I ncidence of Avian Mycoplasmosis in the
region of Batna, Eastern Algeria
Heleili, N.* Mamache, B. and Chelihi, A.
1. Laboratory of microbiology, veterinary department, Hadj Lakhdar University, Batna, Algéria
2. Veterinary surgeon, commune de sefiane 05064, Ngaoues, Batna-Algéria
* Corresponding author email : email@example.com
Veterinary World, 2011, Vol.4(3):101-105 RESEARCH
Avian mycoplasmosis is infectious and contagious disease which affects chicken and turkey as well as many other
species with many economics losses. The absence of data on avian mycoplasmosis in Algeria and the importance of the
poultry breeding in Batna encouraged us to undertake the prevalence of the most pathogenic mycoplasmas in broiler
and layer chickens in this area, Mycoplasma gallisepticum (MG). 143 Mycoplasmas were isolate from 237 samples, at
a rate of 60.33%. MG was isolate at a rate of 21.67% (2.09% in layer hens and 19.58% in broiler chickens). The
serological screening using of breedings showed a sensitivity of 83.10%. This study shows that mycoplasmosis and in
particular MG infection, represent a serious problem in chickens in Algeria in the absence of hygiene conditions and
Key w ords: Mycoplasma gallisepticum, Growth inhibition test, Seroprevalence, Broiler chickens, laying hens,
Mycoplasmosis, Avian Disease.
Avian mycoplasmosis is caused by several
pathogenic mycoplasmas of which Mycoplasma
gallisepticum (MG) and Mycoplasma synoviae (MS)
are the most important. MG causes chronic respiratory
disease of domestic poultry especially in the
management stresses and/or other respiratory
pathogens. Disease is characterized by coryza
conjunctivitis sneezing and sinusitis particularly in
turkeys. It can result in loss of production and
downgrading of meat-type birds and loss of egg
production. MS may cause respiratory disease
synovitis or may result in a silent infection. MG and
MS strains vary in infectivity and virulence and
infections may sometimes be unapparent (Bradbury,
2001; Ley, 2003; OIE, 2008).
The frequent occurrence of unspecified clinical
signs of respiratory disease among poultry flocks in
Batna region and the lack of data on the role of
mycoplasmas in these diseases had encouraged us to
undertake this investigation.
The aim of this study was to evaluate the
prevalence of MG infection in broilers and laying hens
in the region of Batna (Eastern Algeria) using the rapid
slide agglutination test and the isolation and
identification method. Among 237 analyzed samples
MG was isolate at a rate of 21.67% of which 2.09% in
laying hens and 19.58%in broilers.
Material and Methods
Our study was conducted between October 2008
and September 2010 in 23 flocks divided as follows:
laying hens (n = 11), broiler chickens (n = 13). These
farms were divided into 10 districts of the wilaya of
Batna. Birds collected during this investigation were
apparently healthy or with respiratory lesions of lung,
air sacs, or trachea. The number of poultry per farm
varied between 3000 and 5000. Broilers were reared in
the soil over a straw litters. Laying hens were caged. It
should be noted that the density standards, ventilation
and hygiene were exceptionally observed.
Sampling: 148 of blood samples were taken as
69 blood samples from laying hens
79 blood samples from broiler-chickens.
After coagulation during 2 hours at room
temperature, the serum is centrifuged at 1500 tr/min
for 15 min and inactivated at 56°C for 30 min. then the
serum is diluted to 1/5 to reduce non specific and
cross-reactions between MG and MS.
All serum are put in sterile aliquots and
maintained at 4°C until use within the following 48
Bacteriology: For the bacteriological diagnostic, 237
samples of tracheas, lungs and air sacs were taken
from recently dead birds or those dead frozen until use
(Evans et al., 2009).
www.veterinaryworld.org Veterinary World, Vol.4 No.3 March 2011 101
Incidence of Avian Mycoplasmosis in the region of Batna,Eastern Algeria
Culture medium of mycoplasmas: Liquid and solid
Frey`s medium was used for the isolation of
mycoplasmas (Frey et al., 1968). It consist of PPLO
broth base (2.1%) and PPLO agar (1%) (Difco
The media base was enriched with 15% horse
serum; 1%of glucose; and 10% yeast extract.
