INCLUSION BODY HEPATITIS UPDATE: SEROTYPES, CONTROL AND PREVENTION
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2 INCLUSION BODY HEPATITIS UPDATE: SEROTYPES, CONTROL AND PREVENTION Davor Ojkic, DVM, MSc, PhD Animal Health Laboratory University of Guelph Ontario, Canada
3 Animal Health Laboratory Bacteriology Pathology Mycoplasma/ Parasitology Clinical Pathology Virology/ Immunology Toxicology
4 Practitioners & Producers Animal Health Laboratory Ministry of Agriculture U of Guelph Other Universities Public Health Canadian Food Inspection Agency 4
5 Animal Health Laboratory Expertise Diagnosis, monitoring, emergency preparedness 17 veterinarians on staff 70 technical/clerical staff 1,000 test procedures Caseload Distribution 66,400 cases over 1,000,000 tests 5
6 6
7 INCLUSION BODY HEPATITIS UPDATE: SEROTYPES, CONTROL AND PREVENTION Davor Ojkic, DVM, MSc, PhD Animal Health Laboratory University of Guelph Ontario, Canada
8 Outline History Fowl adenovirus (FAdV) Diagnostics Fowl adenovirus infection in broilers/breeders Control/prevention
9 History FAdV Isolations Unintentional isolations VAN DEN ENDE M, DON PA, KIPPS A. The isolation in eggs of a new filterable agent which may be the cause of bovine lumpy skin disease BURMESTER BR, SHARPLESS GR, FONTES AK. Virus isolated from avian lymphomas unrelated to lymphomatosis virus Intentional isolation OLSON NO. A respiratory disease of quail caused by a virus. Proc 54 th Annual Meet US Livestock
10 History - Early Classification McFerran JB, Adair BM. Avian adenoviruses: a review. Avian Pathol. 1977;6(3):
11 History - Previous Classification Genus Aviadenovirus Group I Group II Fowl adenovirus 1-12 Duck adenovirus 1-2 Goose adenovirus 1-3 Hemorrhagic enteritis virus Marble spleen disease virus Chicken splenomegaly virus Group III Egg drop syndrome virus
12 Classification History ICTV ICTV USA USA
13 ICTV Classification Fowl adenovirus A Fowl adenovirus 1 (CELO, 112, Phelps) Fowl adenovirus B Fowl adenovirus 5 (340, TR22) Fowl adenovirus C Fowl adenovirus 4 (KR-5, J-2) Fowl adenovirus 10 (C-2B, M11, CFA20) Fowl adenovirus D Fowl adenovirus 2 (GAL-1, 685, SR48) Fowl adenovirus 3 (SR49, 75) Fowl adenovirus 9 (A2, 90) Fowl adenovirus 11 (380) Fowl adenovirus E Fowl adenovirus 6 (CR119, 168) Fowl adenovirus 7 (YR36, X-11) Fowl adenovirus 8a (TR59, T-8, CFA40) Fowl adenovirus 8b (764, B3)
14 Morphology Icosahedral nm DNA genome Non-enveloped
15 Significance Inclusion body hepatitis (IBH) Hepatitis-hydropericardium syndrome (HHS) Respiratory infections Proventriculitis Gizzard erosions Pancreatitis
16 IBH vs HHS IBH FAdV2, FAdV7, FAdV8, FAdV11 Canada/USA/Americas, Europe, Australia, Asia, Africa HHS FAdV4 Asia, Mexico, Central & South America
17 Diagnosis Post-mortem examination Histologic examination Virus isolation PCR Virus Characterization Serotyping Hexon gene sequencing
18 Post-mortem
19 Histologic Lesions
20 Histologic Lesions
21 Virus Isolation
22 Real-time PCR
23 Genotyping Hexon protein gene fiber hexon
24 Genotyping of reference strains 77.5 FAdV08a E_T8-A FAdV08b E_764 FAdV07 E_ X11 FAdV08b E_B-3A FAdV06 E_CR119 FAdV08a E_TR59 FAdV02 D_SR48 FAdV11 D_380 FAdV02 D_P7A FAdV02 D_685 FAdV03 D_75 FAdV09 D_A-2A FAdV05 B_TR22 FAdV05 B_340 FAdV05 B_IBH-2A FAdV01 A_CELO FAdV04 C_J-2A FAdV10 C_C2B Nucleotide Substitutions (x100) 10 0
