Formation and cultivation ofBorrelia burgdorferi spheroplast-L-form variants

Abstract

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 the organism for a long time (probably with all beta-lactam antibiotics) [corrected] and the cell-wall-dependent antibody titers disappear and emerge after reversion.

Is there a Lyme-like disease in Australia? Summary of the findings to date

“…Alleged L-form variant “rigid” SLOs of Borrelia have been described in other papers, from cultured biopsy and synovial fluid samples from Lyme Borreliosis patients [67], [68], in animal skin sample cultures [69] and in control Borrelia cultures subjected to antispirochaetal agents [70], [71]. However, the SLOs in contaminated cultures observed under electron microscopy have been identified by some researchers [69], [72], [73] as large flagella aggregates from the contaminating bacteria, and therefore not indicative of the presence of Borrelia spirochaetes.…”

“…It is possible that this is what was observed in the cultures conducted by Wills and Barry [66]. However, this does not explain the return of such rigid SLOs to normal, motile spirochaetes after multiple subcultures [66], [67]. SLOs in uncontaminated cultures have been observed by others and can be explained by the flagella passing though filters that block whole bacteria and the flagella then coalesce to form the long SLOs (Doggett, S. pers.…”

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

“…Morphological variants of Bb, some of which are not culturable, are well documented in the medical literature ( Barthold et al , 2010; Hodzic et al , 2014; Kurtti et al , 1987; MacDonald, 2013; Meriläinen et al , 2015; Mursic et al , 1996). These variants may play a role in infection, enabling Bb and other pathogenic spirochetes to evade the immune system ( Döpfer et al , 2012; Menten-Dedoyart et al , 2012; Mursic et al , 1996). Limited Bb growth and non-spiral morphology are thought to be induced by unfavorable environmental conditions ( Brorson et al , 2009), and these features appear to be consistent with our observations.…”

The clinical significance of biochemical failure in the first 2 years in patients treated with neo-adjuvant androgen deprivation and radiotherapy for prostate cancer

“…He notes that these cystic structures, which Bockenstedt et al, observed in their infected animals ( Figure 2 in their study [10]), have been described as a persister mechanism employed by many bacteria, including B. burgdorferi [16][17][18][19][20][21][22][23][24]. The study authors claim that these are not true cysts because they form too fast, appearing in minutes rather than hours or days.…”