BORRELIA-BAKTEERIN ERI MUODOT

[soijuv]

TUTKIMUKSIA KYSTAMUOTOISISTA BORRELIA-BAKTEERISTA

1. RUNSAASTI lisätietoa kystamuotoisista bakteereista.

http://www.lymeinfo.net/medical/LDCysts.pdf

2. MS-taudin ja Alzheimerin taudin yhteys kystamuotoiseen borrelia-bakteeriin:

3. Artikkelissa esitetään hypoteesi, että spirokeetan kystamuodot eivät olekaan lepotilassa olevia, haitattomia, paikallaanpysyviä bakteereita, vaan itseasiassa kykenevät tuhoamaan elimistön soluja. Eri kokoiset kystamuotoiset spirokeetat hermosolujen sisällä saattavat siis selittää suuren osan keskushermostoa tuhoavista sairauksista kuten ALS, Parkinson, Alzheimer jne. Borreliabakteerin kystamuoto olisi siten yhteinen selittävä tekijä näille sairauksille.

4. VIDEO: http://www.youtube.com/watch?v=WozrCFW0mRM

Tri L. Mattman puhui v. 2005 Chicagon Autoimmuunikongressissa borrelibakteerista ja sen vaikeahoitoisesta L-muodosta jota kutsutaan kirjallisuuudessa myös esim. kystamuodoksi, pyöreäksi muodoksi jne. Videolla esitetään esim. kuvia borreliabakteerin tunkeutumisesta punasolujen sisään, borreliabakteerin kystamuotoa MS-potilaan selkäydinnesteestä jne.

Luento kestää n. 50 min.

Tri Mattman kuoli v. 2009.



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Med Hypotheses. 2006;67(4):819-32. Epub 2006 Jul 7.

1. Infection 2001 Dec;29(6):315-9

Association between multiple sclerosis and cystic structures in cerebrospinal fluid.
Brorson O, Brorson SH, Henriksen TH, Skogen PR, Schoyen R.

We previously studied spirochetes (B. burgdorferi) that have converted from spirochetes to cystic forms in CSF in vitro using the same methods as mentioned above [11]. With all these methods used in this study (TEM, AO, DF), the cystic structures observed in the CSF of the MS patients are morphologically similar to cystic forms of spirochetes. We found that cysts which are produced by inoculating B. burgdorferi in CSF at 37 C can be PCR negative using conventional DNA extraction and OspA primers (unpublished observation). This is either because the cyst wall inhibits the entrance to the genome or because the genomes of spirochetes have been changed. We have also to keep in mind that PCR detection of B. burgdorferi spirochetes often may give false-negative results [19).

The positive IgG index associated with MS in our patient cohort proves that the patients had an active inflammatory process in the CNS (Table 1). Inflammatory processes in the b~ and spinal cord of virtually any cause are usually less intense than inflammation in peripheral tissues and some microbiological agents, including spirochetes, provoke a very gentle inflammatory response [20, 21 ]. Considering the nature of MS, this disease could very well be a chronic infection and the clinical picture of MS has repeatedly been confused with neuroborreliosis [22-26). Therefore, we have both microbiological and some clinical support for the hypothesis that the cystic structures found in the CSF of the MS patients may originate from spirochetes which could be the causative agents of MS.

2... the frequency of relapses may be highly dependent on the chosen treatment and the phase of the disease [23−26]. A study of cytomorphic variations of B. burgdorferi isolates from patients with or without antibiotic treatment showed that penicillin can induce membrane-derived vesicles (cysts or spheroblast L-forms) in vivo [27]. This conversion of mobile Borrelia to cystic forms was subsequently observed for ceftriaxone, doxycyclin [17], ciprofloxacin [18] and vancomycin [12] at concentrations achievable in vivo. The fact that B. burgdorferi has the ability to convert (and reconvert) to cystic forms both in vivo and in vitro [1,4−6,10,14,15,21,27,28] may be regarded as an explanation why the infection may be persistent and reactivate. Therefore, it is reasonable to suggest that all germinative forms of the bacterium (and not only the motile form) should be destroyed so that Lyme borreliosis can be treated effectively.

