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. 2022 May 20:9:910799.
doi: 10.3389/fvets.2022.910799. eCollection 2022.

Prevalence of Mycoplasma bovis in Algeria and Characterisation of the Isolated Clones

Yasmine Oucheriah  1 , Nouzha Heleili  1 , Adélie Colin  2 , Catherine Mottet  3 , Florence Tardy  2 , Claire A M Becker  3
Affiliations

Prevalence of Mycoplasma bovis in Algeria and Characterisation of the Isolated Clones

Yasmine Oucheriah et al. Front Vet Sci. .

Abstract

Bovine respiratory disease (BRD) is common in calves in Algeria, but to date, Mycoplasma bovis has never been monitored as a potential etiological agent. Here, to assess the presence (direct detection) and circulation (indirect detection) of M. bovis, broncho-alveolar lavage fluids (BALF) and serum samples were collected from 60 veal calf farms in Algeria. A commercial ELISA kit (ID Screen® ELISA) was used to screen for the presence of specific antibodies against M. bovis in 351 blood sera collected from both diseased and healthy calves, and 69% (241 sera) tested positive. BALFs from the 176 diseased calves were used to screen for M. bovis by real-time-PCR (rt-PCR), and 102 (58%) tested positive. A non-exhaustive set of 53 clones were isolated from 44 calves and further subtyped using polC gene sequencing. No predominant subtype was found, and two clones exhibited a new subtype. Fourteen clones were further characterized by multilocus sequence typing, and results showed a high degree of genetic diversity, with some clones having new alleles and subtypes. The minimum inhibitory concentrations (MICs) of 5 antimicrobials regularly used to treat BRD was determined on 45 clones. Susceptibility profiles showed very broad diversity, confirming the variety of clones actively circulating. We detected clones with high MICs, including increased MICs of enrofloxacin (n = 5). This is the first study to report the presence of M. bovis in Algeria in calves with BRD. This research also finds broad genetic and phenotypic diversity in the actively circulating isolates.

Keywords: Algeria; Mycoplasma bovis; antimicrobial resistance; bovine respiratory disease; genetic diversity.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
M. bovis detection by real-time PCR according to season of sampling. X-axis, season of BALF collection; y-axis, number of calves that tested positive in real-time PCR (target M. bovis) (white bars), total number of calves sampled for each season (black dashes) and number of calves from which clones were isolated (diamonds).
Figure 2
Figure 2
eBURST analysis of the allelic profiles of the Algerian clones (ST4, ST8, ST29, ST188, ST195-198) compared to the closest subtypes found in the PubMLST database. Subtype numbers retrieved from the PubMLST database are indicated in circles, and the new subtypes reported in this study are tagged with an asterisk. Only subtypes close to or common to Algerian isolates are indicated, and their country of origin is color-coded: dark blue, Algeria; light blue, Spain; orange, Israel; light orange, Japan; light green, USA; green, Hungary; red, Switzerland; pink, Lithuania; purple, Canada; lilac, Romania.
Figure 3
Figure 3
Distribution of the MIC profiles of spectinomycin, tylosin and enrofloxacin for 44 clones of M. bovis. X-axis, MICs classes; y-axis, number of clones. Arrows indicate the clinical breakpoints for resistance for Pasteurellaceae, i.e., spectinomycin ≥128 μg/mL, enrofloxacin ≥2 μg/mL, and tylosin ≥32 μg/mL threshold of tilmicosin used (dashed arrow).
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