Infectious Bronchitis: Up Date

Hafez Mohamed Hafez

Institute of Poultry Diseases, Free University Berlin

Knigsweg, 14163 Berlin, Germany
E. mail: hafez@vetmed.fu-berlin.de

Summary
Infectious bronchitis (IB) is an acute, contagious disease of chickens characterised
primarily by respiratory signs. The Infections may also lead to nephritis and egg
production problems. In laying flocks economic losses are due to drop in egg production
and egg quality. The severity of respiratory infections with IB virus can be greatly
enhanced by the presence of other pathogens of the respiratory tract and management
factors. The IB virus may spread through the flock without obvious clinical signs of
disease except a mild cough. In recent years, nephropathogenic strains have become
more common in laying flocks.
Prevention of infectious bronchitis is best achieved through an effective biosecurity
program and proper vaccination. Currently, poultry are almost universally vaccinated
against IBV. However, the multiplicity of serotypes identified around the world presents a
challenge in choosing appropriate strains for vaccination and designing an effective
vaccination program. This means that the efficacy of vaccination program and the
economic impact of the disease in an area will vary over the time depending on the
nature of the virus strains in circulation and the effectiveness of available vaccines to
combat them. Severe outbreaks of IB can occur when a new strain, which is distinct
from the vaccine strain, enters a population. A new variant may emerge if just a few
amino acids are changed during the emergence of a new virus by mutation. However, in
many cases the vast majority of the viral genome remains unchanged, so that existing
vaccines may still control the infection with this new variant. It has also been
demonstrated that "classical" strains of infectious bronchitis virus can act at least as
partial primers for subsequent administration of an inactivated infectious bronchitis
vaccine containing variant and standard strains.
The present paper explores the current situation related to diagnosis and control of
infectious bronchitis.

Introduction
Infectious bronchitis (IB) was first observed in 1930 and has been recognised as a
disease of major economic importance in chickens since that time. Infectious bronchitis
(IB) is an acute, contagious disease of chickens characterised primarily by respiratory

signs. The infection allows secondary invasion of the lungs and air sacs with bacterial
infections such as E. coli, which is mostly accompanied with significant losses due to
condemnations at slaughter. Weight gain and feed conversion efficiency may be
impaired. The Infections may also lead to nephritis and egg production problems. In
laying flocks economic losses are due to drop in egg production and egg quality
(Cavanagh and Naqi, 2003).
IB virus, a single positive-strand RNA virus, is a member of the genus Coronavirus, of
the family Coronaviridae of the order Nidovirales. Coronaviruses were divided into 4
antigenic groups. Infectious bronchitis virus (IBV) was placed in group 3. In recent
studies, it was demonstrated that IBV and Turkey coronavirus (TCV) have a lot of
antigenic and molecular similarities and both were placed in group 3 (Guy, 2000,
Cavanagh et al., 2002).
The virus is enveloped. It is fairly labile, being easily destroyed by disinfectants, sunlight,
heat and other environmental factors. Infectious bronchitis virus has the ability to mutate
or change its genetic makeup either by mutation or recombination (Cavanagh et al.,
1992). As a result, many serotypes have been identified. IB strains vary significantly
from country to country as well as from region to region. Three common serotypes in
North America are the Massachusetts, Connecticut, and Arkansas 99. In Europe various
"Holland variants," usually designated using numbers (D-274, D-212, D-1466), are
recognized. Recently, some strains of IB virus particularly the so-called 793/B or 4/91
serogroup, have been isolated from broiler breeder chickens with clinical signs of
cyanosis, muscular tremors and hyperventilation, leading to rapid death. At postmortem examination one of the most consistent findings was severe tracheitis and a
bilateral pectoral muscle myopathy. This serotype, first reported in the UK in 1992 is now
widespread in Europe and other parts of the world (Gough et al., 1992). In addition,
several strains of IB virus showing a strong affinity for the kidney (nephropathogenic
strains) were isolated (Cumming, 1969) and have become more prevalent in recent
years. Although differences in virulence and tropism exist between strains, primary
replication in the respiratory tract occurs with most IB viruses.
The virus shedding follows in faeces, respiratory discharges and on contaminated
eggshells. The virus is transmitted horizontally by direct contact with infected chickens,
indirectly through ingestion of contaminated feed and water or inhalation contaminated
particles. Transmission from farm to farm is related to movement of contaminated
people, equipment, and vehicles or via airborne transmission and this can occur over
considerable distances. Following infection, chickens may remain carriers and shed the
virus for several weeks. The virus persists in the intestinal tract and is excreted in the
faeces for long periods (Alexander and Gough, 1997). Once the IB virus enters a flock, it
spreads quickly among all the birds in the flock. Therefore, the infection rate within an
infected flock is close to 100%.
The severity of respiratory infections with IB virus can be greatly enhanced by the
presence of other pathogens of the respiratory tract and management factors. The
clinical signs of IB include watery discharge from the eyes and nostrils, sneezing,
snicking, coughing, tracheal rales and gasping. In many cases, however, the IB virus
may spread through the flock without obvious clinical signs of disease except a mild