An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole. INTERNATIONAL MICROBIOLOGY (2004) 7:139?142.

3. Concurrent Neocortical Borreliosis and Alzheimer's Disease Demonstration of a Spirochetal Cyst Form

Annals of the New York Academy of Sciences 1988, 468-470

Alan B. MacDonald, Southampton Hospital, Southampton, NY 11968

A 71 year old man died in Arizona 3 years after the onset of progressive dementia. A diagnosis of probable Alzheimer's disease was based on clinical criteria. The brain was removed at autopsy, frozen (unfixed), and transported to the Department of Pathology, University of California, San Diego, School of Medicine where it was stored at -70 degrees C for further study. The author received the frozen brain and utilized methods previously described (1) for in vitro culture, cytologic, immunohistochemical , and silver impregnation studies. Argyrophilic plaques and neurofibrillary tangles were found in the frontal lobe and hippocampal formation is sufficient number to establish the neuropathologic diagnosis of Alzheimer?s (fig 1 A). Spirochetes were visualized in imprint preparations of freshly thawed frontal lobe cortex with monoclonal antibody H5332, which specifically binds to the outer surface membrane of Borrelia burdorferi (fig.2). Borrelia spirochetes were recovered from cultures of freshly thawed cerebral cortex and hippocampus in Barbour-Stoenner-Kelly medium. An unexpected observation was the identification of cystic forms of the Borrelia spirochete in dark-field preparations of cultured hippocampus, and in imprints of hippocampus using the monoclonal antibody H9724, which binds to class-specific axial filament proteins of Borrelia spirochetes. Oil immersion examination of sections from the hippocampus impregnated with silver disclosed a rare cystic structure (fig. 1B). Previous workers have identified spirochetal cyst forms in cultures of non-pathogenic treponemal spirochetes and have suggested that spirochetes have a complex life cycle. (2-5) Dark-field examination of aged cultures of the B31 reference strain of Borrelia burgdorferi disclosed cystic structures similar to the cysts found in the autopsy brain culture.

The following hypothesis is offered based on these observations. Borrelia spirochetes have a complex life cycle which includes corkscrew shaped forms, uncoiled filamentous forms, L-forms lacking a cell wall, cystic and ameboid forms, and granular forms. These forms may exist as either extracellular or intracellular pathogens. The cystic form of Borrelia burgdorferi may explain the Pick body, which is found in Pick?s disease, and the granular form of Borrelia may explain granulovascular degeneration of nerve cells in the hippocampal formation in Alzheimer?s disease. A cystic form of the Borrelia spirochete would explain the ability of the microbe to persist in the host during a prolonged periodof asymptomatic clinical latency, which spans the period between primary infection and the expression of tertiary manifestations of neuroborreliosis.

3.APMIS 2001 May;109(5):383-8. Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. Gruntar et al.

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 cystinoculated 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.

3. 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.


4. 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.

5. ...For example, B. burgdorferi has the ability to survive in divergent conditions of mammals and ticks by existing in a variety of forms that are ultrastructurally and metabolically distinct. Even in the tick, altered morphologic forms of B. burgdorferi are present [43], but in the mammal, selective pressure from mammalian immune surveillance results in these altered forms becoming more common. These "host adapted" forms generally display altered morphology to varying degrees and are referred to collectively as L-forms or spheroplasts. B. burgdorferi spheroplasts, of which cystic forms and granules are sub-types, have been extensively documented in vitro and in vivo [44?53], both extracellularly and intracellularly [27, 47, 54?57]. Their ability to revert from host-adapted forms back to helical forms under appropriate conditions has been demonstrated in vitro [47, 58, 59].

