Emerging Diseases

in Layer Chicken
 Emerging Disease

 The World Organisation for Animal Health (OIE) defines an

emerging disease as

“A new infection or infestation resulting from the evolution or

change of an existing pathogenic agent, a known infection or
infestation spreading to a new geographic area or population, or a
previously un recognised pathogenic agent or disease diagnosed for
the first time and which has a significant impact on animal or public
health.”

A known or endemic disease is considered to be re-emerging if it

shifts its geographical setting, expands its host range, or
significantly increases its prevalence.
 Introduction
 The nature and intensification of the poultry industry, multiplication of
species, movements of birds, trade of vaccine and environmental changes
causes emergence of certain diseases in recent years

 Both established and emerging poultry diseases, and the pathogens that
cause them, are continual threats to the industry

 Avian influenza
 Avian metapneumovirus
 Infectious laryngotracheitis
 Chicken infectious anemia
 Neoplastic diseases
 Ornithobacterium rhinotrachiale (ORT)
 Eggshell apex abnormalities
 Gallibacterium anatis associated peritonitis
 ETIOLOGY
ORTHOMYXOVIRIDAE

Influenza A Influenza B Influenza C

Infect wide variety of animals Primarily humans occasionally pigs

Wild ducks, chickens, turkeys, and seals but not birds
Pigs, horses, mink, seals & humans

Influenza A

8 gene segments

10 different proteins

Surface protein Internal proteins

HA, NA & Matrix PB1, PB2, PA, nucleoprotein, matrix 1 protein ,

2 protein Nonstructural protein 1 &2
Influenza A Virus
Negative sense RNA Neuraminidase
Single stranded Hemagglutinin
Segmented
M2

PB1
PB2
PA
HA
NP
NA
MA
NS

M1

18 Hemagglutinin subtypes
11 Neuraminidase Subtypes
 Indian scenario of Highly Pathogenic Avian
Influenza
 The H5N1-HPAI was first detected in India in February 2006 in several
commercial farms in Navapur of Nandurbar district in Maharashtra.


 Subsequently, pockets of outbreaks also were reported in backyard

poultry in Madhya Pradesh and Gujarat

 The outbreak was controlled and India regained disease free status in
August 2006

 During the second week of July 2007 HPAI appeared in a small

backyard poultry unit in Manipur

 The incidence could be successfully contained by precise and timely

diagnosis by HSADL, IVRI. India again faced more serious HPAI
outbreak at West Bengal in January 2008
 The disease was rapidly spread mainly in backyard poultry and
ducks in 15 out of 19 districts within a span of one month and
another outbreak was recorded in Tripura in April, 2008 in
backyard ducks and geese

 Though the west Bengal and Tripura outbreaks successfully

contained and India regained disease free status during first
week of November 2008, within two weeks of regaining AIV free
status, AIV again started to hit in Assam and killed thousands
of backyard poultry in last week of November 2008

 Subsequently it was spread to parts of West Bengal and

Mizoram and Sikkim (DADF-2009) and finally controlled

 India regained its AIV free status on October 2009

 Fresh outbreaks of HPAI again sprouted in west Bengal after 15 th
January 2010 (OIE,2010).


 New cases were being reported in Bihar, Sikkim, Jharkhand,

Odessa, Karnataka and Kerala in the year 2011-14 in domestic
poultry, crows, turkeys and ducks.

 As on November 2014; 98 outbreaks have been detected in India,

1, 52, 643 birds have been succumbed to death and 64,51,
985 birds have been destroyed causing huge economic losses,
reckoned to be in excess of Rs 10,000 crore, to the Indian poultry
industry
 Avian Influenza
 Low pathogenic avian influenza (LPAI)
 Associated with mild illness in poultry
 Can evolve into highly pathogenic viruses
 Associated with poultry outbreaks world wide

 High pathogenic avian influenza (HPAI)

