Comparison of culture and PCR to detect Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in ear swabs taken from goats

This study was designed to evaluate the validity of PCR for the direct detection of Mycoplasma (M.) agalactiae and Mycoplasma mycoides subsp. capri (Mmc), as the two species most frequently causing contagious agalactia (CA) in goats. The PCR method was compared with the traditional culture technique to determine which method was most efficient at identifying all auricular carriers present in herds. The samples analyzed were 307 ear swabs taken from goats reared in a CA endemic area. We assessed the validity of each technique to detect each species and agreement between both methods. For each species, the result was taken as true-positive when at least one of the two tests was positive. Of the swabs tested, 246 were scored positive by PCR (235 and 11 for Mmc and M. agalactiae, respectively) and 117 showed a positive culture result (113 for Mmc and 4 for M. agalactiae). 133 of the PCR-positive samples (124 and 9 for Mmc and M. agalactiae, respectively) yielded negative culture results and 4 culture-positive samples tested negative using PCR (2 for each species). Sensitivity and negative predictive values for PCR were 84.62 and 99.32 (for M. agalactiae) and 99.16 and 97.22% (for Mmc) respectively, and for culture were 30.77 and 97.03 (for M. agalactiae) and 47.08 and 36.08% (for Mmc), respectively. PCR proved to be a rapid and sensitive method for the detection of mycoplasmas in the external ear of asymptomatic carriers. Tools such as this are needed to adopt efficient control measures against CA.     

Analysis of the immunoproteome of Mycoplasma mycoides subsp. mycoides small colony type reveals immunogenic homologues to other known virulence traits in related Mycoplasma species

Contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) has been eradicated in the developed world, but it is still present in many countries of sub-Saharan Africa. After initially successful control measures in the 1960s it has been spreading due to a lack of money, fragmentation of veterinary services, uncontrolled cattle movement, insufficient vaccine efficacy and sensitivity of current diagnostic tests.In this study we used two-dimensional polyacrylamide gel electrophoresis followed by immunoblot with sera from MmmSC-infected animals and MALDI-ToF mass spectrometry to identify novel immunogenic proteins as candidate molecules for improved diagnostics and vaccines. We identified 24 immunogens recognized by pooled sera from experimentally infected cattle. Furthermore, a serum from an animal with acute clinical disease as well as severe pathomorphological lesions recognized 13 additional immunogens indicating variation in the antibody responses to CBPP amongst cattle. Most immunogens showed compelling similarity to protein/gene sequences in the two ruminant pathogens M. capricolum subsp. capricolum and M. mycoides subsp. mycoides large colony type both belonging to the mycoides cluster. Three of these proteins, namely glycerol-3-phosphate oxidase, adenylosuccinate synthase, and glyceraldehyde-3-phosphate dehydrogenase, had no compelling homologue in the other distantly related bovine pathogen M. agalactiae. In addition, translation elongation factor Tu, heat shock protein 70, pyruvate dehydrogenase, and FKBP-type peptidyl-prolyl isomerase, which have been found to mediate adhesion to host tissue in other mycoplasmas were shown to be expressed and recognized by sera. These proteins have potential for the development of improved diagnostic tests and possibly vaccines.     

Mycoplasma detection and isolation from one-humped camels (Camelus dromedarius)

In scientific literature, a small amount of information is found concerning mycoplasmosis in camel species. Mycoplasma (M.) arginini, Acholeplasma (A.) laidlawii, and Acholeplasma oculi have been reported to be isolated from these host species. Serologically positive results have been reported for Mycoplasma mycoides subsp. mycoides SC type, Mycoplasma capricolum subsp. capripneumoniae, and M. mycoides subsp. capri. The aims of this study were to detect, isolate, and identify mycoplasmas from camels (Camelus dromedarius). Initially, saliva and ear smears plus conjunctival and vaginal secretions were taken from five female animals, but only conjunctival secretions in three male animals, all belonging to the same farm. An unknown mycoplasma was isolated from one of the vagina samples. Additionally, another unknown and uncultured mycoplasma was detected with molecular biology in the same sample. In the second stage, 23 vaginal secretions were taken from the same farm plus another secretion from a different one. Ten isolates of the same unknown and previously isolated mycoplasma were detected, nine of them recovered from the vagina of female camels. Some mycoplasmas have been related to reproductive disorders; however, there is no evidence that the isolated mycoplasmas are related to such disorders.     

