R E S E A R C H Open Access© The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creati vecommons.org/licenses/by-nc-nd/4.0/. Mahmoodabadi et al. BMC Veterinary Research (2025) 21:340 https://doi.org/10.1186/s12917-025-04793-x BMC Veterinary Research *Correspondence: Hadi Mohebalian mohebalian@um.ac.ir 1Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran 2Faculty of Veterinary Medicine, Department of Pathobiology, Ferdowsi University of Mashhad, Mashhad, Iran 3Faculty of Veterinary Medicine, Department of Clinical Sciences, Ferdowsi University of Mashhad, Mashhad, Iran Abstract Background Canine distemper virus (CDV) is a multisystemic disease in dogs that causes severe neurological signs due to viral replication in neurons and glial cells; this leads to gray matter lesions and demyelination. The resulting inflammation causes additional harm to the affected tissues. This study aimed to investigate the relationship between pro-inflammatory cytokines Tumor Necrosis Factor (TNF-α) and interleukin (IL-6) levels in Cerebrospinal fluid (CSF) and the neurological form of distemper disease in dogs. Results The study included 23 CDV-infected dogs with neurological signs, 10 diseased but CDV-negative (non- infected) dogs, and 12 healthy controls. CSF samples were analyzed using Reverse Transcription Polymerase Chain Reaction (RT-PCR) for CDV sequences and the sandwich enzyme-linked immunosorbent assay (ELISA) method for quantifying IL-6 and TNF-α levels. While a significant increase in CSF cytokine levels was observed between the CDV- infected and healthy controls, no significant differences were detected between the CDV-infected and non-infected diseased groups. Conclusions The elevated levels of IL-6 and TNF-α in the CSF of dogs with nervous distemper suggest their involvement in disease pathogenesis. However, their lack of specificity limits their use as reliable diagnostic biomarkers for distinguishing nervous distemper from other neurological conditions in dogs. Further research is needed to identify more accurate and specific biomarkers for the precise diagnosis and effective management of CDV infection.

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