Pinto bean supplementation modulates gut microbiota and improves markers of gut integrity in a mouse model of estrogen deficiency
To evaluate the effects of PB supplementation and estrogen deficiency on gut microbiota composition, mice were divided into four experimental groups: CON-SES (control diet with sesame oil), CON-VCD (control diet with VCD-induced estrogen deficiency), PB-SES (PB-supplemented diet with sesame oil), and PB-VCD (PB-supplemented diet with VCD-induced estrogen deficiency), with n = 8 per group. Cecal contents were collected at the end of the study and subjected to microbial DNA extraction using the Qiagen PowerFecal Kit (Qiagen, Maryland) following the manufacturer's protocol. The bacterial 16S rRNA gene V3-V4 region was amplified using primers F341 (5'-CCTACGGGNGGCWGCAG-3') and R806 (5'-GGACTACNVGGGTWTCTAAT-3'), which included Illumina adapter sequences and sample-specific barcodes. Amplicons were sequenced on an Illumina MiSeq platform using paired-end 250 bp reads, generating raw FASTQ files for downstream microbiome analysis.