To the uninitiated, it may be tempting to infer that B. burgdorferi cystic forms are degenerative bacterial fragments. This is not the case, since researchers have demonstrated protein synthesis requirements for spirochetal conversion into the spheroplast form [44]. Indeed, it has been unequivocally proven that B. burgdorferi cystic forms are virulent and infectious. Their infectivity, survival under extreme environmental conditions, and ability to revert back to helical forms in vivo have all been demonstrated by inoculation of B. burgdorferi cysts into mice and subsequent recovery of helical spirochetes from the animals [60]. As such, host-adapted forms of B. burgdorferi are considered to be major factors in the relapsing and persistent nature of Lyme disease [61?63].

Just as B. burgdorferi spheroplasts have altered metabolic requirements for growth, so too, do they have unique antibiotic sensitivities, altered surface protein expression, dramatically reduced surface area presented for immune surveillance, and the ability to cause multiple potential problems for PCR analysis. All of the foregoing helps to explain observations of antibiotic resistance, seronegativity, and even frequent PCR negativity in active disease [51, 54, 59, 63, 64]. The failure to address the complexities of the borrelial life cycle in the work by Klempner et al. is a serious error. For example, the fact that cystic forms demonstrate sensitivity to metronidazole while their helical kin are resistant, illustrates the point that B. burgdorferi spheroplasts have altered antibiotic resistance [65]. Attention to these forms during the initial study design might have resulted in different treatment decisions, with consideration that a cell wall-attacking cephalosporin may not have been the ideal antibiotic choice for treatment of cell wall-deficient organisms in patients with late-stage Lyme disease.

In addition, had the authors addressed the intracellularity of B. burgdorferi, this might have broadened their choices of antibiotic therapy. Although the utility of ceftriaxone for Lyme disease has been documented, it has been similarly documented that this agent frequently does not fully eradicate human B. burgdorferi infections [19]. Cephalosporins do not achieve intracellular penetration, a fact that may partially explain well-known treatment failures associated with late stage Lyme disease. Indeed, B. burgdorferi has been documented within a variety of cell types, including but not limited to endothelium, fibroblasts, lymphocytes, macrophages, keratinocytes and synovial cells [17, 51, 54, 66?70]. These findings are critically important since chronic infections are highly dependent on intracellular asylum as a mode of persistence, and localization within eukaryotic cells protects B. burgdorferi from antibiotics [71, 72]. It is particularly surprising that the lead author agreed to use ceftriaxone in this study, since he previously authored a paper on the fibroblast-mediated protection of B. burgdorferi in vitro from concentrations of ceftriaxone achieved in vivo for the treatment of Lyme disease [71]...

http://www.ilads.org/position2.html

44 Alban PS; Johnson PW; Nelson DR. Serum-starvation-induced changes in protein synthesis and morphology of Borrelia burgdorferi. Microbiology 2000;146 (Pt 1):119?27.

45 Bruck DK; Talbot ML; Cluss RG; Boothby JT. Ultrastructural characterization of the stages of spheroplast preparation of Borrelia burgdorferi. J Microbiol. Methods 1995;23:219?28.

46 Benach JL. Functional heterogeneity in the antibodies produced to Borrelia burgdorferi. Wiener Klinische Wochenschrift 1999;111:985?9.

47 Preac-Mursic V; Wanner G; Reinhardt S; Wilske B; Busch U; Marget W. Formation and cultivation of Borrelia burgdorferi spheroplast L-form variants. Infection 1996; 24:218?26.

48 Cluss RG; Goel AS; Rehm HL; Schoenecker JG; Boothby JT. Coordinate synthesis and turnover of heat shock proteins in Borrelia burgdorferi: degradation of DnaK during recovery from heat shock. Infect Immunity 1996;64:1736?43.

49 Kersten A; Poitschek C; Rauch S; Aberer E. Effects of penicillin, ceftriaxone, and doxycycline on the morphology of Borrelia burgdorferi. Antimicrob Agents Chemo1995;39:1127?33.

50 Aberer E; Koszik F; Silberer M. Why is chronic Lyme borreliosis chronic? Clin Infect Dis 1997;25(Suppl 1):S64?S70.

51 Aberer E; Kersten A; Klade H; Poitschek C; Jurecka W. Heterogeneity of Borrelia burgdorferi in the skin. Am J Dermatopathol 1996;18:571?9.

