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Animal Models

Assessment of multi-strain probiotics in regulating diet-induced obesity in Balb/c mice model

International Journal of Obesity (2025)Cite this article

Subjects

Abstract

Background/objectives

This study investigated the efficacy of a novel multi-strain probiotic (MSP), composed of Limosilactobacillus fermentum BAB 7912, Bacillus rugosus PIC5CR, and Bacillus rugosus PIB9CR, in preventing and reverting diet-induced obesity in Balb/c male mice.

Subjects/methods

This study used 8-week-old Balb/c mice. A total of 40 mice were divided into five groups namely control negative (CN), control with obesity (CO), and three treatment groups: microbial consortium treated (MCT), Healthy control 1 (HC1), and Healthy control 2 (HC2). Obesity was induced using a high-fat diet. MSP formulation developed indigenously as part of previous study, was fed to Balb/c mice at different time intervals to study its preventive and ameliorative potential. Animals were dissected for the collection of blood as well as various organs to study the effect of MSP feeding on obesity status. Results were validated using histopathological and metagenomic data.

Results

The CN and other treatment groups gained significant weight at the end of 6 weeks, while no significant weight gain was observed among HC1 group animals that were fed with HFD and MSP together. This highlights the preventive effect of continuous MSP feeding in the HC1 animal group. Initial liver histopathology in the HC1 group revealed enlarged hepatocytes and fat droplets. By week 9, the MCT group, which received MSP with a basal diet, showed liver recovery towards normal, accompanied by body weight improvement from 28.02 ± 0.7 g to 26.18 ± 0.96 g. Metagenomic analysis revealed that MSP treatment increased the relative abundance of health-promoting bacteria, notably Lactobacillaceae (specifically Lactobacillus).

Conclusions

Findings indicated that continuous consumption of MSP contributes significantly in prevention of obesity and associated metabolic disorders. Future studies are needed to explore the mechanisms underlying these effects and to evaluate the potential of MSP for human health.

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Fig. 1
Fig. 2: Body weight of different animal groups.
Fig. 3: Analysis of lipid profile (TC, TG, HDL-C, and LDL-C) in different animal groups.
Fig. 4: Analysis of SGOT & SGPT in different animal groups.
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Data availability

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions, although supplementary data have been provided for the deep understanding of findings to the readers.

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Acknowledgements

Authors are thankful to the School of Science and the School of Pharmacy of RK University for providing the resources and support needed for smooth conduction of the study. First author is also highly thankful towards the Animal Research Facility, Zydus Research Centre, Ahmedabad, for providing the experimental animals. First author is also thankful toward the Govt. of Gujarat for providing the SHODH fellowship (KCG/SHODH/2022-23/202001600004). We also thank Dr. Jigna Kalaria (MD. Pathologist) and Dr. Vishal Bhatt (Chief Operating Officer), Pathology Lab Department of BT Savani Kidney Hospital, Rajkot for conducting histopathology study of animal organs and providing insightful interpretation. I would like to extend my heartfelt thanks to the Gujarat Biotechnology Research Center (GBRC) Gandhinagar for their support in the analysis of the metagenomic study. Special thanks to Ms. Khalida Bloch, Research Scholar, Department of Microbiology, School of Science, RK University, for her invaluable support during the in-vivo study.

Funding

The Govt. of Gujarat, India provided funds under their student startup and innovation policy (SSIP; reference no. RKU/SOS/SSIP/2023-2024/03) that was utilized for procurement of biochemical analysis kits, outsourcing of histopathological analysis of animal organs and metagenomic analysis of fecal samples. Beyond this, no other funding support has been received for this complete study.

Author information

Authors and Affiliations

  1. Postbiotics and Foodomics Lab, Department of Microbiology, School of Science, RK University, Rajkot, Gujarat, India

    Mehul Chauhan, Hina Maniya, Priya Mori & Vijay Kumar

  2. Department of Microbiology, Krishna School of Science, Dr. Kiran and Pallavi Patel Global University (KPGU), Vadodara, Gujarat, India

    Hina Maniya

  3. Department of Health, Nutrition, and Food Sciences, Florida State University, Tallahassee, FL, USA

    Ravindar Nagpal

  4. Department of Pharmacology, School of Pharmacy, RK University, Rajkot, Gujarat, India

    Pravin Tirgar

  5. Department of Microbiology, Bhagwan Mahavir College of Basic & Applied Sciences, Bhagwan Mahavir University, Surat, Gujarat, India

    Vijay Kumar

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Contributions

MC: Conceptualization, data Curation, investigation, methodology, visualization, writing—original draft, writing—review and editing. HM: Investigation, methodology. PM: Methodology. RN: Formal analysis, resources. PT: Resources. VK: Conceptualization, data curation, formal analysis, project administration, supervision, validation, visualization, writing-review and editing. All authors approved the final submission to the International Journal of Obesity.

Corresponding author

Correspondence to Vijay Kumar.

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The authors declare no competing interests.

Ethical approval

The animal study was approved by the Institutional Animal Ethics Committee (IAEC) of the School of Pharmacy, RK University, Rajkot, Gujarat, India (Proposal No. RKCP/Col/Re/22/131).

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Chauhan, M., Maniya, H., Mori, P. et al. Assessment of multi-strain probiotics in regulating diet-induced obesity in Balb/c mice model. Int J Obes (2025). https://doi.org/10.1038/s41366-025-01928-w

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