CURSO : PATOLOGIA AVIAR

AVIAN
METAPNEUMOVIRUS
Different subtypes of the avian metapneumovirus (AMPV A-D)
have been isolated from commercial poultry and wild birds,
which are considered natural reservoirs. These viruses were
shown to not only induce respiratory disease (turkey
rhinotracheitis, swollen head syndrome) but also
reproductive disorders in different avian species. Molecular
methods are used to detect and further characterize AMPV,
although classic virus propagation from field samples is
difficult. Currently, biosecurity in combination with
vaccination is applied in the field to control infection and
reduce the risk from secondary respiratory pathogens.
Avian metapneumovirus (AMPV) causes turkey
rhinotracheitis (or avian pneumovirus infection of
turkeys), an acute respiratory tract infection of
turkeys. It is also associated with swollen head
syndrome (or avian rhinotracheitis) in broilers and
broiler breeders, as well as reproductive disorders,
with a significant drop in egg production in
chickens and ducks. The virus was first detected in
turkeys in South Africa in the late 1970s and has
spread to all the major poultry-producing areas in
the world except for Australia. AMPV has been
detected not only in chickens and turkeys but also
in pheasants, Muscovy ducks, and guinea fowl.
Geese, most other duck species, and possibly
pigeons are suggested to be refractory to disease.
The virus was described first 1978 in South
Africa by S. B. Buys and J. H. Du Perez in
turkeys caused by aMPV-A.
First only in turkeys which is the turkey
rhinotracheitis, over a decade later also in
chickens, which is the swollen head syndrome.
Later type B virus infections were described in
Europe whereby type C infections were indicated
in the upper midwest of the United States by J.
K. A Cook and J. S. Mc Dougall in 1986 who
isolated the causal agent from muscovy ducks
The type D virus was detected 1986 by B.
Giraud in France
Epidemiologic studies provide evidence for the
circulation of AMPV in wild birds, especially
water-associated species. Some outbreaks have
been attributed to vaccine-derived viruses,
which may persist for several months in the
environment. Infection with AMPV is often
complicated by secondary bacterial infections,
leading to high economic losses. In 2001, the
first human metapneumovirus (HMPV) was
isolated and classified as a member of the genus
Metapneumovirus, which causes respiratory
infections in people. Experimental studies
suggest that turkeys also may be susceptible to
HMPV. Complete genome sequencing confirmed
that the genomic organization of HMPV is similar
to that of AMPV. Overall, little is known about
the cross-species pathogenicity of these two
viruses.
Avian metapneumovirus is a member of the family
Paramyxoviridae, genus Metapneumovirus, which
currently comprises the species AMPV and HMPV.

Isolates of AMPV are currently grouped in subtypes A to

D. The sequence of the attachment glycoprotein (G
protein) can be used to subtype different strains,
because it is variable in length and sequence identity
even within AMPV subtypes. Based on the phylogenetic
analysis, it was suggested that the European subtypes A,
B, and D are all more closely related to each other than
to subtype C, which shows more similarity on the
molecular level (sequence identity, genomic
organization, codon usage bias, phylogenetic location)
with HMPV. Whereas European and Asian AMPV C isolates
can be grouped in one genetic sublineage, the other
sublineage comprises the US isolates. The circulation of
subtype D was described in France, but since 1980 has
not been reported again.
 TRANSMISSION AND EPIDEMIOLOGY OF AVIAN METAPNEUMOVIRUS
Wild birds are considered natural
reservoirs for avian metapneumovirus, and
migratory birds may contribute to the
distribution of the virus. A high apparent
prevalence was recently determined
particularly in mallards and American
black ducks. The spread of AMPV appears
to depend on the poultry population
density, standard of hygiene, and
biosecurity. Within or between poultry
flocks, AMPV may spread rapidly
horizontally by direct contact or by
contact with contaminated material
(morbidity rate up to 100%).
AMPV is assumed to be highly contagious.
The enveloped virus is rapidly destroyed
after release from the host to the
environment. Because AMPV affects mainly
ciliated epithelial cells of the upper
respiratory tract, transmission is most
likely to be airborne, especially by
aerosol. Ciliated cells of the reproductive
tract also may be target cells of AMPV.
AMPV C was isolated from eggs of
experimentally infected, SPF turkeys, but
it was suggested that the vertical route
may be short-lived and may play only a
minor role in viral transmission.
Birds appear to shed AMPV for only
a few days after infection. This short
period of shedding suggests that
there is no latency or carrier status
under experimental conditions. There
is evidence that, on farms, AMPV
may persist for longer periods.
Reconvalescent flocks may be
repeatedly reinfected with AMPV
within one fattening period.
 HOST SPECIES, PATHOGENESIS, AND
CLINICAL FINDINGS OF AVIAN
METAPNEUMOVIRUS