52 Angelov L; Dimova P; Berbencova W. Clinical and laboratory evidence of the importance of the tick D. marginatus as a vector of B. burgdorferi in some areas of sporadic Lyme disease in Bulgaria. Eur J Epidemiol 1996;12:499?502.

53 Schaller M; Neubert Ultrastructure of Borrelia burgdorferi after exposure to benzylpenicillin. Infection 1994 22:401?406.

54 Hulinska D; Bartak P; Hercogova J; Hancil J; Basta J; Schramlova J. Electron microscopy of Langerhans cells and Borrelia burgdorferi in Lyme disease patients. Zbl Bakt 1994;280:348?9.

55 Nanagara R; Duray PH; Schumacher HR. Jr. Ultrastructural demonstration of spirochetal antigens in synovial fluid and synovial membrane in chronic Lyme disease: possible factors contributing to persistence of organisms. Hum Pathol 1996;27:1025?34.

56 Hulinska D; Jirous J; Valesova M; Hercogova J. Ultrastructure of Borrelia burgdorferi in tissues of patients with Lyme disease. J Basic Microbiol 1989;29:73?83.

57 MacDonald AB. Concurrent neocortical borreliosis and Alzheimer's disease: Demonstration of a spirochetal cyst form. Ann NY Acad Sci 1988;539:468?70.

58 Brorson O; Brorson SH. Transformation of cystic forms of Borrelia burgdorferii to normal mobile spirochetes. Infection 1997;25:240?6.

59 Brorson O; Brorson. In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium. Infection 1998;26:144?50.

60 Gruntar I, Malovrh T, Murgia R, Cinco M. Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. APMIS 2001;109:383?8.

http://www.ilads.org/position2.html

..Indeed, there have been a number of peer reviewed publications demonstrating persistent infection with B. burgdorferi in humans despite multiple and extended courses of antibiotic therapy [19]. Persistent infection has been demonstrated repeatedly by both polymerase chain reaction (PCR) and histopathology [20-23]. Chronic infection has also been demonstrated by culture despite the well-known difficulties in harvesting B. burgdorferi from Lyme patients, and culture positivity has even been found in patients who are seronegative for the Lyme spirochete [24?30]. In light of such data, it would be illogical to assume that persistent symptoms in chronically ill Lyme disease patients are not related to active infection with B. burgdorferi. Ironically, and in direct opposition to the extensive body of published data, some researchers have attributed chronic symptoms compatible with Lyme disease to alternative vague diagnoses, such as "post-Lyme syndrome," fibromyalgia, or chronic fatigue syndrome [31,32]...


2. Artikkelissa esitetään hypoteesi, että spirokeetan kystamuodot eivät olekaan lepotilassa olevia haitattomia paikallaanpysyviä bakteereita, vaan itseasiassa kykenevät tuhoamaan elimistön soluja. Eri kokoiset kystamuotoiset spirokeetat hermosolujen sisällä saattavat siis selittää suuren osan keskushermostoa tuhoavista sairauksista kuten ALS, Parkinson, Alzheimer jne. Borreliabakteerin kystamuoto olisi siten yhteinen selittävä tekijä näille sairauksille.


Med Hypotheses. 2006;67(4):819-32. Epub 2006 Jul 7.

Spirochetal cyst forms in neurodegenerative disorders,...hiding in plain sight.
Macdonald AB.
St. Catherine of Siena Medical Center, Department of Pathology, 50 Rte 25 A, Smithtown, NY 11787, USA.