Contamination of media was prevented by adding 1%
thallium acetate and 0.5% penicillin G potassium.
Serology: The detection of antibodies against MG
was achieved by the rapid slide agglutination
method.MG antigen was kindly supplied by Dr
Bouchardon.AG, the laboratoire national de
pathologie aviaire (LNPA) of Ploufragan France. The
serum quality was checked using a negative SPF avian
serum, a positive serum against Mycoplasma
25µl of serum and 25µl of antigen are put on a
glass slide previously washed, rinsed and dried. The
antigen and serum are mixed at room temperature of
the reaction is carried out according to Stanley et al.,
Mycoplasma isolation: The reference strain of MG
used is MG ATTC 15302 produced in rabbit and kindly
supplied by the LNPA, France.
About 0.5 g of tissues (tracheas, lungs and air
sacs) are cut in small pieces and ground in a sand bath
containing 5 ml of mycoplasma medium.
0.2 ml of this suspension is inoculated in Frey`s
liquid medium. When a color change is observed in the
liquid medium, another inoculation of a fresh liquid
medium is perform.
Later on an inoculation of Frey`s solid medium
is carried out after a color changing had occurred in the
liquid medium. If no color changing has occurred in
the liquid medium a subculture is done on a solid
medium because it is well known that mycoplasmas
are very slow-growing microorganisms (Kleven,
2008). If Mycoplasma growth is noted on solid
medium, agar blocks containing colonies are
transferred in tubes of liquid medium for
mycoplasmas cloning (Stipkovits et al., 1975).
Positive cultures were characterized by Dienes
staining, biochemical and growth inhibition tests
using MG antiserum produced in rabbit kindly
supplied by Dr Bouchardon.A.G.
Serology: Among 148 tested sera, 123 were positive
representing a rate of infection of 83.10% showing the
strong spread of MG. However, the rate of infection
was slightly higher in broiler-chickens (84.81%) than
in laying hens (81.15%) (Table I).
The highest prevalence (100%) of MG infection
was found in the present study in El Madher, Ain
yagout and Boumia (TableII) Seasonal variation of
prevalence of MG infection was observed in the
present study. The prevalence was higher (91.13%) in
winter season and lower (73.91%) in summer season
According to the age, the highest prevalence of
MG infection was 86.95% in chicks whereas lowest
prevalence was 82.35% in adult (Table IV).
Bacteriolog y: 143 mycoplasmas strains were
isolated from 237 organ samples representing a
positivity rate of 60.33%.
www.veterinaryworld.org Veterinary World, Vol.4 No.3 March 2011 102
Table-1. Serological and bacteriological results of infection Mycoplasma gallisepticum
56 /69 (81.15%)
Table-2. Seroprevalence of Mycoplasma gallisepticum according to the study area
Study area Number of samplesNumber of positive seraRate of SPA %
Incidence of Avian Mycoplasmosis in the region of Batna,Eastern Algeria
31 strains were identified as MG (21.67%). The
MG distribution was as follows: 3 in laying hens and
28 in broiler-chickens representing 2.09% and 19.58%
respectively (Table I).
It should be stated that all flocks either laying
hens or broiler-chickens harbor MG.
For laying hens, in a study Nascimento et al.
(2005) found a rate of infection by MG of about 90%.
A similar study conducted in Bangladesh showed that
the rate of infection varies from 45.1% to 66.5%
(Hossain et al., 2010; Barua et al., 2006). These
results are higher than those in the present study.
This study has revealed a sharp influence of
season on the incidence of avian infection by MG
(Table III). Indeed, rate of infection of 90% has been
noted during winter, compared to that of 73.91%
obtained during summer. These results are in
accordance with those of Sikder et al. (2005), Sarkar et
al. (2005) and Hossain et al. (2010).
The effect of age on the occurrence of
mycoplasmal infection is revealed in the study with a
rate of infection of 69.9% and 48.7% in adult and
young birds respectively (Table II).
