# Extracellular vesicles secretion by Mycoplasma mycoides subsp. mycoides, the etiological agent of contagious bovine pleuropneumonia

Sarah Ganter, Alexandre Villard, Lucia Manso-Silvan, Didier Chevret, Christelle Boulé, Véronique Monnet, Florence Tardy, Patrice Gaurivaud

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Sarah Ganter, Alexandre Villard, Lucia Manso-Silvan, Didier Chevret, Christelle Boulé, et al.. Extracellular vesicles secretion by Mycoplasma mycoides subsp. mycoides, the etiological agent of contagious bovine pleuropneumonia. European Mycoplasma Conference, Mar 2019, London, United Kingdom. pp.47-47. hal-05173786

HAL Id: hal-05173786

https://hal.science/hal-05173786v1

Submitted on 21 Jul 2025

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**Title:** Extracellular vesicles secretion by *Mycoplasma mycoides* subsp. *mycoides*, the etiological agent of contagious bovine pleuropneumonia

**Authors**

Sarah Ganter1, Alexandre Villard1, Lucia Manso-Silvan2, Didier Chevret3, Christelle Boulé4, Véronique Monnet3, Florence Tardy1 and Patrice Gaurivaud1.

**Institution**

1 Université de Lyon, Anses, Laboratoire de Lyon, VetAgro Sup, UMR Mycoplasmoses des Ruminants, 31 avenue Tony Garnier 69364 Lyon cedex 07, France

2 CIRAD-INRA, UMR 1309 ASTRE, F-34398 Montpellier, France

3 PAPPSO, Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France

4 Université Claude Bernard Lyon 1, Centre Technologique des Microstructures, Service « Etudes à façon » EZUS Lyon, Villeurbanne, France

Extracellular vesicles (EV) are nano-sized, membrane-derived, non-self-replicating, spherical structures shed into the extracellular environment by both eukaryotic and prokaryotic cells. In bacteria, EV carry virulence factors like adhesins, toxins and immunomodulatory molecules. They are of importance for the bacteria –host interplay and hence vaccine development. Within the *Mycoplasma* (M.) genus EV have only been described, briefly, in *M. gallisepticum* so far. The aim of this study was to investigate the potential secretion of EV by *M. mycoides* subsp. *mycoides* (Mmm), the etiological agent of contagious bovine pleuropneumonia, and to characterize their membrane protein content.

Several culture conditions were tested to produce EV of Mmm strain Afadé *in vitro*. Vesicles were collected using a classical ultracentrifugation method and observed by
transmission electron microscopy. Different approaches, such as gradient density and
cultures in minimal medium were tested in order to improve EV purification. EV
membrane proteins were extracted with triton x-114 and identified by mass
spectrometry.

*Mmm* was shown to release EV *in vitro*, their size and concentration being dependent on growth conditions. *Mmm* vesicles were produced by living cells and not by cells with decreased viability after stresses. Their release happened in a budding-way from the cell surface as observed by electron miscroscopy. Several virulence factors were identified in EV membranes suggesting that EV could be involved in host-mycoplasma interactions.

*Mmm* can produce *in vitro* extracellular vesicles that carry virulence factors. Whether this also happens *in vivo* has yet to be demonstrated.