25 IBH Outbreaks
26 AHL FAdV Submissions IBH Non-IBH
27 IBH - Clinical Presentation AGE: Range of 6 days to 7 weeks, most are 2 to 3 weeks CLINICAL SIGNS Depressed, dopey birds Reluctant to stand or walk Ruffled feathers Drop dead or in a crouching position (terminal stage) MORTALITY Suddenly increase 1% to 30-40% Increased mortality lasts 4 to 5 days
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29 Epidemiology Vertical Transmission Primary FAdV infection of young breeder flock Reactivation of latent FAdVs in older breeder flocks Introduction of a new, pathogenic FAdVs in a breeder flock Immunosupression/non-pathogenic FAdVs Combination of factors above Horizontal Transmission Field challenge: virulent FAdV serotypes (Ab+ broilers) Field challenge: mixing Ab- and broilers broilers that harbor a virus Immunosupression/non-pathogenic FAdVs Combination of factors above
30 Initial Investigation 487 IBH Cases n= Non-IBH Cases
31 Genotyping IBH Cases 41 Non-IBH Cases n=287 Frequency AB BC MB NS ON PQ SK Province
32 IBH Case Summary VIRUS GENOTYPE STRAIN MATCH ICTV SPECIES FAdV02 P-7A D FAdV08 X-11A a E FAdV08 Stanford E FAdV08 TR-59 E FAdV08 T8-A E FAdV08 Ontario b E FAdV D % IDENTITY # OF ISOLATES % OF IBH CASES
33 IBH Case Summary 3.66% 39.84% 56.50% FAdV02 FAdV08 FAdV11
34 IBH Case Co-detection (n=246) YEAR IBV REOVIRUS IBDV* NDV Total: * No IBDV tests before 2003
35 FAdV: Non-IBH Case Summary VIRUS GENOTYPE STRAIN MATCH ICTV SPECIES % IDENTITY # OF ISOLATES % OF NON- IBH CASES FAdV01 CELO A FAdV02 Merlin D FAdV04 J-2A C FAdV08 TR-59 E FAdV D
36 FAdV: Non-IBH Case Summary FAdV01 FAdV02 FAdV04 FAdV08 FAdV11
37 FAdV: Non-IBH Case Co-detection (n=41) YEAR IBV REOVIRUS IBDV* NDV Total: * No IBDV tests before 2003
38 FAdV Distribution by Province AB BC MB ON PQ SK FAdV02 FAdV08 FAdV11
39 Broiler Breeder Study Determine the serological status of broiler breeders in Ontario flocks by detecting the presence of antibodies to FAdVs Examine shedding of FAdVs in broiler breeder flocks with history of IBH and no history of IBH, by virus isolation and subsequent genotyping
40 Serological Testing 73 broiler-breeder flocks Samples collected before egg production First submission at weeks Second submission at weeks Tests AGID (Antigen: FAdV01-Celo) MNT (FAdV01-Celo, FAdV , FAdV02-P7A, FadV-x11A-like, FAdV08a- Stanford, FAdV08a-TR59, FAdV08a/b, and FAdV )
41 AGID 86.00% 84.00% 82.00% 80.00% 78.00% 76.00% 74.00% 72.00% 70.00% 1st Submission 2nd Submission (n=73, cut-off 20%)
42 MNT 90.00% 80.00% 70.00% 60.00% 50.00% 40.00% 30.00% 20.00% 10.00% 0.00% 1st 2nd (n=73, cut-off 20%)
43 FAdV Shedding Sixteen broiler-breeder flocks 8 with previous history of IBH on farm 8 with no previous history of IBH on farm Virus isolation in cell culture FAdV PCR Genotyping L1 hexon protein gene sequencing
44 Virus Isolation History of IBH wks - reovirus wks - FAdV01 20 wks - FAdV01 & FAdV08a- TR59 45 wks % FAdV03, 92.4% FAdV wks - FAdV08a-TR wks - FAdV01 No History of IBH wks FAdV wks FAdV04 44 wks FAdV08a wks - FAdV04 & FAdV08b wks - reovirus 24 wks - FAdV04 & FAdV wks - FAdV01