Here is proposed a hypothesis that a completely unsuspected biology exists for pathogenic spirochetes, namely that the cystic spirochetal forms (long thought to be static and resting or just a dormant cohort) actually are capable of killing mammalian host cells. At least two "lethal" scenarios are proposed; first, the host cell destruction from the "inside out" by small caliber cystic forms invading the host cell cytoplasm, and second host cell destruction by engulfment of entire host cells by large caliber cystic spirochetal forms. Conventional thinking about spirochetal cyst forms is divided between two polar spheres of influence; one a majority community that completely denies the existence of spirochetal cyst forms, and a second group of academically persecuted individuals who accepts the precepts of such antebellum scientists as Schaudinn, Hoffman, Dutton, Levaditi, Balfour, Fantham, Noguchi, McDonough, Hindle, Steiner, Ingraham, Coutts, Hampp, Warthin, Ovcinnikov, and Delamater. Microscopic images of cystic spirochetes are difficult to ignore, but as has been the case in this century, academic "endowments" have nearly expunged all cystic spirochetal image data from the current textbook versions of what is the truth about the spirochetaceae. If the image database from the last century is obliterated; many opportunities to diagnose will be lost. Variously sized cystic spirochetal profiles within diseased nerve cells explain the following structures: Lewy body of Parkinson's disease, Pick body, ALS spherical body, Alzheimer plaque.

Borrelia infection is therefore a unifying concept to explain diverse neurodegenerative diseases, based not entirely on a corkscrew shaped profile in diseased tissue, but based on small, medium and large caliber rounded cystic profiles derived from pathogenic spirochetes which are..."hiding in plain sight".

PMID: 16828236 [PubMed - in process]

[soijuv]

Kystamuotoisista B garinii -bakteereista tulee liikkuvia tautia aiheuttavia spirokeettoja elimistössä. Tutkimuksessa bakteerit muuntuivat tislatussa vedessä kystamuotoon ja muuntuivat takaisin liikkuviksi spirokeetoiksi kun ne istutettiin hiiriin!

http://www.blackwell-synergy.com/links/ ... 507.x/abs/

Apmis
Volume 109 Page 383 - May 2001
doi:10.1034/j.1600-0463.2001.090507.x
Volume 109 Issue 5


Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo

Igor Gruntar,1 Tadej Malovrh,2 Rossella Murgia3 and Marina Cinco3
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.

[soijuv]

Kystamuotoisista borreliabakteereista on lukemattomia tutkimuksia suurin osa in vitro mutta useita myös in vivo. Esim. A. McDonald on tutkinut aihetta. Viimeisin tutkimus on vuodelta 2006. Hän mm. kuvasi ihmisen selkäydinnesteessä esiintyviä Bb:n kystamuotoja - hän luokitteli muodot kolmeen kategoriaan.

Phillips ym. (1998) mainitsevat tutkimuksessan, että he ovat nähneet viljelyissään useimmiten borreliabakteerin eri kehitysvaiheita. Tutkimuksen mukaan lähes kaikkien tutkimukseen osallistuneiden Elisa oli negatiivinen. 91 % olisi siten diagnosoitu virheellisesti borrelia-negatiiviseksi ilman lisätutkimuksia.


In Vivo Transformation of Cystic Forms of Borrelia to Normal Spirochetes

Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo.

Gruntar I; Malovrh T; Murgia R; Cinco M. 2001.

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.

Cystic Form of Borrelia: Susceptibility to Treatment with Metronidazole

An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to metronidazole.

Brorson O; Brorson S. 1999.

Methodology for Generation of Cystic Forms of Borrelia burgdorferi

A rapid method for generating cystic forms of Borrelia burgdorferi, and their reversal to mobile spirochetes.

Brorson O; Brorson S. 1998.

Transformation of Borrelia Spirochetes to Cystic Forms in Spinal Fluid

In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium.

Brorson O; Brorson S. 1998.

Transformation of Encysted Borrelia Burgdorferi to Normal Spirochetes

Transformation of cystic forms of Borrelia burgdorferi to normal, mobile spirochetes.

Brorson O; Brorson S. 1997.

Confirmatory Study: Transformation of Borrelia to a Cystic Form

Serum starvation-induced cyst formation in Borrelia burgdorferi under defined conditions.

Alban PS; Nelson DR. 1999.

