Hi,

I have been trying to isolate L. borgpetersenii and L.interrogans from kidneys and bladder tissues of wild rats. I have been using BD Difco Leptospira Base EMJH medium, enriched with BD Leptospira enrichment, with 0.1% 5'-Fluorouracil.

I mashed up ~10mg of fresh kidney and bladder samples in 500ul of EMJH and transfer all into another 4ml of EMJH. I do see quite a few leptospires moving immediately after inoculation, however at the second week there are more dead than viable ones. I also tried passaging, but then there will be leptospira observed in the second passage. Cultures were all incubated at 30deg in normal air and without shaking.

May I ask if anyone have faced such difficulties and kindly provide advice on howl to isolate and culture Leptospira from kidney and bladder tissues?

Thanks.

Marcella