A Bacillus subtilis strain with high proficiency for biofilm formation delays the onset of aging-associated morphological defects in cholinergic neurons of C. elegans

Aging is accompanied by progressive functional decline and morphological defects of cholinergic neurons. Some probiotic bacteria with proficiency for biofilm formation in the gut are being discussed as potentially neuroprotective. Here, we screened Bacillus subtilis (B. subtilis) strains for biofilm production, using an in vitro assay. Thereafter, a transgenic Caenorhabditis elegans (C. elegans) strain with its cholinergic neurons expressing green fluorescent protein (GFP) was fed for up to 15 days on either the identified biofilm-producing B. subtilis strain B4U 50 or the control B. subtilis strain 168, with or without addition of a standardized Ginkgo biloba preparation. The morphology of the cholinergic neurons was monitored at several consecutive days by fluorescence microscopy. The occurrence of morphological defects such as knots and ruptures within the axons of cholinergic neurons, as observed with increasing age, was significantly lower in nematodes provided with the B4U 50 strain. Even worms whose diet was switched to B4U 50 later in life showed less damage of their neurons, as compared to worms fed on the control strain. Supplementation with the Ginkgo biloba preparation was neuroprotective in worms fed on either B. subtilis strain, while a stronger beneficial effect was observed upon combined administration of B. subtilis B4U 50 and the Ginkgo biloba preparation. In conclusion, feeding a B. subtilis strain proficient for biofilm formation, B4U 50, was neuroprotective when compared to a diet consisting of control B. subtilis, delaying the onset of aging-associated morphological defects in cholinergic neurons of C. elegans.