Masters Seminar Mvsc Pathology

1. An Updates on Viral Etiology of Gout in
Chicken
Presented By
Bagal Uday Rajendra
M.V.Sc II YEAR
drudayvet11@gmail.com
Department of Veterinary Pathology,
KNP College of Veterinary Science, Shirwal
Maharashtra Animal & Fishery Science University, Nagpur, India.
Masters Seminar – VPP 691
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2. Contents
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01
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02
Contents
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05
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Introduction
What is Gout??
Viral etiology of
Gout
Etiology of Gout
Nephropathogenic
IBV
Avian nephritis
virus
Conclusions &
References
Pathophysiology
of Gout
2

3. Introduction
3

4. What is Gout?
 Gout – Metabolic disorder – Hyperuricemia
 Also, renal diseases decreases the clearance of uric acid from
the blood, which result in acute and chronic hyperuricemia
and the excess uric acid precipitates on either visceral or
articular surfaces
4

5. Non-
Infectious or
Nutritional
Causes
Infectious
or Viral
Causes
Toxins
Etiology
of Gout
 Excess Protein
 Excess Calcium
 Low Phosphorus
 Excess Salt
 Vit A deficiency
 Dehydration
 Nephropathogenic
IBV
 ANV
 IBD
 Mycotoxins
 Nephrotoxic
antibacterials
(Roudybush et al.,1999)5

6. Avian Kidneys
6

7. Purine
Biosynthesis
Pathophysiology of Gout
7

8. Types of Gout
Visceral Gout Articular Gout 8

9. Differences between Visceral & Articular Gout
Visceral gout Articular gout
Onset Acute, But can be chronic Chronic condition
Frequency Very common Rare
Age 1 day and above 4-5 months and above
Sex Both males and females Mostly males
Gross lesions
Kidney • Almost always involved
• Look grossly abnormal with
deposition of white, chalky
precipitates.
• Normal grossly
• May become abnormal with
white urate deposits if the bird
get dehydrated.
Soft tissues • Visceral organs or serosal
surfaces
• Synovium, comb, wattles and
trachea
Joints • Soft tissues around the joints
may or may not be involved.
• Surfaces of muscles, synovial
sheaths of tendon and joints are
involved in severe cases.
• Soft tissues around the joints ,
especially feet.
• Other joints of the legs, wing,
spine, and beak
(Vegad, J.L.(2008) Poultry Diseases 2nd ed.)
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10. Viral Etiology of Gout
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11. Nephropathogenic Infectious Bronchitis
• Infectious bronchitis (IB) is an acute and highly contagious
respiratory disease of chickens characterized by
- Respiratory form
- Reproductive form
- Kidney form
• Listed disease- OIE.
• Worldwide distribution including India.
11

12. Etiology and Structure
Order - Nidovirales
Family - coronaviridae,
Genus - Gammacoronavirus
-Positive sense ssRNA viruse
-Pleomorphic but generally rounded
-Envelope of 90-200 nm in diameter
with club shaped surface
projections(spikes) about 20 nm in
length
(Vegad, J.L.(2008) Poultry Diseases 2nd ed.)12

13. IBV Structural Proteins &Their functions
Structural proteins Functions
• Spike protein (S)
S1 and S2 subunits
Recognizes and bind to the host
receptor
• Membrane Protein (M) Regenerating virion in the cell
• Envelope protein Assembly and morphogenesis of virion
in the cell
• Neucleocapsid Protein (N) Virion structure, replication and
transcription of virus
(Tugba et al .,2017) 13

14. Virus
Sensitivity
Inactivated at 56°C
for 15 -45 min
or at 45°C for 90
min
Inactivated by
70% ethanol, 1%
phenol, 1%
formalin and
iodine
At pH 3.0 - stable
for 14 days
At room
temperature - few
days
14

