Rev. sci. tech. Off. int. Epiz.

, 2000, 19 (2), 425-442

Avian mycoplasmosis (Mycoplasma gallisepticum)

(1) (2)
S. Levisohn & S.H. Kleven
(1) Ministry of Agriculture and Rural Development, Veterinary Services and Animal Health, Kimron Veterinary
Institute, Division of Avian and Aquatic Diseases, P.O. Box 12, Beit Dagan 50250, Israel
(2) Department of Avian Medicine, University of Georgia, Athens, Georgia 30602-48745, United States of
America

Summary
Mycoplasma gallisepticum is t h e most e c o n o m i c a l l y significant m y c o p l a s m a
pathogen of poultry, and has a w o r l d - w i d e distribution. In c o m m o n w i t h other
m y c o p l a s m a s , M. gallisepticum is minute in size w i t h minimal genetic information
and w i t h a total lack of a bacterial cell w a l l . These properties are reflected in a
high degree of interdependence b e t w e e n M. gallisepticum and the host animal,
and in t h e fastidious nature of t h e organism in vitro. Strains of
M. gallisepticum differ markedly w i t h respect t o important biological properties
s u c h as pathogenicity, infectivity, tissue tropism and transmissibility. In addition,
phenotypic variation of major surface antigens o c c u r s at high frequency, w h i c h is
a probable explanation f o r chronic infection by M. gallisepticum despite a strong
immune response.
Infection w i t h M. gallisepticum has a w i d e variety of clinical manifestations, but
even in t h e absence of overt clinical signs, t h e e c o n o m i c i m p a c t m a y be
significant. The most dramatic disease presentation of M. gallisepticum is chronic
respiratory disease in meat-type birds, often as one of several aetiological agents
in a multi-factorial disease complex. Transmission of M. gallisepticum in ovo from
infected breeder birds t o progeny is t h e major route of dissemination of t h e
infection, and is the prime consideration f o r international trade.
In most countries, control p r o g r a m m e s f o r M. gallisepticum are based on
maintaining c o m m e r c i a l breeding stock free of infection. In instances w h e r e
control of M. gallisepticum infection is not feasible, v a c c i n a t i o n , especially w i t h
n e w l y developed live M. gallisepticum v a c c i n e s , is being evaluated as an option.
M a j o r advances in diagnostic methods have been made in r e c e n t years. Control
p r o g r a m m e s have been based on serological methods, w i t h screening f o r
infection usually a c c o m p l i s h e d by the slide plate agglutination (SPA) test o r by
enzyme-linked i m m u n o s o r b e n t assay. Further serological testing and/or
demonstration of t h e presence of t h e organism must be used t o confirm SPA
s u s p e c t e d positive tests. In principle, detection of t h e presence of t h e
M. gallisepticum organism can be by isolation of the organism or detection of the
deoxyribonucleic acid by molecular methods. Polymerase chain reaction
represents a rapid and sensitive alternative t o traditional culture methods, w h i c h
require t i m e - c o n s u m i n g specialised t e c h n i q u e s . The development of molecular
typing methods affords n e w opportunities f o r epidemiological studies and
identification of reservoirs of infection.

Keywords
Avian diseases - Biosecurity - Chronic respiratory disease - In ovo transmission -
Molecular typing - Mycoplasma gallisepticum - Vaccination.

Aetiological agent minute size and total lack of a cell wall ( 1 2 2 , 123). The
presence of only a minimal amount of genetic information
The Mollicutes, more commonly known as mycoplasmas, are accounts for the complex nutritional requirements of these
distinguished phenotypically from other bacteria by their organisms, reflected in an obligate parasitic mode of life, with
426 Rev. sci. tech. Off. int. Epiz., 19 (

a high degree of interdependence between the mycoplasma Variability occurs not only among strains, but within clones of
and the host animal. The fastidious nature of mycoplasmas is a single strain ( 1 0 , 4 0 ) . Continual variability in the expression
also reflected in the culture conditions required in vitro, with of such surface antigens also occurs in vivo (87) and may be a
the consequence that isolation of the organism is usually major factor in the development of clinical disease in addition
performed in specialist laboratories (73). In the absence of a to having a significant impact on the development of
cell wall, organisms are highly pleomorphic and are not serological responses (86). The marked heterogeneity with
usually detectable by standard light microscopy of respect to presentation of the major surface antigens provides
Gram-stained cultures or tissue smears. The typical 'fried egg' a likely explanation of how mycoplasma infections are able to
morphology of the microscopic colonies results from persist in birds despite a strong immune response.
insinuation of the tiny pleomorphic organisms among the
fibres of the mycoplasma agar medium where growth is Resistance to the clinical manifestations of mycoplasma
initiated. Further division of cells, as the colony develops, infection, and positive response to antibiotic therapy for
results in spread onto the surface of the agar (120). In disease require an intact immune system (6, 18). In humans,
addition, since mycoplasmas are not sensitive to antibiotics mycoplasmas are being increasingly implicated as co-factors
that affect cell wall synthesis, these antibiotics may be in new diseases or in diseases of previously unknown
incorporated in culture media used for isolation of aetiology. This has been attributed to improvements in
mycoplasmas, in order to inhibit growth of other bacteria diagnostic techniques and an increase in the susceptible
present in clinical samples (69, 7 3 ) . population of immunodeficient patients (due to human
immunodeficiency virus, cancer chemotherapy, and others)
(6). At present, little is known about the pathogenesis of
Most species of mycoplasma are parasites of animals or plants,
mycoplasmas in immunocompromised poultry. However,
including more than a dozen species which are known to
unusual disease syndromes are often present in flocks that are
infect chickens or turkeys (67). Of these, Mycoplasma
immunocompromised because of infectious or non-infectious
gallisepticum and M. synoviae are. pathogenic for chickens and
agents (129). Complicated infections of multiple aetiologies,
turkeys, while M. iowae affects primarily turkeys, and
probably including one or more avian mycoplasma species,
M. meleagridis only infects turkeys. Mycoplasmas tend to be
are frequently present under commercial conditions and
highly host-specific, although improved diagnostic methods
present a challenge to diagnosis and treatment (68, 6 9 ) .
and precise molecular methods for species identification may
necessitate some re-evaluation of earlier views (6, 137). Avian
mycoplasmas are not known to infect humans or other
mammalian species, although isolations from domestic Description of the disease
animals have been reported sporadically. Mycoplasma
gallisepticum may infect many species of birds, but The most economically significant mycoplasma pathogen of
gallinaceous birds are most susceptible. poultry is M. gallisepticum (74, 9 4 ) . The major consideration
for international trade is the necessity and the ability to
determine the M. gallisepticum status of the imported material
The primary habitats of mycoplasmas are the mucosal (most often hatching eggs or one-day-old chicks). In most
membranes of the respiratory tract, and/or the urogenital cases, the M. gallisepticum status of the progeny chicks is
tract, eyes, mammary glands and joints. Most mycoplasmas determined by the breeder flock from which they are derived.
are considered surface parasites, rarely invading tissues, In this section, some of the factors which affect
although spread to other organs strongly suggests a transitory M. gallisepticum infection of embryos and chicks and the
systemic infection, at the least. Adhesion of mycoplasmas to ability to detect this infection will be discussed. The clinical
host cells is a prerequisite for successful colonisation, and manifestations of M. gallisepticum infection will also be
ensuing pathogenesis (76, 121). New information, relating discussed, since an understanding of the disease is necessary
principally to mycoplasmas infecting humans, suggests that to evaluate policy.
some species may penetrate cells and survive intracellularly
(123). In general, mycoplasma species, and possibly also
Infection with M. gallisepticum may have a wide diversity of
specific strains, exhibit organ and tissue proclivity, although
clinical manifestations, of which chronic respiratory disease
the organism is not necessarily completely excluded from
(CRD) and downgrading of carcasses in meat-type birds are
other organs. Mycoplasma gallisepticum is one of the species of
probably the most dramatic (68, 9 4 ) . Descriptions of gross
mycoplasma that, as primary pathogens, can cause acute and
lesions and histopathology may be found in a recent review
chronic diseases at multiple sites, with wide-ranging
and the references therein (94). Mycoplasma gallisepticum
complications.
infection alone may cause respiratory disease in turkeys, but is
most often mild or sub-clinical in chickens. However,
Mycoplasmas, including M. gallisepticum, have recently been M. gallisepticum is frequently present as one of several
demonstrated to have the ability to vary the expression of aetiological agents in a multi-factorial disease complex
major surface antigens, thus expressing a continually (53, 68). The interactions among M. gallisepticum,
changing 'antigenic profile' to the immune system (123). various respiratory viruses, Escherichia coli, Haemophilus
Rev. sci. tech. Off. int. Epiz., 19 (2) 427