Kinetics of substrate oxidation and hydrogen peroxide production by Mycoplasma mycoides subsp. mycoides Large Colony (LC) type and Mycoplasma mycoides subsp. capri

Mycoplasma mycoides subsp. mycoides Large Colony (LC) type is a pathogen of goats causing contagious agalactia and respiratory disease, found on all continents where small ruminants are kept. It shares close genetic characteristics with M. mycoides subsp. capri. Substrate oxidation by 22 strains of M. mycoides subsp. mycoides LC from nine countries was compared with that of eight strains of M. mycoides subsp. capri from five countries. There was considerable similarity in the substrates used, but substrate saturation coefficients (K_s) varied for different substrates. Substrate utilization patterns and K_s values did not (1) significantly differentiate the LC strains from each other, (2) show any correlation with geographical origin, or (3) distinguish the LC strains from the capri strains. These results support previous studies justifying the reclassification of these subspecies as a single species.     

Detection of Mycoplasma mycoides sub-species mycoides small colony by a specific capture/enrichment monoclonal antibody-based sandwich ELISA

The present study describes the development of a specific Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC) monoclonal antibody (MAb), 6E3, and its application in a sandwich ELISA (sELISA) format. Mab 6E3 reacted only to the 12 MmmSC within the 32 M. mycoides cluster strains and 12 representative strains of other bovine, ovine and caprine associated mycoplasmas examined. A capture/enrichment format of the sELISA that combined MAb 6E3 with a previously developed MAb 3H12 that cross reacted with Mmm Large Colony [Rodriguez, F., Ball, H.J., Finlay, D., Campbell, D., Mackie, D.P., 1996. Detection of Mycoplasma mycoides sub-species mycoides by monoclonal antibody-based sandwich ELISA. Veterinary Microbiology 51, 69–76], retained MmmSC specificity and improved the sensitivity from the 1.2 × 10⁷ cfu/ml for a standard 2 h capture stage sELISA down to as low as 2 cfu/ml for a 72 h capture. A low level of false positives (1%) was observed when this assay was applied to 200 bovine respiratory and milk samples submitted for diagnostic investigation. This simple and specific sELISA provides a suitable assay for screening large numbers of samples for CBPP.     

Changes in Lymphocyte Subsets in the Bronchus‐associated Lymphoid Tissue of Goats Naturally Infected with Different Mycoplasma Species

The distribution of cells containing lysozyme, S‐100 protein, CD3, CD4, CD8, major histocompatibility complex class II antigen and immunoglobulin G (IgG) was analysed in the bronchus‐associated lymphoid tissue (BALT) of goats naturally infected with three Mycoplasma species. This study included the immunohistochemical characterization of the pneumonic lesions of 18 goats (3–5 months old) infected with one of the following Mycoplasma species: M. mycoides ssp. mycoides, Large Colony type (goats no. 1–6), M. mycoides ssp. capri (goats no. 7–12) and M. capricolum ssp. capricolum (goats no. 13–18). Microscopically, infected animals showed a moderate bronchointerstitial pneumonia, characterized by lymphoid hyperplasia of the BALT and infiltration of mononuclear cells in the alveolar walls and airways. The main cellular type in the BALT was represented by CD3⁺ T lymphocytes, and the ratio of CD4⁺:CD8⁺ cells was >2. The BALT showed large germinal centres mainly composed of IgG⁺ B lymphocytes, with numerous S‐100⁺ follicular dendritic cells. The presence of follicular dendritic cells confirmed the high degree of organization of this lymphoid tissue. The immunohistochemical results showed that activated T lymphocytes, particularly in the CD4 subset, and IgG⁺ B cells, play a major role in the immune response of the caprine lung infected with these species of mycoplasmas.     

