Formation and cultivation of Borrelia burgdorferi spheroplast-L-form variants.

 Infection. 1996 May-Jun;24,3:218-26. Erratum in: Infection 1996 Jul-Aug;24,4:335. 
Formation and cultivation of Borrelia burgdorferi spheroplast-L-form variants. Mursic VP, 
Wanner G, Reinhardt S, Wilske B, Busch U, Marget W.
Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität München, Germany.
As clinical persistence of Borrelia burgdorferi in patients with active Lyme borreliosis occurs 
despite obviously adequate antibiotic therapy, in vitro investigations of morphological variants 
and atypical forms of B. burgdorferi were undertaken. In an attempt to learn more about the 
variation of B. burgdorferi and the role of atypical forms in Lyme borreliosis, borreliae isolated 
from antibiotically treated and untreated patients with the clinical diagnosis of definite and 
probable Lyme borreliosis and from patient specimens contaminated with bacteria were 
investigated. Furthermore, the degeneration of the isolates during exposure to penicillin G in 
vitro was analysed. Morphological analysis by darkfield microscopy and scanning electron 
microscopy revealed diverse alterations. Persisters isolated from a great number of patients, 
60-80%, after treatment with antibiotics had an atypical form. The morphological alterations in 
culture with penicillin G developed gradually and increased with duration of incubation. 
Pleomorphism, the presence of elongated forms and spherical structures, the inability of cells 
to replicate, the long period of adaptation to growth in MKP-medium and the 
mycoplasma-like colonies after growth in solid medium, PMR agar, suggest that B. 
burgdorferi produce spheroplast-L-form variants. With regard to the polyphasic course of 
Lyme borreliosis, these forms without cell walls can be a possible reason why Borrelia survive in 
43the organism for a long time, probably with all beta-lactam antibiotics, [corrected] and the 
cell-wall-dependent antibody titers disappear and emerge after reversion

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