 Causes high mortality in domestic poultry
 Subtypes H5 and H7
Cleavage of Hemagglutinin Protein
by Host Proteases
• In LPAI viruses, only trypsin-like proteases
found in the enteric and respiratory tracts
can cleave the HA protein-virus replication
and disease is restricted
• In HPAI viruses, the HA protein can be
cleaved by ubiquitous proteases found in
most cells-virus can replicate systemically
Clinical Signs – Depression and Huddling
Drop in Egg production
Ruffled feathers
Edema of upper eyelid
Swollen blue, cyanotic combs and wattles
Nasal and oral discharge
Diarrhoea and Dehydration
 Clinical Signs
Subcutaneous hemorrhage of shanks
 Clinical Signs – Huddling, Sinusitis and
Respiratory signs

2006
Neurological signs (Nervous signs) Similar to
Newcastle Disease
Avian Influenza
 Avian Influenza - Gross lesions
Haemorrhage in muscle cyanotic wattle, comb, and facial edema

Urate deposit in kidneys Haemorrhages in skin and shank Haemorrhages in intestinal

walls
Lesions in chickens following experimental infection with the Mexican H7N3 HPAI
virus detected at 2 days postinoculation. (a) Prostration and edema of periorbital
tissues; (b) subcutaneous hemorrhage of wattles and comb and conjunctivitis and
swelling of periorbital area; (c) subcutaneous hemorrhages of leg shanks; (d) mucous
in larynx; (e) petechial hemorrhages on breast muscle
 LPAI
LPAI in respiratory disease complex of commercial - H9N2 subtype.

LPAI outbreaks seasonal - summer but in the recent years throughout the
year

Growers respiratory and nervous signs. Mortality 5 to 25 per

In layer, sudden onset of depression, drop in feed intake and respiratory signs
are the first indication.
 Pathology
Gross lesions mainly in respiratory, genital, urinary and digestive system.

Trachea - mucus exudate, fibrinous flacks, mild to sever congestion even

haemorrhages.

Lungs - congestion and edema and few cases consolidation at the insertion of bronchi.

Thoracic and abdominal air sacs -edematous, frothy and contain fibrinous exudates.

Oviduct - severe transmural edema with clear excessive albumin in lumen and relatively
large quantity of thick, white yellow coloured fluid in the abdominal cavity.

Peritonitis with low amount of fibrinous exudates over the ovaries.

Proventriculus - petechial hemorrhages.

Pancreas is enlarged and hardened.

Trachea: Severe acute diffuse mucosal
hemorrhage
LPAI Infected bird
LPAI - Pancreas
Kidneys: Acute diffuse edema and congestion with urate retention
Haemorrhage in proventriculus and sternum
Epicardial and Cloacal haemorrhages
Peritonitis
Peritonitis
 Avian Meta pneumovirus
Family – Paramyxoviridae
Subfamily – Pnemovirinae
Genus – Metapneumovirus
Virus – Avian metapneumovirus

F protein that promotes cell fusion, do

not hemagglutinate

G attachment proteins of these viruses

do not have neuraminidase activity.

Based on molecular analysis of the

genome, aMPV can be classified into
four subtypes: A, B, C and D. India –
Subtype A
 Avian Metapneumo virus
 The virus is able to replicate in the respiratory tract, especially the
upper tissues, resulting in an acute respiratory disease in turkeys and
chickens with significant economic losses, especially if the infection is
associated with secondary pathogens.

 In broilers, aMPV is involved, among other agents, in the swollen head

syndrome (SHS) and is likely to play a role in pathogenesis of
testicular disease.

 Exacerbating factors for APV infection are the common ones for
respiratory diseases and include high ammonia and dust levels in the
atmosphere, overcrowding and intercurrent infections.
Clinical signs a: Control , b: aMPV subtype-B-inoculated bird showing
swelling of periorbital sinuses and the area around the eye (black arrow)
and clear nasal exudate (white arrow).
Immunohistochemistry for aMPV antigens, horseradish
peroxidase method, hematoxylin counterstain.

Nasal cavity Trachea

 ILT – Etiology
Family – Herpesviridae
Subfamily – Alphaherpesvirinae
Genus - Iltovirus
Species – Gallid herpesvirus 1

Electron microscopy - ILTV particles

consisting of a DNA-containing core within
an icosahedral capsid which is surrounded by a
proteinaceous tegument layer and an outer
envelope with incorporated viral glycoproteins.