paragallinarum, and others have been documented ( 4 9 , 6 8 , Embryo mortality, which may occur as a result of infection
107). Vaccine strains of Newcastle disease virus or infectious with M. gallisepticum, has been shown in experimental
bronchitis virus may give marked respiratory reactions in infection studies to be affected by strain differences ( 8 3 , 118).
M. gallisepticum-infected birds, necessitating the use of milder However, the pathogenicity in ovo does not correlate with the
vaccine strains under these conditions (94, 106). The route of pathogenicity of the M. gallisepticum strain in respiratory
exposure and the infectious dose of the microbe, in addition infection of the fowl in vivo ( 8 4 ) . The presence of maternal
to environmental and stress factors, such as temperature and antibodies to M. gallisepticum in the embryonated egg has
ammonia concentration, age and type of birds, are among the been found to reduce the in ovo pathogenicity of infection,
factors that influence mycoplasmosis ( 9 4 ) . Moreover, increasing the probability of survival of the infected embryo
infection with M. gallisepticum may cause a marked reduction (83). However, M. gallisepticum infection may weaken the
in feed consumption efficacy with a significant economic embryo, resulting in difficulty in hatching (pipped embryos)
impact, even in the absence of overt clinical signs. or low-quality chicks.

Loss of production in laying birds may also occur as a result of With the exception of in ovo infection, the upper respiratory
M. gallisepticum infection, and is particularly marked in table tract is generally accepted to be the portal of entry in natural
egg layers infected at peak lay (44). Early infection of pullets infection by M. gallisepticum. The trachea also appears to be
with M. gallisepticum can protect the birds against the the preferential site of infection for most strains of
pathogenic effects of later infection (34). This is the principle M. gallisepticum, although disease signs are usually manifested
underlying the successful use of live M. gallisepticum vaccines in other parts of the respiratory system (air sacs or sinuses).
in layer flocks, as is widely practised in the multi-age layer The trachea serves as a reservoir of infection and is also readily
operations in the United States of America (USA) and to a accessible for sampling the live bird. Levels of
limited extent elsewhere ( 1 9 , 139). Mycoplasma gallisepticum M. gallisepticum in the trachea are highest at the acute stage of
has been implicated in salpingitis and other pathologies of the infection, often before the first serological response is detected
reproductive system, although whether this is the principal or in all the birds ( 8 5 , 144). Detection by culture or other
sole cause of reduction in egg production is not clear ( 3 0 , methods for direct detection of the organism is most
110). successful at this stage of infection ( 5 2 ) . Levels of
7 2
M. gallisepticum in the trachea decrease from > 1 0 to < 1 0
In common with all the pathogenic avian mycoplasmas, colour changing units with progression of the infection. In the
M. gallisepticum is transmitted in ovo. Infection of the embryo absence of exacerbating environmental stress or infection with
by M. gallisepticum and transmission to the progeny (egg other pathogens, the progressive infection in the trachea may
transmission) probably occur as a sequel to acute respiratory be self-limiting (144), but tracheal infection persists in the
infection, due to contiguity of the abdominal air sacs to the presence of humoral or local antibody (66, 144). Tracheal
oviduct (125). Very little information is available about rates levels in chronically infected birds, such as commercial egg
of transmission, and these rates are likely to vary widely under layers several months after infection, may be so low that
different conditions, among individual birds, and at different M. gallisepticum is not detected by usual sampling and culture
times during the same outbreak. The highest frequency of methods. Antibiotic treatment may also markedly reduce
transmission is found during the acute phase of the disease tracheal levels, although non-viable cells may be present,
when M. gallisepticum levels in the respiratory tract reach a which can be detected by molecular methods (26, 5 8 ) .
peak (44, 9 6 ) . In separate studies of experimental
reproduction of the disease in breeder hens, peak Although intercurrent infections and environmental factors
transmission to eggs after respiratory challenge by influence the outcome of M. gallisepticum infection, marked
M. gallisepticum was detected at four weeks post infection in heterogeneity has been observed within the species with
approximately 2 5 % of the progeny (45) and in over 5 0 % at respect to important biological properties. Variability in tissue
three to six weeks (130). At eight to fifteen weeks after tropism is indicated by isolation of an encephalitic form
infection, transmission levels were approximately 3 % (125) or of M. gallisepticum from turkeys (22, 136). An
approximately 5% at twenty to twenty-five weeks post M. gallisepticum variant with proclivity for the cloaca
infection (45). However, some individual birds showed produced a persistent low level serological response (102). A
consistently high transmission rates throughout the test strain of M. gallisepticum with proclivity for the eye,
period. During chronic infection under field conditions, producing unilateral enlargement of the eyeball, has been
transmission to eggs is likely to occur at much lower levels. reported (119), in addition to a natural outbreak of
Transmission of M. gallisepticum from infected chicks at hatch keratoconjunctivitis in layer chickens, apparently caused by
is a major source of infection whereby even low levels of M. gallisepticum (109). Mycoplasma gallisepticum has been
transmission to the egg result in widespread infection among demonstrated to be the aetiological agent of conjunctivitis in
the progeny. However, at low transmission rates, detection of free-ranging house finches (Carpodacus mexicanas) and
the infection by sampling embryos or one-day-old chicks American goldfinches (Carduelis tristis) in the eastern USA
requires a very large sample size to be statistically feasible. and Canada ( 9 1 , 99). The M. gallisepticum isolates from song
428 Rev. sci. tech. Off. int. Epiz., 19

