Van Thi Thu Hao, Gor Mian-Chee, Anwar Arif, Scott Peter C, Moore Robert J
School of Science, RMIT University, Bundoora, Victoria 3083, Australia.
Scolexia Pty. Ltd., Moonee Ponds, Victoria 3039, Australia.
Vet Microbiol. 2017 Aug;207:226-230. doi: 10.1016/j.vetmic.2017.06.022. Epub 2017 Jun 29.
Spotty liver disease (SLD) causes significant egg production losses and mortality in chickens and is therefore a disease of concern for some sectors of the poultry industry. Although the first reports of the disease came from the United States in the 1950s it is only recently that the organism that causes the disease was identified, isolated, and characterised as a new bacterial species, Campylobacter hepaticus. The first isolations of C. hepaticus were from the livers and bile of SLD affected birds. Isolates could only be recovered from samples that had a monoculture of C. hepaticus in the tissues, as a selective culturing method has not yet been developed. In non-selective growth conditions the slow growing C. hepaticus is quickly outgrown by many other members of the chicken microbiota. Therefore, it is currently not possible to use a culturing approach to evaluate C. hepaticus carriage in tissues, such as the gastrointestinal tract (GIT), that also carry complex microbial populations. As it is suspected that birds become infected via the faecal-oral route it is important that pathogen carriage in the GIT is investigated. In the present study, a specific and sensitive quantitative real-time PCR assay, based on the glycerol kinase gene of C. hepaticus, was developed. The assay facilitated the detection and quantification of C. hepaticus in tissue samples from clinical cases of SLD. It was shown that in infected birds C. hepaticus colonises the small intestine, increasing in abundance from duodenum to ileum, and is at highest levels within the ceaca. C. hepaticus was also readily detected in cloacal swabs, indicating that thecl-oral infection.
斑点肝病(SLD)会导致鸡的产蛋量大幅下降和死亡率上升,因此是家禽行业某些部门关注的一种疾病。尽管该疾病的首次报告于20世纪50年代来自美国,但直到最近才鉴定、分离并将引起该疾病的病原体表征为一种新的细菌物种——肝弯曲杆菌。肝弯曲杆菌的首次分离是从受SLD影响的禽类的肝脏和胆汁中获得的。由于尚未开发出选择性培养方法,只有从组织中肝弯曲杆菌为单一培养物的样本中才能分离出菌株。在非选择性生长条件下,生长缓慢的肝弯曲杆菌会很快被鸡微生物群的许多其他成员超过。因此,目前不可能使用培养方法来评估肝弯曲杆菌在诸如胃肠道(GIT)等也携带复杂微生物群体的组织中的携带情况。由于怀疑禽类是通过粪口途径感染的,因此研究GIT中的病原体携带情况很重要。在本研究中,基于肝弯曲杆菌的甘油激酶基因开发了一种特异性和灵敏的定量实时PCR检测方法。该检测方法有助于检测和定量SLD临床病例组织样本中的肝弯曲杆菌。结果表明,在受感染的禽类中,肝弯曲杆菌定殖于小肠,从十二指肠到回肠丰度增加,在盲肠中含量最高。在泄殖腔拭子中也很容易检测到肝弯曲杆菌,这表明存在粪口感染。