Newcastle Disease Outbreak in vaccinated Broiler Breeders and Layers

The need for a new vaccine

mf_rois@yahoo.co.id
August 15th 2011

Road Map
1. Newcastle Disease Virus

2.
3.

Classification

The reason for ND vaccine breaks

Vaccination against ND

Pathotypes vs genotypes Vaccination trials with local ND vaccine Challenge Serology

Newcastle Disease
Taxonomy: Family: Paramyxoviridae Genus: Avulavirus Species: Avian Paramyxovirus 1 (Newcastle Disease virus) 1926 by Kraneveld in Buitenzorg, Indonesia 1926 by Doyle in Newcastle, UK

Newcastle Disease Virus

Fusion protein Hemaglutinin & Neuraminidase protein Matrix protein

RNP complex RNA NP Nucleoprotein P Phosphoprotein L Polymerase

Newcastle Disease Virus

F HN
P L

2 antigenic determinants
Classification of NDV Serotype Pathotype Genotype

N RNA

NDV replication in the cytoplasma

Transmission
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.

Incubation period
4-6 days

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

Clinical signs
VVND pathotype 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

Gross lesions
Haemorrhagic lesions in intestine: proventriculus, cecal tonsil and small intestines Airsacculitis Egg yolk in abdominal cavity Haemorrhagic lesions of ovaries Vasodilations of blood vessels, e.g. brain

Haemorrhagic lesions and congestion in trachea

Pathogenesis
At least 236 species are susceptible Chickens are highly susceptible Ducks and geese --> few or no clinical sings

Current Field Observations

Revaccination: 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 drop

ND IB Shell Problems Egg Drop Pale eggs Misshappen eggs Respiratory signs Affects interior quality Shell-less eggs Mortality + + + + + + + + + + Coryza EDS + + + + + + MG + + + +/-

+/-* +/-

*Feed intake problems

Infectious Bronchitis

Newcastle Disease

Newcastle Disease in Broiler Breeder and Layer Pullets

Newcastle Disease in Laying Birds

ND Virus dissemination 3. Brain

2. Lung 2. Airsac 1. Initial infection 3. Liver

2. Viremia

3. Proventiculus

3. Intestines

ND - Phases of infection Replication Viremia Replication

Live vaccine

Killed vaccine

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

The vaccine homolgous with the challenge virus reduced oral shedding significantly more than the heterologous vaccines. NDV vaccines formulated to phylogenetically closer to the potential outbreak viruses may provide better 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 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 1946 - Mesogens Roakin (1948), Komarow, Herts, Mukteswar - Lentogens B1 introduced in 1950 Lasota introduced in 1952 Killed 1970 -1980 - longer immunity, adjuvants

Available ND vaccines in the market

Genotype I

Genotype II

Immunogenicity

Safety

Immunogenicity efficacious

Safety mild

ND vaccine efficacy
ND vaccine technology is more than 50 years old.

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

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.

Common Interpretation of ND titres

Doubling Dilutions 1 2 4 8 16 32 64 128 256 512 1024

Log Titres 0 1 2 3 4 5 6 7 8 9 10 No immunity

Interpretation Maternal Immunity Live Prime Vaccine Poor Immunity Protection Against Mortality, No Protection Against Egg Production Drop Live Booster Vaccine

Field Challenge and Seroconversion Post Infection

Killed Booster Vaccine

2048

11

Pathotypes
Before RNA Isolation & DNA Sequencing of the fusion cleavage site based on virulence in poultry

Pathotype ICPI
Viscerotropic velogenic Neurotropic velogenic Mesogenic Lentogenic Asymptomatic > 1.5 2.0 > 1.5 2.0 > 0.7 1.5 0.2 0.7 0.0 0.2

IVPI
2.0 3.0 2.0 3.0 0.0 0.5 0.0 0.0

MDT
< 60 hours < 60 hours > 60 h 90 hours > 90 hours > 90 hours

Genotyping based on F-gene Sequence

F HN
P L

2 antigenic determinants

N RNA

Genotypes of NDV

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

University of Maryland

Only few published genome sequences of NDV isolated from Ind

Since the late 80s Indonesian NDV belong to genotype VII

Strain cockatoo/Indones ia/14698/90 ID-1/88 moluccan/Indone sia/904/87 chick/Indonesia/ W3/89 Bali-1/07 Host cockatoo cockatoo NA chicken chicken
Year of isolation

Sequence complete genome partial F partial M &F partial F partial F

Length (nt)

Genotype

F cleavage site RRQKRF RRQKRF RRQKRF RRQKRF RRQKRF

Virulence* V V V NA V

ICPI 1.84 NA NA NA 1.77

GenBank No.

1990 1988 1987 1989 2007

15192 325 1195 395 315

VII VII VII VII VII

AY562 985 EU604 253 AY444 497 AF503 645 AB42 6628

* V, virulent .