birds comprise a unique biotype according to biological while the eggs are still in the hatchery. As mentioned above,
properties and molecular typing (92). egg transmission occurs at the highest levels during the early
stages of infection, and this is the critical period with respect
So-called variant strains, characteristically differing from to the status of the exported material. Negative serological
prototype M. gallisepticum by reduced virulence, results in the breeder flock prior to shipment of the progeny
transmissibility and antigenicity, were first described in the are a good indication that the flock was free of
early 1970s ( 1 5 0 ) . More recently, similar strains were M. gallisepticum when the eggs were set in the hatchery
recognised in turkeys (5). In experimental infection studies, twenty-one days earlier.
variant strains of M. gallisepticum were often difficult to
reisolate from individual birds, but persisted in the flock and Diagnosis of M. gallisepticum infections in poultry breeder
were particularly problematic with respect to diagnosis ( 2 9 , flocks is often performed in the absence of overt clinical signs,
62). Although variant strains were originally regarded as and screening for infection is usually accomplished by the
exceptional and unusual phenomena, these now appear to be slide plate agglutination (SPA) test with commercial stained
extreme and stable examples of a spectrum of variable antigen. Slide plate agglutination is highly efficient in
biological and antigenic properties within the detecting immunoglobulins of the immunoglobulin M (IgM)
M. gallisepticum species. Important examples of 'atypical' class, which are the earliest response to mycoplasma infection
M. gallisepticum strains are the live vaccine strains F, ts-11 and (64). Although the test is rapid, highly sensitive and relatively
6/85 which will be discussed in detail at the end of this paper.
inexpensive, care must be taken to perform the test according
Each of the vaccine strains differs markedly from the others
to the instructions of the antigen manufacturer, including
and from fully pathogenic M. gallisepticum, in biological
appropriate control sera (73). Sets of positive control sera
properties such as infectivity, transmissibility, persistence and
from chickens infected by contact with M. gallisepticum or
immune response, in addition to genetic and antigenic
M. synoviae and negative control sera are currently available
markers which can be detected in molecular diagnostic tests.
from the Poultry Diagnostic Research Center, University of
Georgia, Athens, USA. The greatest disadvantage of the SPA
test is low specificity, with false positive reactions and

Diagnostic methods cross-reactions encountered relatively frequently. Serological

detection of M. gallisepticum may be complicated by
co-infection of flocks with M. synoviae, due to serological
Whatever method is employed for mycoplasma control, rapid
cross-reactions between the two mycoplasma species (4). In
and precise diagnostic methods are of paramount importance.
addition, non-specific serological reactions are frequently
Major advances in diagnostic methods have been made in
detected after use of inactivated vaccines ( 4 3 ) . Further
recent years, and when validated by field experience, are
serological testing and/or demonstration of the presence of the
reflected in the periodic updates of standard procedures.
organism must be used to confirm positive or suspected
Information may be found in publications supporting the
National Poultry Improvement Plan (NPIP) (3) and in the positive SPA tests. In some cases, marking of birds which are
Manual of Standards for Diagnostic Tests and Vaccines of the sampled may be desirable to facilitate repeat testing of those
Office International des Epizooties (OIE) ( 7 3 ) as well as in which give suspicious serological reactions.
peer-reviewed publications that relate to specific methods.
Traditionally, the test of choice for confirmatory serology has
The basis for control programmes has centred on serological been haemagglutination-inhibition (HI), which can be
methods, both screening and confirmatory, with reactors performed with fresh culture of a haemagglutinating test
confirmed by isolation of the organism. In international trade, strain of M. gallisepticum (73) or with standardised preserved
the M. gallisepticum status o f chicks (or embryos, in the case of antigen (3). Diagnostically significant titres in the HI test may
fertile eggs), is usually determined by the testing and not be detected until three or more weeks after infection.
certification of the breeder flock of origin. Diagnosis is made However, the test is highly specific, even to the level of
on a flock basis, and the presence of one or more infected differentiation among strains (70). The major factors in
birds in the flock sample constitutes an infected flock. Factors support of alternative methods are the delay in development
which affect the accuracy of the testing procedures are the of HI antibodies, the strain specificity which may result in lack
sample size, the time period between tests, and the timing of of detection of variant M. gallisepticum strains and the
the test with respect to shipment of the chicks. General technical problems which may be encountered in producing
guidelines of the NPIP require testing of 1 0 % of the flock (a high titre specific HI antigen and in performing the test.
minimum of 3 0 0 birds) before the onset of egg production
and every sixty to ninety days thereafter (3). Regulations and Commercial enzyme-linked immunosorbent assay (ELISA)
policy instituted by breeder companies relating to flocks from
kits are widely available and are increasingly used for
which fertile eggs are exported are often more stringent,
serological confirmation ( 5 9 ) . Marked differences may be
mandating serological screening at more frequent intervals
found among the different manufacturers, and care should be
(for instance, every two weeks) in order to obtain test results
taken to use a product which has been validated with a wide
 Rev.sci. tech. Off. int. Epiz., 19 (2) 429

