Lyme disease is a serious bacterial infection caused by a tick bite and affects humans and animals.
This page contains citations and complete abstracts for medical and scientific articles from the National Institutes of Health (NIH), National Library of Medicine (NLM) MEDLINE database about persistence or relapse of Lyme disease infection despite "appropriate" or "conventional" antibiotic treatment. Citations are sorted by date within categories with particularly significant portions highlighted in bold red lettering. Links to free online full text articles are indicated by bold lime lettering.
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Could the following researchers have discovered reasons for the survival of the Lyme disease bacteria in some patients even though seemingly adequate antibiotics were used for treatment?
Note: Most of the citations or abstracts below were gleaned from articles found by the following MEDLINE "search" link:
TITLE:
Conversion of Borrelia garinii cystic forms to motile
spirochetes in vivo.
AUTHORS:
Gruntar I, Malovrh T, Murgia R, Cinco M.
AUTHOR AFFILIATION:
Institute of Microbiology and Parasitology, Veterinary Faculty,
Ljubljana, Slovenia. gruntaig@mail.vf.uni-lj.si
SOURCE:
APMIS 2001 May;109(5):383-8
ABSTRACT:
Cystic forms (also called spheroplasts or starvation forms) and their
ability to reconvert into normal motile spirochetes have already been
demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of
this study was to determine whether motile B. garinii could develop from
cystic forms, not only in vitro but also in vivo, in cyst-inoculated
mice. The cysts prepared in distilled water were able to reconvert into
normal motile spirochetes at any time during in vitro experiments,
lasting one month, even after freeze-thawing of the cysts. Motile
spirochetes were successfully isolated from 2 out of 15 mice inoculated
intraperitoneally with cystic forms, showing the infectivity of the
cysts. The demonstrated capacity of the cysts to reconvert into motile
spirochetes in vivo and their surprising resistance to adverse
environmental conditions should lead to further studies on the role and
function of these forms in Lyme disease.
PMID: 11478686
TITLE:
Serum-starvation-induced changes in protein synthesis and morphology
of Borrelia burgdorferi.
Full text article:
Microbiology -- Alban et al. 146 (1): 119
AUTHORS:
Alban PS; Johnson PW; Nelson DR
AUTHOR AFFILIATION:
Department of Biochemistry, Microbiology, and Molecular Genetics,
University of Rhode Island, Kingston 02881, USA.
SOURCE:
Microbiology 2000 Jan;146 ( Pt 1):119-27
ABSTRACT:
It has been demonstrated previously that motile Borrelia burgdorferi
cells transform into non-motile cyst-forms when incubated for several
weeks in BSKII (a complex medium) lacking rabbit serum.
B.
burgdorferi cells cannot synthesize fatty acids de novo and serum is
thought to provide a source of fatty acids and lipids. When B.
burgdorferi cells were serum-starved in defined RPMI medium, -90% of
the cells formed spherical cysts within 48 h. Cyst formation was
inhibited by tetracycline.
Cyst opening and recovery of vegetative
cells was rapidly induced by the addition of either BSKII or rabbit
serum. The percentage of viable cells recovered from cysts ranged
from 2.9% to 52-5%. Viability was inversely proportional to cyst age.
Protein synthesis by B. burgdorferi during serum starvation was
examined by labelling cells with Tran35S-Label and analysing the
labelled proteins by two-dimensional gel electrophoresis and
fluorography. The synthesis of over 20 proteins was induced during
serum starvation. Western blots of proteins from vegetative cells and
cysts probed with sera from either B. burgdorferi-infected humans or
monkeys revealed that several cyst proteins were antigenic.
These
data suggest that cells of B. burgdorferi, although possessing a
small genome and extremely limited biosynthetic capabilities, rapidly
respond to conditions of serum starvation by inducing changes in
protein synthesis and cell morphology. This study may help explain
how cells of B. burgdorferi can survive periods of nutrient
deprivation in different hosts and host tissues.
TITLE:
A rapid method for generating cystic forms of Borrelia burgdorferi, and
their reversal to mobile spirochetes.
