Roadmap for Diagnosis and Control of Important Poultry Diseases

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Roadmap for Diagnosis and Control of Important
Poultry Diseases

• Dr. J.M.Kataria
• Director
• National Institute of Animal Health
• Department of Animal Husbandry, Dairying and
  Fisheries
• Ministry of Agriculture, Govt.of India

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Indian Poultry Industry

• Egg production -Ranked 4th
• Contributes - Rs.29000 crores to GNP
• Potential tool
• - To fight poverty
• - Malnutrition
• - Unemployment
• Comprises - Small large commercial farms
  Network of hatcheries
• Supporting Allied industries
• - Compound feed
• - Equipments
• - Machinery
• - Pharma and Biologicals

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• Poultry Diseases can be classified as
• Ever-existent Diseases.
• E.g. Bacterial pathogens, ND etc.
• 2. Emerging diseases.
• E.g. CIA, ARV infections, IBH-HPS etc.
• 3. Re-emerging Diseases.
• E.g. IB, MD etc.
• 4. Exotic Diseases.
• E.g. AI

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NEWCASTLE DISEASE

• Highly contagious
• Dreaded disease recorded through out the world
• Virus belongs to family paramyxoviridae
• Three pathotypes-velogenic, mesogenic and
  lentogenic
• Over 250 species of birds are susceptible
• Pathotyping of isolates from outbreaks is
  important
• Fusion-gene is of prime importance associated
  with virulence
• Virulent has two basic amino acid at the cleavage
  site
• Low pathogenic has one basic amino acid

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LESIONS OF RANIKHET DISEASE IN PIGEONS AND
GUINEA FOWL
Pigeons affected with NDV Showing characteristic
signs of Nervous lesions like torticollis.
Guinea fowl showing nervous Lesions like
lameness.
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• Diagnosis
• PM lesions
• Histopathology
• Serological Tests ( HI, SNT, ELISA)
• RT-PCR

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RT-PCR

• Diagnosis of all the three pathotypes and also
  for specific amplification and differentiation

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Direct Quantitative Real Time RT-PCR Pathotyping
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Infectious Bursal Disease

• Severe immunosuppressive disease caused by IBD
  virus, a member of birnaviridae family
• Two serotypes
• Classical, very virulent and variant pathotypes
• A region of interest is VP2 having hyper variable
  region

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GROSS LESIONS OF IBD INFECTIONS IN CHICKENS
HAEMORRHAGES IN THIGH MUSCLE
HAEMORRHAGES IN BURSA
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Infectious Bursal Disease (IBD)
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• Diagnosis
• Signs lesions
• Histopathology
• AGID
• Immunofluorescence test
• VN test
• ELISA
• RT-PCR

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RE analysis of IBDV

• Differentiation of very virulent IBDV from
  classical and variant viruses

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• Control
• Avoid immuno suppression
• Role of MA
• Intermediate strain vaccine
• Inactivated vaccine in breeders
• Thermostable vaccine

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CHICKEN INFECTIOUS ANAEMIA Aplastic
anaemia Generalized lymphoid atrophy
Clinical disease Haemorrhages Increased
mortality (secondary complications)
Multifactorial Diseases
Key Haemorrhagic syndrome CIAV Aplastic
anaemia syndrome Role Gangrenous dermatitis
Blue wing disease Circular single stranded DNA
virus belonging to circoviridae family
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GROSS LESIONS IN SPF CHICKS EXPERIMENTALLY
INFECTED WITH CIAV
1
2
3
4
5
A
B

• A Normal organs of control chick (thymus,
  
• longitudinal section of femur bones, bursa,
  spleen and liver) (12 days old) B
• Severe atrophic thymus,
• Longitudinal section of femur bones showing
  pale bone marrow,
• Atrophic bursa,
• Atrophic spleen and
• Pale liver of CIAV infected chick (12 DPI)

'A A control chick showing normal sized thymus
(12 DPI) 'B CIAV infected chick showing
severe atrophy of thymus (12 DPI)
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• Diagnosis Control
• Signs lesions
• Histopathology
• Virus isolation ( in MDCC-CU 147 )
• Immunofluorescence test
• immunofluorescence or immunoperoxidase
  staining
• No effective vaccine

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MAREKS DISEASE

• Lymphomatous and neuropathic
• Alphaherpesvirus
• Characterized by pleomorphic lymphocytic
  infiltration, tumors and arteriosclerosis.
• Though very virulent MD virus was reported in
  European countries but in India it has not yet
  been isolated from any infected flock.

