Aymée Luiza, Nogueira Di Azevedo Maria Isabel, de Souza Pedrosa Juliana, Loria de Melo Juliana Dos Santos, Carvalho-Costa Filipe Anibal, Lilenbaum Walter
Laboratory of Veterinary Bacteriology, Biomedical Institute, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil.
Laboratory of Epidemiology and Molecular Systematics, Oswaldo Cruz Institute, Rio de Janeiro, Brazil.
Vet Microbiol. 2022 May;268:109413. doi: 10.1016/j.vetmic.2022.109413. Epub 2022 Mar 30.
Bovine Genital Leptospirosis (BGL) is an important reproductive disease. The main agents are Sejroe strains, particularly the Hardjo genotypes from Leptospira interrogans and L. borgpetersenii. Although other Sejroe strain, L. santarosai genotype Guaricura, has been frequently isolated from asymptomatic and slaughtered cattle, even from vaginal fluid samples, the role of this strain as real agent of BGL remains uncertain. This study aimed to reinforce L. santarosai strain Guaricura as an important BGL agent, through genetic characterization of a uterine isolate from a live subfertile cow. Urine, cervicovaginal mucus (CVM) and uterine fragment (UF) were collected. In a set up field laboratory, urine, CVM and UF were immediately seeded in T80/40LH medium with antimicrobial cocktail STAFF. Cultures were subcultured in T80/40LH without cocktails, stored at 29ºC and weekly examined. DNA from urine, CVM and UF samples were submitted to PCR targeting lipL32 and secY genes. One leptospiral isolate was recovered from uterine sample; it was serogrouped as Sejroe (titre 25,600) and secY sequencing revealed high genetic similarity with L. santarosai strains from Guaricura serovar. The isolation of this strain from uterus of a live subfertile cow represents substantial evidence that L. santarosai strain Guaricura indeed plays an important role as a BGL agent.
牛生殖器钩端螺旋体病(BGL)是一种重要的生殖疾病。主要病原体是 sejroe 菌株,特别是问号钩端螺旋体和博氏钩端螺旋体的 Hardjo 基因型。尽管其他 sejroe 菌株,即圣塔罗莎钩端螺旋体 Guaricura 基因型,经常从无症状和屠宰的牛中分离出来,甚至从阴道液样本中分离出来,但该菌株作为 BGL 的真正病原体的作用仍不确定。本研究旨在通过对一头活体亚生育能力母牛子宫分离株的基因特征分析,强化圣塔罗莎钩端螺旋体 Guaricura 菌株作为重要 BGL 病原体的地位。收集了尿液、宫颈阴道黏液(CVM)和子宫片段(UF)。在一个现场实验室中,立即将尿液、CVM 和 UF 接种到含有抗菌鸡尾酒 STAFF 的 T80/40LH 培养基中。培养物在不含鸡尾酒的 T80/40LH 中传代培养,保存在 29℃,每周检查一次。将尿液、CVM 和 UF 样本的 DNA 进行针对 lipL32 和 secY 基因的 PCR 检测。从子宫样本中分离出一株钩端螺旋体;血清分组为 sejroe(滴度 25,600),secY 测序显示与 Guaricura 血清型的圣塔罗莎钩端螺旋体菌株具有高度遗传相似性。从一头活体亚生育能力母牛的子宫中分离出该菌株,充分证明圣塔罗莎钩端螺旋体 Guaricura 菌株确实作为 BGL 病原体发挥着重要作用。