The envelope contains five major glycoprotein

spikes (gB, gC, gD, gX, gK) are the major
immunogens of Laryngotracheitis virus.

Virus (ILTV) strains are antigenically homogenous, however vary in

Vary in virulence
Global distribution of ILT as of 2013
 Incidence of ILT in Namakkal Region of Tamil Nadu

Year Tracheitis ILT

No. of Farms No. of birds No. of Farms No. of birds
2009- 10 156 820 27 78
2010- 11 327 1666 109 543
2011-12 472 2064 252 1018
2012-13 318 1541 554 3128
2013-14 268 1327 926 4792

Srinivassan P Balachandran C Gopalakrishna Murthy T R Saravanan S Pazhanivel N

Mohan B and Murali Manohar B. 2012. Pathology of infectious laryngotracheitis in
commercial layer chicken. Indian Vet. J.88 (8): 75-78.

Gowthaman V, Singh SD, Dhama K, Barathidasan R, Mathapati BS, Srinivasan P,

Saravanan S, Ramakrishnan MA. 2014. Molecular detection and characterization of
infectious laryngo tracheitis virus (Gallid herpesvirus-1) from clinical samples of
commercial poultry flocks in India. Virus Dis., 25:345–349
 ILT- Clinical signs
 Clinically, the disease may appear in three forms -Peracute , acute, and
chronic or mild.
 In peracute form, sudden onset and rapid spread. Mortality may be as high
as 70%.

 In acute form, the onset of illness is slower and respiratory signs may extend
over some days before deaths are seen. Mortality may range from 10-30%.

 Chronic or mild ILT may be seen among survivors of the above forms of the
disease, although some outbreaks themselves may be entirely mild. The
mild form is the most commonly seen type and is called “silent, vaccinal, or
almond-shape eye” ILT.

 The disease spread very slowly and the path of infection through a cage
house may be apparent as a result it persisted for 2 to 6 weeks period in
each flock.

 High environmental temperatures (35 C) cause higher mortality from LTV

infection, hence during summer the incidence is more compared to other
seasons.
ILT – Peracute Infected birds are lethargic, have swollen eye-lids
followed by ocular discharge.
ILT – Peracute Birds exhibit mouth breathing, coughing and emit
respiratory noises.
Difficult Respiration
ILT Peracute - Clots of blood may be coughed up and can be
found on the floor and walls of the house.
ILT – Acute mucoid nasal discharge, gasping and swollen eyelids.
ILT – Chronic : conjunctivitis, swelling of the infraorbital sinuses
(almond shaped eyes), and nasal discharge.
ILT – Body condition and Liver lesion
ILT – Egg boundness
 ILT- Tracheal lesion

The trachea at the bottom of the image is least affected, while the one at the top of the image
is most affected. The mucosal surface of each organ is stippled by varying degrees of bright
red hemorrhage.
Trachea showing diffuse haemorrhage in the affected birds.
ILT – Tracheal lesions
ILT- Tracheal lesion
Fibrinous tracheitis
Air sac and Lung lesions
ILT – Tracheal and Lung lesions
ILT – Bursal enlargement in growers
Cytology - Tracheal exudate showing inclusion
 ILT - Trachea

Diffuse lymphocytic aggregation,

Hyperplasia, lymphcytic infiltration degeneration of the epithelium and
and oedema haemorrhages

Lumen contain fibrin and debris Necrosis and soughing of epithelium

ILT- Tr achea: Intr anuclear inclusions and
syncytia for mation
ILT – syncytia with inclusion
Syncytial cell (arrow) with intranuclear inclusion bodies (open
arrows).
ILT – Inclusion by Phloxine tartrazin stain
 ILT- Lung