45 IBH Broiler Study Target: 50 IBH cases Fresh and formalinized tissue samples Histologic examination Virus isolation Genotyping Polymerase chain reaction IBDV CAV
46 Breeder Flock Age Breeder age at hatch Percentage N/A 6.62% weeks 2.21% weeks 8.82% weeks 3.68% weeks 10.29% weeks 18.38% weeks 16.18% 30 weeks or younger 33.82%
47 History of IBH Frequency Past IBH n % N/A No Yes N/A No Yes
48 Broiler Ages n 52 Mean 15.3 Frequency Broiler Age (d)
49 Mortality (%) 25 n Range % Mean 3.051% 20 Frequency
50 Condemnations Normal Fit (Mean=1.321, SD=2.234) 30 Frequency n Range % Mean 1.321%
51 Genotyping Frequency n % FAdV FAdV02 P7-A FAdV FAdV FAdV FAdV02 P7-A FAdV08a Stanford FAdV
52 Mortality by Genotype FAdV % FAdV % FAdV %
53 IBDV Vaccination Frequency n % N/A No Yes N/A No Yes
54 IBDV Genotyping IM Amino Acid Substitutions (x100) * SA NC171 Del E 9109 Del A GLS IBD Blen STC D78 2 Bursine Lukert SP_04_02 Bursine Plus 0 * was from IBDV-vaccinated flock
55 Chicken Anemia Virus Th Th Th Th Lu Li BF k br CAV Vaccine CAV SPAFAS 2 Amino Acid Substitutions (x100) 0 Percent Identity CAV Vaccine Intervet CAV SPAFAS br k BF Li Lu Th Th T h T h
56 Surveillance for viruses in broiler flocks in Ontario ( ) Surveillance for viruses in broiler flocks in Ontario Avian adeno-associated virus Avian influenza Chicken anemia virus Fowl adenovirus Infectious bursal disease virus Avian encephalomyelitis virus Avian reovirus Chicken astrovirus/avian nephritis virus Infectious bronchitis virus Infectious laryngotracheitis virus Newcastle disease virus 15 birds per flock Blood Cloacal swabs (3 pools of 5 swabs per pool) Caecal tonsils (3 pools of 5 swabs per pool) 56
57 Frequency Fowl adenovirus 250 VI, PCR & AGID EXPOSED Negative FLOCK STATUS FLOCK STATUS n % EXPOSED Negative
58 n= _FAdV08a_Stanford _FAdV08a_Stanford _FAdV08a_Stanford FAdV08a_Stanford FAdV08a_T8-A, ATCC VR-830 FAdV08b_764 FAdV08_Esurient FAdV07_x11a, ATCC VR-835 FAdV07_x11 FAdV08b_B3-A, ATCC VR-832 FAdV06_CR119 FAdV08a_TR _FAdV08a_TR _FAdV08_TR _FAdV08_TR _FAdV08a_TR _FAdV08a_TR _FAdV08a_TR _FAdV _0024._FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV11 FAdV11_1047 FAdV02_P7-A, ATCC VR-827 FAdV02_ _FAdV02_ _FAdV02_ _FAdV02_ _FAdV02_685 FAdV03_75-1A-1 ATCC# VR831 FAdV09_A-2A ATCC VR-833 FAdV05_IBH-2A, ATCC VR-828 FAdV01_CELO, ATCC VR _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV _FAdV01 FAdV04_J2-A, ATCC# VR829 FAdV10_C-2B, ATCC VR-834 FAdV08a_Stanford FAdV08a_TR59 FAdV11 FAdV02 FAdV Amino Acid Substitution per 100 residues 10 0
59 FAdV Genotyping Summary VIRUS # GENOTYPED PROPORTION FAdV % FAdV % FAdV08a_Esurient % FAdV08a_Stanford % FAdV08a_TR % FAdV % TOTAL: % 59
60 Conclusions Broilers - FAdV infection widespread Breeders - FAdV seroconversion correlated with flock age Broilers from younger breeder flocks more likely to get IBH IBH primary disease FAdV02, FAdV08, FAdV11 primary IBH viruses FAdV04 not involved in IBH in Canada
61 Control Clean-barn disease - breeder seroconversion Litter transfer Vaccination Live (Australia) Inactivated Licensed Autogenous Ontario Limited licence FAdV08/FAdV11 IBH outbreaks sporadic
62 Virology Section Animal Health Laboratory
63
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