The isolation of Leptospira is challenging, since the bacteria of this genus are susceptible to adverse environmental conditions and may not remain viable for extend periods in urine samples. This study attempted to develop and evaluate a simple and practical method to isolate leptospires from bovine urine samples. A culture medium for sample transport, named Leptospira Transport Medium (LTM), was described and validated using reference serovars of Leptospira spp. in addition to autochthonous strains isolated in Brazil. We evaluated LTM in the field, by collecting 215 urine samples from slaughtered cattle and immediately seeding them in LTM and Fletcher's medium, used as control. The cultures were sent to a laboratory within 10 days for further processing. Moreover, 16S PCR was also performed on the urine samples directly to detect Leptospira DNA. Using LTM enabled 52 isolates (24.2%) to be obtained in pure culture, and contamination was only observed in 15/215 samples (7.0%). Regarding the samples in Fletcher's medium, 10 (4.6%) isolates were obtained. With 16S PCR performed in the urine samples, 31 samples (14.4%) were determined to be positive. LTM was developed and used in a simple and practical way and can significantly improve the isolation of leptospires from urine samples, as well as being highly useful in remote areas, not only in Brazil but also in other countries where few easily accessible laboratories are available. Furthermore, LTM can be prepared by laboratories and provided to veterinarians and technicians for urine collection in the field.

钩端螺旋体的分离具有挑战性，因为该属细菌容易受到不利环境条件的影响，并且在尿液样本中可能无法长期存活。这项研究试图开发和评估一种简单实用的方法从牛尿液样品中分离钩端藤。描述了一种用于样品运输的培养基，称为钩端螺旋体运输介质（LTM），并使用钩端螺旋体的参考血清对它们进行了验证spp。除了在巴西分离出的本地菌株。我们在野外评估了LTM，方法是从屠宰牛身上收集215个尿液样本，然后立即将它们播种到LTM和Fletcher培养基中，用作对照。将培养物在10天内发送到实验室进行进一步处理。此外，还直接对尿液样本进行了16S PCR检测钩端螺旋体脱氧核糖核酸。使用LTM可以在纯培养物中获得52个分离株（24.2％），并且仅在15/215个样品中观察到了污染（7.0％）。关于在Fletcher培养基中的样品，获得了10个（4.6％）分离株。通过对尿液样本进行16S PCR，确定了31个样本（14.4％）为阳性。LTM的开发和使用以简单实用的方式进行，可以显着改善钩端螺旋体与尿液样本的分离，并且不仅在巴西而且在其他国家/地区都非常容易获得实验室，在偏远地区非常有用。此外，LTM可以由实验室制备，并提供给兽医和技术人员以在野外收集尿液。