cough. The mortality is usually very low, unless complicated by other factors such as E.
coli, Mycoplasma gallisepticum, Mycoplasma synoviae, immunosuppression, poor air
quality and high stocking density (McMartin, 1993).
Mortality may occur in young birds due to respiratory or kidney failure. Chickens infected
with nephropathogenic IB viruses excrete watery droppings, resulting in wet litter
(Cumming, 1969). In recent years, nephropathogenic strains have become more
common in laying flocks. These strains may cause a high mortality during the infection
or long time after as a result of kidney damage that progress to urolithiasis (Brown et al.,
1987). After apparent recovery, chronic nephritis can produce sudden death some time
later, especially in brown birds.
In the neonatal, susceptible chick, infection can lead to permanent damage to the
developing oviduct resulting in the occurrence in flocks of non-laying hens, the so-called
false layers (Crinion, et al., 1971). Infection of laying hens with IB virus can result in a
drop in egg production and the production of pale, thin-shelled, rough shelled and
misshapen eggs, this may occur without respiratory signs. Hatchability may also be
reduced. The negative influence of IB on egg quality may persist for many weeks (6 8
weeks) or months after production has recovered. A 5 - 10% decrease in egg production
may occur in flocks affected with the new IB virus strain 4/91, with some flocks suffering
drops of up to 50%. Pale eggs can appear 2 to 5 days after exposure to the virus. The
occurrence of pale eggs can persist for several weeks. In addition, the infection is mostly
accompanied with reduced internal egg quality in form of a serous thinning of the thick
albumin "watery whites"(McMartin, 1993).
In infected chickens respiratory tract lesions include mucous or thick exudate in the
trachea. The mucus tends to accumulate in the lower part of the trachea and a mucus
plug is sometimes found near the bronchi. In addition, the air sacs appear cloudy. In
case of infection with nephrotropic strain, the kidneys of affected birds are pale, mottled,
and can be 2 to 3 times their normal size. Urates are common and can be identified
easily in the kidneys and ureters at necropsy. In layers impacted oviducts, ruptured ova,
internal layers (egg peritonitis) are constant findings. Infection of very young chicks may
result in the development of cystic right oviducts.
Infectious bronchitis is difficult to differentiate from many of the other respiratory
diseases. Clinical signs are indicative, but not diagnostic and confirmation requires the
direct detection or isolation of IB virus or indirectly by detection of antibodies.
Using conventional direct methods many limitations can influence the efficacy of the
laboratory diagnosis. In cases of direct detection using electron microscopy,
immunofluorescence or immunoperoxidase a sufficient amount of antigen in the samples
is necessary and required. For the virus isolation in chicken embryos or in tracheal
organ cultures, beside a long time need to isolate the virus also the short shedding
duration is a further limited factor (Gelb and Jackwood, 1998) Recently, molecular
biological methods have become increasingly applicable to the diagnosis of infectious
bronchitis. The availability of these methods can overcome the limitations of traditional
approaches in the diagnosis (Jackwood et al., 1992; Handberg et al., 1999). The
currently available PCR does not, however, distinguish between vaccine and field strains
of this genotype and the vaccinal virus strain could be detected for long time
(Zimmermann and Hafez, 2001).

