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Anticlostridial agent 8‐hydroxyquinoline improves the isolation of faecal bifidobacteria on modified Wilkins–Chalgren agar with mupirocin

Novakova, J., Vlkova, E., Salmonova, H., Pechar, R., Rada, V., Kokoska, L.
Letters in applied microbiology 2016 v.62 no.4 pp. 330-335
8-hydroxyquinoline, Bifidobacterium, Clostridium, agar, feces, humans, selective media
The need for suitable selective cultivation media for the isolation of Bifidobacterium spp. continues to be a real concern in the field of intestinal microbiology. Isolation of bifidobacteria from human and animal faecal samples using selective agar plating may be problematic especially in samples with increased clostridial counts than bifidobacterial counts. Due to the absence of anticlostridial agents in existing selective media, clostridia can displace bifidobacteria resulting in incorrect estimation of their counts. Therefore, we supplemented the existing selective medium ‘modified Wilkins Chalgren agar with mupirocin’ (MWM) with 90 mg l⁻¹ of 8‐hydroxyquinoline (8HQ), which was recently proved to act selectively against clostridia. The newly composed ‘modified Wilkins–Chalgren agar with 8HQ’ (MWMQ) was tested on pure bifidobacterial and clostridial strains, their mixtures, and using faecal samples of mammalian origin; its selectivity was evaluated by genus‐specific identification of isolates. The results demonstrated that the presence of 8HQ in this agar eliminated the growth of nonbifidobacterial strains on MWMQ compared to that on MWM, whereas the recovery of bifidobacterial counts was at satisfactory levels. In conclusion, MWMQ could be recommended for bifidobacterial isolation from human and animal faeces especially when bifidobacteria are not numerically dominant and there are chances of clostridial contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: Routine isolation of bifidobacteria from mammalian faeces does not use a reliable selective agar with an anticlostridial agent. Overgrowth of clostridia may result in incorrect estimation of bifidobacterial counts. Thus, in order to improve the selectivity of existing media for bifidobacterial isolation, we chose the modified Wilkins–Chalgren agar with mupirocin and supplemented it with 8‐hydroxyquinoline (8HQ), a molecule that shows anticlostridial activity without affecting the growth of bifidobacteria. This newly composed medium showed enhanced selectivity and specificity compared to the original medium and therefore, can be recommended for the isolation of bifidobacteria from mammal faeces.