On post-hatch day 21, 120 chicks from each incubation group (control group, TML group, and TMH group) were randomly selected and further subdivided into three subgroups (thermal neutral [TN], cold stress [CS], and HS) to be used in the thermal stress experiment (for a duration of 7 days). For 7 days, TN subgroups were kept at 21°C, CS subgroups were exposed to 16°C, and HS subgroups were exposed to 35°C. On days 1, 4, and 7 of the thermal stress experiments (post-hatch days 22, 25, and 28), nine chicks were randomly selected from each subgroup and their Tb and BW were recorded. After 1 day of thermal stress, ileum samples were collected from each subgroup and snap-frozen for subsequent total RNA extraction. Then, the chicks exposed to 4 and 7 days of thermal stress were humanely euthanized to collect jejunum and ileum samples (about 1 cm), which were maintained in a 10% neutral buffered formalin solution for histological morphometric evaluation.

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