Effects of Psychobiotic and Prebiotic Administration on Gut Microbiota and Depression-Like Behaviors in Mice

Gülçin Yılmaz^1, and Ayhan Dağ²

¹Institute of Health Sciences, Department of Nutrition and Dietetics, Lokman Hekim University, Ankara, Turkey

²Faculty of Health Sciences, Department of Nutrition and Dietetics, Lokman Hekim University, Ankara, Turkey

Cite this paper:
Gülçin Yılmaz and Ayhan Dağ. Effects of Psychobiotic and Prebiotic Administration on Gut Microbiota and Depression-Like Behaviors in Mice. Journal of Food and Nutrition Research. 2026; 14(5):126-132. doi: 10.12691/jfnr-14-5-1

Abstract

Depression is a multifactorial mental disorder in which the gut–brain axis and gut microbiota have been suggested to play important roles. This study aimed to investigate the effects of psychobiotic (Bifidobacterium breve CCFM1025) and prebiotic (inulin) interventions on gut microbiota composition and depression-like behaviors in a chronic unpredictable mild stress (CUMS) mouse model. A total of 24 male C57BL/6 mice were randomly divided into four groups (n = 6): control, CUMS, CUMS + psychobiotic, and CUMS + prebiotic. Depression-like behaviors were induced using a 5-week CUMS protocol. The psychobiotic group received Bifidobacterium breve CCFM1025 (10⁹ CFU/day), while the prebiotic group received inulin (66 mg/kg) via oral gavage. Behavioral assessments were performed using the forced swim test (FST), tail suspension test (TST), and sucrose preference test (SPT). Gut microbiota composition was analyzed using 16S rRNA gene sequencing. Statistical analyses were conducted using Kruskal–Wallis and Mann–Whitney U tests. The results showed that both psychobiotic and prebiotic interventions significantly improved depression-like behaviors compared to the CUMS group, as indicated by reduced immobility time in FST and increased sucrose preference (p < 0.05). However, no statistically significant differences were observed in overall gut microbiota diversity or composition among the groups (p > 0.05). In conclusion, psychobiotic and prebiotic interventions improved behavioral outcomes in a CUMS-induced depression model; however, these effects were not accompanied by significant changes in gut microbiota composition. Therefore, the observed behavioral improvements may not be directly mediated by alterations in microbial diversity and may instead involve alternative mechanisms. These findings should be interpreted with caution due to the limited sample size and lack of functional analyses. These findings highlight the need for future studies incorporating detailed mechanistic investigations to better understand the underlying pathways.

Keywords:
depression microbiota behavioral changes psychobiotic

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References:

