Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (13): 2620-2628.doi: 10.3864/j.issn.0578-1752.2023.13.015

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles    

Establishment of a Canine Experimental Infection Model with a H3N2 Subtype Canine Influenza Virus

HUANG Cheng1(), YANG LongFeng2, SUN Peng3, CHENG HuiMin1, YANG ZhiYuan1, LIN Jian1, ZHU HongWei3, LIU LiXin1, SUN HouMin3, LI JiaFeng3, ZHAO JiCheng1, DUAN HuiJuan1, PAN Jie1, LIU YueHuan1()   

  1. 1 Institute of Animal Husbandry and Veterinary Medicine, Beijing Municipal Academy of Agricultural and Forestry Sciences, Beijing 100097
    2 Yanqing District Animal Disease Prevention and Control Center of Beijing Municipality, Beijing 102100
    3 YEBIO Bioengineering Co., Ltd of Qingdao, Qingdao 266113, Shandong
  • Received:2022-03-28 Accepted:2022-08-02 Online:2023-07-01 Published:2023-07-06
  • Contact: LIU YueHuan

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Abstract:

【Objective】 In this research, an experimental animal infection model of canine influenza virus (CIV, H3N2 subtype) was established to better understand the pathogenesis of canine influenza, so as to lay the foundation for vaccine efficacy evaluation. 【Method】 26 beagles aged 6-13 months with negative CIV Haemagglutination Inhibition (HI) antibody (HI<1﹕10) were selected, three beagles of which were challenged by 1mL nasal spray of PBS, and 23 beagles of which were challenged by 1mL nasal spray of H3N2 CIV (A/canine/China/Huabei-170607/2017(H3N2), HB strain for short) with 5 groups (10, 103, 105, 106, and 107 50% EID50) as 3, 5, 5, 5, and 5 beagles each group, respectively. Clinical symptoms, lung lesions, histopathological changes of lung, calculating the proportion of consolidation mass, HI antibody titer and virus shedding were examined at 14 days after virus challenge together with three control beagles. 【Result】 3 beagles inoculated with a dose of 10 EID50 H3N2 CIV did not show any clinical symptoms, gross lesions in lungs and histopathological changes, the consolidation rate was 0%, and the virus shedding was not detected. The geometric mean titer (GMT) of HI antibodies was 1﹕15.9. However, 4/5 of beagles inoculated with a dose of 103 EID50 H3N2 CIV showed the clinical symptoms and virus shedding, such as puffing, runny nose and cough. 2/5 of beagles showed light lung consolidation, whose rate was 1.4%. The GMT of HI antibodies was 1:320. All beagles (5/5) infected with a dose of 105 EID50 H3N2 CIV showed clinical symptoms at day 5 after challenge, such as runny nose and cough, virus shedding, lung consolidation, and widened alveolar septum, and the consolidation rate was 4.2%. The GMT of HI antibodies was 1﹕2 940.7. 5/5 of beagles infected with a dose of 106 EID50 all showed severe clinical symptoms at day 4 after challenge, such as cough and elevated body temperature, virus shedding, obvious pathological features in the lungs, and the GMT of HI antibodies was 1﹕2 228.7. The clinical symptoms appeared earlier 1 day than that in 105 EID50 dose infection group, and the degree of lung consolidation increased. The lung consolidation ratio was 17.9%. 5/5 of beagles infected with doses of 107 EID50 all showed severe clinical symptoms at day 3 after challenge which appeared earlier 1 day than that in 106 EID50 dose infection group, virus shedding and widened alveolar septum in the lungs, the GMT of HI antibodies was 1﹕2 940.7, and the consolidation rate of lung was 29.0%. The control beagles did not show any clinical symptoms, gross lesions and histopathological changes in lungs, the virus shedding was not detected, and GMT of HI antibodies was <1﹕10. 【Conclusion】 The dose of 106 EID50 was the minimum virus inoculation dose that could cause obvious pathogenesis in beagles. An experimental animal infection model of CIV subtype H3N2 in beagles was established.

Key words: beagles, canine influenza virus, H3N2 subtype, lung, infection model

Table 1

Experimental grouping"

组别 Group 数量 Numbers 剂量 Dosage (mL) 病毒含量（EID50）Virus titer 攻毒方式 Challenge routine
A 3 1 0 喷雾Nasal spray
B 3 1 10 喷雾Nasal spray
C 5 1 103 喷雾Nasal spray
D 5 1 105 喷雾Nasal spray
E 5 1 106 喷雾Nasal spray
F 5 1 107 喷雾Nasal spray

Table 2

Clinical symptom score criteria"

症状
Symptom		 程度
Degree		 分数
Score
咳嗽
Cough		 无Nothing 0
出现Presence 2
鼻汁
Runny nose		 无Nothing 0
轻度Mild 1
严重Severe 2
精神状况
Mental status		 健活Alive 0
沉郁Depression 1

Fig. 1

Body temperature variation"

Fig. 2

Clinical symptom scores"

Table 3

Proportion of the lung consolidation"

组别 Group A B C D E F
病毒含量（EID50）Virus titer 阴性对照 Non-challenged group 10 103 105 106 107
肺脏实变比例平均值
Mean value of lung consolidation ratio		 0 0 1.4% 4.2% 17.9% 29.0%

Fig. 3

Gross lesions of the experimental beagles’ lungs"

Fig. 4

Histopathology of the experimental beagles’ lungs"

Table 4

Results of virus shedding and HI antibody titers"

组别 Group A B C D E F
病毒含量（EID50）Virus titer 阴性对照 Non-challenged group 10 103 105 106 107
病毒分离 Virus isolation 0/3 0/3 4/5 5/5 5/5 5/5
HI抗体效价GMT HI antibody GMT <1:10 1:15.9 1:320 1:2940.7 1:2228.7 1:2940.7
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