spectrum of field samples and strains. The recommended (131), but results should be treated with caution due to the
ELISA kits have excellent sensitivity and specificity, but possibility of non-specific reactions and the relatively low
transitory non-specific reactions may still occur, for similar sensitivity of the test for IgG. Antibodies to M. gallisepticum
reasons to those occurring in the SPA test (4, 4 3 ) . Potential have been detected in chicken bile by various serological tests,
improvements in ELISA specificity may result from the use of including the indirect immunoperoxidase assay, which may
a blocking ELISA utilising an M. gallisepticum-specific also be an option for progeny chicks (9).
monoclonal antibody ( 2 8 ) , utilisation of highly purified
antigens ( 1 0 3 ) or recombinant antigen, as suggested for In principle, the presence of the M. gallisepticum organism can
M. synoviae ELISA (108). However, increasing the specificity be confirmed by isolation in mycoplasma media or by
presents the risk of decreasing the ability to detect all
detection of the specific deoxyribonucleic acid (DNA).
M. gallisepticum strains.
Isolation is still considered the 'gold standard', but the
existence of circumstances where M. gallisepticum may be
Some possible pitfalls exist in the dependence on serological present but cannot be isolated even by the most skilful
testing for determination of flock status for M. gallisepticum. techniques, is now fairly well accepted. Detailed methods for
Temporal development of antibodies has been described in culture and identification of M. gallisepticum may be found in
experimentally infected poultry (4, 6 4 ) and is presumed to the OIE Manual and other texts ( 6 9 , 7 3 , 74). The ability of
follow a similar course in field infection. However, some flock
culture media to support the growth of M. gallisepticum
treatments, such as the use of certain antibodies, may affect
should be confirmed by testing with a low passage isolate.
the development of the immune response. In experimental
Identification of M. gallisepticum and differentiation from
infection trials, fewer serological responses were found in
other mycoplasma isolates is usually based on immunological
M. gallisepticum-infected chickens or turkeys treated with
methods, most frequently immunofluorescence, requiring
antibiotics than in the M. gallisepticum-infected non-
specific antisera that are not available commercially. An
medicated groups (54, 5 5 , 5 6 ) . Immune suppressive agents,
alternative method for identification is the use of DNA-based
infective or non-infective, may delay the onset of a detectable
tests, using specific or universal mycoplasma tests ( 7 8 , 8 0 ) .
serological response to infection (129). Several studies have
reported problems in detecting M. synoviae infection using
Polymerase chain reaction (PCR) represents a rapid and
the SPA test, primarily in turkeys (113). Although it is not
sensitive alternative to traditional culture methods which
clear to what extent, if at all, a similar phenomenon occurs
require specialised media and reagents and are
with M. gallisepticum ( 1 1 4 ) , some laboratories have
time-consuming (57). A major advantage of the
introduced the ELISA test as a supplementary or alternative
implementation of M. gallisepticum-PCR technology is that
screening method. An ELISA kit which detects both
the investment in training and equipment can be exploited for
M. gallisepticum and M. synoviae antibodies in a single
diagnosis of an increasingly wide range of poultry diseases for
reaction may be used for preliminary screening.
which PCR is now one of the tests of choice.

Serological testing of progeny chicks to determine

M. gallisepticum status is sometimes desired, particularly Results of the PCR test can be obtained in one or two days, as
when access to the parental flock is limited or when questions opposed to the usual one to three weeks for isolation and
arise about the testing methods used at the place of origin. identification of M. gallisepticum. Equally important is the
Immunoglobulin G passes from the maternal circulation into ability to obtain accurate PCR results in the presence of mixed
the yolk of the egg, with the pattern of rise and fall of infection with several species of mycoplasma, contamination
antibodies following that of the serum, but delayed by five to by secondary bacterial infections, and inhibition of growth by
six days, the time required for the maturation of the egg (48, antibiotics, antibodies or other host factors. In particular, the
128). Transferred IgM has also been detected at low levels in problem of co-infection with saprophytic mycoplasmas that
the yolk, and more significant levels in the egg white (145). grow more rapidly than M. gallisepticum in enrichment
However, only maternal IgG, derived from the egg yolk, is cultures is a major impediment to isolation. Detection of DNA
present in the circulation of the chick (128). This passes from from non-viable organisms, for instance after antibiotic
the yolk to the embryo during the last five or six days of treatment, is a possible drawback to the PCR method (58).
embryo development and to the chick for approximately two
days post hatching, at which times serum levels should peak. The availability of a commercial kit for M. gallisepticum-PCR
Catabolism of maternal antibody occurs during the first two was a major impetus to the introduction and acceptance of the
weeks post hatching, with declining antibody titres. Synthesis PCR technology as a supplementary diagnostic method for
of new antibodies in the chick, both IgG and IgM, begins in M. gallisepticum. An additional advantage of the commercial
the first week of life, and is expected to reach adult levels at M. gallisepticum-PCR kit is the ability to differentiate between
four to six weeks of age ( 8 1 , 128). The ELISA is usually standard strains of M. gallisepticum and the F vaccine strain.
recommended for testing of yolk samples in fertile or Recently, increased interest has been shown in alternative,
non-fertile eggs, and testing of maternal antibodies in the non-commercial PCR tests that are less expensive and
chick ( 1 7 ) . Reactions to the SPA test may also be present somewhat more rapid than the commercial kit (78). All PCR
430 Rev. sci. tech. Off. int. Epiz., 19 (2)

methods require specialised and precise technical skills in a In birds sacrificed for sampling or in fresh carcasses, after
dedicated laboratory. Due to the high sensitivity of the test, necropsy, or from dead-in-shell or pipped embryos, isolation
care must be taken to avoid false positive reactions due to of M. gallisepticum may be performed successfully from a
extraneous DNA, but this can be prevented or detected by variety of organs, usually from the respiratory or reproductive
appropriate controls. A critical control in the use of PCR for tract (2, 7 3 , 9 4 ) . Isolation has been successful from the brain
diagnosis is the inclusion of an internal control to avoid 'false or the eye of fowl with relevant clinical signs (22, 1 0 9 ) , in
negative' results due to the presence of inhibitory substances addition to bile of infected birds (9). Sampling of carcasses for
in the reaction mixture. Amplification of the internal control isolation, including those that have been frozen, may be
amplicon, which can be readily differentiated from the target problematic due to the presence of bacterial contamination or
DNA amplicon, indicates that there are no inhibitors of the lysis of cells which liberates inhibitory substances. Sampling
PCR reaction. Recently, an intrinsic control was developed for for PCR must also assure that conditions are such that no
ari M. meleagridis PCR test (104), but such a control is not in degradation of the DNA occurs by intrinsic or environmental
routine use for M. gallisepticum-PCR testing. factors.

