除湿利肺消毒散对鸡传染性支气管炎的预防和治疗效果探究

doi:10.16431/j.cnki.1671-7236.2025.01.044

摘要/Abstract

摘要： 【目的】探究复方中药制剂除湿利肺消毒散(简称XDS)对鸡传染性支气管炎病毒(IBV)人工感染试验鸡的预防和治疗作用。【方法】将25枚9日龄SPF鸡胚随机分为5组,经尿囊腔分别接种不同剂量的XDS(1.33、0.665、0.332、0.166、0 g/0.1 mL)与IBV病毒液(10³ EID₅₀)混合液,对照组使用无菌PBS代替XDS。37 ℃孵育36 h,收集尿囊液用于病毒定量。将60只试验鸡随机分为预防组、治疗组、模型组和对照组,每组15只。前3组试验鸡在15日龄通过滴鼻点眼接种0.1 mL 10⁵ EID₅₀/0.1 mL的IBV(gx2021/12-z株),对照组以相同的方式接种同体积无菌PBS。预防组鸡自12日龄通过饮水以含XDS生药量2.66 g/(d·只)给药3 d,治疗组鸡自感染后1 d以相同途径和剂量给药5 d,对照组和模型组鸡不给药。试验期间监测试验鸡死亡率、体重增长情况及临床症状,并在感染后3、7、14 d采集各组鸡肾脏及气管组织,通过苏木精、伊红(HE)染色组织后,用于病理组织学观察,通过实时荧光定量PCR检测各组鸡气管中免疫相关基因(Toll样受体7(TLR7)、干扰素调节因子7(IRF7)、α-干扰素(IFN-α)和IFN-β)、促炎性细胞因子基因(肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6和IL-8)的表达水平及气管和肾脏的病毒载量。【结果】给药浓度0.332、0.665和1.33 g/0.1 mL组鸡胚尿囊液病毒载量均显著低于对照组(P＜0.05),且病毒载量随给药剂量增大而降低。预防组和治疗组鸡全部存活,而模型组鸡死亡率为20%。在临床症状方面,与模型组相比,预防组及治疗组中XDS可有效缓解IBV感染导致鸡的气管啰音、张口呼吸等临床症状;在感染后14 d,模型组鸡体重显著低于对照组(P＜0.05),而其他各组鸡体重与对照组均无显著差异(P＞0.05)。HE染色结果显示,IBV感染后7 d模型组鸡气管纤毛上皮细胞脱落、坏死,而治疗组鸡气管黏膜层完整性较好;IBV感染后14 d模型组鸡肾小管结构破坏,而治疗组鸡肾脏未见明显病理改变,预防组气管及肾脏损伤程度介于模型组和治疗组之间。在病毒载量方面,实时荧光定量PCR结果显示,预防组鸡气管及肾脏的病毒载量在感染后3和14 d显著低于模型组(P＜0.05),治疗组鸡气管及肾脏的病毒载量在观察期内均显著低于模型组(P＜0.05)。预防组和治疗组鸡气管免疫相关基因TLR7、IFN-α表达水平在感染后3 d显著高于模型组(P＜0.05);IBV感染后7 d,预防组鸡气管TLR7基因表达水平显著高于模型组(P＜0.05),治疗组鸡气管基因TLR7、IRF7、IFN-α、IFN-β表达水平均显著高于模型组(P＜0.05)。在IBV感染后3和7 d预防组和治疗组鸡气管TNF-α、IL-1β、IL-6、IL-12基因表达水平均显著低于模型组(P＜0.05)。【结论】除湿利肺消毒散可有效缓解IBV感染鸡的临床症状,促进鸡体重增长,降低感染鸡气管及肾脏病毒载量并减轻组织病理损伤,维持鸡体内的免疫稳态,对鸡传染性支气管炎具有较好的预防和治疗作用。

