Abstract：Bordetella bronchiseptica (Bb) is the main pathogen of respiratory infectious in rabbits (Oryctolagus cuniculus). To investigate the effect of cold stress on the infection of Bb in rabbits, 90 New Zealand laboratory rabbits (45-day-old healthy) were randomly divided into 5 groups, injected intranasally with 0.5 mL 4×109 CFU/mL Bb (group: Bb/RT, Bb/cold) or phosphate buffer saline (PBS) alone (group: PBS/RT, control /RT, PBS/cold). Rabbits in Bb/RT group and PBS/RT group (20±2 ℃) were kept in different cages in the same room. Bb/cold group and PBS/cold group (6±2 ℃) were kept in different cages in the same room, while control/RT group (20±2 ℃) were kept, separately. Three rabbits of each group were randomly sampled at 7 h, 10 h, 1 d, 3 d, 7 d and 12 d post-infection. The concentration of Bb in the respiratory tract (nose, trachea, lungs) , heart, liver, and kidney were tested by TaqMan fluorescence quantitative PCR. The changes of rabbit interleukin (IL-10), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ) mRNA were also determined by SYBR-Green reverse transcription (RT-PCR). The results indicated that Bb can spread through the air to rabbits, Bb was detected in nasal cavity of rabbits in both PBS/RT group and PBS/cold group at 7 h. Cold stress had no significant effect on Bb proliferation in the nasal cavity of infected rabbit. The Bb concentration in the nasal cavity of Bb/RT group did not differ significantly from Bb/cold group, PBS/RT group did not differ significantly from PBS/cold group at any time point examined. Bb proliferation in trachea of infected rabbits which exposed to cold stress was faster in the first 3 days. The concentration of Bb in Bb/RT and Bb/cold group was close but significantly higher than PBS/RT group and PBS/cold group (P<0.01) at 12 h, Bb/RT group higher than PBS/RT group (P<0.05) at 1 d and 3 d; there was no significant difference for all infected groups after the 7 d. None of Bb was detected in lungs, livers, kidneys, and hearts of all rabbits. Based on 2-ΔΔCt transformation, IL-10, IFN-γ and TNF-α were found to increase in expression in the blood of infected rabbits. The expression of IL-10 was up-regulated, but there was no significant change. The expression levels of IFN-γ and TNF-α changed among sampling time and peaked at 3 d. However, there was no significant difference in the expression change of IL-10, IFN-γ and TNF-α between different infected groups at the same sampling time. Cold stress could promote the propagation of Bb in trachea of rabbits within 3 d after exposure. Nasal cavity and trachea were the main organs that Bb invades and colonizes. Cold stress did not significantly effect the cytokines expression in Bb infected rabbits. This study provides a technical basis for the prevention and control of Bb infection.
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