AUTHORS:
Brorson O; Brorson SH
AUTHOR
AFFILIATION:
Department of Microbiology, Vestfold Sentralsykehus, Tonsberg, Norway.
SOURCE:
APMIS 1998 Dec;106(12):1131-41
ABSTRACT:
Mobile Borrelia burgdorferi were transferred to distilled water (10(6)
per ml). The cultures were observed by dark field microscopy (DFM),
interference contrast microscopy (ICM) and transmission electron
microscopy (TEM).
95% of the spirochetes were converted to cysts
after 1 min, and after 4 h no normal mobile borreliae were observed.
When transferred to growth medium (BSK-H), the cysts became smaller
and more irregular, and were filled with organic substances. After 1
day, 1-5 thin structures sprouted from the cysts. They continued to
grow in both length and thickness until they attained a normal
spirochetal structure. Finally, these new-born spirochetes detached
from the cysts, by which time their mobility had become normal.
The
present method for producing large amounts of cystic forms of B.
burgdorferi is well suited for further studies of this unique
microbe.
TITLE:
In vitro conversion of Borrelia burgdorferi to cystic forms in spinal
fluid, and transformation to mobile spirochetes by incubation in BSK-H
medium.
AUTHORS:
Brorson O; Brorson SH
AUTHOR
AFFILIATION:
Dept. of Microbiology, Vestfold Sentralsykehus, Tonsberg.
SOURCE:
Infection 1998 May-Jun;26(3):144-50
ABSTRACT:
The purpose of this study was to examine the structural alterations of
Borrelia burgdorferi when exposed to spinal fluid.
Normal, mobile
spirochetes were inoculated into spinal fluid, and the spirochetes
were converted to cysts (spheroplast L-forms) after 1-24 h. When
these cystic forms were transferred to a rich BSK-H medium, the cysts
were converted back to normal, mobile spirochetes after incubation
for 9 to 17 days.
The cultures were examined by dark field microscopy
(DFM), interference contrast microscopy (ICM) and transmission
electron microscopy (TEM). When neuroborreliosis is suspected, it is
necessary to realize that B. burgdorferi can be present in a cystic
form, and these cysts have to be recognized by microscopy. This study
may also explain why cultivation of spinal fluid often is negative
with respect to B. burgdorferi.
TITLE:
Transformation of cystic forms of Borrelia burgdorferi to normal,
mobile spirochetes.
AUTHORS:
Brorson O; Brorson SH
AUTHOR
AFFILIATION:
Dept. of Microbiology, Ulleval University Hospital, Oslo, Norway.
SOURCE:
Infection 1997 Jul-Aug;25(4):240-6
ABSTRACT:
The purpose of this study was to evaluate the behaviour of Borrelia
burgdorferi under controlled conditions.
The occurrence of cystic
forms of Borrelia burgdorferi in vitro was noted, and these cysts
were able to be transformed to normal, mobile spirochetes.
B.
burgdorferi was cultivated in a commercial culture medium without
serum. The spirochetes multiplied only slowly in this medium, and
transformation to encysted forms was observed after 1 week. When
these cysts were transferred to the same culture medium with rabbit
serum, the encysted forms developed into regular, mobile spirochetes
after 6 weeks, and their regeneration time was normal. Examination of
these cysts in the transmission electron microscope revealed
transverse fission inside the cysts.
It is probable that similar
phenomena may occur in vivo under conditions unfavourable for
spirochetes. These observations may help to explain why diagnosis and
treatment of B. burgdorferi infections in humans can be difficult.
TITLE:
Heterogeneity of Borrelia burgdorferi in the skin.
AUTHORS:
Aberer E; Kersten A; Klade H; Poitschek C; Jurecka W
AUTHOR AFFILIATION:
Department of Dermatology, University of Vienna,
Austria.
SOURCE:
Am J Dermatopathol 1996 Dec;18(6):571-9
ABSTRACT:
The reliability of various in vitro techniques to identify
Borrelia burgdorferi infection is still unsatisfactory.