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DIAGNOSIS

• Clinical signs
• Histopathology
• Virus isolation
• Membrane immunofluorescent staining for MD tumor
  associated surface antigen
• Positive immunohistochemical staining for pp 38
  antigen

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CONTROL

• live attenuated vaccines
• Cell-associated or cell-free (lyopholised) live
  virus or HVT
• Oil adjuvant whole cell preparations and
  especially gB expressing fowl pox and
  baculoviurses elicit protective immune responses.
  
• A DNA vaccine containing the infectious BAC 20
  clone of serotype 1 MDV was tested.
• In India -HTV (FC-126) or SB-1 or in combination
  of the both.
• Unlike the European countries, Rispen strains not
  used in India .

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Infectious Bronchitis

• Characterized by severe respiratory symptoms
  reproductive disorder and nephritis
• Variant- High mortality in young chicks so far
  20 IBV variants identified worldwide and the
  little are no cross protection
• New IBV like viruses have been isolated from
  turkeys
• Spike protein gene (S-1) is of prime importance
  having hypervariable region

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• Appearance of new variants
• High rate of evolution
• Genetic recombination
• Insertions
• Mutations
• Exhibit antigenic drift
• An accurate and rapid serotype determination is
  an important factor in controlling IBV

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Avian Reovirus ARV- Double stranded RNA having
10 segmentsS-gene coding for ?-protein is of
prime interest

• MANIFESTATIONS OF REOVIRUS
• JOINT
• Arthritis/tenosynovitis
• Ruptured gastrocnemius tenders
• Slipped tendon
• RESPIRATORY TRACT INFECTION
• Respiratory disease (Fahey Crawley 1954)
• DIGESTIVE TRACT INFECTION
• Osteoporosis
• Cloacal pasting
• Hepatitis
• HEART INVOLVEMENT
• Pericarditis
• Hydropericardium
• BURSAL ATROPHY

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LESIONS OF CHICKENS AFFECTED WITH AVIAN REOVIRUS
Swollen foot pad and hock joint in infected chick
Reovirus infected chicks with stunted growth
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Diagnosis Control

• Signs lesions
• Histopathology
• Demonstration of Viral antigens
• Serological test (AGPT, FAT, ELISA)
• Live and Attenuated Vaccines
  available
• No vaccination in India at Present

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Avian Influenza

• HPAI arises from LPAI viruses.

• Mortality Morbidity very high.(100)
• Multiple visceral organ failure.
• Nearly 24 outbreaks till 2004 have been reported.
• How to differentiate b/w HPAI LPAI ?
• DEFINITION OF EU
• An infection of poultry caused by an
  influenza A virus that has an intravenous
  pathogenicity index in 6-week-old chickens gt1.2 /
  Any infection with influenza A viruses of' H5 or
  H7 subtype for which nucleotide sequencing has
  demonstrated the presence of multiple basic amino
  acids at the cleavage site of the haemagglutinin.

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Clinical Symptoms

• Dependant on virulence of virus, species
  affected
• and other environmental conditions

• HPAI
• Often no clinical signs other than sudden death
• Possible signs of illness
• Soft-shelled eggs
• Depression
• Cyanosis of wattle and combs
• Diarrhea
• Loss of appetite
• Respiratory distress

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Diagnosis

• Signs lesions
• Histopathology
• Viral Isolation Demonstration (ECE)
• Serological test (HI)
• RT-PCR

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• Vaccination
• Vaccine is a valuable tool in the control and
  elimination of avian influenza
• Vaccine alone is unlikely to lead to a successful
  eradication however vaccination combined with
  stamping out and adequate surveillance will
  likely lead to eradication in less time.
• Strategic vaccination in birds, if accompanied by
  appropriate surveillance will reduce the amount
  of virus excreted and lead to less viral exposure
  for humans
• Vaccine, if used, must be produced in accordance
  with OIE guidelines
• Inactivated vaccines are the commonly used
  vaccines throughout the world, at present.

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AVIAN MYCOPLASMA

• Caused by Mycoplasma gallisepticum (MG) and M.
  synoviae (MS).
• C R D characterized by coryza, conjunctivitis,
  sneezing, and by sinusitis
• loss of production and downgrading of meat-type
  birds, and loss of egg production

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DIAGNOSIS

• Serological Test
• Isolation in media
• Detection of their DNA
• FAT
• Immunoperoxidase tests

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CONTROL

• Both killed vaccines (bacteria) and live vaccines
  are currently in commercial use.
• A temperature sensitive (ts) clone derived from
  Australian M. synoviae field isolate produced by
  chemical mutagenesis was tested as live vaccine.
• Bio-security measures.

