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A novel MLSA allelic profile 'A15' of Mycoplasma mycoides subsp mycoides in Niger.

ABSTRACT: Mycoplasma mycoides subsp. mycoides (Mmm) is the aetiological agent of contagious bovine pleuropneumonia (CBPP). The aim of the present study was to identify the profiles of the Mmm strains isolated in Niger using the 'Multilocus Sequence Analysis' (MLSA) typing technique based on polymorphism analysis of housekeeping and non-coding genes. The investigation was conducted on samples (n=22) comprising of lung tissues, lymph node and pleural fluids. Following classical PCR, Mmm positive amplicons (n=6) were identified. These positive amplicons were then amplified using eight loci of the PG1 reference strain (LocPG1-0001, Loc-PG1-0103, Loc-PG1-0287, Loc-PG1-0431, Loc-PG1-0489, Loc-PG1-0523, Loc-PG1-0710 and Loc-PG1-0827). Sequencing followed by the determination of the profile of each strain by the combination of the allele numbers revealed three different MLSA profiles namely; A11, E01 and A15. The profiles A11 and E01 were previously identified. The novel profile identified in this study was named profile A15. The difference was detected while comparing sequences of non-coding loci. This novel profile was named 'A15' according to the similarities with African reference strain profile 'A00' at the seven loci level (loc-0103, loc-0287, loc-0431, loc-0489, loc-0523, loc-0710 and loc-0827). For CBPP control measures, identification and molecular characterization of Mmm strains is very important. Thus, the use of MLSA technique is relevant to identify profiles of Mmm circulating in Niger. Other countries where CBPP is still endemic are encouraged to use a MLSA scheme to address this issue and, most importantly, to rapidly trace back the origin of outbreaks, which will help reduce the transmission and spread of the disease. In addition, mapping the profiles of strains circulating in each of the countries of the sub-region is necessary for effective control of CBPP.

SUBMITTER: Yansambou MS

PROVIDER: S-EPMC8136944 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS), also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical ?(1->6)-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan) and associated with an antigenic switch constitutes a finely tuned mechanism that may be involved in virulence.

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A novel MLSA allelic profile 'A15' of Mycoplasma mycoides subsp mycoides in Niger.

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