15. 1930s
1940s
1950s
2000
2010
Text Here
You can simply
impress your
audience and add a
unique zing
Classical IB
first
reported in
USA
Nephropathogenic
IBV Strains
reported in
Australia
Uterotropic
IBV Strains
IBV Strains
causing deep
muscle
myopathy
1960s
1990s
Enterotropic
IBV Strains
Proventriculus
pathogenic
IBV Strains
QX IBV
Strains water
belly/ascites
syndrome in
layers
History
15

16. Strains of IBV
16

17. 17

18. ( Jackwood, M.W. 2012) 18

19. Nephropathogenic Strains
• Australian T strain
• Holte Strain
• Gray Strain
• M 41
• 793B
• 4/91
• IS 722
(Chandra 1987)
• The Mass (Gen Bank accession number HM179146) and 793B types were the most common IBV types
reported in India since 1991.
• Around the year 2000 in INDIA , visceral gout and nephritis was observed in birds less than 2 wk of age
and several nephropathogenic strains of IBV were identified including India/PDRC/Pune/9/99.
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20. • By the air, it takes just few of the virus particles to infect a bird.
• Spread by people and virus contaminated substances ( fomites)
• Carriers
• Not vertically transmitted
• Incubation period - 18-36 hrs.
Spread of the disease
20

21. Pathogenesis of Virus
Kidney or
Oviduct
DisseminationViraemia
Upper
Respiratory
Tract
IBV
• The virus is primarily epitheliotropic and replicates in many epithelial cells, including
those of respiratory tract, kidney and gonads producing lesions and in alimentary
tract, many times with little pathological effect.
(Ignjatovic et al., 2000).
21

22. • Respiratory System :-
 Loss and thickening of the cilia as well as thickening of the lamina propria due to the hyperplasia and the
infiltration of lymphoid cells also necrosis of epithelial cells. Hyperplasia of goblet cells and subsequent
catarrhal inflammation. Focal catarrhal pneumonia may also be noted.
(Cavanagh & Naqi 1997, Dhinakaraj & Jones 1997, Benyeda et al., 2010).
• Kidneys –
 Replication in the proximal convoluted tubules , distal convoluted, collecting tubules.
(Chong and Apostolov, 1982; Owen et al., 1991; Chen et al., 1996)
 The virus causes granular degeneration, vacuolation and desquamation of the tubular epithelium with
massive infiltration of heterophils in the interstitium in acute stages of the disease. In the chronic phase
interstitial lymphocytic nephritis
(Albassam et al.,1986).
Conti…..
22

23. Clinical signs
Abnormal respiratory sounds
 Gasping
 Sneezing
 Watery nasal discharge
 Facial swelling
Respiratory form
23

24. Reproductive form
The most common form damages the fully functional oviduct during
laying and results in reduced egg production and quality.
• Drop in egg production may sometimes be more than 50%
• Loss of egg quality, poor albumin and soft shelled.
Thin and Watery poor quality albumin Thin shelled eggs 24

25. Nephritic form
Young growing birds of 3-6 weeks of age are mostly affected
• Depression,
• Ruffled feathers
• Wet droppings
• Increased water intake
• Mortality may reach up to 30%
(Lee et al.,2004)
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26. Post mortem lesions
Respiratory Tract lesions
26

27. Urogenital lesions
27

28. Microscopic lesions
gouty tophi, surrounded by foreign body macrophages, are scattered throughout the kidney.
Hematoxylin and Eosin | 256x
28

29. Morbidity and Mortality
Morbidity
22
23
24
25
26
27
28
29
30
31
IB nephritis Respiratory
complex
Mortality
29

30. Incidence and Distribution
30

31. Prevalence of IBV in Different Regions in India
(Reddy ,2016)
Region Sample processed IBV isolations Prevalence%
Central 166 88 53.01
North 86 41 46.89
South 246 106 43.90
East 21 17 80.95
Total 521 252 48.37
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32. Prevalence of Nephropathogenic IBV in India
• Bayry, et al., (2005) described the emergence of a nephropathogenic avian
infectious bronchitis (IBV) in 1- to 2- week- old broiler chicks in unvaccinated
flocks in India. The Indian IBV isolate exhibited a relatively high degree of
sequence divergence with reference strains.
• Gaba, et al., (2010) isolated Infectious bronchitis virus from kidney tissue samples
collected from birds with visceral gout. The isolate was having sequence homology
with 4/91 and 793/B strains of virus.
32