关键词: 鸡传染性支气管炎; 除湿利肺消毒散; 预防; 治疗

Abstract: 【Objective】 The aim of this study was to investigate the preventive and therapeutic effects of the compound traditional Chinese medicine preparation Chushi Lifei Xiaodusan (XDS) on chickens artificially infected with Infectious bronchitis virus (IBV). 【Method】 Twenty-five 9-day-old SPF chicken embryos were randomly divided into five groups and inoculated via the allantoic cavity with different doses of XDS (1.33, 0.665, 0.332, 0.166, and 0 g/0.1 mL) mixed with IBV virus solution (10³ EID₅₀). The control group received sterile PBS instead of XDS. After incubation at 37 ℃ for 36 hours, allantoic fluid was collected for viral quantification. Sixty experimental chickens were randomly divided into prevention,treatment,model and control groups,with 15 chickens in each group.Chickens in the first three groups were inoculated intranasally and intraocularly with 0.1 mL of IBV (gx2021/12-z strain) at 15 days old at a dose of 10⁵ EID₅₀/0.1 mL,while chickens in control group received an equal volume of sterile PBS.Chickens in prevention group received XDS in drinking water at a dose of 2.66 g/d per chicken starting from 12 days old for 3 days.Chickens in treatment group received XDS in the same manner and dose for 5 days starting from 1 day post-infection.Chickens in control and model groups did not receive XDS.Throughout the experiment,the mortality rate,weight gain and clinical symptoms of the chickens were monitored.Trachea and kidney tissues were collected 3,7 and 14 days post-infection to stain with hematoxylin and eosin for histopathological observation, measure the expression of immune-related genes (TLR7,IRF7,IFN-α and IFN-β) and pro-inflammatory cytokine genes (TNF-α,IL-1β,IL-6 and IL-8) using Real-time quantitative PCR,and to determine the viral load in trachea and kidney. 【Result】 The viral loads in the allantoic fluid of chicken embryos in the 0.332, 0.665, and 1.33 g/0.1 mL dosage groups were significantly lower than those in the control group (P＜0.05), and the viral loads decreased with increasing dosage. All chickens in prevention and treatment groups survived,while the chickens in model group had a mortality rate of 20%.In terms of clinical symptoms,compared to model group,XDS effectively alleviated clinical symptoms such as tracheal rales and open-mouth breathing in prevention and treatment groups. 14 days post-infection,the body weight of chickens in model group was significantly lower than that in control group (P＜0.05),whereas no significant differences were observed among the other groups (P＞0.05).HE staining results showed that 7 days post-infection,tracheal ciliated epithelial cells of chickens in model group exhibited shedding and necrosis,while chickens in treatment group maintained better tracheal mucosal integrity. 14 days post-infection,chickens in model group displayed damaged renal tubule structures,whereas chickens in treatment group showed no obvious pathological changes. The extent of tracheal and renal damage in prevention group was intermediate between that of model group and treatment group. In terms of viral load,Real-time quantitative PCR results indicated that the viral load in the trachea and kidneys of chickens in prevention group was significantly lower than that of model group 3 and 14 days post-infection (P＜0.05).The viral load of chickens in treatment group was consistently lower than that of model group throughout the observation period (P＜0.05).The expression levels of immune-related genes TLR7 and IFN-α of chicken trachea were significantly higher in prevention and treatment groups than that in model group 3 days post-infection (P＜0.05). 7 days post-infection,TLR7 gene expression level of chicken trachea were significantly higher in prevention group,and TLR7,IRF7,IFN-α and IFN-β gene expression levels of chicken trachea were significantly higher in treatment group compared to model group (P＜0.05).The expression levels of pro-inflammatory cytokine genes TNF-α,IL-1β, IL-6 and IL-12 of chicken trachea were significantly lower in prevention and treatment groups than that in model group 3 and 7 days post-infection (P＜0.05). 【Conclusion】 XDS effectively alleviated clinical symptoms,enhanced weight gain,reduced viral load in the trachea and kidney,mitigated pathological damage in trachea and kidneys of chicken,and maintained immune homeostasis in chickens infected with IBV.It had good preventive and therapeutic effect on infectious bronchitis of chicken.

Key words: infectious bronchitis; Chushi Lifei Xiaodusan; prevention; treatment

中图分类号:

S859.7

杨帆, 周俊, 李绮婷, 林巧儿, 梁维佳, 蔡仕楷, 覃丽梅. 除湿利肺消毒散对鸡传染性支气管炎的预防和治疗效果探究[J]. 中国畜牧兽医, 2025, 52(1): 481-490.

YANG Fan, ZHOU Jun, LI Qiting, LIN Qiaoer, LIANG Weijia, CAI Shikai, QIN Limei. Preventive and Therapeutic Effects of Chushi Lifei Xiaodusan on Infectious Bronchitis in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 481-490.

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