This study aimed to assess the impact of seasonal thermal stress on oxidative stress, immune response, and stress hormones of lactating dairy cows in subtropical regions with different levels of temperature-humidity index (THI). A total of 32 healthy lactating Holstein dairy cows experienced 4 seasons (8 cows/season). The physiological parameters were categorized into low THI (LTHI, THI = 42.97 ± 0.95) in winter, moderate THI (MTHI, THI = 61.84 ± 0.42) in spring and autumn, and high THI period (HTHI, THI = 86.09 ± 0.23) in summer. The blood samples were collected twice in each season to measure oxidative stress, inflammatory and hormonal parameters. Our results showed THI had a positive correlation with the rectal temperature (R ² = 0.821, P < 0.001) and respiratory rate (R ² = 0.816, P < 0.001). Dry matter intake, milk yield and fat percentage also significantly differed among groups (P < 0.05). Compared with the MTHI group, the LTHI group exhibited a significant increase in malondialdehyde (MDA) level (P < 0.001), and the HTHI group displayed a significant increase in levels of cortisol, interleukin (IL)-10, IL-1β and tumor necrosis factor-α (P < 0.001). Opposite changes in serum endotoxin and immunoglobulin G levels were observed with the increasing THI (P < 0.001). LTHI notably increased the triiodothyronine level, although the thyroxine level was reduced by LTHI and HTHI compared with the MTHI group. In conclusion, LTHI and HTHI conditions may induce different degrees of oxidative stress, inflammation response, and stress hormone imbalances on lactating dairy cows, therefore environmental management is necessary for the health of dairy cows in extreme weather conditions.

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