33. Conti…
• Two isolates of infectious bronchitis virus (IBV) from field outbreaks in 2008 and 2010
were identified in India.
- (India/LKW/56/IVRI/08) --------------- Massachuchate
- (India/NMK/72/IVRI/10 ) ------------------ 4/91 and 7
(Sumi et al.,2012)
• Gola et al.(2017) diagnosed IB in 9 broiler and 1 layer chicken flock in Uttarakhand
state of India by isolation of virus in Embryonated chicken egg from tissue samples.
The necropsy and histopathological examination indicated involvement of
Nephropathogenic IBV.
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34. Diagnosis
34

35. Conti…
35

36. Avian nephritis virus ( ANV)
 ANV is a recently emerged virus and the most recently identified member of the
avian astroviruses.
( Smyth.,2017)
 Avian nephritis virus (ANV) infection is associated with growth depression and
kidney lesions in young chickens and has been detected in many avian species,
including turkeys, ducklings and pigeons.
( Bidin et al.,2011).
 One day old chick are the most susceptible to the diseases but some of the
characteristic lesions develop in older birds up to the four weeks of age.
( Imada et al.,1997)
36

37. History
• Astroviruses were first described by Madeley & Cosgrove (1975) as the cause of
gastroenteritis in infants.
• ANV was first isolated from rectal contents of normal broiler chicks
(Yamaguchi et al., 1979).
• ANV typically causes histological changes in the kidneys, although viral antigens can be
detected in the liver, spleen, pancreas, kidney, jejunum, and rectum. (Imada et al., 1979,
1983).
• Experimental infections demonstrated that ANV primarily results in a subclinical
disease (Imada et al., 1979, 1983; Maeda et al., 1979; Yamaguchi et al., 1979; Jordan and
Pattison, 1996)
37

38. Etiology and Structure
(Madeley & Cosgrove,2002)
Family- Astroviridae
Genus- Avastrovirus
Small round, non-enveloped viruses,
28 to 30 nm in diameter with +
sense, ssRNA virus.
Characteristic five or six-pointed star-
like surface projections.
In 2000 the viral genome was
completely sequenced and the virus
was re-classified as a new member of
the family Astroviridae.
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39. Strain Classification
VIRUS STAINS References
ANV-1 G-4260 Imada et al.,2000
ANV-1 EF91-276M
( UK 1984)
Todd et al., 2009
ANV-1 (china 2009) Zhao et al., 2011
ANV -2 Imada et al., unpublished
ANV-3 19 (Netherland) De wit et al., 2011
39

40. Spread
• By direct or indirect contact
• Infection can be transmitted by oral administration of virus
to day-old birds
• Egg transmission may also occur
• Virus is consistently isolated from the kidneys or feces
during the first 10 days after infection
40

41. Pathogenesis
Entry
The virus first replicates in the Enterocytes of the jejunum
Mild to severe diarrhea develops
Viraemia
The virus colonize in several organs and replicates in the kidney
Replicates in the tubular epithelial cells causes degeneration of tubular epithelial cells , tubulonephrosis,
and also interstitial Nephritis 41

42. Clinical Signs
• Runting shunting syndromes and uneven flock
performance-
1.Hatch chicks failure to grow
2.Diarrhea due to enteritis
3.Leg weakness
4.Weak feathering
5.Chicks hurdle for heat
( Smyth.,2017)
42

43. Post mortem lesions
 Enteric tract-
Dilated caeca containing yellow, frothy contents and
gaseous fluid, loss of tone and hyperemia of the
intestinal tract.
43