A recent innovation in diagnosis is the development Several rapid methods for extraction of DNA from
of molecular typing methods for differentiation of mycoplasma cells have been used for sample preparation for
M. gallisepticum strains. The most commonly used molecular M. gallisepticum-PCR (58, 7 8 , 7 9 ) . Samples for PCR are often
pooled (three to five tracheal swabs per PCR reaction) to
typing method is random amplification of polymorphic DNA
increase sample size and reduce the cost of testing. However,
(RAPD), a PCR-based technique which gives a unique strain
pooling of samples may increase the possibility of inhibition
fingerprint (35, 4 1 ) . This technique is in routine use in a few
by substances that may be present in the mucus o r other
specialist laboratories, and readily distinguishes among the
tissue fluids, thus decreasing the sensitivity of the PCR test.
live M. gallisepticum vaccine strains and the field strains
W h e n pooling large numbers of samples, purification of the
present in natural infection ( 2 1 , 75). The RAPD technique has
DNA may be necessary using standard methods or rapid
also been used for molecular tracking of spread of infection
commercial kits.
among flocks and from putative reservoirs of infection in
commercial poultry (20, 8 8 , 8 9 ) . The technique requires a
Testing for the presence of M. gallisepticum in the
high degree of technical expertise, and gives a satisfactory
embryonated eggs or progeny chicks by culture is not
degree of reliability and reproducibility within each
recommended as a routine method for determining the status
laboratory, but only to a limited degree between different
of the flock. Low levels of in ovo transmission necessitate the
laboratories. Thus, the precision necessary for construction of
sampling of a large number of embryos. Mycoplasma
a database of strain-specific genomic fingerprints has not yet
gallisepticum can be isolated with relatively high frequency
been achieved. A major impediment to widespread
from pipped eggs from infected flocks (77), and preliminary
application is the necessity to perform RAPD on pure cultures
results suggest that this may also be a recommended sample
of M. gallisepticum isolates. This requires specialised skills in
site for PCR (A. Ramirez and S. Levisohn, unpublished
addition to lengthening the time required to obtain results.
findings).
Current research is attempting to develop rapid molecular
methods for specific detection of M. gallisepticum biotypes,
such as the live vaccine strains, as has been successful for the F
vaccine strain (63).
Epidemiology
Sampling, sample transport and processing for Mycoplasma gallisepticum infection occurs naturally in
M. gallisepticum testing are highly critical stages in diagnosis. chickens and turkeys. Transmission of M. gallisepticum
Sampling for M. gallisepticum in live birds is usually from the infection occurs by the following two major routes:
trachea or choanal cleft. In a comparative study, isolation rates
were higher from the latter site, with less stress for the bird - vertically (in ovo), from an infected breeder flock to the
(16). During the acute phase of infection, between twenty and progeny
thirty individual samples for isolation are usually sufficient, - horizontally, by direct or indirect contact of susceptible
whereas a larger sample size may be necessary at the chronic birds with infected carriers or contaminated debris.
stage of infection. Swabbing technique, including the type of
swab used and prewetting before sampling, may affect the Important characteristics of M. gallisepticum that affect the
success of isolation, especially when relatively few organisms epidemiology of infection are the relatively stringent host
are present ( 1 5 2 , 153). Isolation of M. gallisepticum has been specificity and the frequent occurrence of asymptomatic
successfully performed from the cloaca in experimentally infection. Molecular diagnostic methods for highly sensitive
infected chickens, although this may be attributable to an and specific detection of M. gallisepticum have been
unusual tissue proclivity in the M. gallisepticum strain used developed recently, and these are important tools for studying
(102). the epidemiology of M. gallisepticum infection. Mycoplasma
Rev. sci. tech. Off. int. Epiz., 19 (2) 431

gallisepticum can now be detected with greater efficiency, in a Isolation of M. gallisepticum from the respiratory tract of
wide variety of clinical samples. In addition, molecular typing ducks has been reported, with no apparent clinical signs (7).
methods for M. gallisepticum strains allow identification of Mycoplasma gallisepticum was also isolated from embryonated
specific strains of M. gallisepticum, as discussed in the section duck eggs from the infected flock. The apparent source of
on diagnosis. infection was infected chickens which were raised together
with the ducks. Experimental infection of specific-
In addition to domestic poultry, M. gallisepticum has been pathogen-free (SPF) ducks with M. gallisepticum resulted in
reported fairly frequently in other gallinaceous birds, either colonisation but only limited respiratory signs ( 6 0 ) .
free-ranging or maintained in captivity, and sporadically in Mycoplasma gallisepticum and M. synoviae also were isolated
birds of other genera ( 9 4 ) . Natural infection with from geese, apparently infected by contact with infected
M. gallisepticum has been reported in a variety of game birds, chickens, and from deadrin-shell goose embryos from the
in some cases with marked clinical signs. However, evaluation infected flock (8). Recently, M. gallisepticum was isolated from
of the extent of this phenomenon is difficult, due to difficulties the air sac in fattening geese with typical signs of CRD
in isolating M. gallisepticum in the presence of high levels of (S. Levisohn, unpublished findings). Molecular typing of the
saprophytic mycoplasmas (24, 9 4 ) . In some cases, the source M. gallisepticum isolates from geese gave the same RAPD
pattern as that found in M. gallisepticum isolates from an
of infection appears to be M. gallisepticum present in
outbreak in a broiler-breeder flock in the same region,
nearby domestic poultry. Retrospective testing by
although the geese and the chickens were not in contact at any
M. gallisepticum-PCR of stored clinical samples from
time.
partridges and pheasants in the United Kingdom with upper
respiratory disease demonstrated evidence for a higher rate of
infection than was found by isolation (15). This supported the In examining clinical samples, care must be taken to
theory that M. gallisepticum is a major cause of disease in these differentiate between M. gallisepticum and the closely related
birds, which also serve as a potential source of infection for species M. imitans, which is relatively frequently isolated
domestic poultry. Experimental infection of red-legged from ducks and geese (12). Some early reports of
partridges (Alectoris rufa) with M. gallisepticum demonstrated M. gallisepticum, prior to recognition of the M. imitans
the occurrence of pathogenic effects and persistent carriage in species, may have been misidentified. A useful tool for
this host (37). differentiation of M. gallisepticum and M. imitans is
an M. gallisepticum-PCR restriction fragment length
Mycoplasma gallisepticum has been implicated as the causative polymorphism test which exploits a unique sequence
agent in an outbreak of conjunctivitis in free-ranging house difference in the 16S ribosomal ribonucleic acid (rRNA) genes
finches and American goldfinches, and in a case of of the two species (57).
conjunctivitis in a blue jay (Cyanocitta cristata) that was
housed in a cage previously occupied by afflicted house
finches (91, 99). Initially observed in 1 9 9 4 , the infection is The clinical significance of M. imitans in chickens and turkeys
now nearly ubiquitous in wild finches over the entire eastern has not yet been established, although a potential for
range of these birds in the USA and Canada (36, 100). To pathogenicity has been demonstrated in experimental
investigate the molecular epidemiology of this outbreak, DNA infection studies. Mixed infection of M. imitans with
fingerprints of isolates were made by RAPD. Mycoplasma infectious bronchitis virus in chickens (38) or rhinotracheitis
gallisepticum isolates from songbirds (representatives of all virus in turkey poults (39) indicates a synergism between
three host species) examined from 1 9 9 4 to 1 9 9 6 in eleven these agents. Experimental infection of SPF ducks with
States were compared with vaccine and reference strains and M. imitans resulted in colonisation of the respiratory tract, but
with contemporary isolates from commercial poultry. All no respiratory signs (61). Mycoplasma imitans was isolated
M. gallisepticum isolates from songbirds had RAPD banding only sporadically from experimentally infected SPF chickens,
patterns identical to each other but different from other strains and no pathogenic effects were reported. In experimental
and isolates tested. The unique finch RAPD pattern is infection of partridges, colonisation by M. imitans was lower
consistently found in all isolates from these birds (36, 9 2 ) . and pathogenicity less than that found for M. gallisepticum
These results indicate that isolates of M. gallisepticum from (37).
conjunctivitis in songbirds are all of the same molecular type,
which suggests a single source. The RAPD typing results gave Horizontal infection of M. gallisepticum occurs readily by
no indication that infection is shared between songbirds and contact with infected birds, most probably by the airborne
commercial poultry. A slow spread to chickens has been route with organisms excreted from the respiratory system of
observed under experimental conditions, but only mild infected birds. Experimental infection with M. gallisepticum is
clinical signs are detected ( 1 1 1 , 132) and no natural infection often performed by aerosol presentation of the inoculum, a
of the unique finch biotype has been detected in domestic method which is highly efficient and believed to be similar to
poultry (92). the natural mode of infection (14). In an attempt to
432 Rev. sci. tech. Off. int. Epiz., 19