Population sizes and frequency of Malassezia pachydermatis at skin and mucosal sites on healthy dogs

Skin and mucosal carriage of Malassezia pachydermatis was studied in 20 healthy pet dogs of various breeds and in 20 kennelled beagles. Using swabs, anal carriage was detected in 10 pet dogs and 11 beagles and the nose, mouth, prepuce and vulva were shown to be infrequently colonised. M pachydermatis was isolated from the external ear canal of 11 beagles and two pet dogs; both the population sizes and frequency of isolation were significantly (P<0·05) greater in the beagles. The yeast was infrequently isolated from the axilla and groin in low numbers using contact plates and detergent scrub samples but was often cultured from the lower lip and the dorsal interdigital spaces; isolation frequencies and population sizes in the two groups of dogs were not significantly different. These results demonstrate that the anus, external ear canal and lip and interdigital skin of healthy dogs are frequently colonised by M pachydermatis.     

Seroprevalence of and risk factors for Mycoplasma mycoides subspecies capri infection in small ruminants in Northern Jordan

During the period from January 2002 to December 2003, serum samples were collected from 104 small ruminant flocks consisting of 18 sheep flocks, 27 goat flocks, and 59 mixed flocks containing both sheep and goats in northern Jordan. Only female sheep and goats were sampled. At least five females aged over 2 years per flock per species were sampled and examined for Mycoplasma mycoides subspecies capri using the latex agglutination test. To increase the chances of detecting positive flocks, sick or older ewes were sampled. Specific information was obtained using a questionnaire to identify potential risk factors for M. mycoides subsp. capri seropositivity in small ruminants. The true flock-level seroprevalences of M. mycoides subsp. capri were 34%, 32%, and 38% in small ruminants (sheep and goats), sheep, and goats, respectively. Differences between flock-level seroprevalences in sheep and goats were not significant (p = 0.7). Multivariable logistic regression analysis of 21 production and health management practices showed four to be associated with M. mycoides subsp. capri seropositivity including flocks which were grazed and fed concentrate supplement (OR = 4.6), improper cleaning of milking utensils (OR = 4.7), buying new animals to replace culled ones (OR = 0.3), and treating against helminths when clinical signs of helminth infections appear (OR = 0.4).     

Kinetics of substrate oxidation and hydrogen peroxide production by Mycoplasma mycoides subsp. mycoides Large Colony (LC) type and Mycoplasma mycoides subsp. capri

Mycoplasma mycoides subsp. mycoides Large Colony (LC) type is a pathogen of goats causing contagious agalactia and respiratory disease, found on all continents where small ruminants are kept. It shares close genetic characteristics with M. mycoides subsp. capri. Substrate oxidation by 22 strains of M. mycoides subsp. mycoides LC from nine countries was compared with that of eight strains of M. mycoides subsp. capri from five countries. There was considerable similarity in the substrates used, but substrate saturation coefficients (K_s) varied for different substrates. Substrate utilization patterns and K_s values did not (1) significantly differentiate the LC strains from each other, (2) show any correlation with geographical origin, or (3) distinguish the LC strains from the capri strains. These results support previous studies justifying the reclassification of these subspecies as a single species.     

Characterisation of protein and antigen variability among Mycoplasma mycoides subsp. mycoides (LC) and Mycoplasma agalactiae field strains by SDS-PAGE and immunoblotting

Mycoplasma mycoides subsp. mycoides (LC) (Mmm LC) and Mycoplasma agalactiae are the most important mycoplasma species involved in the contagious agalactia syndrome. A total of 25 field strains from Spain and the two type strains were analysed by SDS-PAGE and immunoblotting. Two polyclonal antisera (PAbs) raised against a pool of strains of each mycoplasma species were used. The results revealed a high degree of protein variability among the field strains. The type strain of Mmm LC appeared to be representative of the field strains of this species, whereas this was not the case with the M. agalactiae type strain. Whereas M. agalactiae is known to possess a gene family regulating surface antigen diversity, there is a need to study the mechanisms used byMmm LC to generate antigenic variability in more detail.     

In vitro antimycoplasmal activity of six Jordanian medicinal plants against three Mycoplasma species

The in vitro effect of six Jordanian traditional medicine plant methanolic extracts were tested against 32 isolates of Mycoplasma species; Mycoplasma mycoides subsp. mycoides LC (6), Mycoplasma capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions in Jordan. All Mycoplasma species showed susceptibility to Artemisia herba-alba and Artemisia arborescens with MIC ranges from 3.125–12.5 mg/ml. Allium sativum and Punica grantum showed limited activity against some Mycoplasma isolates. Olea europea and Citrullus colocynthis showed no in vitro activity against any of the Mycoplasma species tested. Artemisia herba-alba and Artemisia arborescens may therefore be useful for the treatment of mycoplasma infections. The authors are British Civil Servants and as such their work is subject to British Crown Copyright. This means the exclusive copyright for the article cannot be transferred.     

