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environments may help reduce external variables. Secondly, our hypothesis that Pediococcus acidilactici
CCFM6432 alleviates depressive symptoms by reducing systemic inflammation is based on functional
predictions of the gut microbiota and preclinical results. Further research is needed to provide evidence on
the specific interactions between gut metabolites (such as lactic acid), circulating inflammatory factors, and
brain immune status. Additionally, the measurement of lactic acid in this study was conducted using a non-
targeted metabolomics approach, without distinguishing between D- and L-lactic acid. Future studies
should aim to identify the specific molecular configuration of lactic acid responsible for the observed effects.

In conclusion, the utilization of probiotics for mitigating mood disorders holds considerable potential. This
study is the first to report the clinical efficacy of a single strain of Pediococcus acidilactici in the adjunctive
treatment of depression and its regulatory effects on the gut microbiota of patients. Future investigations
should prioritize exploring optimal intervention dosages and elucidating the pharmacological mechanisms
of probiotics, aiming to contribute to the advancement of innovative antidepressant strategies.

DECLARATIONS
Authors’ contributions
Conceptualization, writing - original draft, investigation and funding acquisition: Tian P
Investigation, data curation, visualization: Yang H
Methodology, resources: Hang F
Funding acquisition: Wang G
Resources: Mao X
Formal analysis, methodology, validation: Jin X
Writing - review and editing, resources and supervision: Zhao J

Availability of data and materials
All data are available in the main text. Other raw data that support the findings of this study are available
from the corresponding author upon reasonable request.

Financial support and sponsorship
This work was supported by the National Key R&D Program of China (2023YFC2506004), National Natural
Science Foundation of China (32201988, 32394051), Special Fund for Science and Technology Program of
Jiangsu Province (BM2022019), the Fundamental Research Funds for the Central Universities
(JUSRP123047), and the Program of Collaborative Innovation Centre of Food Safety and Quality Control in
Jiangsu Province.

Conflicts of interest
All authors declared that there are no conflicts of interest.

Ethical approval and consent to participate
The human trial was conducted in accordance with the Declaration of Helsinki and approved by the
Medical Ethics Committee of Yixing People’s Hospital (reference number: WXMHCIRB2023LLky002).
Informed consent was obtained from all participants, and details of the ethical statement and trial design are
registered with the Chinese Clinical Trial Registry (ChiCTR2300071025).

Consent for publication
Not applicable.