Abstract
BackgroundThe genetic diversity of Mycoplasma agalactiae (MA) isolates collected in Spain from goats in an area with contagious agalactia (CA) was assessed using a set of validated and new molecular typing methods. Validated methods included pulsed field gel electrophoresis (PFGE), variable number of tandem repeats (VNTR) typing, and Southern blot hybridization using a set of MA DNA probes, including those for typing the vpma genes repertoire. New approaches were based on PCR and targeted genomic regions that diverged between strains as defined by in silico genomic comparisons of sequenced MA genomes.ResultsOverall, the data showed that all typing tools yielded consistent results, with the VNTR analyses being the most rapid method to differentiate the MA isolates with a discriminatory ability comparable to that of PFGE and of a set of new PCR assays. All molecular typing approaches indicated that the Spanish isolates from the endemic area in Murcia were very diverse, with different clonal isolates probably restricted to separate, but geographically close, local areas.ConclusionsThe important genetic diversity of MA observed in infected goats from Spain contrasts with the overall homogeneity of the genomic background encountered in MA from sheep with CA in Southern France or Italy, suggesting that assessment of the disease status in endemic areas may require different approaches in sheep and in goats. A number of congruent sub-typing tools are now available for the differentiation of caprine isolates with comparable discriminatory powers.
Highlights
The genetic diversity of Mycoplasma agalactiae (MA) isolates collected in Spain from goats in an area with contagious agalactia (CA) was assessed using a set of validated and new molecular typing methods
Strain typing by pulsed field gel electrophoresis (PFGE) Four restriction endonucleases, SmaI, MluI, KpnI and BstZI, were selected that yielded interpretable PFGE banding patterns on our set of isolates
Typing tools already validated with MA mostly isolated from ovine, such as variable number of tandem repeats (VNTR) or PFGE assays [9,13,24], were shown to be useful to discriminate MA collected in goats from a common geographical origin
Summary
The genetic diversity of Mycoplasma agalactiae (MA) isolates collected in Spain from goats in an area with contagious agalactia (CA) was assessed using a set of validated and new molecular typing methods. CA is listed by the World Organization for Animal Health and is responsible for significant economic loss, with its severity and expression being dependent on several factors, including the host species, the aetiological agent, the production system and the environmental conditions [1,2,3,4]. In this context, defining the sanitary status of herds or regions with respect to CA is essential, and very challenging. Far management strategies have been the most satisfactory method for controlling this disease [4]
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