ND Lokal Genotype 7

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Transcript of ND Lokal Genotype 7

mf_rois@yahoo.co.id

August 15August 15thth 2011 2011

Newcastle Disease Outbreak Newcastle Disease Outbreak in vaccinated Broiler Breeders and Layersin vaccinated Broiler Breeders and Layers

The need for a new vaccineThe need for a new vaccine

Road Map

1.Newcastle Disease Virus

- Classification

2.The reason for ND vaccine breaks

3.Vaccination against ND

- Pathotypes vs genotypes- Vaccination trials with local ND vaccine- Challenge- Serology

Taxonomy:Family: ParamyxoviridaeGenus: AvulavirusSpecies: Avian Paramyxovirus 1 (Newcastle Disease virus)

1926 by Kraneveld in Buitenzorg, Indonesia1926 by Doyle in Newcastle, UK

Newcastle DiseaseNewcastle Disease

Newcastle Disease VirusNewcastle Disease VirusFusion protein

Hemaglutinin & Neuraminidase protein

Matrix protein

RNP complex• RNA• NP Nucleoprotein• P Phosphoprotein• L Polymerase

FHN

MP

L

N

RNA

Newcastle Disease Virus

2 antigenic 2 antigenic determinantsdeterminants

Classification of NDV

• Serotype• Pathotype• Genotype

NDV replication in the cytoplasma

• Air-borne or carried on contaminated shoes of workers, visitors, tyres, equipment, feed sacs, egg crates, wild birds.

• Mainly aerosol.

• Improperly cleaned houses and hospital pens are sources of NDV.

Transmission

Incubation period•4-6 days

Susceptibility:Susceptibility: (1)Hosts; natural outbreaks occur in chickens, other

domestic birds. Duck and geese no clinical signs.

(2)Breed; Layers are more susceptible than broilers. (3)Sex; male and females are equally susceptible.

in grower: breeder male higher density ND.(4)Age; resistance increases with age. (5) Season; ND is more common in the cold–dry as

well as cold-humid periods, season changes and windy periods of the year.

•Severe respiratory signs

•Oedema eyes and head

•Green diarrhoea

•Muscular tremors

•Torticollis

•Paralysis of legs and wings

•Opisthotonos

•Mortality up to 100 %

in susceptible birds

Clinical signsVVND pathotype

• Haemorrhagic lesions in intestine:

proventriculus, cecal tonsil and small intestines

• Haemorrhagic lesions and congestion in trachea

• Airsacculitis

• Egg yolk in abdominal cavity

• Haemorrhagic lesions of ovaries

• Vasodilations of blood vessels, e.g. brain

Gross lesions

•At least 236 species are susceptible

•Chickens are highly susceptible

•Ducks and geese --> few or no clinical sings

Pathogenesis

Current Field Observations

RevaccinationRevaccination: Yes, but wait - till acute phase has passed- till feed intake has been stabilized- with killed vaccine- antibiotic may help to prevent secondary infection

INSANITY :

DOING THE SAME THING OVER AND OVER AGAIN, EXPECTING DEFFERENT RESULT

( ALBERT EINSTEIN 1879 – 1955 )

Differential Diagnosis for egg fault/production Differential Diagnosis for egg fault/production dropdrop

ND IB Coryza EDS MG

Shell Problems + + - + +

Egg Drop + + + + +

Pale eggs + + - + -

Misshappen eggs - + - - -

Respiratory signs + + + - +

Affects interior quality

- + - - -

Shell-less eggs +/-* - - + -

Mortality +/- - - - +/-

*Feed intake problems

Infectious Bronchitis Newcastle Disease

Newcastle Disease in Broiler Breeder and Layer PulletsNewcastle Disease in Broiler Breeder and Layer Pullets

Newcastle Disease in Laying BirdsNewcastle Disease in Laying Birds

ND Virus disseminationND Virus dissemination

1. Initial infection

2. Viremia

2. Airsac

3. Proventiculus 3. Intestines

2. Lung

3. Brain

3. Liver

ND - Phases of infection

Replication

Viremia

Replication

Live vaccineLive vaccine Killed vaccineKilled vaccine

Perivascular cuffing of lymphocytes and plasma cells in brain (IHC)

…The vaccine homolgous with the challenge virus homolgous with the challenge virus reduced oral shedding significantly reduced oral shedding significantly more than the heterologous vaccines. NDV vaccines formulated to phylogenetically closer to the potential outbreak viruses may provide better may provide better ND control by reducing virus transmission ND control by reducing virus transmission from infected birds.

