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Barbour‐Stoenner‐Kelly II (BSK‐II) and BSK‐H media were used for cultivation and isolation of fastidious Borrelia species ‐ the causative agents of Lyme borreliosis. Culture media have a limited shelf life and require adequate storage. Our goal was to assess how the growth of Borrelia would be affected by prolonged storage of media and inadequate storage conditions (BSK‐H stored at +4 °C for 2.5 years and BSK‐II stored at −20 °C for 11 years). Growth of different Borrelia afzelii, Borrelia garinii, Borrelia lusitaniae and Borrelia valaisiana strains was assessed during 2 weeks of incubation at 33 °C. Monitored parameters included cell count per mL, morphology and motility. The results of this study have shown weaker growth of borrelia strains in BSK‐H at +4 °C (median final cell number of 1.5 × 106/mL) than in BSK‐II at −20 °C (median final cell number of 7.75 × 106/mL) and in fresh BSK‐H media (median final cell number of 8.95 × 106/mL). Duration of storage of media had no impact on Borrelia morphology and motility. Our results indicate that temperature of −20 °C is optimal for long‐term storage of medium, BSK‐II stored for 11 years provided effective support to growth of Borrelia and may be employed for cultivation. [ABSTRACT FROM AUTHOR]

The aim of the present study was to assess the possible use of a modified medium, prepared in the laboratory using the constituents of Barbour-Stonner-Kelly (BSK) medium and medium 199 as base, for the culture of Borrelia strains, comparing the growth of individual strains in this medium and in the BSK-H medium, and the protein profile and antigenic characteristics of Borrelia proteins expressed in these media. A qualitative evaluation of growth of Borrelia species was made with acceptable results (morphology and motility), but during a quantitative evaluation using the three main genospecies of Borrelia, the better results were obtained with a B. burgdorferi sensu stricto strain. The modified medium did not enable the growth of a B. afzelii strain. The protein profile and antigenic characteristic of the expressed proteins in the modified medium were studied with satisfactory results. These results suggest the modified medium as an alternative for the cultivation of Borrelia strains, with some limitations, in poorly-resourced laboratories.