School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
Vet Microbiol. 2013 Oct 25;166(3-4):429-37. doi: 10.1016/j.vetmic.2013.06.026. Epub 2013 Jul 9.
The presence of Helicobacter spp. was examined in the liver and in different regions of the gastrointestinal tract (GIT) including the stomach, 3 cm above ileum, ileum, caecum, colon and rectum of 10 ringtail possums (RTPs) and 3 koalas using a combination of microscopy, culture and PCR. Helicobacter was detected in the distal end of the GIT of 7 of 10 RTPs by direct PCR and in all (10/10) RTPs by nested PCR. Five 'S' shaped isolates with bipolar sheathed flagella were isolated from the lower bowel of 3 of the 10 RTPs. 16S rRNA sequence analysis of these 5 isolates confirmed them as potentially novel Helicobacter species. No Helicobacter species were cultured from the koalas, however Helicobacter DNA was detected, in the majority of liver and/or stomach samples of the three koalas and in the colonic region of one koala, using nested PCR. The 16S rRNA gene was sequenced directly from DNA extracted from the homogenised livers and mucus scrapings of the stomach from koala 1 and were confirmed to be Helicobacter species. Based on histopathological examination of sections from the liver and intestine no evidence of infection could be related to the presence of helicobacters in either the RTP or koala. Based on our results, it is possible that diet may influence the detection of Helicobacter species; however this required further investigation.
采用显微镜检查、培养和 PCR 相结合的方法，研究了 10 只长尾袋貂（RTP）和 3 只考拉的肝脏和胃肠道（GIT）不同部位（胃、空肠 3cm 处、空肠、盲肠、结肠和直肠）是否存在螺旋体属。直接 PCR 检测到 10 只 RTP 中有 7 只 RTP 的 GIT 远端存在螺旋体，巢式 PCR 检测到所有 10 只 RTP 均存在螺旋体。从 3 只 RTP 的下消化道分离出 5 株具有双极鞘鞭毛的“S”形分离株。这 5 株分离株的 16S rRNA 序列分析证实它们可能是新的螺旋体属。考拉未培养出螺旋体，但使用巢式 PCR 检测到 3 只考拉的大多数肝和/或胃样本以及 1 只考拉的结肠区域存在螺旋体 DNA。直接从考拉 1 的肝脏和胃粘液刮取物中提取的 DNA 中直接测序 16S rRNA 基因，确认它们为螺旋体属。根据肝和肠切片的组织病理学检查，在 RTP 或考拉中，均未发现与螺旋体存在相关的感染证据。基于我们的结果，饮食可能会影响螺旋体属的检测；但这需要进一步研究。