Pleomorphism In Borrelia: Spheroplast L-Form Variants Without Cell Walls

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

Mursic VP; Wanner G; Reinhardt S; Wilske B; Busch U; Marget W. 1996.

In Vivo Morphologic Variants of Borrelia Burgdorferi in Skin Biopsies Heterogeneity of Borrelia burgdorferi in the skin.

Aberer E; Kersten A; Klade H; Poitschek C; Jurecka W. 1996.

Survival of Encysted Borrelia Following Incubation with Antibiotics

Effects of penicillin, ceftriaxone, and doxycycline on morphology of Borrelia burgdorferi.

Kersten A; Poitschek C; Rauch S; Aberer E. 1995.

In Vivo Discovery of Borrelia Cysts in Erythema Chronicum Migrans

Electron microscopy of Langerhans cells and Borrelia burgdorferi in Lyme disease patients.

Hulinska D; Bartak P; Hercogova J; Hancil J; Basta J; Schramlova J. 1994.

"The form of spirochetes was unusual, i.e. cyst-like (Fig.5). The surface membrane of cyst of Bb was antigenically different, negative with McAb H 5332 , positive with lectin WGA in the IEM method (Fig.7)
in contrast to the material of the spirochetes inside the cyst, which was positive with McAbH 5332 in the outer envelope on cross section

In Vivo Findings: Observation of Borrelia Cysts in Autopsy of Human Brain

Concurrent neocortical Borreliosis and Alzheimer?s disease; demonstration of a spirochetal cyst form.

Alan B. MacDonald. 1988.

Early Observations of Spirochetes and Granular Forms

The infective granule in certain protozoal infections, as illustrated by the spirochaetosis of Sudanese fowl.

Andrew Balfour, M.D. 1911

A.McDonaldin sivulta: http://66.102.9.104/search?q=cache:i5Z7 ... cd=7&gl=fi

Spirochete Life Cycle?

Empirical Data:

Cystic Borrelia forms (At least three categories are proposed)

Figure1. "Primary" type cysts ? Rapidly reversible to motile corkscrew shaped forms: Cystic Borrelia rapidly emerge from the motile corkscrew shaped forms as a response to "stressful conditions" in their environment. Figure 1 Cystic Forms of Borrelia burgdorferi (American

Type Culture Collection 35210), darkfield image 1000x, original, Alan MacDonald, MD unpublished, Photograph date 1988

Figure 3 Cystic Borrelia burgdorferi from cultured human spinal fluid, 1000x original magnification, Oligreen stain for single strand DNA, Western Blot positive for antibodies to Borrelia burgdorferi in spinal fluid. Oligreen stained preparation ( demonstrates single strand DNA ? green signal ? within the otherwise "empty cyst". 1000x magnification.

Figure 4 Cystic Borrelia burgdorferi from spinal fluid culture for comparison, showing internal DNA granular foci, 1000x magnification, Oligreen stain for single strand DNA Figure 5 Cystic Borrelia burgdorferi fromspinal fluid culture, for comparison, showing a dense "nucleoid-like profile", 1000x magnification, Oligreen stain for single strand DNA. Tertiary ("Chronic" or "Latency" type) Borrelia cystic forms containing internal granular structures of variable caliber. Figure 6 Borrelia Burgdorferi strain B31, from Rocky Mountain Laboratory, NIAID, NIH,

Hamilton Montana, Conventional corkscrew and Concurrent Cystic forms, 1000x magnification, Oligreen stain (Invitrogen), Alan MacDonald, MD, unpublished image, 2006, Note: These cystic forms are NOT detected in conventional Acridine orange or in Picogreen (Invitrogen ) stains for double strand spirochetal DNA, and therefore would be "invisible" with standard detection modalities. These profiles overturn the concept that only during times of stress will you expect to find Cysts of Borrelia. These cyst forms developed in BSK culture medium during log phase normal growth of the reference strain of Borrelia burgdorferi. http://66.102.9.104/search?q=cache:i5Z7 ... cd=7&gl=fi


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A Proposal for the Reliable Culture of Borrelia burgdorferi from Patients with Chronic Lyme Disease, Even from Those Previously Aggressively Treated