In the present study, MG has been isolated at a
rate of 21.67% (Table V). This rate is higher than to
those found by Helail, 1998 (11.89%).
The results of mycoplasma isolation from
different organs showed that air sacs are the main site
of multiplication of this microorganism from dead
birds (90%). Similar result was also obtained by
Helail, 1998 (36.29%) and Shaker, 2005 (92%). Lungs
are the second site of isolation (71.42%) followed by
MG isolation rate from tracheas (62.96%) is
higher than to those obtained by Paakpinyo and
Sasipreeyajan, 2007; Feberwee et al., 2005 and
Gharaibeh and Al Roussan, 2008 (44.66%, 33% and
Results obtained in our study revealed that the
isolation rate of MG from the lungs is higher than in
others studies (shaker, 2005; Helail, 1998).
The discordance observed between serology and
isolation may be attributed to different factors such as
the existence of tissular inhibitors of mycoplasma
growth (Boussetta et al., 1997), the use of dry swabs
reducing the viability of microorganisms (Zain et al.,
1995), the absence of localization of mycoplasmas
The serological test in this study, for instance the
rapid serum agglutination (RSA) may lack specificity
and/or sensitivity. Therefore, its use is strongly recom-
mended for monitoring flocks rather than for testing
individual birds. For this reason we had tried to
establish the test sensitivity and specificity under our
laboratory conditions by using fresh sera. The latter
had been decomplemented in order to ovoid cross
reactions between MG and MS and diluted to 1/5. The
evaluation of the RSA test was validated using known
positive and negative control sera. In our study the size
of tested samples is similar to that reported by several
others ranging between 10 and 30 birds (Boucetta et
al., 1997; Kermorgant, 1998 and Sabir, 2003).
This study has revealed a very high prevalence
of Mycoplasma gallisepticum infection either in
laying hens or in broiler-chickens in Batna region. Our
results are higher than those of similar studies.
For instance, Thai et al. (2009) and
Papanikolaou (2002) found a rate of infection of
37.83% and 21% respectively. However, our findings
are slightly in accordance with those of Wunderwald et
al. (2002), Sarkar et al. (2005) and Hossain et al.
(2007) with a rate of infection of 75%, 58.90% and
Concerning broiler-chickens, 84.81% of the
flocks are MG infected (Table I). This value is greatly
higher than results obtained by Baruta et al. (2001) and
Aimeur et al. (2010) (1.25%, 30%) However, our
results are in accordance with those obtained by
orajaka et al. (2002) and mainly those of Evans et al.
(2009) with a positivity rate of 64.9% and 100%
www.veterinaryworld.org Veterinary World, Vol.4 No.3 March 2011 103
Table-3. Seasonal seroprevalence of infection Mycoplasma gallisepticum
Season Tested seraPositive seraNegative seraRate of positive sera
Table-4. Seroprevalence of Mycoplasma gallisepticum in chickens according to age
AgeNumber of samplesPositive cases Negative cases
Incidence of Avian Mycoplasmosis in the region of Batna, Eastern Algeria
11. Frey M L, Hanson R P and Anderson D P. (1968): A medium
for the isolation of avian mycoplasmas. Am. J. Vet.Res. 29:
Ghoraibeh S and Al Roussan D. (2008): the use of molecular
techniques in isolation and characterization of Mycoplasma
gallisepticum from commercial chickens in Jordan. Int. J. Of
Gordon R.F (1979)Pathologies des volailles. Maloine S. A.
Helail S S S. (1998): Evaluation of mycoplasma antigens
used for the detection of mycoplasma infection in chicken.
Thesis presented for master degree, Cairo Univ. Egypt, pp.45.
Hossain K M M, Ali M Y and Haque M I. (2007):
Seroprevalence of mycoplasma gallisepticum infection in
chicken in the greater rajshahi district of bangladesh. Bangl.
J. Vet. Med. 5: 09–14.
Hossain K M M, Hossain Md T, Yamato I. (2010):
Seroprevalence of Salmonella and Mycoplasma
gallisepticum Infection in Chickens in Rajshahi and
Surrounding Districts of Bangladesh. Int. J.Biol. 2, 74-80.