Turkeys, chickens, and also ducks were shown

to develop clinical signs with different
subtypes of avian metapneumovirus. Whereas
AMPV A and B are associated with clinical signs
in chickens and turkeys, AMPV C of the North
American lineage affects mainly turkeys,
ducks of Asian and European lineage, and
other commercial poultry less frequently,
which may be subclinical infections.
Pheasants, Muscovy ducks, and guinea
fowl kept in captivity may show signs,
whereas geese, most other duck species,
and pigeons are thought to be refractory
to disease. Viral contact was also
demonstrated for farmed ostriches. The
American lineage of AMPV has been
detected in sea gulls, sparrows, and
additional wild bird species.
AMPV induces an acute, highly contagious
infection of the upper respiratory tract.
The main target cells are epithelial cells,
but macrophages also may carry the virus.
Infection leads to clumping and loss of
cilia, which allows secondary pathogens to
invade. Furthermore, studies indicate an
immunosuppressive potential of the virus,
which supports the replication of
coinfecting pathogens. The clearance of
AMPV coincides with the induction of AMPV
antibodies and the disappearance of
clinical signs
Avian metapneumovirus affects all age
groups, although younger birds seem to be
more susceptible. In fattening turkeys the
upper respiratory tract is predominantly
affected, whereas in laying hens only a
mild respiratory infection with a drop in
egg production (up to 70%) and egg quality
has been seen.

Coughing associated with lower respiratory

tract involvement may lead to prolapse of
the uterus in laying turkeys.
Typical respiratory signs of avian
metapneumovirus in young turkeys include:
serous ocular and nasal discharge
frothy eyes
conjunctivitis
At later stages, signs include
mucopurulent, turbid nasal
discharge; plugged nostrils; swollen
infraorbital sinuses; and snicking,
sneezing, coughing, or tracheal
rales. These respiratory signs are
accompanied by depression,
anorexia, and ruffled feathers.
The incubation period is 3–7 days. The
mortality may be 1%–50% depending on age
and constitution of the flock as well as
secondary infections. Birds without
secondary infections with good
constitution may recover within 7–10 days.
However, in birds with secondary
infections and under poor management,
the disease may be prolonged and
exacerbated by airsacculitis, pericarditis,
pneumonia, and perihepatitis.
Infection in chickens and pheasants is less
clearly defined and may not always be
associated with clinical signs. AMPV is
associated with swollen head syndrome in
chickens. This condition is characterized
by swelling of the peri- and infraorbital
sinuses, frothy eyes, nasal discharge,
torticollis, and opisthotonos due to ear
infection. Typically, <4% of the flock is
affected, although respiratory signs may
be widespread. Mortality is rarely >2%. In
broiler breeders and commercial layers,
egg production and quality are frequently
affected.
AMPV C infection of ducks may lead to
respiratory signs and a drop in egg
production (40%–85%) as well as poor egg
shell quality. Clinically, signs may wane
after 9–12 days if no secondary infections
complicate the disease in ducks.