44. Conti…
 Kidney lesion
• Marked swelling, minute pin point
hemorrhages and paleness of the kidneys
along with yellowish white to chalky white
urate deposition on the serosal surface.
44

45. Microscopic lesions
• Necrosis and degeneration of the epithelial cells
of the proximal convulated tubules with
infiltration of granulocytes and interstial
lymphocyte in kidneys.
• The extent of urates deposition was variable in
nature and was in different forms i.e. amorphous
form in early age, varying sized foci with needle
shaped crystalline pattern or clear spaces in later
stage of the birds.
(Bulbule et al., 2013)
Urate crystals in tubules and interstitial
nephritis by H & E. 45

46. Urates deposition in Renal tubules H&E x240 46

47. Morbidity and Mortality
• A report by Bulbule et al., 2013 shows severe outbreak of gout due to Astrovirus
infection in India in 2011-12 which causes 40% mortality and 50-60 % morbidity due to
Runting and Shunting syndromes.
Mortality
40%Morbidity
60%
Astrovirus
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48. Prevalence of ANV in India
• Bulbule et al., (2013)- Demonstrated the presence of CAstV in 41.7% of cases and a mixed
infection of CAstV and avian nephritis virus (ANV) in 36.4% of cases by qRT-PCR analysis
from kidney samples of gout-affected birds.
• Ghodasara et al., (2015)- Isolated and detected Avian Nephritis Virus (ANV) by RT-PCR from a
field outbreaks of gout in commercial broiler in Raipur and Chhattisgarh districts of M.P.
• Gowthaman et al., (2015)- Investigated 21 broiler flocks at different parts of India with clinical
signs of nephritis, diarrhea, uneven and shunted growth out of 21 flocks, 2 flocks were found to
be positive for ANV in RT-PCR assay and further BLAST analysis revealed that the ANV of
Indian origin was closely related to the ANV -1 strain.
48

49. Diagnosis
• Gross and microscopic lesions
• Isolated by inoculation of suspected material (kidney or rectal contents) in the yolk sac
of SPF chick embryos and in chick kidney cells.
• RT PCR , quantitative RT-PCR of kidney or gut content samples.
These tests are designed to detect multiple strains and allow quick differentiation from
other viruses.
• Serologic diagnosis can be made using serum neutralization test or ELISA tests, but
these may detect only a limited number of strains of ANV because of its high antigenic
diversity.
49

50. Infectious Bursal Disease
• 1st described – 1962 – Avian nephrosis
• Gumboro Disease- 1st recognized in Gumboro dist. of Delaware state in USA.
• It is an acute and highly contagious viral disease of immature chickens
• It is caused by Birnavirus and have two serotypes.
• Serotype 1 is pathogenic.
• IBD is also a cause of gout in chicken
• Were the kidneys are swollen and may contain urate deposits, these deposits are due to
blocking of the ureters by a swollen bursa.
(Vegad, J.L.(2008) Poultry Diseases 2nd ed.)
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51. Kidney lesions in IBD
Enlarged Bursa with Kidney Urates
51

52. Conclusion
• Mass and 793/B stains of IB were the predominant till 1991 in India
• Nephropathogenic strains are found to be predominant in the outbreaks of infectious
bronchitis in India in last decade.
• Astrovirus are proved be cause of nephritis and gout in poultry. In India astrovirus has
been confirmed in many outbreaks of visceral gout in young chicken.
• While diagnosing, treating and designing prevention and control strategies for gout in
poultry due consideration has to be given to viral etiology as well.
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53. References
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Dis, 30, 468-476.
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Pathol., 40(2): 173-177.
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gout in commercial broilers in India. Avian Pathol. 2013, 42, 464–473.
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5. Chen, B.Y., Hosi, S., Nunoya, T. and Itakura, C. (1996). Histopathology and immunohistochemistry of renal lesions due to infectious bronchitis virus in
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astrovirus in chicken and turkeys with enteric and locomotion disorders.Avian Pathology, 40, 453–461.
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Microbiology 53: 239–252.
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