circulation are deposited in the superficial epithelial and glandular cells of the
oviduct.
Chicks frequently receive up to 3 weeks of protection from
From the oviduct, IgG is transferred into the maturing oocyst and accumulates in the yolk sac.
maternal antibodies allowing their immune system to mature to
Ig produced locally in the oviduct likely constitutes an insignificant proportion of the transferred
Ig.
a level capable of protecting them if exposed to a potentially
The developing chick acquires maternal IgG from the yolk sac beside IgA and IgM are
transferred via the amniotic fluid. The developing embryo swallows IgA- and IgM-containing
amniotic fluid.
harmful virus or bacteria.
The transfer of IgG begins during the first week of embryonation but occurs most predominantly
during the last 3 days before hatching. The transfer from the yolk continues after hatch.
Peak levels of maternal IgG in the circulation of the newly hatched chick are reached around 2-3
days of age.
Maternally derived antibodies decline linearly in the recipient and become undetectable after 2-5
weeks.
Types of poultry vaccines
1
Live
2
Inactivated
3
Subunit
4
DNA
5
Recombinant
5
Types of vaccines, continued
Methods of Vaccine-Application
6. Toxoid vaccines
These vaccines are used when a bacterial toxin is the main cause of
illness. When the immune system receives a vaccine containing a
harmless toxoid, it learns how to fight off the natural toxin. The immune
system produces antibodies that block the toxin. E.g Vaccines against
diphtheria and tetanus.
7.Gene deleted vaccines
• Oral/ drinking water
– IBV, NDV, IBDV
• Spray
– IBV, NDV, ILT
• Eye drop
– IBV, NDV, ILT, M. gallisepticum
These are genetically engineered vaccines which involve the removal or
mutation of virulence gene of the pathogen
• Wing web
8.Peptide vaccine
• Intramuscular/Subcutaneous
These are the subunit vaccine prepared by chemical synthesis of short
immunogenic peptides.
– Poxvirus, AE, CAV (live)
– ND, IBV, IBD inactivated and/or Reovirus combinations
– Fowl Cholera, Infectious Coryza, Salmonella spp….
197
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Scheme of immunization
Vaccination programs
• Primary vaccination
• One dose vaccines (BCG, measles, mumps,
rubella, yellow fever)
• Multiple dose vaccines (polio, DPT, hepatitis B)
• Booster vaccination
• To maintain immunity level after it declines
after some time has elapsed (DT, MMR
199
Interpretation of vaccination titers
Outline
1- Intensity of Response:As indicated by the Mean Titer.
i.e. above Baseline Titers” : vary according to type of bird , age , vaccine
type , vaccination program, and other factors.
2- Uniformity of Response:As indicated by the % CV. i.e. Is the vaccine actually getting to the all birds
or not.
% CV
Uniformity
Less than 30 %
Excellent
From 30-50 %
Good
Greater than 50 %
Need to Improve






Introduction
Types of poultry vaccines
Methods of vaccine administration
Vaccination programs
Causes of vaccination failure
Interaction between different vaccines
Causes of vaccine failure
• Poor cold chain maintenance (storage and transportation)
• Vaccine reconstitution
• Vaccine administration
Conclusions
Take home message:
• Health status of the birds
• Human factor/ personnel
• Post vaccination
Goal: to prevent entry of diseases into your
flocks
Plan: develop and implement a common sense
disease prevention program
34
11/1/2015
THANK YOU
Questions
????
35