Lung- Fibrinous bronchopneumonia

Lung – Mild pulmonary congestion

Lung – Fibrinous pneumonia with dense cellular

infiltrate of mononuclear cells and heterophils
Indirect immunoperoxidase test . (b) tracheal lumen: to the left a syncytium
without staining, and to the right a syncytium with focal staining); (c) tracheal
lumen: partial staining of syncytium cytoplasm; (d) tracheal lumen: staining of
nuclei from a disrupted syncytium ; (e) same as (d) but at higher magnification.
ILT - Immunohistochemistry
ILT – Egg inoculation
CAM showing multinucleated cells and intranuclear
inclusions bodies (arrows).
CAM : Epithelial cells exhibited prominent eosinophilic intranuclear inclusions and
formation of syncytia typical of ILTV infection. Inset: ILTV inclusion bodies at
higher (40x) magnification demonstrate peripheralization of hromatin.
Combined infection of ILT and FP: ILT - amphophilic intranuclear
inclusions (thin arrows) and FP - acidophilic haloed intracytoplasmic
inclusions (white block arrows) are shown.
Amplification of partial glycoprotein- G gene (US4 gene) from the
tissue samples yielded expected product size of 589 bp
Visualization of the 647-bp PCR product from the Thymidine kinase gene
of ILTV by Agarose gel electrophoresis (1.5%) after staining with ethidium
bromide Lane-1: molecular marker; Lane 2-7 : ILTV Field Isolates Lane 8 :
ILT Vaccine (Merile)
 Chicken Anemia Virus
Family: Circoviridae
Genus: Gyrovirus
Genome: Single stranded circular DNA
Envelope: Non
Morphology: Icosahedral particle of 22-26 nm

Viral Proteins
VP1 is the only capsid protein
VP2 has phosphatase activity and participate
in the capsid formation as a chaperon by
allowing VP1 to fold
VP3 or apoptotin induces cell death by
apoptosis
Replication : Lymphoid cells
Serotype: 1
Transmission: Vertical and horizontal
 Chicken infectious anemia
 Chicken infectious anemia virus is the causative agent of an immunosuppressive
disease of young chickens.

 The clinical symptoms are generally observed at 10 to 14 d of age and include

increased mortality, anemia, thymic atrophy, subcutaneous hemorrhages and
gangrenous dermatitis.

 According to the presenting signs and lesions, the disease has been variously
called anaemia-dermatis syndrome, blue wing disease, infectious anaemia and
Haemorrhagic syndrome

 Despite the lack of clinical disease, older birds infected with CIAV have been
found to have a decreased immune response as evidenced by poor vaccine
response and increased severity of other infections
Pale carcass and internal organs
Gangrenous Dermatitis
Muscular and visceral
haemorrhages
Pale enlarged liver with focal
necrosis
Ulceration of proventriculus and
gizzard mucosa
Atrophy of lymphoid or gans
Pale and mottled kidneys
Pale yellowish bone marrow
CAV and concurrent infections
 Bone marrow

Normal - bone marrow with abundant severe bone marrow hypoplasia and
cells of the erythrocytic and granulocytic complete aplasia, fully depletion of the
series (H&E , 400x) erythrocytic and granulocytic series,
both accompanied by space occupying
adipocytic replacement (H&E , 400x)
Thymus with indistinct cortex with moderate depletion of
lymphoid follicles (H&E X40 and 400X)
Histopathology – Bursa and Spleen-
Lymphoid depletion
a. Subcutaneous haemorrhage and oedema with cell swelling of the overlying epidermis in the skin of the
thigh region.
b: Haematopoietic (red) bone marrow with CAV inclusions (arrow) in haemocytoblast.
c: Hypoplastic bone marrow with CAV inclusions (arrows) in haemocytoblast.
 Marek’s disease
MDV – enveloped double stranded
DNA virus
Originally Gamma herpes virus But
now Alpha herpes virus (Genome)

Three serotypes
Serotype 1 – Pathogenic or oncogenic
virus with attenuated strains – 4
pathotypes
Mild (MMDV), Virulent (VMDV), Very
Virulent (VVMDV) and Very Virulent
plus (VV + MDV)

Serotype 2 – Nonpathogenic strain s

Serotype 3 – Herpes viruses of turkey
 Marek’s Disease – Current situation
 Marek's disease virus (MDV) is a highly cell-associated, lymphotropic
alphaherpesvirus that causes Marek's disease (MD) in chickens. Clinical signs
include immunosuppression, polyneuritis, and T-cell lymphoma formation in the
visceral and ectoderm-derived tissues.