Using a high-power resolution videomicroscope and
staining with the borrelia genus-specific monoclonal
flagellar antibody H9724, we identified borrelial
structures in skin biopsies of erythema chronicum
migrans (from which borrelia later was cultured), of
acrodermatitis chronica atrophicans, and of morphea. In
addition to typical borreliae, we noted stained structures
of varying shapes identical to borreliae found in a
"borrelia-injected skin" model; identical to
agar-embedded borreliae; and identical to cultured
borreliae following exposure to hyperimmune sera and/or
antibiotics. We conclude that the H9724-reactive
structures represent various forms of B. burgdorferi
rather than staining artifacts. These "atypical" forms of
B. burgdorferi may represent in vivo morphologic
variants of this bacterium.
TITLE:
Formation and cultivation of Borrelia burgdorferi spheroplast-L-form
variants [published erratum appears in Infection 1996 Jul-Aug;24(4):335]
AUTHORS:
Mursic VP; Wanner G; Reinhardt S; Wilske B; Busch U; Marget W
AUTHOR
AFFILIATION:
Max von Pettenkofer-Institut, Ludwig-Maximilians-Universitat Munchen,
Germany.
SOURCE:
Infection 1996 May-Jun;24(3):218-26
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.
TITLE:
Effects of penicillin, ceftriaxone, and doxycycline on morphology of
Borrelia burgdorferi.
AUTHORS:
Kersten A; Poitschek C; Rauch S; Aberer E
AUTHOR
AFFILIATION:
Department of Dermatology, University of Vienna, Austria.
SOURCE:
Antimicrob Agents Chemother 1995 May;39(5):1127-33
ABSTRACT:
Antibiotic therapy with penicillin, doxycycline, and ceftriaxone has
proven to be effective for the treatment of Lyme borreliosis.
In some
patients, however, it was noticed that borreliae can survival in the
tissues in spite of seemingly adequate therapy.
For a better
understanding of this phenomenon, we investigated the different modes
of degeneration of Borrelia burgdorferi suspensions during a 96-h
exposure to various antibiotics. By dark-field microscopy and
ultrastructural investigations, increasing blebbing and the gradual
formation of granular and cystic structures could be followed during
the exposure time. Although antibiotic concentrations at the MIC at
which 90% of organisms are inhibited after 72 h were 80% or even
greater, motile organisms were still present after incubation with
penicillin and doxycycline but not after incubation with ceftriaxone.
By transmission electron microscopy, intact spirochetal parts, mostly
situated in cysts, were seen up to 96 h after exposure with all three
antibiotics tested.
According to experiences from studies with other
spirochetes it is suggested that encysted borreliae, granules, and
the remaining blebs might be responsible for the ongoing antigenic
stimulus leading to complaints of chronic Lyme borreliosis.
TITLE:
Ultrastructure of Borrelia burgdorferi after exposure to
benzylpenicillin.
AUTHORS:
Schaller M; Neubert U
AUTHOR
AFFILIATION:
Dermatologische Klinik, Ludwig-Maximilians-Universitat, Munchen,
Germany.
SOURCE:
Infection 1994 Nov-Dec;22(6):401-6
ABSTRACT:
The aim of this study was to investigate the morphological changes of
Borrelia burgdorferi associated with penicillin treatment.
An isolate
of B. burgdorferi from an erythema migrans lesion was cultivated in
BSK II medium and exposed to increasing concentrations (0.0625 mg/l-2
mg/l) of penicillin G for 5 days. The in vitro minimal inhibitory
concentration (MIC) was determined to be 0.5 mg/l by broth dilution
method. The morphological structures of untreated spirochetes, as
well as their characteristic ultrastructural changes when exposed to
penicillin, were observed by electron microscopy.
The following
alterations were discovered: (i) Numerous outer sheath blebs at a
penicillin concentration of 0.0625 mg/l. (ii) A characteristic
irregular waveform of the borrelial cells and complete loss of the
outer sheath at a penicillin concentration of 0.125 mg/l. (iii) The
presence of "spheroplasts" at the same concentration. (iv) Structural
changes of the protoplasmic cylinder complex which showed an
irregular pattern at a penicillin concentration of 0.125 mg/l. (v)
Disruption of the protoplasmic cylinder complex into several parts at
penicillin concentrations of 0.25 mg/l and 0.5 mg/l. (vi) Severe
cytolysis at penicillin concentrations of 1 mg/l and 2 mg/l.