Mycoplasma bovis and otitis in dairy calves in the United Kingdom

Signs of severe otitis media in 20% of dairy calves on one farm were associated with Mycoplasma bovis infection, based on isolation from the external ear canal and nares. Affected calves seroconverted to M. bovis and no other significant bacteria were isolated. Infection was considered likely to have originated from cows in the milking herd based on evidence of seroconversion and detection of infection in a milk sample. M. bovis infection should be considered when investigating otitis problems in calves.     

Recovery of Mycoplasma agalactiae from the ears of goats experimentally infected by the intramammary route

The role of inapparent carriers of Mycoplasma agalactiae and the strategies used to colonise the external ear canal in goats remain unclear. This study examined the ability of M. agalactiae to colonise the ears of goats infected experimentally by the intramammary route. The right mammary glands of 15 lactating goats were inoculated with 10(10) colony forming units (cfu) of M. agalactiae. The goats were randomly assigned to three groups of five animals each and sampled at slaughter at 5, 15 or 45 days post-infection (dpi). A further four goats served as uninfected controls. Right and left ear swabs were collected for detection of M. agalactiae by culture before and after sacrifice. M. agalactiae was detected in 19/20 (95%) ear swabs from goats sampled at 15 and 45dpi, whereas all ear swabs collected before inoculation, ear swabs collected from the group sampled at 5dpi and ear swabs from control goats at the time of sacrifice were negative for M. agalactiae. Blood samples collected at 6, 12, 24, 48 and 72h post-infection for detection of M. agalactiae by culture were also negative. There were differences in the antigenic profiles of isolates recovered from the ears compared to the 7MAG strain used to inoculate the animals and most isolates from the mammary gland, milk and supramammary lymph nodes.     

Field trial of a combined vaccine against caprine contagious agalactia: humoral immune response in lactating goats

Two vaccines against Mycoplasma agalactiae and Mycoplasma mycoides subsp. mycoides (large colony type) were developed using inactivated strains selected in previous characterization studies. Formaldehyde and phenol were used as the inactivating agents for vaccines A and B, respectively. Aluminium hydroxide plus purified saponin (Quil-A) were added to both vaccines as adjuvant. The field trial was designed to evaluate the specific humoral immune response to the two mycoplasma species in lactating goats over a period of 7 months. The vaccines were tested on 120 goats randomly assigned to three groups of 40 animals each. Two groups received two injections of vaccine A or B respectively, and a third group remained in the herd as control. Antibody titres determined by ELISA indicated a significant difference between both vaccines and the control group over a 6-month period. Immunoblotting assays also revealed the production of antibodies against the two mycoplasma species. Further field trials are underway to evaluate the efficacy and protection conferred to the animals by these specific antibodies.     

Molecular Epidemiology of Contagious Bovine Pleuropneumonia in Tanzania Based on Amplified Fragment Length Polymorphism and Pulsed‐field Gel Electrophoresis Analysis

The genetic diversity of 60 field strains of Mycoplasma mycoides ssp. mycoides, small colony type (M. mycoides SC), comprising 56 isolates from cattle in Tanzania, one from Kenya, two from Botswana and one from Portugal, as well as the type (PG1^T) and vaccine (T₁‐SR49) strains, was investigated. The strains were analysed for variations in the EcoRI and Csp6I restriction sites in the genomic DNA using the amplified fragment length polymorphism (AFLP) technique, and variations in the BamHI restriction sites using pulsed‐field gel electrophoresis (PFGE). Six AFLP types were detected among the analysed strains. The AFLP profiles of the type and vaccine strains were indistinguishable from each other. Indistinguishable AFLP profiles were found for 55 Tanzanian field strains, one of them isolated in 1990 and the other 54 isolated in 1998/1999, although one strain isolated in 1999 showed a different profile. Strains from different countries revealed different AFLP profiles. Six PFGE types were detected among the analysed strains, with all the 56 Tanzanian field strains displaying indistinguishable PFGE profiles. Strains from different countries revealed different PFGE profiles, and so did the type and vaccine strains. The strong genomic homogeneity among M. mycoides SC strains associated with outbreaks of contagious bovine pleuropneumonia in different regions of Tanzania suggests that the outbreaks of the disease in the 1990–99 period might have been caused by a single epidemic clone. Moreover, this study has demonstrated that AFLP and PFGE are potential tools for molecular epidemiological studies of M. mycoides SC infections.     