 Although MD has been controlled by vaccination for over 30 years, it continues to

be a serious threat to the health and welfare of poultry, and there is growing
evidence that intensive use of vaccines is driving the virus to increasing
virulence

 The visceral or ‘acute’ form of MD became the dominant form and causing
higher mortality than the ‘classical’ slower-developing neural form.

 Visceral form commonly noticed from 20 wk to 72 wk (more common up to 45

wk)

 Morbidity and mortality in laying hens due to MD ranged from 0 to 60% or even
greater, with losses of up to 75 % being common.
Marek’s Disease – Current situation
(Sep 2009 to Sep 2013)
S.No Oncogenic virus Number
of flock
1. MDV 68
2. LLV 14
3. REV 24
4. MDV +REV 03
5. MDV +LLV 24
6. LLV +REV 21
7. MDV+LLV+REV 06
8. Total 160
MDV Pathogenesis
Immunosuppression in Marek’s Disease
 MDV can induce two phases of immunosuppression. Primary
immunosuppression occurs temporarily, immediately after systemic infection
with MDV, due to lytic infection of B cells.

 In contrast, the secondary immunosuppression, which is characterized by

functional deficiencies of T cells, starts about 2 weeks after infection, and
continues for a long period .

 MDV can induce apoptosis of peripheral CD4 + T cells and down-regulate

the expression of CD8 + T cells in infected chickens.

 These immunomodulations may be central to the immunosuppression of T

cells induced by MDV, and it is important for understanding the pathogenesis
of MD to clarify the mechanism of these immunomodulation.
MD- Comb thickening and necrosis
MD - Emaciation
 Visceral form of MD
(Liver enlargement with granular appearance)
 Visceral form of MD
(Liver diffusely enlarged)
 Visceral form of MD
( Liver enlargement with focal to multiple nodules )
 Visceral form of MD
(Liver parenchyma replaced by tumor nodules )
 Visceral form of MD
(Mild Proventriculus thickening)
 Visceral form of MD
( Diffuse Proventriculus thickening)
 Visceral form of MD
(Proventriculus mucosa with nodules)
 Visceral form of MD-
(Heart with lymphoma)
 Visceral form of MD
(Pericardium and Heart with lymphoma)
 Visceral form of MD
(Lung with lymphoma)
 Visceral form of MD
(Spleen enlargement and rupture)
 Visceral form of MD
(Spleen enlargement and nodule formation)
 Visceral form of MD
(Intestine and Mesentery showing lymphoma)
 Visceral form of MD
(Intestine showing lymphoma)
Visceral form of MD
(Kidney enlargement)
 Visceral form of MD
(Ovary atrophy and Ascites)
 Visceral form of MD
(Ovary with cauliflower like growth)
 Visceral form of MD
(Ovary with cauliflower like growth)
 Visceral form of MD
(Ovary with cauliflower like growth and muscle tumor)
 Visceral form of MD
(Lesions in visceral organs)
MD in a 14 week old flock
Visceral form of MD in Desi bird
Gross and histopathologic of
MDV. A. Gross anatomical change,
"a" to "e" were heart, lung, liver,
proventriculus, intestinal tract,
respectively, and tumointestinal
tract and mesentery;

B. "f" to "n" refer to bursa of

fabricius, lung, liver, skeletal
musclers, ovary, spleen, kidney,
proventriculus, cardiac muscle.
Arrows show that pleomorphic
lymphocytes infiltrated and formed
tumors (h, i, m, n), and other
organs were infiltrated by
lymphocytes extensively and their
organizations were completely
destroyed (f, g, j, k, i).
ORT
Very small colonies (1-2 mm), non-haemolytic,
Gram-negative, highly pleomorphic rod
greyish white, opaque, convex with butyrous
odour
 Species affected

 Avian species such as chicken, duck, goose, guinea fowl, ostrich,

pigeon, quail and turkey

 Infections caused by Ornithobacterium rhinotracheale (ORT), are

not promptly recognized. Because either the isolation of ORT is

difficult or investigators are less well-versed with ORT other than the

pathogens like Avibacterium paragallinarum etc.