Hudson P, et.al.(2006): Identification of avirulenc associated
determinant, dihydrolipoamide dehydrogenase (lpd), in
Mycoplasma gallisepticum through in vivo screening of
transposon mutants. Infect. Immun. 74: 931-939.
Kempf I, et.al.(1998): Mycoplasmose à Mycoplasma
gallisepticum: réalisation d'un modèle expérimental rôle de
l'ammoniac comme facteur d'exacerbation. Avian Pathol. 17:
Kermorgant P. (1999): Les mycoplasmoses aviaires:
enquête sérologique réalisée en Bretagne en 1998. Thèse de
docteur vétérinaire Faculté de Médecine Vétérinaire Nantes,
Kleven S H. (2008): Control of Avian Mycoplasma
Infections in Commercial Poultry. Avian Dis. 52: 367–374.
Ley D H. (2003): mycoplasma gallisepticum infection. In:
diseases of poultry. Saif Y.M., Barnes.H.J., Glisson J.R.,
Fadly A.M., Mc Dacgald L.R. and Swane D.E. eds Iowa
State University Press, Ames, Iowa, USA 41-52.
Nascimento E R, et.al.(2005): Eradication of Mycoplasma
gallisepticum and M. synoviae from a chicken flock by
antimicrobial injections in eggs and chicks. Acta.Sci.Vet. 33:
Nunoya TT, et.al.(1995): Occurrence of keratoconjun-
ctivitis apparently caused by Mycoplasma gallisepticum in
layer chickens. Vet. Pathol. 32: 11-18.
Orajaka L J E, Okoye J O A, Oboegbulem S I. (2002):
Seroepidemiologic survey of mycoplasmal infections in
native and exotic chickens in Nsukka District of South
Eastern Nigeria. J.Sustainable.Agric.Environ. 4: 77-82.
Pakpinyo S and Sasipreeyajan J. (2007): Molecular
characterization and determination of antimicrobial resistance of
Mycobacterium gallisepticum isolated from chickens.
Veterinary Microbiol. 125: 59–65.
Papanikolaou J N. (2002): Epidemiological investigation of
Mycoplasmas, M. gallisepticum, M. synoviae and M.
meleagridis in poultry flocks in Greece. Journal of the
hellenic veterinary medical society, 53: 297-303.
especially MG in some sites after initial infection
(Takase et al., 2000; Kempf et al., 1998), the difficulty
of isolating mycoplasmas after some antibiotics
treatment (Aimeur et al., 2010).
The authors are thankful to the Laboratory of
microbiology, Institute for Animal Science and
Agricultural Sciences, University Batna, Algeria and
Dr Bouchardon.A.G from the laboratoire national de
pathologie aviaire (LNPA) of Ploufragan, France for
providing some facilities to conduct the present
1.Aimeur R, Bouaziz O., Kabouia R, Bererhi H. (2010):
Prévalence de Mycoplasma gallisepticum et de Mycoplasma
synoviae dans les élevages avicoles de l'Est Algérien. Rev.
Med. Vet. 161(3): 141-145.
Amin MM and Jordan FTW. (1979): Infection of the chicken
with a virulent or avirulent strain of Mycojplasma
gallisepticum alone and together with Newcastle disease
virus of E. coli or both. Vet Microbiol. 4: 35-45.
Amin MM, Sidique MAB and Rahman MM. (1992):
Investigation on chronic respiratory disease in chickens:
part-II. BUA Res. Progress. 6: 519-526
Barua S R., Prodhan A M, Islam S and Chowdhury S. (2006):
Study on Mycoplasma Gallisepticum in Chickens in Selected
Areas of Bangladesh. Bangl. J. Vet. Med. 4: 141–142.
Baruta D A, Ardoino S M, Marengo M L. (2001): A survey in
infections caused by Mycoplasma synoviae and Mycoplasma
gallisepticum in broiler chickens slaughtered in General Pico
using the Slide Quick Agglutination technique. Ciencia.Vet.