[1]	Zhu, F., Tu, H., & Chen, T. (2022). The microbiota–gut–brain axis in depression: the potential pathophysiological mechanisms and microbiota combined antidepression effect. Nutrients, 14(10), 2081.
[2]	Gonzalez Olmo, B. M., Butler, M. J., & Barrientos, R. M. (2021). Evolution of the human diet and its impact on gut microbiota, immune responses, and brain health. Nutrients, 13(1), 196.
[3]	Jach, M. E., Serefko, A., Szopa, A., Sajnaga, E., Golczyk, H., Santos, L. S., ... & Sieniawska, E. (2023). The role of probiotics and their metabolites in the treatment of depression. Molecules, 28(7), 3213.
[4]	Huang, F., & Wu, X. (2021). Brain neurotransmitter modulation by gut microbiota in anxiety and depression. Frontiers in cell and developmental biology, 9, 649103.
[5]	Kraimi, N., Lormant, F., Calandreau, L., Kempf, F., Zemb, O., Lemarchand, J., ... & Leterrier, C. (2022). Microbiota and stress: a loop that impacts memory. Psychoneuroendocrinology, 136, 105594.
[6]	Radford-Smith, D. E., & Anthony, D. C. (2023). Prebiotic and probiotic modulation of the microbiota–gut–brain axis in depression. Nutrients, 15(8), 1880.
[7]	Trzeciak, P., & Herbet, M. (2021). Role of the intestinal microbiome, intestinal barrier and psychobiotics in depression. Nutrients, 13(3), 927.
[8]	Wei, C. L., Wang, S., Yen, J. T., Cheng, Y. F., Liao, C. L., Hsu, C. C., ... & Tsai, Y. C. (2019). Antidepressant-like activities of live and heat-killed Lactobacillus paracasei PS23 in chronic corticosterone-treated mice and possible mechanisms. Brain research, 1711, 202-213.
[9]	Gibson, G. R., Hutkins, R., Sanders, M. E., Prescott, S. L., Reimer, R. A., Salminen, S. J., ... & Reid, G. (2017). Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nature reviews Gastroenterology & hepatology, 14(8), 491-502.
[10]	Tian, P., Chen, Y., Zhu, H., Wang, L., Qian, X., Zou, R., ... & Chen, W. (2022). Bifidobacterium breve CCFM1025 attenuates major depression disorder via regulating gut microbiome and tryptophan metabolism: A randomized clinical trial. Brain, behavior, and immunity, 100, 233-241.
[11]	Magalhães-Guedes, K. T. (2022). Psychobiotic therapy: method to reinforce the immune system. Clinical Psychopharmacology and Neuroscience, 20(1), 17.
[12]	Qin, Y. Q., Wang, L. Y., Yang, X. Y., Xu, Y. J., Fan, G., Fan, Y. G., ... & Li, X. (2023). Inulin: Properties and health benefits. Food & function, 14(7), 2948-2968.
[13]	Tian, P., O'Riordan, K. J., Lee, Y. K., Wang, G., Zhao, J., Zhang, H., ... & Chen, W. (2020). Towards a psychobiotic therapy for depression: Bifidobacterium breve CCFM1025 reverses chronic stress-induced depressive symptoms and gut microbial abnormalities in mice. Neurobiology of stress, 12, 100216.
[14]	Steru, L., Chermat, R., Thierry, B., & Simon, P. (1985). The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology, 85(3), 367-370.
[15]	Chen, L., Faas, G. C., Ferando, I., & Mody, I. (2015). Novel insights into the behavioral analysis of mice subjected to the forced-swim test. Translational psychiatry, 5(4), e551-e551.
[16]	Gross, M., & Pinhasov, A. (2016). Chronic mild stress in submissive mice: Marked polydipsia and social avoidance without hedonic deficit in the sucrose preference test. Behavioural brain research, 298, 25-34.
[17]	Wu, M., Tian, T., Mao, Q., Zou, T., Zhou, C. J., Xie, J., & Chen, J. J. (2020). Associations between disordered gut microbiota and changes of neurotransmitters and short-chain fatty acids in depressed mice. Translational Psychiatry, 10(1), 350.
[18]	Yang, X., Wang, G., Gong, X., Huang, C., Mao, Q., Zeng, L., ... & Xie, P. (2019). Effects of chronic stress on intestinal amino acid pathways. Physiology & behavior, 204, 199-209.
[19]	Jin, S., Lu, Y., Zuo, Y., Xu, Q., Hao, Y., Zuo, H., ... & Li, Y. (2025). Akkermansia muciniphila ameliorates chronic stress-induced colorectal tumor growth by releasing outer membrane vesicles. Gut microbes, 17(1), 2555618.
[20]	Schroeder, B. O., Birchenough, G. M., Ståhlman, M., Arike, L., Johansson, M. E., Hansson, G. C., & Bäckhed, F. (2018). Bifidobacteria or fiber protects against diet-induced microbiota-mediated colonic mucus deterioration. Cell host & microbe, 23(1), 27-40.
[21]	Liu, P., Liu, Z., Wang, J., Wang, J., Gao, M., Zhang, Y., ... & Zhang, K. (2024). Immunoregulatory role of the gut microbiota in inflammatory depression. Nature communications, 15(1), 3003.
[22]	Ju, S., Shin, Y., Han, S., Kwon, J., Choi, T. G., Kang, I., & Kim, S. S. (2023). The gut–brain axis in schizophrenia: the implications of the gut microbiome and SCFA production. Nutrients, 15(20), 4391.
[23]	Hamamah, S., Aghazarian, A., Nazaryan, A., Hajnal, A., & Covasa, M. (2022). Role of microbiota-gut-brain axis in regulating dopaminergic signaling. Biomedicines 2022, 10, 436.
[24]	Li, H., Xiang, Y., Zhu, Z., Wang, W., Jiang, Z., Zhao, M., ... & Ho, C. S. (2021). Rifaximin-mediated gut microbiota regulation modulates the function of microglia and protects against CUMS-induced depression-like behaviors in adolescent rat. Journal of neuroinflammation, 18(1), 254.
[25]	Nguyen, T. Q., Martínez-Álvaro, M., Lima, J., Auffret, M. D., Rutherford, K. M., Simm, G., ... & Roehe, R. (2023). Identification of intestinal and fecal microbial biomarkers using a porcine social stress model. Frontiers in Microbiology, 14, 1197371.
[26]	Fung, T. C., Vuong, H. E., Luna, C. D., Pronovost, G. N., Aleksandrova, A. A., Riley, N. G., ... & Hsiao, E. Y. (2019). Intestinal serotonin and fluoxetine exposure modulate bacterial colonization in the gut. Nature microbiology, 4(12), 2064-2073.