Abstract
Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease of domestic goats. The exact distribution of CCPP is not known but it is present in Africa and the Middle East and represents a significant threat to many disease-free areas including Europe. Furthermore, CCPP has been recently identified in Tajikistan and China. A typing method with an improved resolution based on Multi-Locus Sequence Analysis (MLSA) has been developed to trace new epidemics and to elucidate whether the recently identified cases in continental Asia were due to recent importation of Mccp. The H2 locus, a polymorphic region already in use as a molecular marker for Mccp evolution, was complemented with seven new loci selected according to the analysis of polymorphisms observed among the genome sequences of three Mccp strains. A total of 25 strains, including the two new strains from Asia, were analysed by MLSA resulting in the discrimination of 15 sequence types based on 53 polymorphic positions. A distance tree inferred from the concatenated sequences of the eight selected loci revealed two evolutionary lineages comprising five groups, which showed good correlation with geographic origins. The presence of a distinct Asian cluster strongly indicates that CCPP was not recently imported to continental Asia. It is more likely that the disease has been endemic in the area for a long time, as supported by historical clinical descriptions. In conclusion, this MLSA strategy constitutes a highly discriminative tool for the molecular epidemiology of CCPP.
Highlights
Contagious Caprine Pleuropneumonia (CCPP) is a devastating disease of goats included in the list of notifiable diseases of the World Organisation for Animal Health (OIE)
Direct amplification and sequencing from clinical material is possible even in cases where concomitant bacteria or antibiotic therapy hamper Mycoplasma capricolum subsp. capripneumoniae (Mccp) isolation, as was demonstrated in this work by analysis of a clinical sample from the Tajik outbreak. All these advantages had already been taken into consideration by the authors when developing an initial tool for Mccp typing based on the H2 locus [8]
The objective was to evolve from a single locus sequence typing method to the analysis of multiple molecular markers in order to provide greater resolution by considerably increasing variability
Summary
Contagious Caprine Pleuropneumonia (CCPP) is a devastating disease of goats included in the list of notifiable diseases of the World Organisation for Animal Health (OIE). The first description of the disease dates back to 1873, in Algeria [1]. Capripneumoniae (Mccp) was only isolated and characterised a century later, in 1976 [2]. This may be explained by several factors. Mccp is one of the most fastidious mycoplasmas to grow in vitro and cultures are often overgrown by concomitant bacteria, hampering its isolation. Mccp may be difficult to identify because it belongs to the Mycoplasma mycoides cluster, a group of five closely
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