standardise the conditions for M. gallisepticum challenge, a Introduction of M. gallisepticum by free-flying wild birds that
special chamber for experimental aerosol infection has been penetrate the premises of a poultry farm is often proposed as a
designed (143). method of transmission. Mycoplasma gallisepticum and
M. synoviae have been demonstrated by specific PCR tests in
wild birds, including some captured in the area of
No published evidence is available to demonstrate the role of M. gallisepticum-infected poultry flocks, but no conclusive
airborne transmission of M. gallisepticum under field evidence for a role in transmission has been found
conditions, although circumstantial evidence is often found. (S. Levisohn, unpublished findings).
However, transmission of enzootic pneumonia in swine,
caused by M. hyopneumoniae, is likely to be similar to the Molecular typing methods for the identification of
situation with respect to M. gallisepticum in commercial M. gallisepticum strains are now being used for
poultry operations. Early studies demonstrated the influence epidemiological studies, helping to identify the link between
of the location of a farm on disease status, since the distance outbreaks occurring on different premises. Evidence has been
from the source of infection was identified as the major risk found for infection by the same strain in different flocks
factor for infection of swine with M. hyopneumoniae (47, 133). within the same poultry production company (20) or among
The risk of infection rises as the proximity of susceptible flocks in the same geographic area, including those in
animals to the source of infection increases, and as the density different poultry sectors ( 8 8 , 8 9 ) . Molecular typing of
of the animal population in the area increases. A recent study M. gallisepticum isolates has provided conclusive evidence for
conclusively demonstrated the presence of airborne presence of M. gallisepticum vaccine strains in non-vaccinated
M. hyopneumoniae on farms where acute respiratory disease flocks (90) (S.H. Kleven, unpublished findings).
was present (134). The organism was detected with a sensitive
PCR test of air samples in the pig houses where disease
problems were common.
Public health and international
Delineation of the lateral spread of M. gallisepticum from an trade implications
introduced infected bird within small populations of chickens No public health or zoonotic issues are associated with
was studied by following appearance of M. gallisepticum M. gallisepticum.
serological reactions in contact birds (101). A multi-phased
pattern of transmission was found, with variation in reaction Since M. gallisepticum is transmitted by embryonated (fertile)
among birds. Increasing the population density increased the eggs, buyers or importers of day-old breeder replacement
rate of spread. Using M. gallisepticum-PCR or isolation to chicks or hatching eggs should be provided assurance that the
detect infection, the spread of M. gallisepticum from actively flocks from which the eggs originated are free of infection in
infected turkey poults to susceptible pen mates can be order to prevent dissemination of the disease. Sellers should
detected within four to seven days, although serological provide copies of negative serological test results from serum
responses were not detected until approximately two weeks drawn one month or less before the eggs were produced. In
after the birds were placed in contact (S. Levisohn, the case of international exports, such test certificates should
unpublished findings). Live M. gallisepticum vaccines are be provided by an official agency of the exporting
transmitted more slowly among birds in the same pen, and government, such as the NPIP in the USA. For further
between pens, although the biological basis for this difference confidence in test results, results of PCR tests taken one week
is not known (65, 9 3 ) . or less before the hatching eggs were produced may be
provided. Fresh or frozen poultry meat products or infertile
eggs produced for human consumption are not ordinarily
Many M. gallisepticum outbreaks in breeder flocks maintained
considered risks for M. gallisepticum infection.
under conditions of biosecurity appear to be attributable to
human factors. Some of the possible means by which this may
occur in the specific cases can be identified by careful analysis
of traffic patterns between flocks and behaviour patterns of Prevention and control
the poultry farm personnel. Investigations into the survival In most countries, control programmes for M. gallisepticum
time of M. gallisepticum on materials found in the poultry are based on maintenance of freedom of infection in
house environment demonstrate the resilience of this commercial breeding stock. Voluntary M. gallisepticum
organism. Mycoplasma gallisepticum could be reisolated from control programmes in the USA are administered by the
several inert materials after two days, and survived on human United States Department of Agriculture under the NPIP;
hair for three days (23). Early studies demonstrated the ability testing provisions and protocols are provided in an official
of M. gallisepticum to survive for as long as nine days in water publication (3). Mycoplasma gallisepticum is included in the
containing biological material ( 1 0 % mycoplasma broth) European Communities Council Directive (90/539/EEC)
(117). which stipulates conditions concerning trade within the
Rev. sci. tech. Off. int. Epiz., 19 (2) 433

European Community and imports from third countries of In cases where control of M. gallisepticum infection is not
poultry and hatching eggs ( 3 1 ) . feasible, vaccination with either inactivated or live vaccines
may be an option. Vaccination as a control measure was first
The basic concepts of the NPIP are as follows: suggested by Adler (1) and was put into practice in the 1960s
by Luginbuhl et al. ( 9 8 ) and by Fabricant (34). Vaccination is
a) a high level of biosecurity of breeder flocks, with
most commonly used in commercial egg pullets to be placed
production in single-aged, all-in/all-out farms
on multi-age commercial egg production sites, and in some
b) routine monitoring by serological testing backed up by instances, in broiler breeder pullets. The subject of
rapid and specific confirmatory tests M. gallisepticum vaccination has recently been reviewed by
c) immediate slaughter of infected breeding flocks to prevent Whithear (139). With any M. gallisepticum vaccine,
transmission of mycoplasma to the progeny. vaccination before exposure to wild-type infection is essential.

