Search

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 × 10 6 /mL) than in BSK-II at -20 °C (median final cell number of 7.75 × 10 6 /mL) and in fresh BSK-H media (median final cell number of 8.95 × 10 6 /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., (© 2020 APMIS. Published by John Wiley & Sons Ltd.)

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.

Modified Kelly-Pettenkofer (MKP) medium is one of the several media used for isolation and cultivation of Borrelia. The aim of the study was to assess whether particular Borrelia species (B. afzelii, B. garinii, and B. burgdorferi sensu stricto) have the ability to grow in MKP medium stored at +4 °C for periods for 1 month up to 1 year, and how prolonged storage may influences Borrelia growth and morphology. The growth of Borrelia was evaluated after 5 days of incubation at 33 °C: cell count per mL, morphology, and motility were assessed. The results of this study showed that the duration of storage of MKP medium had statistically significant influence on growth of B. afzelii (p = 0.021) and B. garinii (p = 0.004), but not on growth of B. burgdorferi sensu stricto (p = 0.204), whereas duration of storage of the medium had no impact on Borrelia morphology and motility. The results of the study indicate that medium stored for more than 1 and up to 12 months supports Borrelia growth., (© 2013 APMIS. Published by John Wiley & Sons Ltd.)

Broth microdilution and macrodilution assays were used to determine minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of six antimicrobial agents (ceftriaxone, cefuroxime sodium, azithromycin, amoxicillin, doxycycline and amikacin) for nine European human isolates of Borrelia burgdorferi sensu stricto (s.s.). Strains were obtained from patients diagnosed with Lyme borreliosis in Slovenia. Modified Kelly-Pettenkofer medium with a final inoculum of 10⁵ Borrelia cells/mL and incubation periods of 72 h and of 3 weeks and 6 weeks were used in the determination of MICs and MBCs, respectively. Observed MICs indicated that all isolates were susceptible to all the tested antimicrobial agents with the exception of amikacin. Cefuroxime sodium (MIC₉₀=0.063 mg/L), azithromycin (MIC₉₀=0.22 mg/L) and ceftriaxone (MIC₉₀=0.25 mg/L) displayed the lowest MICs, followed by amoxicillin (MIC₉₀=1 mg/L) and doxycycline (MIC₉₀=2 mg/L); no strain was susceptible to amikacin (MIC₉₀=256 mg/L). MBCs after incubation for 3 weeks and 6 weeks were determined for amoxicillin (MBC₉₀=32 mg/L), doxycycline (MBC₉₀)=32 mg/L) and amikacin (MBC₉₀=1024 mg/L) and were found to be high (but not defined) for azithromycin (MBC₉₀>0.88 mg/L), cefuroxime sodium (MBC₉₀>4 mg/L) and ceftriaxone (MBC₉₀>4 mg/L). In determination of borrelial susceptibility to antimicrobial agents, intrinsic low susceptibility or methodological factors could result in low in vitro susceptibility of individual strains. This study is the first report on the antibiotic susceptibility of a series of European human isolates of B. burgdorferi s.s., (Copyright © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.)

Human activities such as deforestation, urbanization, and environmental pollution lead to a reduction in the spatial boundary between wild animals, domestic animals and humans. These activities increase the risk for the emergence of pathogens from the sylvatic cycle in the population of domestic animals and humans. Foxes are recognized as potential reservoirs for a number of bacterial pathogens of medical and public health concern. The aim of the present study was to investigate the prevalence and spatial distribution of bacterial tick-borne pathogens from the Anaplasmataceae family, Borrelia burgdorferi sensu lato (s.l.), Rickettsia spp., Coxiella burnetii, Francisella tularensis, Bartonella spp., in the red fox population from Serbia and to discuss the obtained results from the epidemiological point of view. Legally hunted red foxes (Vulpes vulpes) from 14 localities in Serbia were included in the study and spleen samples from 129 animals were tested with conventional PCR assays for the presence of bacterial tick-borne pathogens. DNA of Candidatus Neoehrlichia sp. (FU98), Borrelia burgdorferi sensu stricto, Borrelia lusitaniae, and Borrelia garinii was detected in 6 (4.7%), 1 (0.8%), 2 (1.6%) and 1 (0.8%) animals, respectively. Co-infection by Candidatus Neoehrlichia sp. (FU98) and B. garinii was detected in one animal. All samples were negative for other tested bacterial tickborne pathogens. The results of the present study indicate the potential role of foxes in natural cycles of Candidatus Neoehrlichia sp. (FU98) and causative agents of Lyme borreliosis in the investigated areas. Further research is required to elucidate the role of foxes in the epidemiology of these and other tick-borne zoonotic pathogens in the Republic of Serbia. [ABSTRACT FROM AUTHOR]