If you want to truly understand something, try to change it.

Kurt Lewin (1890 -1947)

Newcastle Disease….• Prevention:

(1) Control and reduce movement Control and reduce movement of visitors, workers, equipment and vehicles. (2) Change clothes and boots between flocks & houses. (3) Properly dispose sick and dead birds. Sick birds should be culled and slaughtered immediately. (4) Clean, disinfect and fumigate house and equipment between flocks. Leave the house empty and sealed for 21 days. (5) Vaccination with live vaccines, monitored by serology- detection of the presence and titre level of antibodies in serum.

ND vaccines (history)

Live – 1946Live – 1946- Mesogens- MesogensRoakin (1948), Komarow, Herts, Mukteswar

- Lentogens- LentogensB1 – introduced in 1950Lasota – introduced in 1952

Killed – 1970 -1980Killed – 1970 -1980- longer immunity, adjuvants

Immunogenicity

Available ND vaccines in the market Available ND vaccines in the market

Immunogenicityefficacious

Safetymild

Safety

Genotype I

Genotype II

ND vaccine technology is more than 50 years old.

Seed virus can only protect against Seed virus can only protect against mortalitymortality, but cannot prevent replication, dissemination and shedding.

ND vaccine efficacy

Serotype

Pathotypes

Genotypes

Serotypes

•HN (HA activity)

•1 Serotype (Cross reaction in serology and protection against mortality)

• Relationship between antigenicity and efficacy is still unclear.

Doubling Dilutions

Log Titres

1 02 14 28 316 4

32 564 6

128 7256 8512 9

1024 10

2048 11

Field Challenge and Seroconversion Post Infection

Killed Booster Vaccine

Interpretation

No immunity Maternal Immunity

Poor Immunity

Live Prime Vaccine

Live Booster VaccineProtection Against Mortality, No Protection Against Egg

Production Drop

Common Interpretation of ND titresCommon Interpretation of ND titres

Pathotype ICPI IVPI MDT

Viscerotropic velogenic

> 1.5 – 2.0 2.0 – 3.0 < 60 hours

Neurotropic velogenic

> 1.5 – 2.0 2.0 – 3.0 < 60 hours

Mesogenic > 0.7 – 1.5 0.0 – 0.5 > 60 h – 90 hours

Lentogenic 0.2 – 0.7 0.0 > 90 hours

Asymptomatic 0.0 – 0.2 0.0 > 90 hours

PathotypesBefore RNA Isolation & DNA Sequencing of the fusion cleavage site – based on virulence based on virulence in poultryin poultry

FFHN

MP

L

N

RNA

GenotypingGenotyping based on F-gene Sequence

2 antigenic 2 antigenic determinantsdeterminants

Total: 15,192 nucleotides with 82.7 % identity to ND Lasota

University of Maryland

Genotypes of NDV

Strain Host Year of isolation

Sequence Length (nt)

Genotype F cleavage site

Virulence* ICPI GenBank No.

cockatoo/Indonesia/14698/90

cockatoo 1990 complete genome

15192 VII RRQKRF V 1.84 AY562985

ID-1/88 cockatoo 1988 partial F 325 VII RRQKRF V NA EU604253

moluccan/Indonesia/904/87

NA 1987 partial M & F

1195 VII RRQKRF V NA AY444497

chick/Indonesia/W3/89

chicken 1989 partial F 395 VII RRQKRF NA NA AF503645

Bali-1/07 chicken 2007 partial F 315 VII RRQKRF V 1.77 AB426628

Only few published genome sequences of NDV isolated from Indonesia

Since the late 80’s Indonesian NDV belong to genotype VII

* V, virulent .