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Molecular Biological Techniques in Disease
Diagnosis

• RE analysis of whole genome or variable region
• Polymerase Chain Reaction
• Highly sensitive, quick and confirmative
• Used to detect the virus from decomposed tissue
• Diagnosis and differentiation
• Nucleic acid hybridization
• Types of probes
• Hot
• Cold- Non-hazardous, can be stored for longer
  duration
• Detection and differentiation
• In situ hybridization
• Dot-blot
• Riboprobes- for detection of RNA viruses

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• Nucleotide sequencing
• Cycle sequencing by Sangers method
• Maxim-Gilbert method
• Molecular epidemiology
• Evaluation of viruses
• Differentiation of strains
• Complete genome analysis

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Vaccines for the Future

• Genetically engineered bacteria viruses- holds
  great promise for revolutionizing health
  management of poultry.
• Development of multivalent single dose vaccines.
• Improvements in routes of administration to
  decrease stress and
• increase immune effectiveness. Eg. Multivalent
  vaccines by oral or drinking water.
• RNA viruses particularly IBDV and IBV change
  their epitopes.
• Quick identification of such epitopes and develop
  vaccines.
• Practicing in-ovo vaccination
• Development of food-based vaccines.
• Reduce the cost of vaccine production.
• Vaccines for rural poultry.

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IN-OVO VACCINATION

• Chickens develop some immunologic maturity before
  
• hatching.
• Vaccination in 18-day day embryos
• Vaccine virus deposited in amniotic fluid
• Embryos acquired infection via the respiratory
  tract.
• No-appreciable adverse effect on hatchability or
  weight gain.
• ADVANTAGES
• Delivery of a uniform dose of vaccine in eggs.
• Reduced labour cost associated with post-hatch
  vaccination.
• Early post hatch resistance to field outbreaks.

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IN-OVO VACCINATION

• Mareks Disease
• Quite useful ability of embryos vaccinated chick,
  to resist an early post hatch challenge
• Maternal antibody interfere with HVT vaccination
• Infectious bursal disease
• No B-cells immunologic tolerance
• Moderate type vaccine viruses are pathogenic
• Low virulence virus
• - No detectable lesions or
• - Transient lesion in BF
• Newcastle Disease
• - ND-B1 routinely used post hatch vaccination
• - Lethal for embryos.
• Chemical treatment reduced virulence for embryos.

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VACCINES FOR VILLAGE CHICKENS

• Village flocks require special vaccines
• Transportation and storage without refrigerators
• Thermostable vaccines
• Individual application is impossible
• Through feed and water
• Extremely cheap
• Small dose pack size
• Test for vaccine evaluation
• Local chickens
• Challenge organisms of local origin

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FOOD BASED VACCINES

• Could be useful for commercial chickens
• Spray on feed
• Concentrate of vaccine coated pellet
• Stresses associated with handling for
  vaccination,
• spray vaccination.
• Vaccine must be robust, capable of infecting
  through digestive tract.
• All attenuated vaccines not suitable for food
  vaccines e.g. fowl pox

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• Roadmap for Control
• 1. Surveillance of high risk diseases
• Sero-monitoring
• Molecular epidemiology
• Isolation identification and characterization of
  etiological agents
• Monitoring of diseases in other Avian species
• Formation of data bank on important diseases
• Link between state, SAUs, central, and private,
  laboratories with the referral laboratory
• Immediate reporting of diseases to refer
  laboratory

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• 2. Use of uniform vaccination schedule
• 3. Evaluation of the commercial vaccines from
• market
• Development of diagnostics for quick, precise
  diagnosis and evaluation of commercial
  diagnostics.
• Restriction of immunosuppressive agents.
• - Testing of poultry feeds for mycotoxins
• - Immunosuppressive potential of vaccines.
• 6. Implementation of strict Bio-security in farm
  and
• hatchery level with facilities for
  incineration.

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7. Quarantine measures to be strengthened. 8.
Restriction of poultry Trade in disease affected
areas. 9. Development of human resources
specialized in Avian diseases. 10. Initiation of
poultry disease eradication programme (E.g.
Ranikhet Disease)
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• Thank you for your kind attention

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