Culture media for differential isolation of Lactobacillus casei Shirota from oral samples

This study aimed to develop a solid culture medium for differential isolation of the probiotic strain Lactobacillus casei Shirota (LcS) and for selective cultivation of lactobacilli present in oral samples.Type strains of lactobacilli and isolates from commercial probiotic products were inoculated onto modified de Man Rogosa Sharpe agar (termed ‘LcS Select’), containing bromophenol blue pH indicator, vancomycin and reducing agent L-cysteine hydrochloride for differential colony morphology development. L. casei Shirota cultured on the novel medium produced distinctive colony morphologies, different from other lactobacilli tested.LcS-characteristic colonies were recovered on LcS Select medium from samples of saliva and tongue plaque following a four-week probiotic intervention study. The viable count of presumptive LcS colonies correlated with those isolated on a non-commercial lactitol-LBS-vancomycin agar (LLV) developed for a selective isolation of LcS from faeces.The novel LcS Select medium proved suitable for differential isolation of the probiotic strain L. casei Shirota from oral samples containing mixed microbial populations. It can also be used for selective growth of vancomycin-resistant lactobacilli.There are few available culture media that are sufficiently selective to enable isolation of probiotic strains from mixed populations. LcS Select medium provides a cheaper, yet effective tool in this context.

► ‘LcS Select’ medium is suitable for isolation of L. casei Shirota from oral samples.
► ‘LcS Select’ is a novel formulation and is simple to prepare.
► The medium allows development of LcS-specific colony morphologies.
► LcS Select supports a differential growth of vancomycin resistant lactobacilli.
► The agar is suitable for quantification of LcS from samples in intervention studies.