The increasing interest in paraprobiotics has raised concerns about their stability and performance throughout their shelf life. This study investigated whether Bifidobacterium animalis BB-12 ( BB )-derived paraprobiotics ( BB P) retain their ability to restore gut microbiota (GM) balance and fecal metabolites in rats fed a high-fat diet (HFD) after various storage times. BB P was produced through ohmic heating (OH) processing, followed by freeze-drying and storing at −20°C, 4°C, and 25°C for 4 months. Flow cytometry (FC) was used to analyze BB P stability, while their biochemical, metabolomic, and microbiomic functionality were studied in vivo. Results showed that BB Ps stored at −20°C in powder form exhibited significantly higher stability. Administration of both fresh and stored BB Ps alleviated HFD-induced metabolic disorders. Compared with the HFD group, BBP administration at the start of storage (IP) declined serum total cholesterol and triglycerides by approximately 28% and 20%, respectively, and raised HDL by 22%. Furthermore, it significantly decreased serum AST and ALT activities by 16% and 38%, respectively. Additionally, it alleviated HFD-induced dysbiosis in rats by enhancing GM microbial diversity and the abundance of beneficial bacteria, such as Bacteroides and Akkermansia , even after prolonged storage. 1 H-NMR spectroscopy revealed enhanced levels of amino acids and short-chain fatty acids (SCFAs), with acetate and propionate concentrations increasing by up to 81% and 157%, respectively, while also indicating decreased fecal sugars and alcohols, confirming improved gut metabolic activity. Overall, BB Ps retained substantial bioactivity and partially maintained their ability to restore HFD-induced dysbiosis throughout storage, although functionality declined slightly over time.
Yolmeh et al. (2026) studied this question.