Discrimination between Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capricolum using PCR-RFLP and PCR.

In this study, the dihydrolipoyl dehydrogenase (lpdA) gene was used to distinguish Mycoplasma mycoides subsp. capri (Mmc) from Mycoplasma capricolum subsp. capricolum (Mcc), two of four Mycoplasma species that cause contagious agalactia in sheep and goats. After alignment of nucleotide sequences of both species, specific primer sets were designed from unchanging and variable gene segments. The first primer set LPD-C1-F/LPD-C1-R was used to amplify a 911 bp fragment that was subsequently co-digested with FastDigest PstI, SspI, EcoRI and ClaI enzymes. The PCR-RFLP profiles differentiated the two mycoplasma species. The second primer set was used to distinguish Mmc from Mcc by single tube PCR. Both methods were further applied to identify 54 isolates collected from dairy herds from different provinces in Sardinia. The results of this study showed that PCR-RFLP and PCR could be used in routine diagnosis for rapid and specific simultaneous discrimination of Mmc and Mcc.

Fatal transmission of contagious caprine pleuropneumonia to an Arabian oryx (Oryx leucoryx).

Contagious caprine pleuropneumonia (CCPP) is an infectious respiratory disease mainly affecting domestic goats. As CCPP has never been documented in grazing antelopes (subfamily hippotraginae), they were not considered susceptible. Mycoplasma capricolum subspecies capripneumoniae (Mccp) was isolated from pleural liquid collected during the necropsy of a severely emaciated Arabian oryx with mild nasal discharge. The Mccp isolate was then genotyped using a multilocus sequence scheme; the sequence type was identical to the Mccp strain previously identified in a sand gazelle from a nearby enclosure. This case shows for the first time that members of the hippotraginae subfamily, here the Arabian oryx, can be affected by CCPP. In addition, genotyping shows that the oryx was most probably infected, at a distance, by sand gazelles.

Fatal outbreak of Mycoplasma capricolum pneumonia in endangered markhors.

A pneumonia outbreak reduced the numbers of a wild population of endangered markhors (Capra falconeri) in Tajikistan in 2010. The infection was diagnosed by histologic examination and bacteriologic testing. Mycoplasma capricolum subsp. capricolum was the sole infectious agent detected. Cross-species transmission from domestic goats may have occurred.

Molecular detection of a mixed infection of Goatpox virus, Orf virus, and Mycoplasma capricolum subsp. capripneumoniae in goats.

The current study investigated an outbreak of mixed infection with Goatpox virus (GTPV), Orf virus (ORFV), and Mycoplasma capricolum subsp. capripneumoniae (MCCP) that occurred on a Chinese goat farm, with a case fatality rate of 60.2%. The observed clinical signs were ecthyma and accelerated respiration with frequent coughing. Specific fragments of the p32 gene of GTPV, B2L gene of ORFV, and 16S ribosomal RNA gene of MCCP were synchronously amplified by polymerase chain reaction (PCR) from the tissues of 12 dead goats. The PCR products were cloned, sequenced, and aligned with related reference sequences in GenBank for further identification of the pathogens. The present study reports a mixed infection with GTPV, ORFV, and MCCP in goats.

Detection of contagious caprine pleuropneumonia in East Turkey.

A study was implemented to investigate the presence of contagious caprine pleuropneumonia (CCPP) in East Turkey. This study was based on clinical surveillance in the field, surveillance at regional slaughterhouses and regular submission of suspected lesions to regional laboratories. The results showed that the agent of CCPP, Mycoplasma capricolum subspecies capripneumoniae (Mccp), could be detected by culture and specific polymerase chain reaction from 37.5% (12/32) of lung samples taken from goats of ten different herds. This agent was also isolated from two of 13 sheep samples (one from the lung and the other from a nasal swab). Mycoplasma capricolum subsp. capripneumoniae was isolated in pure culture and characterised at a finer molecular level. The East Turkish isolate was found to be closely related to another strain of Turkish origin, as well as to Mccp strains isolated in Tunisia. The isolation of Mccp from sheep lung lesions brings the strict host-specificity of this pathogen into question. It may also indicate that Mccp presents a risk for wildlife in the region. Such results, the authors believe, demonstrate that adequate risk assessments should be undertaken in Turkey and neighbouring countries.

[Reclassification of the four China isolated strains of the pathogen for contagious caprine pleuropneumonia].

Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp). The aims of this study were to identify 4 Chinese isolated strains employing molecular methods and to determine the appropriate subspecies classification of these strains. Three genome fragments (A, B and C) from each strain were amplified and then transformed into plasmids. The inserted fragments were sequenced and analyzed by comparison with six members of the Mycoplasma mycoides cluster. Cleavage of the PCR products of the 4 strains with PstI yielded three fragments 548, 420 and 128bp in length, just like strain F38. The other M. mycoides cluster members had only 2 fragments of 428 and 128bp. Homology analysis of fragment B indicated that the 4 strains exhibited 99.5% homology with Mccp reference strain F38, 98.9% with M. capricolum subsp. Capricolum (Mcc) strain California Kid, and only 95.4% with Mmc strain ZZ. In fragment C, the 4 strains had 67.4% - 67.6% homology with Mmc PG3, 95.1% -98.6% with Mcc strains 8601-50 and California Kid, 99.6% - 99.8% with Mccp strains 97097ET, Gabes and F38. The analysis revealed that 4 pathogeny strains, 87001, 87002, 367, 1653, isolated from China are more closely related to Mccp than to Mcc. Therefore the pathogeny of CCPP in China should be reclassified as Mccp.

Isolation of Mycoplasma capricolum-like strains from chickens.

Members of the genus Mycoplasma infect a wide range of hosts, but individual Mycoplasma species tend to exhibit a considerable degree of host specificity. We characterized Mycoplasma strain 700, isolated from a kidney of a layer hen in Spain and Mycoplasma strains ULB-A and ULB-B, isolated from the air sac and from the bile of stunted broiler chickens in Slovenia. The serologic examination showed that these three strains are antigenically unrelated to all of the recognized Mycoplasma species of avian origin, but closely related to the ruminant mycoplasma Mycoplasma capricolum subspecies capricolum (M. capricolum). The comparison of their 16S rRNA gene sequences with the sequence of M. capricolum (California kid) revealed 99.66% sequence identity for the strain 700 and 99.59% identity for strains ULB-A and ULB-B. Moreover, the predicted DnaK sequences of the M. capricolum-like strains, isolated from chickens, were identical to DnaK sequences of M. capricolum. Comparison of their dnaK gene sequences with M. capricolum showed 99.64% sequence identity for strain 700 and 99.27% identity for strains ULB-A and ULB-B. In the flock from which M. capricolum-like strains ULB-A and ULB-B were isolated, the majority of chickens (83% of the chickens examined) raised antibodies reacting with M. capricolum antigens. Notably, the infection of chickens with M. capricolum-like strains represents an unusual exception to the range of Mycoplasma species host specificity.