White Chick Syndrome Associated

with Chicken Astrovirus in

Ontario, Canada
Kathleen E. Longa, Rachel M. Ouckamab, Alex Weiszc,
Marina L. Brashd, and Davor Ojkićd

American Association of Avian Pathologists

July 24, 2017

aMaple Leaf Foods, New Hamburg, Ontario, Canada.

bMaple Lodge Hatcheries, Port Hope, Ontario, Canada.
cGuelph Poultry Veterinary Services, Guelph, Ontario, Canada.
dAnimal Health Laboratory, University of Guelph, Guelph, Ontario, Canada.
 WCS = white chick syndrome

History and Overview CAstV = chicken astrovirus

ANV = avian nephritis virus

▪ WCS observed sporadically in Ontario for past 30 years (1,5)

▪ Repeated investigations into etiology; no cause identified
▪ Virus isolation
▪ Direct electron microscopy
▪ Bacterial and mycoplasma culture
▪ Hepatitis E PCR, fowl adenovirus PCR

▪ 2011-2012: samples sent to Ireland, Dr. Daniel Todd

▪ Positive for CAstV by PCR and ELISA

▪ 2012: instituted PCR tests for CAstV and ANV in Ontario

▪ This is a historical overview of WCS cases in Ontario, Canada from 2009 to 2016
▪ Three companies with seven commercial broiler hatcheries
▪ Note that cases vary in availability of flock results and diagnostic testing performed

AAAP 2017 2
 Materials and Methods
▪ Case = broiler breeder flock hatching
characteristic progeny
▪ Postmortem and histology of 5 to 8 chicks
▪ Control chicks in 12 cases
▪ CAstV and ANV real-time reverse-
transcriptase PCR
▪ Liver +/- additional tissues (11)
▪ Capsid nucleotide sequencing evaluation
of positive samples and phylogenetic
analysis (12)
▪ In situ hybridization with a riboprobe
capable of reproducing RSS (9)
▪ CAstV positive livers with histologic lesions

AAAP 2017 3
WCS Results from 2009 to 2016
Total number of WCS cases from 2009 to 2016 64 cases

Average broiler breeder flock age of lay at onset 38 weeks of age

Range of broiler breeder flock ages of lay at onset 27 to 57 weeks of age

Percent of cases with egg production drop 12.5%

Average egg production drop in affected flocks 13.4%

Range of egg production drops in affected flocks 7% to 21%

Average hatchability drop 19.4%

Range of hatchability drops 0% to 68.4%

AAAP 2017 4
 Number of WCS Cases per Year
Annual Provincial Case Incidence Rate

22
25 Total of 64 cases from 2009 to 2016 20%
18.3%

Annual Case Incidence Rate

18%
Number of WCS Cases

20 16%
14%
13
15 10.8% 12%
10%
10 7 8%
5 5 5.8% 5
4 6%
4.2% 4.2% 3 4.2%
5 3.3% 4%
2.5%
2%
0 0%
2009 2010 2011 2012 2013 2014 2015 2016
Year
AAAP 2017 5
 Frequency of WCS Cases in
Individual Broiler Breeder Barns
35
30 29
Number of Barns

25
20
15
10
10
5 3
1
0
1 2 3 4
Number of WCS Cases

Affected flocks were not always in consecutive placements

AAAP 2017 6
AAAP 2017 7
Normal liver, 1 day old chick Affected liver, 1 day old chick

8
Affected liver, 1 day old chick Affected liver, 1 day old chick
Normal kidney, 1 day old chick Affected kidney, 1 day old chick

Normal spleen, 1 day old chick Affected spleen, 1 day old chick 9
 CAstV and ANV Real-time RT-PCR
Test and CAstV CAstV Dead CAstV Chick CAstV Chick CAstV Chick Control Flock
Tissue Chick Liver in Shell Liver Kidney Jejunum Cloacal Swab CAstV
positive/
21/23 6/6 16/16 16/16 13/14 4/12
tested
percent
91% 100% 100% 100% 93% 33%
positive

Test and ANV Chick ANV Dead in ANV Chick ANV Chick ANV Chick Control Flock
Tissue Liver Shell Liver Kidney Jejunum Cloacal Swab ANV
positive/
0/21 0/4 0/15 0/15 0/15 0/9
tested
percent
0% 0% 0% 0% 0% 0%
positive