TITLE:
Analysis of Borrelia burgdorferi membrane architecture by
freeze-fracture electron microscopy.
AUTHORS:
Radolf JD; Bourell KW; Akins DR; Brusca JS; Norgard MV
AUTHOR
AFFILIATION:
Department of Internal Medicine, University of Texas Southwestern
Medical Center, Dallas 75235.
SOURCE:
J Bacteriol 1994 Jan;176(1):21-31
ABSTRACT:
Freeze-fracture electron microscopy was used to investigate the
membrane architectures of high-passage Borrelia burgdorferi B31 and
low- and high-passage isolates of B. burgdorferi N40. In all three
organisms, fractures occurred almost exclusively through the outer
membrane (OM), and the large majority of intramembranous particles
were distributed randomly throughout the concave OM leaflet. The
density of intramembranous particles in the concave OM leaflet of the
high-passage N40 isolate was significantly greater than that in the
corresponding leaflet of the low-passage N40 isolate. Also noted in
the OMs of all three organisms were unusual structures, designated
linear bodies, which typically were more or less perpendicular to the
axis of the bacterium. A comparison of freeze-fractured B.
burgdorferi and Treponema pallidum, the syphilis spirochete, revealed
that the OM architectures of these two pathogens differed markedly.
All large membrane blebs appeared to be bounded by a membrane
identical to the OM of B. burgdorferi whole cells; in some blebs, the
fracture plane also revealed a second bilayer closely resembling the
B. burgdorferi cytoplasmic membrane. Aggregation of the lipoprotein
immunogens outer surface protein A (OspA) and OspB on the bacterial
surface by incubation of B. burgdorferi B31 with specific polyclonal
antisera did not affect the distribution of OM particles, supporting
the contention that lipoproteins do not form particles in
freeze-fractured OMs.
The expression of poorly immunogenic,
surface-exposed proteins as virulence determinants may be part of the
parasitic strategy used by B. burgdorferi to establish and maintain
chronic infection in Lyme disease.
TITLE:
The fate of Borrelia burgdorferi, the agent for Lyme disease, in mouse
macrophages. Destruction, survival, recovery.
AUTHORS:
Montgomery RR; Nathanson MH; Malawista SE
AUTHOR AFFILIATION:
Department of Internal Medicine, Yale University
School of Medicine, New Haven, CT 06510.
SOURCE:
J Immunol 1993 Feb 1;150(3):909-15
ABSTRACT:
The macrophage is a known reservoir for a number of
infectious agents, and is therefore a likely candidate
site for persistence of Borrelia burgdorferi, the Lyme
spirochete. We report that unopsonized B. burgdorferi
enter macrophages rapidly, resulting mainly in
degradation but occasionally in apparent intracellular
persistence.
We studied uptake of spirochetes by
macrophages by simultaneously labeling infected cells
with antibodies to B. burgdorferi and with sequential
components of the endocytic pathway, and we examined
optical sections (0.5-1.0 micron in thickness) of these
cells by confocal fluorescence microscopy at multiple
time points after infection. We found that only 5 min of
incubation at 37 degrees C were required for nearly
100% of B. burgdorferi to enter a lysosomal
glycoprotein-positive compartment, whereas 60 min
were required for 90% of the spirochetes to appear in a
cathepsin L-positive compartment under the same
conditions. We also labeled infected living cells with
acridine orange to distinguish live from killed
intracellular organisms. Although the large majority of
spirochetes within a given cell were dead, we saw
occasional live ones up to 24 h (the longest interval
examined) after all extracellular organisms had been
lysed in distilled water.
Moreover, we can reculture
spirochetes from macrophages after infection.
Persistence of spirochetes within macrophages provides
a possible pathogenetic mechanism for chronic or
recurrent Lyme disease in man.
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Last updated on 3 April 2002 by
Art Doherty
Lompoc, California
doherty@utech.net