Genomic polymorphism of Mycoplasma flocculare revealed by a newly developed multilocus sequence typing scheme.

Mycoplasma flocculare is genetically closely related to M. hyopneumoniae, the etiologic agent of porcine enzootic pneumonia, and is frequently isolated with this second species. In this article, we report on the development of the first multilocus sequence typing (MLST) scheme for M. flocculare, based on three genes (adk, rpoB and tpiA). In total, 5022 bp of sequence were analyzed. MLST was used to characterize seven M. flocculare isolates and the reference strain. Eight distinct sequence types were defined, showing the great intraspecies variability of M. flocculare, and the high discriminatory power of the new typing method. The relative contribution of recombinations to the genomic evolution of M. flocculare was revealed by calculating the index of association (IA: 0.0185). This MLST scheme is now available for the acquisition of new knowledge on M. flocculare epidemiology via an online database comprising the DNA sequences of each allele, available at http://pubmlst.org/mflocculare/.

Pathogenicity study of Mycoplasma hyorhinis and M. flocculare in specific-pathogen-free pigs pre-infected with M. hyopneumoniae.

Mycoplasma (M.) hyopneumoniae is the initiator agent of the porcine respiratory disease complex (PRDC) and the etiological agent of enzootic pneumonia. M. hyorhinis and M. flocculare are also found in extensive gross pneumonia-like lesions, but their role is not known. We investigated the pathogenicity of M. hyorhinis and M. flocculare in specific-pathogen-free pigs pre-infected or not with M. hyopneumoniae. Mono-inoculated pigs with M. flocculare showed no clinical signs, hematological changes or macroscopic lesions upon necropsy. Mono-inoculated pigs with M. hyorhinis showed, overall seven days after inoculation, an increase in mean temperature with increases in white blood cell (monocyte) counts and in concentrations of pig major acute phase protein, whereas the average daily weight gain (ADWG) decreased compared with non-infected animals. M. hyorhinis was detected in serous membranes (polyserositis) but not in bronchi. Co-infected pigs with M. hyopneumoniae and M. hyorhinis or M. flocculare showed lower ADWG during the third week of the experiment and higher haptoglobin concentrations in contrast to pigs only mono-infected with M. hyopneumoniae. In pigs co-infected with M. hyopneumoniae and M. hyorhinis, it was interesting to observe that (i) M. hyorhinis was detected in bronchi of six pigs, (ii) M. hyopneumoniae was detected in polyserositis and (iii) there was a slight delay in the production of anti-M. hyopneumoniae IgG. The extent of pneumonia was not statistically different between groups. These results suggest that mycoplasmal associations appear to induce an additive effect and increase the inflammatory status in pigs, probably involving in the impairment of the immune system.

Novel Streptococcus suis Sequence Type 834 among Humans, Madagascar.

Two cases of meningitis caused by Streptococcus suis occurred in Madagascar, 1 in 2015 and 1 in 2016. We report the characterization of the novel sequence type, 834, which carried the mrp+/sly+/epf+ virulence marker and a mutation G→T at position 174, leading to a substitution mutS1 to mutS284.

Multilocus sequence typing of Mycoplasma hyorhinis strains identified by a real-time TaqMan PCR assay.

A real-time TaqMan PCR assay based on the gene encoding the protein p37 was developed to detect Mycoplasma hyorhinis. Its specificity was validated with 29 epidemiologically unrelated M. hyorhinis strains (28 field strains and one reference strain) and other mycoplasma species or with other microorganisms commonly found in pigs. The estimated detection limit of this qPCR assay was 125 microorganism equivalents/µl. The same 29 epidemiologically unrelated M. hyorhinis strains and four previously fully sequenced strains were typed by two portable typing methods, the sequencing of the p37 gene and a multilocus sequence typing (MLST) scheme. The first method revealed 18 distinct nucleotide sequences and insufficient discriminatory power (0.934). The MLST scheme was developed with the sequenced genomes of the M. hyorhinis strains HUB-1, GDL-1, MCLD, and SK76 and based on the genes dnaA, rpoB, gyrB, gltX, adk, and gmk. In total, 2,304 bp of sequence was analyzed for each strain. MLST was capable of subdividing the 33 strains into 29 distinct sequence types. The discriminatory power of the method was >0.95, which is the threshold value for interpreting typing results with confidence (D=0.989). Population analysis showed that recombination in M. hyorhinis occurs and that strains are diverse but with a certain clonality (one unique clonal complex was identified). The new qPCR assay and the robust MLST scheme are available for the acquisition of new knowledge on M. hyorhinis epidemiology. A web-accessible database has been set up for the M. hyorhinis MLST scheme at http://pubmlst.org/mhyorhinis/.