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Lyme borreliosis is caused by the spirochete  Borrelia burgdorferi  sensu lato, a fastidious bacterium that replicates slowly and requires special conditions to grow in the laboratory. Borrelia isolation from clinical material is a golden standard for microbiological diagnosis of borrelial infection. Important factors that affect  in vitro  borrelia growth are temperature of incubation and number of borrelia cells in the sample. The  aim  of the study was to assess the influence of temperature on borrelia growth and survival by evaluation and comparison of growth of 31 different borrelia strains at five different temperatures and to determine the influence of different inoculums on borrelia growth at different temperatures. Borreliae were cultured in the MKP medium; the initial and final number of spirochetes was determined by dark field microscopy using Neubauer counting chamber. The growth of borrelia was defined as final number of cells/mL after three days of incubation. For all three  Borrelia  species, the best growth was found at 33°C, followed by 37, 28, and 23°C, while no growth was detected at 4°C ( P< 0.05). The growth of  B .  afzelii  species was weaker in comparison to the other two species at 23, 28, 33 and 37°C ( P <0.05), respectively. There was no statistically significant difference between the growth of  B .  garinii  and  B .  burgdorferi  sensu stricto at 28, 33, and 37°C ( P >0.05), respectively. Inoculum had statistically significant influence on growth of all three  Borrelia  species at all tested temperatures except at 4°C.

Comparison of Growth of Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi Sensu Stricto at Five Different Temperatures