Nucleotide identity (%) of complete genome

Sukorejo/ 019
Banjarmasin/ 010 Sukorejo/019 Cockatoo/90 92.0

Cockatoo /90
92.3 92.0

LaSota
82.6 82.7 85.5

Ulster
85.1 84.6 86.7

PhyLMV 42
84.7 84.2 85.9
Genotype I II III IV V VI VII Class I

Class II

S. Samal & T. Prajitno

Alignments of F cleavage sites

Banjarmasin/010 Gianyar/013 Sragen/014 Kudus/017 Kudus/018 Sukorejo/019 Makassar/003 Bali/020 Linggasari/050 Parungkuda/058 Kisaran/069 LaSota Phy.LMV42 GRRQKRFI ........ ........ ........ ........ GRRRKRFI ...R.... ...R.... ...R.... ...R.... ...R.... .G.QG.L. .GKQG.L.

ICPI 1.71 1.92 1.91 1.92

Genetic variability - NDV

Early (I, II, III; IV)

and recent (V, VII, VIII) lineages

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

Genotypes of NDV
VII

V,VI, VIII III-IV

Vaccine strains

II

R&D Vaksindo

ND-affected countries in the world and its genotypes

VII

V VII

VII, VIII, IX

VII

VII

R&D Vaksindo

Inactivated ND vaccine (Genotype VII)

ND HI Titer dengan Antigen yang berbeda

Antigen Lasota Vs ND Genotype 7
12.00 11.00 10.00 9.00 8.00 7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00

GMT

Vaksinasi
21 w 8.58 7.58 8.22 6.88 23 w 9.00 7.75 8.60 7.35

Vaksinasi
25 w 8.95 8.40 8.16 7.06 28 w 10.47 8.40 8.40 7.68

Challenge
29 w 9.55 8.25 7.75 6.75 31 w 9.26 9.37 9.35 9.05 32 w 8.87 8.67 8.94 9.13

ND Ls Ag LS ND Ls Ag ND Gen 7 ND Gen 7 ag Ls ND Gen 7 ag Nd Gen 7

ND HI Titer dengan Antigen yang berbeda #trial 2 Antigen Lasota Vs ND Genotype 7

12.00 11.00 10.00 9.00 8.00

GMT

7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00

Vaksinasi

Challenge

Ag LS Ag ND Gen 7

48 w 8.59 7.93

50 w 9.52 8.88

53 w 9.20 8.35

55 w 9.41 9.18

56 w 10.55 9.40

Control bird (SPF): Diarrhea Conjunctivitis Tracheitis Swollen spleen Hemorrhagic proventiculus Hyperemia etc.

ETIKET Label

Contoh program vaksin untuk kontrol ND

Week
0

Day
2-4 ND-IB Live ND L ND-IB Live

Vaksin

Dosis

Aplikasi
1 ds Tetes Mata kanan 0,2cc Injeksi sc 1 ds Tetes Mata Kiri 1 ds Cekok 1 ds Cekok 1 ds Tetes Mata Kanan 1 ds Tusuk Sayap 0,5 cc Injeksi sc 0,5 cc Injeksi Dada Kanan 0,5 cc Injeksi Dada Kiri 1 ds Tetes Mata Kiri 0,5 cc Injeksi Dada Kiri 0.5 cc IM 0,5 cc Injeksi Dada Kanan 1 ds Tetes Mata Kiri 0,5 cc Injeksi Dada Kanan 0,5 cc Injeksi Dada Kiri 0,5 cc IM

Jenis
ND apatho-IB H-120

1 2 3

10 14 22-23

ND Clone -IB H120 IBD M IBD M ND Lasota

Gumboro Intermediate plus Gumboro Intermediate plus ND Live Pox ND-AI (H5NI) Coryza I (Alum) ND/IB Kill Variant ND-IB Live AI (H5NI) ND L ND/EDS/IBmulti ND-IB Live Coryza II (Alum) AI (H5NI) ND L

35

ND-AI Trivalen/Tetra ND-IBmulti ND Lasota-IB M41 Bivalent ND Genotype 7 ND/EDS/IBmulti ND Lasota-IB M41 Trivalen/Tetra Bivalent

6 10

42 70

11 13 15 16 18 19

80

105 112

In a Nutshell
1. Egg production drop main issue in breeders and layers 2. HI antigens suggests protection, but is not. 3. Diagnosis and sound vaccination program is important 4. Current ND vaccines protects against clinical signs, but cant prevent shedding and spread 5. Emergency live ND vaccination in face of challenge protects against clinical signs, but severely affects egg production 6. 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 date 8. Trial with inactivated ND vaccine (Genotype VII) proved against mortallity and egg production drop

Thank You

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

Author Unknown