In general, purchasers of fertile hatching eggs or day-old Inactivated bacterins have proved to be efficacious in
poults or chicks require certification that the source breeding prevention of respiratory signs and lesions in chickens ( 5 1 ,
flock is free of M. gallisepticum infection. 124, 148, 149), and have been demonstrated to be beneficial
in reducing egg production losses and egg transmission (44,
However, poultry production has increased dramatically in 4 5 , 5 1 ) . However, other studies have shown minimal or no
many parts of the world, resulting in the construction of large effectiveness. Bacterins have had a negligible effect in reducing
multi-age production complexes, usually for commercial egg M. gallisepticum populations in the upper respiratory tract
production, but occasionally also for breeding stock or even (66) and are generally felt to be of minimal value in long-term
broilers. In addition, large populations of poultry are often control of infection on multi-age production sites. Since they
reared in relatively small geographic areas, sometimes with are inactivated, bacterins are considered safer than live
mixed avian species or mixed types of commercial poultry M. gallisepticum vaccines; however, bacterins have the
maintained in close proximity. In such instances, disadvantages of high cost and the need to individually
maintenance of freedom of infection in all flocks may be vaccinate each bird. Two doses are considered more effective
difficult or impossible. Therefore, suitable antibiotic than a single dose, but this is seldom practised because of the
medication may be used to alleviate clinical signs and reduce expense.
production losses or egg transmission. In some situations
vaccination may be an option. Three strains of live M. gallisepticum vaccine are available in
different countries of the world, namely: F strain, 6/85 and
Mycoplasma gallisepticum is known to be susceptible to ts-11, although not all countries officially permit the use of
several antibiotics, including macrolides, tetracyclines, live vaccines.
fluoroquinolones and others ( 1 3 , 8 2 , 9 5 ) , but is resistant to
penicillins or other antibiotics which act by inhibiting cell wall The origin of the F strain is somewhat unclear, but the strain
biosynthesis. Mycoplasma gallisepticum may develop was probably isolated by Yamamoto and Adler (146) and later
resistance against commonly used antibiotics (154). utilised as a vaccine (34, 9 8 ) . The F strain is considered to be
Antibiotic medication has been used in the treatment of an M. gallisepticum strain of low to moderate virulence and
respiratory disease (46, 5 5 , 5 6 , 1 3 5 ) , to prevent egg transmissibility (65), but is capable of inducing respiratory
production losses ( 1 1 , 1 1 6 ) , and to reduce egg transmission signs in broilers ( 1 2 7 ) and is considered too virulent for use as
(115, 151). Although suitable antibiotic medication may a vaccine in broilers (126) or turkeys (97). The strain persists
reduce the severity of clinical signs and lesions, it may also in the upper respiratory tract for the life of the flock (65).
significantly reduce populations of M. gallisepticum in the Mycoplasma gallisepticum isolates from turkeys with clinical
respiratory tract (26, 5 4 ) . However, medication should not be respiratory disease have been determined to be F strain in
relied upon to eliminate M. gallisepticum infection from an origin (90). Vaccination with the F strain has been shown to
infected flock and should be regarded as a method for be effective in preventing egg production losses in commercial
short-term amelioration of signs and economic effects, rather layers ( 1 9 , 2 7 , 105). An important characteristic of F strain is
than as a long-term solution to the problem. the ability to induce resistance against infection by wild-type
or challenge infections. In laboratory studies, the F strain
In instances where the use of infected breeder flocks for egg significantly reduced populations of a challenge strain in the
production is necessary, egg dipping in an antibiotic solution upper respiratory tract (25, 8 5 ) , effectively displaced infection
may be used to reduce egg transmission (50, 112). Methods of with a challenge strain in pen trial studies (72), and displaced
egg dipping and antibiotic injection to control M. meleagridis wild-type M. gallisepticum in a multi-age commercial egg
egg transmission in turkey eggs (42) may be suitable for production site (71). The F strain vaccine can be administered
elimination of M. gallisepticum egg transmission. Egg heating, by several routes including intraocular and intranasal, and by
an alternative method of reducing egg transmission, has also coarse spray. Vaccine is generally administered at eight to
been practised (147). fourteen weeks of age, but can be administered as early as two
434 Rev. sci. tech. Off. int. Epiz., 19

weeks or less if chicks are at risk of exposure to wild-type the vaccine strain on multi-age production sites. This
infection before eight weeks. characteristic could be used as a tool for
M. gallisepticum eradication from such sites. The F strain
Development and characterisation of the 6/85 strain of clearly has a strong ability to displace wild-type
M. gallisepticum live vaccine were described by Evans (32, M. gallisepticum strains and has been used to displace a field
33). The 6/85 strain is a modified M. gallisepticum strain strain of M. gallisepticum on a multi-age commercial egg
originating in the USA which is avirulent for chickens and production site (71). Unfortunately, when vaccination on this
turkeys, is not easily transmitted from bird to bird, and farm was discontinued, F strain continued to cycle from flock
induces resistance against challenge with virulent to flock; eradication of M. gallisepticum was therefore not
M. gallisepticum. The vaccine has been approved for achieved. In a pen trial study, F strain demonstrated the
marketing in more than ten countries, in addition to the USA. ability to displace a virulent challenge strain when vaccinated
The 6/85 vaccine is administered by spray, does not induce an birds were placed with chickens which had been previously
antibody response, and can be detected in the upper challenged with R strain. However, neither ts-11 nor 6/85
respiratory tract for four to eight weeks after vaccination (32, exhibited an ability to displace the virulent challenge strain
93). The vaccine has been used primarily for prevention of under the conditions of this study. In contrast, when ts-11
egg production losses in commercial layers in the USA. was used to vaccinate replacement pullets on a site previously
populated with F strain, displacement of F with the ts-11
The development and characteristics of the ts-11 strain occurred in all vaccinated flocks. When vaccination was
M. gallisepticum vaccine strain have been described by discontinued, M. gallisepticum could no longer be detected on
Whithear et al. ( 1 4 0 , 1 4 1 , 142). The strain was developed by the farm (138). Although displacement has not been
chemical mutagenesis of an M. gallisepticum isolate from extensively studied under field conditions, field data suggest
Australia and was selected as a temperature-sensitive mutant. that use of the 6/85 strain over several years on multi-age
The strain is avirulent for chickens and turkeys, has a low farms has resulted in production complexes that are
propensity to spread from bird to bird, eficits a slow serologically negative for M. gallisepticum, suggesting that
development of circulating antibody, and induces protection displacement has occurred (K. Honneger, personal
to M. gallisepticum challenge. The ts-11 strain persists for the communication). Field experience and unpublished data
life of the flock in the upper respiratory tract and induces suggest that when field challenge occurs with highly virulent
long-lived immunity (139). Growing pullets are vaccinated by M. gallisepticum strains, flocks may become infected with
intraocular administration to each bird. The commercial wild-type M. gallisepticum (N. Ikuta and S.H. Kleven,
vaccine based on the ts-11 strain is registered in Australia (the unpublished findings). In such cases, the use of F strain may
country of origin) and the USA, with registration approved or
be necessary on the site for at least one production cycle, after
pending in approximately twenty other countries world-wide.
which the milder 6/85 or ts-11 strains could be employed.

Both the ts-11 and 6/85 strains are characterised by lack of

virulence and very poor ability to spread from bird to bird. Vaccination of turkeys has not been shown to be feasible.
Various types of chickens and turkeys kept in close contact However, restricted use of the 6/85 strain in commercial
across a wire barrier with vaccinated chickens for several turkeys has produced positive assessments (R.P. Chin,
months did not become infected with either strain, although K. Honneger and S.H. Kleven, unpublished findings),
evidence of very slight spread was found in chickens kept in although no reports have been published on this use of the
direct contact with ts-11 vaccinated birds (93). Because of vaccine. No reports exist of successful use of
these superior safety characteristics of avirulence and low M. gallisepticum vaccines in broilers.
potential for unintended spread to nearby unvaccinated
flocks, both 6/85 and ts-11 are considered to be preferable to Although vaccination is considered an alternative means of
F strain. Techniques for identification of specific control of M. gallisepticum, especially in commercial layers,
M. gallisepticum strains from field outbreaks have the use of stock which is free from M. gallisepticum is
demonstrated that both of these strains present significantly
preferable whenever possible.
less risk of infecting nearby poultry flocks than F strain.
Therefore, unless the wild-type is highly virulent, these strains
are the preferable choices when M. gallisepticum vaccination is
necessary.

