Nishigaki Alice, Previdelli Renato, Alexander James L, Balarajah Sharmili, Roberts Lauren, Marchesi Julian R
Royal Veterinary College, 4 Royal College Street, London NW1 0TU, UK.
Division of Digestive Diseases, Department of Metabolism, Digestion and Reproduction, Imperial College London, St. Mary's Hospital, London W2 1NY, UK.
Animals (Basel). 2023 Sep 4;13(17):2807. doi: 10.3390/ani13172807.
Stool sampling is a useful tool for diagnosing gastrointestinal disease in veterinary medicine. The sub-clinical disease burden of spp. in cattle can become significant for farmers. However, current methods of faecal sampling in a rural setting for diagnosis are not consistently sufficient for the preservation of spp. in faeces. This study evaluated the use of a commercial stool storage kit for bacterial preservation in cow faecal samples compared to unpreserved stools placed into refrigeration at different time-points. A stool sample was collected per-rectum from one apparently healthy Holstein-Freisen cow. The sample was weighed and aliquoted into two sterile Falcon tubes and into two commercial kit tubes. The aliquots were then placed into refrigeration at 4 °C at 0, 24, and 96 h after processing. One commercial kit tube was not aliquoted and remained at ambient temperature. After 2 weeks, DNA was extracted from the samples and analysed using endpoint PCR, revealing a sub-clinical infection with spp. The bacterium was best preserved when the stool was stored in the commercial kit at ambient temperature and re-homogenised immediately prior to DNA extraction. The unpreserved stool did not maintain obvious levels of spp. after 24 h at ambient temperature. This commercial kit should be considered for use in the diagnosis of salmonellosis in cattle.
粪便采样是兽医学中诊断胃肠道疾病的一种有用工具。牛体内 spp. 的亚临床疾病负担对养殖户来说可能相当严重。然而，目前在农村环境中用于诊断的粪便采样方法，在保存粪便中的 spp. 方面并不始终充分。本研究评估了一种商用粪便保存试剂盒在保存奶牛粪便样本中的细菌方面的用途，并与在不同时间点放入冰箱的未保存粪便进行了比较。从一头明显健康的荷斯坦 - 弗里生奶牛经直肠采集一份粪便样本。称取样本重量后，将其分装到两个无菌 Falcon 管和两个商用试剂盒管中。分装后的样本在处理后 0、24 和 96 小时放入 4°C 的冰箱中。一个商用试剂盒管未分装，保持在室温下。两周后，从样本中提取 DNA 并使用终点 PCR 进行分析，发现存在 spp. 的亚临床感染。当粪便保存在室温下的商用试剂盒中，并在提取 DNA 前立即重新匀浆时，细菌保存效果最佳。未保存的粪便在室温下放置 24 小时后， spp. 水平未保持明显。在牛沙门氏菌病的诊断中应考虑使用这种商用试剂盒。
