Influence of MKP medium stored for prolonged periods on growth and morphology ofBorrelia afzelii, Borrelia garinii, and Borrelia burgdorferisensu stricto

Abstract

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.

A Practical Approach to the Diagnosis of Lyme Borreliosis: From Clinical Heterogeneity to Laboratory Methods

“…Although in vitro cultivation of Borrelia from clinical samples represents the golden standard for proving an active infection, this method cannot be routinely used for diagnosis as it is time consuming and has low clinical sensitivity ( 54 , 164 ). Borrelia burgdorferi sensu lato culture can be obtained from various tissues and body fluids with variable yield using dedicated media, such as the modified Kelly-Pettenkofer medium (MKP), the Barbour-Stoenner-Kelly II (BSK-II) medium, and the commercially available BSK-H medium ( 165 , 166 ). Borrelia cultivation from clinical samples is mostly successful from skin biopsy when compared to blood and CSF cultures ( 165 , 167 ).…”

“…Borrelia burgdorferi sensu lato culture can be obtained from various tissues and body fluids with variable yield using dedicated media, such as the modified Kelly-Pettenkofer medium (MKP), the Barbour-Stoenner-Kelly II (BSK-II) medium, and the commercially available BSK-H medium ( 165 , 166 ). Borrelia cultivation from clinical samples is mostly successful from skin biopsy when compared to blood and CSF cultures ( 165 , 167 ).…”

Diagnostic Tools for Borrelia Assessment in Humans

“…Borrelia burgdorferi sensu lato can be cultivated from various tissues and body fluids in modified Kelly-Pettenkofer medium (MKP), Barbour-Stoenner-Kelly II (BSK-II) medium, and commercially available BSK-H medium [3,18]. The yield of Borrelia culture from clinical samples is usually low with a higher rate of positivity from skin biopsy compared to blood and CSF cultures [18]. Borrelia cultivation and isolation is a time-consuming and demanding procedure.…”

“…Borrelia cultivation and isolation is a time-consuming and demanding procedure. Successful cultivation can take up to 12 weeks [18]. In addition, preparation of culture media is demanding and expensive.…”

Comparison of growth and morphology of Borrelia burgdorferi sensu lato in BSK‐H and BSK‐II media stored for prolonged periods

“…The price of commercial media is quite high, while preparation of culture media “in‐house” is demanding and requires sterile working conditions due to the susceptibility of borrelia strains to nonstandard conditions and contaminants (13, 16). All media are enriched with supplements that have limited shelf life and require frequent preparation of new batches and adequate storage in order to provide effective support of growth of borrelia strains in vitro (10–12, 16).…”