As a result of great heterogeneity of IB virus, serotyping and genotyping using

haemagglutination inhibition (HI) or virus neutralization (VN) and PCR of isolates may
produce results which are not in agreement. The HI test can be subject to problems with
cross reactivity. The VN test is very specific and may result in viruses with very similar
amino acid sequences behaving like unrelated strains. With genotyping, using universal
oligonucleotides may not bind to new variant types, leading to false negatives. It is
currently accepted that virus isolation and primary characterisation, followed by
serotyping/genotyping, are the recommended procedures for IB diagnosis.
The detection of antibodies can be achieved using haemagglutination inhibition tests
(HI), Agar gel precipitation (AGP) or enzyme-linked immunosorbent assays (ELISA) and
neutralization tests. The tests differ in their sensitivity and specificity (De Wit et al.,
1997). With the widespread use of vaccines for the prevention of diseases in commercial
poultry, serology has become of limited value in diagnosing IB infections of chickens. It
is essential for routine serological testing to compare two sets of serum samples. One
collected at onset of clinical disease and the second at 3 to 4 weeks later. It is also
important to mention that none of the available methods can differentiate between the
antibody responses to a field challenge from that to vaccination. In addition, a marked
increase in antibody using HI with a given IB serotype it cannot be assumed that the
field challenge virus was of the same or a closely related serotype. When birds are
immunized with one serotype and subsequently challenged with another serotype the
antibody response tends to be much higher to the antigen used in the initial vaccination.
Brown and Bracewell (1988) suggested that HI-test may be more strain-than serotypespecific.
Prevention of infectious bronchitis is best achieved through an effective biosecurity
program. Good management procedures include cleaning and disinfection of the poultry
house, proper ventilation, repopulating with day old chicks and strict isolation are very
important. The second line of defence is proper vaccination. Vaccines have been used
to control the infectious bronchitis infections since the 1950s. Currently, poultry are
almost universally vaccinated against IBV. Most vaccines used worldwide are standard
Massachusetts strains of the IB virus, e.g., H120, H52, Ma5 and M41.In some European
countries, strains D274, D1466, and 4/91, D274, D1466, and 4/91 (793/B) are used.
(Cavanagh and Naqi, 2003). Both live and inactivated virus vaccines are used. Infectious
bronchitis vaccination programs in broilers involve the use of modified live vaccines.
Vaccination of layers and breeder flocks involve administering a series of live vaccines
and progressively increasing the aggressiveness of the route of vaccination (i.e., start
with water administration and progress to fine particle spray) and strain of vaccine
(highly attenuated, H120) to less attenuated, H52). Evidence that some vaccines
increased in virulence after back-passage in chickens was reported by Hopkins and
Yoder (1986). Good vaccination practices are especially important when administering
live infectious bronchitis vaccines. The virus is relatively fragile and can easily be
inactivated if proper vaccination procedures (e.g. protection from sunlight, removal of
sanitizers from water used for mixing/administration, use of skim milk stabilizer, etc.) are
not followed. In breeders and some layer flocks an additional vaccination using
inactivated oil-emulsion vaccine is used prior to onset of egg production to stimulate the
antibody production. Inactivated infectious bronchitis vaccines do not stimulate local
(IgA) and cell-mediated (T-cells) immunity as effectively as modified live virus vaccines.

That is why some companies will continue to vaccinate layers and breeder flocks during
the production using a live vaccine.
The multiplicity of serotypes identified around the world presents a challenge in choosing
appropriate strains for vaccination and designing an effective vaccination program. This
means that the efficacy of vaccination program and the economic impact of the disease
in an area will vary over the time depending on the nature of the virus strains in
circulation and the effectiveness of available vaccines to combat them. Severe
outbreaks of IB can occur when a new strain, which is distinct from the vaccine strain,
enters a population. A new variant may emerge if just a few amino acids are changed
during the emergence of a new virus by mutation. However, in many cases the vast
majority of the viral genome remains unchanged, so that existing vaccines may still
control the infection with this new variant. It has also been demonstrated that "classical"
strains of infectious bronchitis virus can act at least as partial primers for subsequent
administration of an inactivated infectious bronchitis vaccine containing variant and
standard strains. In aim to establish an efficient vaccination program against infectious
bronchitis, it is essential to identify the prevalent serotypes in the region and to
determine the cross-protective potential of available vaccines. Although no reasonable
combination of infectious bronchitis vaccine strains provides full protection against all
heterologous challenges, it is both undesirable and unnecessary to consider developing
a new live vaccine for each new emerging strain, rather than to use a combination of
strains, which are able to protect against a wide variety of serotypes protect types.
There are combinations, which are able to provide broad protection. Recent studies,
showed that for e.g. the use of the two live attenuated IB vaccine strains 4/91 and Ma5,
resulted in considerably broaden protection against challenge with a wide variety of
antigenically different strains (Cook et al., 1998a).

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