Brugh M
U.S. Department of Agriculture, Athens, Georgia 30605.
Avian Dis. 1988 Oct-Dec;32(4):695-703.
A combination of in vitro and in vivo selection procedures was used to examine the possibility that certain mildly pathogenic field isolates of avian influenza (AI) virus may contain minority subpopulations of highly pathogenic virus. Two mildly pathogenic H5N2 isolates, A/chicken/New Jersey/12508/86 (NJ12508) and A/chicken/Florida/27716/86 (FL27716), recovered from chickens epidemiologically associated with urban live-bird markets, were cloned in trypsin-free chicken embryo fibroblast cultures. Selected clones were inoculated intranasally and intratracheally (IN/IT) into specific-pathogen-free laying hens, and virus reisolated from the hens that died was serially passed in hens by IN/IT inoculation. Several highly pathogenic reisolates were recovered from hens infected with the cloned NJ12508 or FL27716 virus. A highly pathogenic NJ12508 reisolate killed 19 of 24 IN/IT-inoculated hens, and a FL27716 reisolate killed all 24 inoculated hens; signs and lesions were typical of fowl plague. In contrast, uncloned NJ12508 stock virus killed 1 of 24 hens and FL27716 stock virus killed 4 of 24 hens, and neither produced the complete spectrum of lesions associated with fowl plague. Recovery of highly pathogenic viruses from these isolates demonstrates the coexistence of pathogenically distinct subpopulations of virus. Competition for dominance among such subpopulations could explain the variable pathogenicity of some AI viruses.
采用体外和体内选择程序相结合的方法,来检验某些低致病性禽流感(AI)病毒的野外分离株可能含有高致病性病毒的少数亚群这一可能性。从与城市活禽市场有流行病学关联的鸡群中分离出的两株低致病性H5N2分离株,即A/鸡/新泽西/12508/86(NJ12508)和A/鸡/佛罗里达/27716/86(FL27716),在无胰蛋白酶的鸡胚成纤维细胞培养物中进行克隆。挑选出的克隆株经鼻内和气管内(IN/IT)接种到无特定病原体的产蛋母鸡中,从死亡母鸡中重新分离出的病毒通过IN/IT接种在母鸡中连续传代。从感染克隆的NJ12508或FL27716病毒的母鸡中获得了几株高致病性的重新分离株。一株高致病性的NJ12508重新分离株致使24只经IN/IT接种的母鸡中有19只死亡,一株FL27716重新分离株致使所有24只接种母鸡死亡;症状和病变均为典型的禽瘟表现。相比之下,未克隆的NJ12508原种病毒致使24只母鸡中有1只死亡,FL27716原种病毒致使24只母鸡中有4只死亡,且两者均未产生与禽瘟相关的完整病变谱。从这些分离株中获得高致病性病毒,证明了病毒致病性不同的亚群共存。这些亚群之间争夺主导地位的竞争可以解释一些禽流感病毒致病性的差异。