Mycoplasmas isolated from the respiratory tract of cattle and goats in Tanzania

A microbiological study of the mycoplasma flora in the respiratory tracts of cattle and goats in selected regions of Tanzania is described. In the examination of cattle, mycoplasmas were isolated from 60 (17.8%) of the 338 examined lung samples, 8 (47.1%) of the 17 lymph nodes, 4 (13.3%) of the 30 pleural fluid samples and 4 (3.9%) of the 103 nasal swabs examined. All the isolates were identified as Mycoplasma mycoides subsp. mycoides, Small Colony type except for one isolate from pleural fluid which was identified as Mycoplasma arginini. M. mycoides subsp. mycoides, Small Colony type was isolated from samples originating from Dodoma, Iringa, Mbeya, Morogoro and Shinyanga regions where outbreaks of contagious bovine pleuropneumonia had been reported. In the examination of goats, mycoplasmas were isolated from 54 (34.0%) of the 159 examined lung samples, 41 (18.1%) of the 226 nasal swabs and 4 (40.0%) of the 10 pleural fluid samples. The species demonstrated were Mycoplasma capricolum subsp. capripneumoniae, M. mycoides subsp. mycoides, Small Colony type Mycoplasma ovipneumoniae and M. Capricolum subsp. arginini. The isolation of M. capripneumoniae in the Coast and Morogoro regions confirmed the presence of contagious caprine pleuropneumonia in the regions.     

Mycoplasmas isolated from goats and sheep with pleuropneumonia in Saudi Arabia

Summary Pleuropneumonia of goats and sheep has been known to cause serious economic losses in many countries including Saudi Arabia. A preliminary investigation was carried out on 163 goats and 130 sheep which showed clinical symptoms or post-mortem lesions of pleuropneumonia. Pathological samples from the respiratory tract of live or slaughtered goats and sheep (183 and 160 samples respectively) were cultured for mycoplasmas. Of 343 mycoplasma cultures examined 211 (61·5%) isolates were characterised biochemically. A total of 116 isolates were serologically identified asM. mycoides subsp.mycoides, M. ovipneumonia, M. conjunctivae, M. agalactiae subsp.agalactiae, M. putrifaciens andM. arginini. The remaining mycoplasma isolates were not identified. The significance of the identified isolates and their probable pathogenic effect on the respiratory tract of goats and sheep are discussed. Their pathogenicity needs to be established experimentally before initiating a control programme in the Kingdom.

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Resumen Se llevó a cabo una investigación preliminar en 163 cabras y 130 ovejas las cuales mostraron síntomas clínicos o lesiones postmortem de pleuroneumonía contagiosa. Se cultivaron en medios especiales muestras patológicas del tracto respiratorio de cabras y ovejas vivas (183 y 160 muestras respectivamente. De 343 cultivos examinados, 211 (61·5%) aislamientos se caracterizaron bioquímicamente. Un total de 116 aislamientos se caracterizaron serológicamente comoM. mycoides, subsp.mycoides, M. ovipneumonia, M. conjuntivae, M. agalactiae subsp.agalactiae, M. putrifaciens yM. arginini. Los restantes micoplasmas aislados no fueron identificados. Se discute el significado de los aislamientos identificados y su posible efecto patógeno en el tracto respiratorio de cabras y ovejas. Es necesario establecer la patogenicidad experimentalmente antes de iniciar un programa de control en el Reino.

