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<p>Cold stress is a significant challenge to buffalo health; however, the molecular mechanisms underlying their adaptation remain insufficiently explored. This study employed quantitative real-time PCR (qRT-PCR) to evaluate the relative gene expression of Toll-like receptors (<i>TLR1</i>–<i>TLR10</i>) in peripheral blood mononuclear cells (PBMCs) of Murrah buffalo calves (<i>n</i> = 6) exposed to graded ambient temperatures (24&#xa0;°C, 21&#xa0;°C, 18&#xa0;°C, and 15&#xa0;°C). Progressive cooling induced notable physiological changes, including a gradual decline in rectal temperature and respiration rate. Transcriptional activation was first detected at 21&#xa0;°C for <i>TLR4</i>, <i>TLR6</i>, and <i>TLR8</i>, and expanded to include all examined TLR genes at 18&#xa0;°C. Peak expression occurred at 15&#xa0;°C, with <i>TLR4</i>, <i>TLR6</i>, and <i>TLR8</i> demonstrating the most pronounced upregulation. Segmented regression analysis indicated a potential transcriptional intensification between 18&#xa0;°C and 21&#xa0;°C for several genes, although these observations should be interpreted cautiously due to the limited sample size. Among the targets, <i>TLR5</i> exhibited the strongest inverse correlation with ambient temperature. Collectively, these findings provide insights into the buffalo-specific innate immune response to cold exposure and identify several TLRs as promising molecular indicators for monitoring cold stress resilience.</p>

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Targeted gene expression of toll-like receptors during cold stress in buffalo calves

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月11日

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  • Vansh Sharma,
  • Priyambada Kumari,
  • Brijesh Yadav,
  • Reetu Kumari,
  • Shanker K. Singh,
  • Arun K. Madan,
  • Manish Tiwari

摘要

Cold stress is a significant challenge to buffalo health; however, the molecular mechanisms underlying their adaptation remain insufficiently explored. This study employed quantitative real-time PCR (qRT-PCR) to evaluate the relative gene expression of Toll-like receptors (TLR1–TLR10) in peripheral blood mononuclear cells (PBMCs) of Murrah buffalo calves (n = 6) exposed to graded ambient temperatures (24 °C, 21 °C, 18 °C, and 15 °C). Progressive cooling induced notable physiological changes, including a gradual decline in rectal temperature and respiration rate. Transcriptional activation was first detected at 21 °C for TLR4, TLR6, and TLR8, and expanded to include all examined TLR genes at 18 °C. Peak expression occurred at 15 °C, with TLR4, TLR6, and TLR8 demonstrating the most pronounced upregulation. Segmented regression analysis indicated a potential transcriptional intensification between 18 °C and 21 °C for several genes, although these observations should be interpreted cautiously due to the limited sample size. Among the targets, TLR5 exhibited the strongest inverse correlation with ambient temperature. Collectively, these findings provide insights into the buffalo-specific innate immune response to cold exposure and identify several TLRs as promising molecular indicators for monitoring cold stress resilience.