AAAP 2017 10
 Capsid Amino Acid Sequencing

▪ All field samples clustered in CAstV group B, subgroup Bii

▪ Field samples from all provinces shared between 93.0% and 100.0%
amino acid similarity
▪ Field samples from an outbreak in 2015 had 95.4% to 100% amino acid
similarity
▪ In situ hybridization results for CAstV in chick liver were negative
▪ Despite up to 98.1% amino acid similarity between field sequences and the
probe sequence
▪ Possibly due to true antigenic differences or time when samples were
collected

AAAP 2017 11
Phylogenetic Tree of Partial Capsid Gene Sequences
JN582311_CAstV-VF06-7/8
JN582306_CAstV-1010
JN582305_CAstV-11522
HQ330506_CAstV-11672
JN582327_CAstV-11672
JN582308_CAstV-VF06-1/2
JN582310_CAstV-VF06-7/5
Subgroup B i
JN582307_CAstV-VF06-1/1
JN582309_CAstV-VF06-1/4
JN582328_CAstV-FP3
15-047770-0003_ON-MapleLeaf
15-047771-0004_ON-MapleLeaf
15-044251-0004_ON-MapleLeaf
JN582316_CAstV-VF08-3
JN582314_CAstV-VF07-4/2
JN582315_CAstV-VF08-29
JN582313_CAstV-VF06-7/3
JF832365_CAstV-4175
JF414802_CAstV-GA2011
JN582312_CAstV-05V150/152/154
15-054558-0003_ON_Horizon
15-048550-0006_ON-MapleLeaf
15-049148-0004_ON-Horizon
15-047767-0003_ON-MapleLeaf
KC618324_CAstV-PDRC/1804 Subgroup B ii
KC618323_CAstV-PDRC/447
JX945856_CAstV-PDRC/264
KJ621032_CAstV-VRDC/CAstV/NZ/VHINP-6
KJ621033_CAstV-VRDC/CAstV/NZ/VHINP-7
JX945861_CAstV-PDRC/573
JX945860_CAstV-PDRC/542
KJ621028_CAstV-VRDC/CAstV/SZ/VHINP-2
JX945867_CAstV-PDRC/589
JX945864_CAstV-PDRC/579
KR052479_CAstV-PL/G059/2014
KT886453_CAstV-CAstV/Poland/G059/2014 Subgroup A iii
JN582321_CAstV-VF08-46
JN582324_CAstV-VF08-18/7
JN582323_CAstV-VF08-54 Subgroup A ii
JN582320_CAstV-VF08-60
JN582319_CAstV-VF08-56
JN582318_CAstV-P22-18.8 Subgroup A i
JN582317_CAstV-612
44.2

40 35 30 25 20 15 10 5 0 AAAP 2017 12
Amino Acid Substitution per 100 residues
 Discussion
▪ Egg production drops:
▪ Previous reports: 0% (7), 3 to 10% (13), 5 to 20% (2), up to 27% (8)
▪ 1/8 – production drop 7%
▪ 6/8 – production drop between 10 and 20%
▪ 1/8 – production drop of 21%
▪ Hatchability drops:
▪ Previous reports: 4 to 5% (7); average 29% and up to 68% (13); hatchability as
low as 40% (2); hatchability as low as 19.7% (8)
▪ Here, ranged from 0% to 68.4% hatchability drop; average drop of 19.4%
▪ Severe losses due to undersized eggs previously reported (8)
▪ Did not occur in these cases
▪ Otherwise, very few clinical signs in broiler breeders
▪ Prolonged feed clean-up time
▪ Mild, non-specific signs may not be associated with WCS

AAAP 2017 13
Discussion & Future Opportunities
▪ Predictive diagnostic testing
▪ None registered/available in Canada
▪ Ongoing work to understand usefulness of American ELISA tests
▪ Would allow us to understand pullet and breeder flock immunity to target
interventions
▪ Better understand epidemiology of WCS
▪ Causes(s) of apparent outbreaks
▪ Control biosecurity or management factors that can increase risk
▪ Darkling beetle control (10)
▪ Control strategies
▪ Some have trialed litter transfer – discouraged due to pathogen risks
▪ Astroviruses difficult to grow in sufficient quantities for vaccine production
▪ Recombinant baculoviruses trialed for other CAstVs (3,9)
▪ No commercial or autogenous vaccines in use at this time
▪ Cost of targeted vaccination would be financially justifiable (4)
AAAP 2017 14
 Acknowledgements

▪ Dr. Holly Sellers

▪ Dr. Emily Martin
▪ Dr. Karen Burns-Grogan
▪ Animal Health Laboratory,
University of Guelph
▪ Poultry Diagnostic &
Research Center,
University of Georgia

AAAP 2017 15
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