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Résumé On sait que la pleuropneumonie du mouton et de la chèvre cause de sérieuses pertes économiques dans beaucoup de pays, dont l'Arabie Saoudite. Une recherche préliminaire a été entreprise sur 163 chèvres et 130 moutons montrant des sympt?mes cliniques ou des lésions à l'autopsie. Des prélèvements du tractus respiratoire de chèvres et de moutons vivants ou abattus (respectivement 183 et 160 échantillons) ont été mis en culture pour la recherche de mycoplasmes. Sur les 343 cultures de mycoplasmes examinées, 211 souches (soit 61,5%) ont été caractérisées au point de vue biochimique. 116 souches ont pu être identifiées sérologiquement comme étantM. mycoides sub.mycoides, M. ovipneumoniae, M. conjunctivae, M. agalactiae sub.agalactiae, M. putrefaciens etM. arginini. Les autres souches de mycoplasmes n'ont pas été identifiées. La signification des souches identifiées et leur effet pathogène probable sur le tractus respiratoire des chèvres et des moutons sont discutés. Leur pouvoir pathogène demande à être expérimentalement établi avant de mettre en place un programme de contr?le de la maladie dans le Royaume.

     

Comparison of Sampling Procedures for Isolating Pulmonary Mycoplasmas in Cattle

Three sampling procedures were compared to determine the optimal technique for isolating mycoplasmas in cattle with respiratory diseases. The prevalence of mycoplasmas isolated from these animals is also reported. In the first group, bronchoalveolar lavage (BAL) and nasal swab cultures were compared with the corresponding lung cultures from cattle necropsied for fatal respiratory diseases (n = 20). In a second group, nasal swabs were compared with corresponding BAL cultures in living animals with recurrent respiratory pathologies (n = 49). There was complete agreement between the paired BAL and lung cultures. In contrast, nasal cultures were not representative of the mycoplasmas present in the lower respiratory airways. The relative sensitivity and specificity of the nasal swab technique compared to BAL in living animals confirmed that the nasal swab cultures were not predictive of lower respiratory airway pathogens, such as Mycoplasma bovis. BAL is considered to be the best method for isolating M. bovis in cattle with respiratory diseases as it combines reliability and feasibility under field sampling conditions. In the present study, Mycoplasma dispar (43%) and M. bovis (29%) were mainly isolated in mixed infections. This confirms the need to search for mycoplasmas in routine examinations and to take them into account in therapeutic strategies for respiratory diseases in cattle.     

Eradication of elephant ear mites (Loxoanoetus bassoni) in two African elephants (Loxodonta africana)

Elephant ear mites, not previously described in North America, were eradicated in two African elephants (Loxodonta africana) after six otic instillations of ivermectin at 2-wk intervals. The microscopic examination of a clear, mucoid discharge collected from the external ear canals of two wild-born African elephants housed in a New York State zoo for 25 yr revealed live mites (Loxoaneotus bassoni). The cytologic examination demonstrated no evidence of inflammation or infection. Both elephants were asymptomatic with normal hemograms and serum chemistry panels. A diagnosis of otoacariasis was made. Each elephant was treated six times with 5 ml of 1% ivermectin syrup instilled in each ear canal once every 2 wk. Microscopic examinations of clear mucus collected from each elephant's ear canals 9 days after the first instillation of ivermectin were negative for any life stages of ear mites. Microscopic examinations of mucus collected from both elephants' ear canals at 6, 11, and 16 wk, as well as annually post-treatment for 7 yr, confirmed eradication of the ear mites. The L. bassoni ear mite was first identified in the external ear canals of wild, asymptomatic, lesion-free, African elephants culled in Kruger National Park in South Africa. However, a new species in the same genus of mites (Loxoanoetus lenae) was identified at the necropsy of an 86-yr-old Asian elephant (Elephas maximus) living in a circus in Australia. The autopsy revealed a marked, ballooning distension of bone around the left external acoustic meatus, suggestive of mite-induced otitis externa, as seen in cattle infested with ear mites (Raillieta auris). Elephant health care providers should identify the prevalence of, and consider treatment of, elephants in their care infested with ear mites, given the possible risk for adverse health effects.     

Isolation of Mycoplasma meleagridis from Free‐ranging Birds of Prey in Germany

In this study tracheal swabs and air sac biopsies of 68 raptors of different species that were found injured or debilitated in Germany were investigated for the occurrence of mycoplasmas. Mycoplasma meleagridis, Mycoplasma falconis, Mycoplasma buteonis, Mycoplasma gypis and five mycoplasma isolates not identified so far could be isolated from 32 (47 %) birds. Mycoplasma meleagridis could be detected in five birds. These birds did not show clinical signs or histopathological alterations in air sac biopsies related to the infection.