Genotype

II

I

IV

V

VI

VII

III

Class I

Class II

Nucleotide identity (%) of complete genome

Sukorejo/019

Cockatoo/90

LaSota Ulster Phy-LMV 42

Banjarmasin/010

92.0 92.3 82.6 85.1 84.7

Sukorejo/019 92.0 82.7 84.6 84.2

Cockatoo/90 85.5 86.7 85.9

S. Samal & T. Prajitno

GRRQKR↓FI......↓........↓........↓........↓..GRRRKR↓FI...R..↓.....R..↓.....R..↓.....R..↓.....R..↓...G.QG.↓L..G.QG.↓L..GKQG.↓L..GKQG.↓L.

Banjarmasin/010 Banjarmasin/010 Gianyar/013 Gianyar/013 Sragen/014 Sragen/014 Kudus/017 Kudus/017 Kudus/018 Kudus/018 Sukorejo/019Sukorejo/019Makassar/003 Makassar/003 Bali/020 Bali/020 Linggasari/050Linggasari/050Parungkuda/058Parungkuda/058Kisaran/069Kisaran/069LaSotaLaSotaPhy.LMV42Phy.LMV42

Alignments of F cleavage sites ICPI1.711.921.911.92

Genetic variability - NDV

Early (I, II, III; IV)

and recent (V, VII, VIII) lineages

(Virus Res, 120:36-48, 2006)

VII

V,VI, VIII

III-IV

II

I R&D Vaksindo

Vaccine strains

Genotypes of NDV

ND-affected countries in the world and its genotypesND-affected countries in the world and its genotypes

VII

VII, VIII, IXV

VII

VII

VII

R&D Vaksindo

Inactivated ND vaccine (Genotype VII)

Vaksinasi Challenge

ND HI Titer dengan Antigen yang berbeda

Vaksinasi

ND HI Titer dengan Antigen yang berbeda#trial 2

Vaksinasi Challenge

Control bird (SPF):DiarrheaConjunctivitisTracheitisSwollen spleenHemorrhagic proventiculusHyperemiaetc.

ETIKET Label

Week Day Vaksin Dosis Aplikasi Jenis0 2-4 ND-IB Live 1 dsTetes Mata kanan ND apatho-IB H-120 ND L 0,2ccInjeksi sc 1 10 ND-IB Live 1 dsTetes Mata Kiri ND Clone -IB H120 2 14 Gumboro Intermediate plus 1 dsCekok IBD M 3 22-23 Gumboro Intermediate plus 1 dsCekok IBD M ND Live 1 dsTetes Mata Kanan ND Lasota 5 35 Pox 1 dsTusuk Sayap ND-AI (H5NI) 0,5 ccInjeksi sc ND-AI 6 42 Coryza I (Alum) 0,5 ccInjeksi Dada Kanan Trivalen/Tetra

10 70 ND/IB Kill Variant 0,5 ccInjeksi Dada Kiri ND-IBmulti ND-IB Live 1 dsTetes Mata Kiri ND Lasota-IB M41

11 80 AI (H5NI) 0,5 ccInjeksi Dada Kiri Bivalent13 ND L 0.5 ccIM ND Genotype 7

15 105 ND/EDS/IBmulti 0,5 ccInjeksi Dada Kanan ND/EDS/IBmulti

16 112 ND-IB Live 1 dsTetes Mata Kiri ND Lasota-IB M41 Coryza II (Alum) 0,5 ccInjeksi Dada Kanan Trivalen/Tetra

18 AI (H5NI) 0,5 ccInjeksi Dada Kiri Bivalent19 ND L 0,5 ccIM

Contoh program vaksin untuk kontrol ND

1. Egg production drop – main issue in breeders and layers2. HI antigens suggests protection, but is not.3. Diagnosis and sound vaccination program is important4. Current ND vaccines protects against clinical signs, but cant prevent shedding and spread5. Emergency live ND vaccination in face of challenge protects against clinical signs, but severely affects egg production6. ND outbreaks in vaccinated poultry flocks are due to the emergence of antigenic variants (Genotype VII). Genotype VII vaccine is needed.7. ND vaccine seed is out of date8. Trial with inactivated ND vaccine (Genotype VII) proved against mortallity and egg production drop

In a NutshellIn a Nutshell

If nothing ever changed, there'd be no butterflies. Author Unknown

Thank You