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Comparative Study
. 2016 Jun 16;11(6):e0157706.
doi: 10.1371/journal.pone.0157706. eCollection 2016.

Comparison of Growth of Borrelia Afzelii, Borrelia Garinii, and Borrelia Burgdorferi Sensu Stricto at Five Different Temperatures

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Free PMC article
Comparative Study

Comparison of Growth of Borrelia Afzelii, Borrelia Garinii, and Borrelia Burgdorferi Sensu Stricto at Five Different Temperatures

Gorana Veinović et al. PLoS One. .
Free PMC article

Abstract

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.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Growth of Borrelia afzelii (Ba) strains at 4, 23, 28, 33, and 37°C. Median (■), P25 (♦) and P75 (▲) are expressed as number of cells/mL.
Fig 2
Fig 2. Growth of Borrelia garinii (Bg) strains at 4, 23, 28, 33 and 37°C. Median (■), P25 (♦) and P75 (▲) are expressed as number of cells/mL.
Fig 3
Fig 3. Growth of Borrelia burgdorferi sensu stricto (Bbss) strains at 4, 23, 28, 33 and 37°C. Median (■), P25 (♦) and P75 (▲) are expressed as number of cells/mL.
Fig 4
Fig 4. Growth of Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi sensu stricto at different temperatures expressed as number of Borrelia cells/mL and presented as median (■), P25 (♦) and P75 (▲); A: growth at 4°C, B: 23°C, C: 28°C D: 33°C and E: 37°C.

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Grant support

This study was supported by the Ministry of Higher Education, Science and Sport of Slovenia (grant no. P3-0083).
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