 Impact of ORT
 Clinical signs such as depression, gasping, severe dyspnoea mucus
discharge and weakness

 Associated with high economic losses in poultry due to mortality and

condemnation rates, a drop in egg production or a decrease of the
performance results

 The severity of clinical signs, duration of the disease are influenced by

many environmental factors such as poor management, inadequate
ventilation, high stocking density, poor litter conditions, poor hygiene
or concurrent infections
 Triggering effect

 Newcastle disease virus and to a lesser extent Infectious Bronchitis

virus showed triggering effects on the ORT infection in chickens

 Not only viruses but also bacteria such as Escherichia coli and
Bordetella avium were able to trigger the ORT infection
ORT
Histopathology

Trachea
 Hypertrophy of goblet
cells and deciliation

 Desquamation of epithelium
with moderate haemorrhage
 Egg shell apex abnormality (EAA)
 The EAA are characterized by a roughened shell surface, shell thinning,
increased translucency, cracks and breaks.

 The abnormalities were confined to a region up to approximately 2 cm from

the apex of the egg, and in most cases there was a very clear demarcation
zone and increased translucency visible at candling.

 Egg production losses including the loss of eggs due to breakage of soft-
shell eggs, increased number of downgraded eggs and increased labour
costs due to the selection of eggs with EAA and cleaning of the facilities due
to broken eggs were the main contributors to the economic impact of this
eggshell pathology.

 Decrease hatchability in breeder chicken

 Identification of EAA

 The identification of EAA was based on the outer eggshell

characteristics and egg candling in combination with eggshell
strength measurement.

 Eggs with EAA are characterized by an altered shell surface, which

is confined to the top cone of the egg.

 At candling a clear demarcation zone separates the altered eggshell

from the normal part and the abnormal eggshell shows increased
translucency.

 Egg shell strength is decreased in the affected flocks

 Cause of EAA
 Mycoplasma synoviae is considered the second most important
mycoplasma affecting chickens.

 It causes respiratory diseases subclinical respiratory tract infection and

synovitis in chicken

 Certain strains of MS alone or in combination with IBV cause the

eggshell apex abnormalities

 M. synoviae may affect the composition and concentration of eggshell

matrix proteins in the uterine fluid, which are needed for the regulation of
the growth of calcite during eggshell calcification .

 M. synoviae may also affect ciliary motility in the oviduct, which could
lead changes in the uterine fluid content affecting the deposition of
calcium carbonate crystals.
Normal egg shell layers
A: SEM image of an unaffected eggshell showing the inner membranes (A), the
mammillary knob layer (B) and part of the palisade layer (C).

b: SEM image of an abnormal apical eggshell. The mammillary knob layer is

absent and only part of the palisade layer is present: inner membranes (A),
palisade layer (B), vertical layer (arrow) and cuticule (arrowhead). Note also the
increased density of the inner membranes.
Shell breaks in the translucent area
 Gallibacterium anatis associated peritonitis
 Avian pathogenic Escherichia coli (APEC) has typically been incriminated as
the causative agent of laying hen peritonitis.

 However, evidence suggests that Gallibacterium anatis may also play a role in
the pathogenesis of this disease and their importance is likely to be
underestimated due to overgrowth by other bacterial species and the difficulties
of identification.

 G. anatis is a Gram negative, non-motile, encapsulated, usually β-hemolytic

coccobacillus of the Pasteurellaceae family that forms grayish, round,
semitransparent colonies.

 This bacterium is known to be a member of the normal upper respiratory tract

and lower reproductive tract of chickens but has also been implicated in
peritonitis and salpingitis in commercial laying hens leads to decreased egg
production

 The gross pathological lesions of reproductive disorders due to gallibacterium

infection cannot be distinguished from infections with E. coli.
Gallibacterium anatis associated peritonitis
 Conclusion
 The true prevalence of many emerging diseases are not known. Since we
live in a “global village”, we cannot afford to be complacent about the
tremendous economic, social and public health burden of these diseases.

 The critical steps in addressing any emerging disease are bioexclusion,

surveillance and biocontainment.

 Eradication requires that there is an effective means of detecting,

containing and preventing dissemination of the disease causing agent.

 The success of any eradication program hinges on early detection

through thorough and diligent surveillance and this is possibly where we
are not adequately prepared to handle emerging diseases.
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