Warble flies (Hypoderma spp.) cause large financial losses to the livestock sector on the Qinghai-Tibet Plateau, but little is known about the molecular adaptations that enable them to survive at high elevations. A comparative proteomic analysis was conducted on third-instar larvae from three important species—H. bovis, H. sinense, and the endemic H. pantholopsum—was motivated by this knowledge gap. By integrating mass spectrometry and two-dimensional differential in-gel electrophoresis (2D-DIGE), a total of 54 differentially expressed proteins were identified, which were enriched in stress-response biomarkers such as heat shock protein HSC72 and HSP60, and predominantly exhibited acidic properties with isoelectric points ranging from 3.78 to 6.76, and differentially expressed (P<0.05). According to functional profiling, the main molecular functions were binding (39.4%) and catalytic activity (37.9%), with cellular and metabolic processes receiving a significant amount of attention (30.0% each). Only two proteins were found to be conserved across species, which was a startling discovery in contrast to the 41.5% of differential proteins that were specific to the H. bovis and H. pantholopsum pair. Comparative proteomic analysis demonstrated the upregulation of pathways associated with protein homeostasis and energy metabolism in H. pantholopsum, a species highly adapted to high-altitude habitats. Phylogenetic analysis of HSC72 uncovered functional conservation, indicating an adaptive response molded by long-term evolutionary processes. Collectively, our findings identify potential targets for future species-specific control strategies against hypodermosis on the Qinghai-Tibet Plateau, while also unraveling key molecular mechanisms that underpin the adaptation of these parasitic larvae to the extreme high-altitude environment.

To Cite This Article: Wang S, Li Z, Meng R, Duo H, Zhang X, Shen X, Li J, Lin Y, Caidan D, Zeng X, Chen W, Almutairi MH and Fu Y, 2026. Comparative proteomics reveals molecular adaptations to high-altitude stress in three Hypoderma species. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2026.008