Assessing Gut Microbiome Changes in Mice

A total of 132 fecal samples were collected from female mice (n = 66) of the C57Bl/6 strain (Charles River, Calco, LC, Italy) and included in the experimental set reported in Figure 1. All procedures for animal housing and maintenance were performed in compliance with previous studies [46 (link),47 (link),48 (link)]. The age of the animals at the start of the study was between 8 and 12 weeks. The mice were monitored daily, and fecal samples were collected every 15 days for the S100B assay and 16S amplicon sequencing analysis. In order to test the effect of Pentamidine (PTM), a S100B inhibitor, as indicated in Figure 2, PTM was administered intraperitoneally (4 mg/kg) to 25 mice (50 fecal samples) vs. 28 untreated controls (as a part of the previous experimental set involving a total of 66 animals), following a previously established protocol [47 (link)]. Additionally, in a pilot test, 300 µL of a solution (phosphate-buffered saline—PBS) containing 200 µg of S100B (Sigma, St. Louis, MO, USA) or of vehicle (phosphate-buffered saline—PBS) was orally administered, respectively, to 3 and 2 C57Bl/6 mice. After 15 days, fecal samples (n = 10) were collected and analyzed for microbiota biodiversity.

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Romano Spica V., Valeriani F., Orsini M., Clementi M.E., Seguella L., Gianfranceschi G., Di Liddo R., Di Sante G., Ubaldi F., Ria F., Esposito G, & Michetti F. (2023). S100B Affects Gut Microbiota Biodiversity. International Journal of Molecular Sciences, 24(3), 2248.

Publication 2023

C57Bl/6 strain

Animals Assay C57bl mice Fecal Female Mice Microbiota Pentamidine Phosphate River Saline Sequencing analysis Strain

Corresponding Organization : Libera Università Maria SS. Assunta

Other organizations : Istituto Zooprofilattico Sperimentale delle Venezie, Sapienza University of Rome, University of Padua, University of Perugia

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Variable analysis

independent variables

Administration of Pentamidine (PTM), a S100B inhibitor, to 25 mice vs. 28 untreated controls
Oral administration of a solution containing 200 µg of S100B or vehicle (phosphate-buffered saline - PBS) to 3 and 2 C57Bl/6 mice, respectively

dependent variables

S100B assay
16S amplicon sequencing analysis
Microbiota biodiversity

control variables

Female mice of the C57Bl/6 strain (Charles River, Calco, LC, Italy)
Animal housing and maintenance procedures
Age of the animals (8-12 weeks)

controls

Positive control: Oral administration of a solution containing 200 µg of S100B to 3 C57Bl/6 mice
Negative control: Oral administration of vehicle (phosphate-buffered saline - PBS) to 2 C57Bl/6 mice

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