Facultad de Ciencias Veterinarias. UNLPam.
Behbahan G G N, et.al.(2005): Isolation and detection of
Mycoplasma gallisepticum by polymerase chain reaction
and restriction fragment length polymorphism. Iranian J.Vet.
6 (2): 35-41.
Boussetta M, Chaouachi N, Mlik B. (1997): Etude
sérologique et bactériologique des mycoplasmoses aviaires
dans la région du Cap Bon en Tunisie. Rev.Elèv.Med.Vet..
Pays Trop. 50: 93-96.
Bradbury JM. (2001): Avian Mycoplasmosis. In: Poultry
Diseases.5th ed. Jordan.D. and Raragher T. EDS. W.b.
Sanders? London, UK, 178-193.
Evans J D, Thornton D L, Branton SL. (2009): Diagnosis of
Mycoplasma gallisepticum from a broiler breeder flock:
comparison of three diagnostics methods. International
Journal of Poult Sci. 8: 104-107.
Feberwee A., et.al.(2005): Comparison of Culture, PCR, and
Different Serologic Tests for Detection of Mycoplasma
gallisepticum and Mycoplasma synoviae Infections. Avian
www.veterinaryworld.org Veterinary World, Vol.4 No.3 March 2011 104
Table-5. I solation of Mycoplasma gallisepticum from different type of sampled
Organsamples samplesPositive digitoninRate of mycoplasmas Rate of MG
Incidence of Avian Mycoplasmosis in the region of Batna,Eastern Algeria
27. Poveda J B, et.al.(1990): An Epizootiological Study of
Avian Mycoplasmas in Southern Spain. Avian Pathol. 19:
Rozina A. 2000: studies on the development of a vaccine
using indigenous strains of Mycoplasma gallisepticum.
Thesis of doctor of philosophy degree. Department of
microbiology, University of Karashi, Pakistan.
Sabir J. (2003): Enquête sérologique sur Mycoplasma
gallisepticum et Mycoplasma synoviae dans les élevages de
poules pondeuses. Thèse de Docteur Vétérinaire, IAV
Hassan II, Maroc, (2003), 110 pages.
Sarkar S K, et.al.(2005): Sero-Prevalence of Mycoplasma
gallisepticum Infection of Chickens in Model Breeder Poultry
Farms of Bangladesh. Int.J. Poult. Sci. 4: 32-35.
Sikder A J, et.al. (2005): Seroprevalence of Salmonella and
Mycoplasma gallisepticum Infection in the Six Model
Breeder Poultry Farms at Patuakhali District in Bangladesh.
Int.J. Poult. Sci. 4: 905-910.
Stanley W A, et.al.(2001): Monitoring Mycoplasm a gallisepticum
and Mycoplasma synoviae infection in breeder chickens after
treatment with enrofloxacin. Avian Dis. 145: 534-539.
33. Stipkovits L, El-ebeedy A A, Kisary J and Varga L.
(1975): Mycoplasma infection of geese I. incidence of
Mycoplasmas and Acholeplasmas in geese. Avian pathoL. 4:
Takase K, et.al.(2000): Serological monitoring on layer
farms with specific pathogen free chickens. J.Vet.Med.Sci.
Thái T H, Đ ?c N N, Giáp N V, Hýõng C T T. (2009): Sero -
Prevalence of Mycoplasma gallisepticum Infection in
Chicken Ross 308 and Colour ISA Raised in Some Provinces
in The North of Vietnam. T?p chí Khoa h?c và Phát tri?n,
T?p 7: 306 – 313.
Villate D, 2001. Maladies des volailles. Éditions France
agricole, 2 édition, pp399.
Wunderwald C and Hoop R K. (2002): Serological
monitoring of 40 Swiss fancy breed poultry flocks. Avian
Pathol. 31: 157–162.
Zain Z M, Bradbury J M. (1995): The influence of type of
swab and laboratory method on the recovery of Mycoplasma
gallisepticum and Mycoplasma synoviae in broth medium.
Avian pathol. 24: 707-716.
www.veterinaryworld.org Veterinary World, Vol.4 No.3 March 2011 105