Although it was suggested that mice may

be susceptible to AMPV C replication and
develop LUNG lesions, other studies using
another AMPV strain did not successfully
establish an infection.
Macroscopic lesions depend on the course
of infection, especially on secondary
bacterial infections, and are most
prominent on days 4–10 after infection.
Gross lesions induced after experimental
infection are due to rhinitis, tracheitis,
sinusitis, and airsacculitis. Infected birds
may be free of gross lesions. Serous to
turbid mucus may be observed in the nasal
cavity, nasal turbinates, trachea, and in
infraorbital sinuses. During the course of
infection, the secreted mucus turns from
clear and serous to turbid and purulent.
Nonspecific signs of inflammation, such as
swelling and hyperemia of the mucosa and
excessive mucus, can be seen in the upper
respiratory tract and in the air sacs. If secondary
bacterial infections are present, copious
inflammatory exudates are found in the
respiratory tract. In addition, pneumonia,
pericarditis, perihepatitis, splenomegaly, and
hepatomegaly are seen. In the reproductive
tract of laying turkeys, lesions can include egg
peritonitis, ovary and oviduct regression, folded
shell membranes in the oviduct, and misshapen
eggs. Some infected birds are free of gross
lesions.
Microscopic examination of the upper
respiratory tract, including the secondary
bronchi during the first 2 days after AMPV
infection, reveals loss of cilia, increased
glandular activity, congestion, and mild
mononuclear infiltration of the submucosa. The
most pronounced microscopic lesions are found
in the mucosa or the nasal turbinates, which
may be the most suitable tissue for microscopic
evaluation. Harderian glands and lacrimal
glands may also show infiltration of
lymphocytes and formulation of lymphoid
follicle-like structures in the interstitial tissue
and around the secondary collecting ducts. The
peak of microscopic lesion development in
turkeys is expected 3–6 days after infection; it
may be shorter in chickens
Obtaining samples from the upper respiratory tract
of birds in the early stages of the disease is
extremely important when attempting avian
metapneumovirus isolation. Especially in broiler-
type chickens, samples should be taken before the
sixth day after infection. Once clinical signs are
obvious, the isolation of replicating AMPV may not
be successful. The most suitable samples for AMPV
detection are tracheal and choanal swabs. Tracheal
organ cultures prepared from turkey or chicken
embryos, or 1- to 2-day-old chicks, are the most
sensitive for primary isolation of most AMPV
subtypes. Ciliostasis may occur within 7 days of
AMPV A and B but not subtype C inoculation or
after passages.
The virus has also been isolated after the
inoculation of 6- to 8-day-old
embryonated chicken or turkey eggs via
the yolk sac route and identified by
electron microscopy, virus neutralization
test, or molecular techniques. Cell
cultures have not proved successful for
the primary isolation of the virus.
However, once the virus has been isolated
and adapted in the systems above, it will
grow in a variety of avian and mammalian
cultures, inducing a cytopathic effect.
Reverse transcriptase PCR (RT-PCR), as
well as real-time RT-PCR, tests targeting
the F, N, or G gene of AMPV have been
developed. Some systems are
commercially available, and these
techniques are widely used to detect virus
in clinical material, particularly
respiratory swabs. Samples may also be
submitted on FTA cards for molecular
diagnosis. Some nested RT-PCR tests have
been constructed so that the subtype as
well as the identity of virus can be
determined from the clinical sample.
Because of difficulties in isolation and
identification of AMPV, serologic assays have been
developed to confirm infection in commercial
chickens and turkeys. A number of commercial
ELISA kits are available and are commonly used,
but other techniques, including virus neutralization
and indirect immunofluorescence tests, have also
been used. Both acute and convalescent serum
samples should be submitted for analysis (paired
serum samples from affected flocks). Although
ELISA systems that use either subgroup A or B
strains as antigens detect antibodies to both of
these subgroups because of some cross-reactivity,
the homologous antigen should be used for the
efficient detection of subgroup C. The subtype
specificity of the applied test may result in limited
or no detection of other subtypes or new, emerging
AMPV strains that do not cross-react
Paramyxoviruses (particularly Newcastle disease,
Avian Avulavirus-1 (AAV-1) and AAV-3), infectious
bronchitis virus, and influenza viruses may cause
respiratory disease and egg production problems in
chickens and turkeys that closely resemble AMPV
infection. These viruses can be differentiated on the
basis of morphology, hemagglutinating and
neuraminidase activity, and molecular
characteristics. A wide range of bacteria
and Mycoplasma spp can cause signs very similar
to those of AMPV. These agents are frequently
present as secondary opportunistic pathogens and
may mask the presence of the AMPV.
Good management practices can significantly
reduce the severity of avian metapneumovirus
infection, especially in turkeys; in particular,
optimal ventilation, stocking densities,
temperature control, litter quality, and biosecurity
have a positive influence on the outcome of the
disease. The virus is sensitive to lipid solvents,
stable at pH 3–9, and may be easily inactivated at
temperatures above 50° C. Disinfectants such as
quaternary ammonia, ethanol, iodophors, phenol
derivatives, as well as bleach may be used to
reduce the viability of AMPV. Some success in
reducing disease severity by controlling secondary
bacterial infections with antibiotics has also been
reported.
Both live and inactivated vaccines are
available for immunization of chickens and
turkeys and are widely used in countries
where the disease is endemic. Studies
suggest that maternal antibodies may
partially interfere with vaccine virus
replication but overall do not provide
sufficient protection against AMPV
infection. A vaccination program should
plan for the first immunization as soon as
possible after hatching and may even be
applied in the hatchery. It is crucial to
achieve a homogenous state of
immunization per flock and farm by
application of an adequate vaccine dose to
all birds.
Live vaccines, which may be applied
by spray or drinking water in the field,
stimulate both local respiratory and
systemic immunity, and cross-
protection between subtypes may
occur. But live vaccines may induce
only short-lived protection, especially
for grow-out of toms, because of the
fast decline of local immunity. Thus,
repeated revaccination of turkeys is
common practice. There is, however,
a risk of reversion of the live vaccine
strains to more virulent variants.
Inactivated AMPV vaccines are often used for
booster immunization of layer and breeder flocks
after priming with live vaccines. Whereas
inactivated vaccines alone induce only partial
protection against AMPV infection, the most
efficient and long-lasting protection is achieved by
a combined prime-boost vaccination program. This
program comprises repeated priming with live-
attenuated vaccines and booster immunization
with inactivated adjuvanted vaccines. As
experimentally shown, in ovo vaccination may also
be a promising strategy for effective, early
induction of an immune response. Besides live
attenuated and classical inactivated vaccines,
some genetically engineered viruses, including
recombinant vectored vaccines, have been
designed and tested under experimental
conditions. These have induced partial protection
and need further development.
TAREA

• Anorexia
• Vacuna inactivada
• Macrófagos
• proteína de matriz
• Glicoproteínas
• PCR de transcriptasa inversa
• Agente casual
 BIBLIOGRAFIA

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