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Infections caused by  Mycoplasma  species belonging to the  ‘mycoides  cluster’ negatively affect the agricultural sector through losses in livestock productivity. These  Mycoplasma  strains are resistant to many conventional antibiotics due to the total lack of cell wall. Therefore, there is an urgent need to develop new antimicrobial agents from alternative sources such as medicinal plants to curb the resistance threat. Recent studies on extracts from  Solanum aculeastrum  and  Piliostigma thonningii  revealed interesting antimycoplasmal activities hence the motivation to investigate the antimycoplasmal activities of constituent compounds. The CH 2 Cl 2 /MeOH extracts from the berries of  S. aculeastrum  yielded a new β-sitosterol derivative ( 1 ) along with six known ones including; lupeol ( 2 ), two long-chain fatty alcohols namely undecyl alcohol ( 3 ) and lauryl alcohol ( 4 ); two long-chain fatty acids namely; myristic acid ( 5 ) and nervonic acid ( 6 ) as well as a glycosidic steroidal alkaloid; (25R)-3β- O -α- L -rhamnopyranosyl-(1→2)- O -[α- L -rhamnopyranosyl-(1→4)]-β- D -glucopyranosyloxy-22α-N-spirosol-5-ene ( 7 ) from the MeOH extracts. A new furan diglycoside, (2,5- D -diglucopyranosyloxy-furan) ( 8 ) was also characterized from the CH 2 Cl 2 /MeOH extract of stem bark of  P. thonningii . The structures of the compounds were determined on the basis of spectroscopic evidence and comparison with literature data. Compounds  1, 3, 4, 7 , and  8  isolated in sufficient yields were tested against the growth of two  Mycoplasma mycoides  subsp.  mycoides  ( Mmm ), two  M. mycoides. capri  ( Mmc ), and one  M. capricolum capricolum  ( Mcc ) using broth dilution methods, while the minimum inhibitory concentration (MIC) was determined by serial dilution. The inhibition of  Mycoplasma in vitro  growth was determined by the use of both flow cytometry (FCM) and color change units (CCU) methods. Compounds  4  and  7  showed moderate activity against the growth of  Mmm  and  Mmc  but were inactive against the growth of  Mcc . The lowest MIC value was 50 μg/ml for compound  7  against  Mmm . The rest of the compounds showed minimal or no activity against the strains of  Mycoplasma mycoides  tested. This is the first report on the use of combined FCM and CCU to determine inhibition of  in vitro  growth of  Mycoplasma mycoides . The activity of these compounds against other bacterial strains should be tested and their safety profiles determined.

Antimycoplasmal Activities of Compounds from Solanum aculeastrum and Piliostigma thonningii against Strains from the Mycoplasma mycoides Cluster