Steinparzer Romana, Mair Tamara, Unterweger Christine, Steinrigl Adi, Schmoll Friedrich
Institute for Veterinary Disease Control, Austrian Agency for Health and Food Safety (AGES), Robert Koch Gasse 17, 2340 Mödling, Austria.
University Clinic for Swine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, Veterinaerplatz 1, 1210 Vienna, Austria.
Vet Sci. 2021 May 25;8(6):90. doi: 10.3390/vetsci8060090.
spp. cause the zoonotic disease leptospirosis, which occurs in numerous mammalians worldwide. Isolation is still important for serotyping and genotyping of , which in turn is essential for epidemiological surveillance of leptospirosis and the development of diagnostic tests and vaccines. However, isolation of from clinical specimens is inherently insensitive. This study was conducted to examine the influence of selective agents, sample filtration, sample pH and the use of phosphate buffered saline (PBS) buffer for sample storage to improve the success of cultivation and isolation of serovar Icterohaemorrhagiae from swine urine. EMJH (Ellinghausen McCullough, Johnson and Harris) medium including the selective agents sulfamethoxazole, trimethoprim, amphotericin, fosfomycin and 5-fluorouracil (STAFF) increased the success of isolation from spiked swine urine samples. Sample filtration yielded only negative results. Isolation in EMJH-STAFF was successful from swine urine with a density as low as 10 /mL, and urine with pH ≤ 7 impaired the cultivation rate. Cultivation and isolation were not improved by the addition of PBS to spiked urine samples prior to storage for 24 h at 4 °C. The results of the study demonstrate that cultivation and isolation of leptospires from swine urine can be improved by enhanced methods.
某些物种会引发人畜共患疾病钩端螺旋体病，该病在全球众多哺乳动物中都有发生。分离培养对于钩端螺旋体的血清分型和基因分型仍然很重要，而这反过来对于钩端螺旋体病的流行病学监测以及诊断测试和疫苗的研发至关重要。然而，从临床标本中分离钩端螺旋体本身就不敏感。本研究旨在考察选择性试剂、样品过滤、样品pH值以及使用磷酸盐缓冲盐水（PBS）缓冲液进行样品储存对提高从猪尿液中培养和分离出血清型黄疸出血型钩端螺旋体成功率的影响。含有磺胺甲恶唑、甲氧苄啶、两性霉素、磷霉素和5-氟尿嘧啶（STAFF）等选择性试剂的EMJH（埃林豪森-麦卡洛、约翰逊和哈里斯）培养基提高了从加标的猪尿液样本中分离钩端螺旋体的成功率。样品过滤仅得到阴性结果。在EMJH-STAFF培养基中，从密度低至10 /mL的猪尿液中成功分离出钩端螺旋体，且pH≤7的尿液会降低培养率。在4℃下将加标尿液样本储存24小时之前添加PBS并不能提高培养和分离效果。该研究结果表明，通过改进方法可以提高从猪尿液中培养和分离钩端螺旋体的成功率。