An important characteristic of M. gallisepticum vaccines is the

ability to induce resistance to infection from wild-type
challenge, resulting in 'displacement' of wild-type strains with
Rev. sci. tech. Off. int. Epiz., 19 (2) 435

Mycoplasmose aviaire [Mycoplasma gallisepticum)

S. Levisohn & S.H. Kleven

Résumé
Mycoplasma gallisepticum est un mycoplasme aviaire de distribution mondiale.
C'est celui dont le pouvoir pathogène a les c o n s é q u e n c e s économiques les plus
graves. À l'instar des autres m y c o p l a s m e s , M. gallisepticum est d'une taille
minuscule, il est doté d'une information génétique minime et il est t o t a l e m e n t
dépourvu de paroi bactérienne. C'est ce qui explique l'interdépendance étroite
entre M. gallisepticum et l'animal hôte, ainsi que les difficultés d'isoler
l'organisme in vitro. Les souches de M. gallisepticum offrent des différences très
nettes au regard de propriétés biologiques importantes telles que le pouvoir
pathogène, l'infectiosité, le tropisme tissulaire et la transmissibilité. De plus, la
variation phénotypique des principaux antigènes de surface est très fréquente ;
c'est probablement l'a raison pour laquelle M. gallisepticum induit une infection
chronique en dépit de l'existence d'une forte réponse immunitaire.
L'infection due à M. gallisepticum se manifeste par une grande variété de signes
cliniques, mais même lorsqu'elle est inapparente, l'impact économique peut être
important. La forme clinique la plus grave est une maladie respiratoire chronique
chez les volailles destinées à la consommation ; M. gallisepticum est l'un des
agents responsables de ce syndrome multifactoriel. La transmission in ovo de
M. gallisepticum entre les sujets reproducteurs contaminés et leur progéniture
est la principale voie de propagation de l'infection et un sujet de préoccupation
majeur pour les échanges internationaux.
Dans la plupart des pays, les programmes de lutte contre
M. gallisepticum consistent à préserver de l'infection les élevages de sujets
reproducteurs. Dans les cas où il est impossible de mettre en œuvre une
prophylaxie sanitaire, on a recours à la v a c c i n a t i o n , notamment à l'aide de
nouveaux v a c c i n s à mycoplasmes vivants.
Des progrès majeurs ont été accomplis ces dernières années en matière de
diagnostic. La prophylaxie se fonde sur des programmes de dépistage
sérologique utilisant l'épreuve d'agglutination sur lame ou des épreuves
immuno-enzymatiques. Les réactions positives à l'épreuve d'agglutination sur
lame doivent être confirmées par d'autres épreuves sérologiques et/ou par des
tests démontrant la présence du mycoplasme. En principe, on peut soit isoler
M. gallisepticum, soit identifier son acide désoxyribonucléique à l'aide de
méthodes moléculaires. L'amplification en chaîne par polymérase est une
méthode alternative, à la fois plus rapide et plus sensible que les méthodes
traditionnelles de mise en culture qui font appel à des t e c h n i q u e s spécialisées et
longues. Les méthodes r é c e m m e n t développées de typage moléculaire offrent de
nouvelles perspectives pour les études épidémiologiques et l'identification des
réservoirs d'infection.

Mots-clés
Biosécurité - Maladie respiratoire chronique - Maladies aviaires - Mycoplasma
gallisepticum - Transmission in ovo - Typage moléculaire - Vaccination.
436 Rev. sci. tech. Off. int. Epiz., 1912)

Micoplasmosis aviar (Mycoplasma gallisepticum)

S. Levisohn & S.H. Kleven

Resumen
Mycoplasma gallisepticum, un patògeno presente en el mundo entero, es el
micoplasma que afecta con mayores repercusiones económicas a las aves de
corral. Al igual que otros micoplasmas, M. gallisepticum es un organismo
diminuto, carente por completo de pared celular bacteriana y dotado de un
volumen mínimo de información genética. En la práctica, estas propiedades se
traducen en un grado muy alto de interdependencia entre M. gallisepticum y sus
huéspedes y lo convierten en un organismo muy exigente para su cultivo in vitro.
Las diversas cepas de M. gallisepticum presentan sensibles diferencias en
cuanto a ciertas propiedades biológicas de importancia, como la patogenicidad,
la infectividad, la transmisibilidad o el tropismo por ciertos tejidos. Por otra parte,
los principales antígenos de superficie experimentan con gran frecuencia
cambios fenotípicos, lo que seguramente explica los casos de infección crónica
por M. gallisepticum incluso en presencia de una intensa respuesta inmunitaria.
La infección por M. gallisepticum puede expresarse clínicamente de formas muy
diversas, pero aun en ausencia de signos clínicos puede tener efectos
económicos significativos. La más grave manifestación clínica de la infección por
M. gallisepticum es la enfermedad respiratoria crónica en aves de engorde, que a
menudo se presenta asociada a otros agentes etiológicos, dando lugar a un
complejo patológico multifactorial. La transmisión de M. gallisepticum in ovo (de
reproductores infectados a su progenie) constituye el mecanismo básico de
propagación de la enfermedad y el principal factor que hay que tener en cuenta
de cara al comercio internacional.
En la mayoría de los países, los programas de lucha contra M. gallisepticum están
centrados en mantener libres de infección a las bandadas reproductoras de las
explotaciones industriales. Para los casos en que no sea factible controlar la
infección, se está evaluando la alternativa de las vacunaciones, utilizando sobre
todo una nueva generación de vacunas preparadas a partir de organismos vivos.
En los últimos años se han realizado notables progresos en lo que concierne a los
métodos de diagnóstico. Los programas de control han venido basándose en la
aplicación de técnicas serológicas en campañas de detección, sobre todo la
prueba de seroaglutinación en placa y el ensayo inmunoenzimático. Para
confirmar los posibles resultados positivos a la prueba de seroaglutinación es
preciso aplicar nuevas pruebas serológicas y/o demostrar la presencia del
microorganismo, lo que en principio es posible ya sea por aislamiento del
organismo o detectando su A D N por métodos moleculares. La amplificación en
cadena por la polimerasa representa una alternativa rápida y sensible a los
tradicionales métodos de cultivo, que requieren la aplicación de técnicas lentas y
laboriosas. La aparición de métodos moleculares de tipificación abre nuevas
oportunidades para realizar estudios epidemiológicos o descubrir reservorios de
la infección.
Palabras clave
Enfermedad respiratoria crónica - Enfermedades aviares - Mycoplasma gallisepticum -
Seguridad biológica - Tipificación molecular - Transmisión in ovo - Vacunación.
Rev. sci. tech. Off. int. Epiz., 19 (2) 437

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