S. E. Phillips, L. H. Mattman, D. Hulinska, H. Moayad

Infection 26 (1998) 364-367

http://www.cbc.ca/ideas/features/Aids/phillips.html

....." RESULTS
Of the 47 patients with chronic Lyme disease, 43 (91%) cultured positive for B. burgdorferi. while 23/23 (100%) of
the controls cultured negative. Many of the cultures were clearly spirochetes when examined under light microscopy (Figures 1-3). Immuno-electron microscopy and Osp A PCR confirmation provided additional confirmatory evidence as to the identity of the spirochetes (Figures 4-7). The slide cultures consistently demonstrated the fastest and most abundant yields. With this technique, placement in the Coplin jar allows for varying gradations of oxygen tension. Sometimes spirochetal growth can be seen after as little as 20 h. appearing as a band near the upper end of the smear.

DISCUSSION
An attempt to culture B. burgdorferi from the blood of previously aggressively treated chronic Lyme disease patients seemed at first a monumental task. Before undertaking this effort, we therefore had to be as sure as possible that the organisms were indeed present in the blood of these patients. As a first step, we scrutinized a report where B. burgdorferi had been cultured from the blood of patients with early untreated disease. From this group of patients it had been noted in follow-up that subsequent blood cultures became routinely negative after antibiotic therapy, despite 71% of the patients remaining symptomatic [9]. Three possibilities readily come to mind for the explanation of this paradox: either 1) the infection is cleared, but a post-infectious process continues, or 2) the organism is cleared from the blood rapidly. but finds a pathogenic harbor elsewhere, or 3) the organism is maintained in the blood in an altered state which cannot be cultured on routine media.

In response to the first possibility, the notion of a post-Lvme syndrome has countless flaws. A post-infectious syndrome could not explain the observation that patients with "post-Lyme" or "post-Lyme fibromyalgia" responded to re-treatment with antibiotics, only to relapse with its discontinuation [13-15]. With the advent of PCR, antigen capture, and the benefit of those rare successful culture experiments even in the face of prior "curative treatment" [3-8]. the notion of "post-Lyme" should have been dismissed long ago. In response to the second possibility, given the common finding of circulating immune complexes with Lyme disease, we thought this unlikely [16]. Thus we were left with the third and most logical possibility. Specifically, we chose to pursue the organism in its cell wall-deficient state, i.e. L-forms, as previously reported [17].

Although L-forms will complex with fluorescent antibody to B. burgdorferi, only as they revert to classic parent forms can the typical spirochetal morphology be seen. There has been a considerable spectrum of cell wall deficiency demonstrated in our laboratory. B. burgdorferi may exist in various forms depending on its environment. In addition to the spirochetal form, we have demonstrated its growth both as amorphous L-forms and rounded giant L-bodies which have been previously described as cystic forms [11, 18]. As B. burgdorferi reverts from cell wall deficiency with the rebuilding of its cell wall, classic spirochetal forms can be seen. Most often, in our cultures, B. burgdorferi can be seen in varying stages of reversion, i.e. some L-dependent spirochetal forms within an L-form colony.

The L-form variants, osmotically fragile by nature, require precise conditions to grow in culture. our medium and methodology are specifically designed for the fostering of cell wall-deficient organisms and their reversion to classic parent forms. In most instances, the methods must be followed precisely. Even small variations produce no growth. For example, 2% yeast extract instead of 1% is inhibitory. or if the yeast extract is autoclaved with the rest of the medium instead of separately. that too will be inhibitory. However, there is one aspect of B. burgdorferi's growth characteristics which we found to be remarkably non-fastidious. The organism can be easily grown throughout a wide range of pH, from 6.8-7.8. This explains the different ratios of NaHC03 used in the various types of culture mediums. We are still not sure about the optimal pH for culture. Future research will address that question more specifically.

It should be noted from this study that currently accepted standards for serologic diagnosis seem to be inadequate. Only a small minority of participants in the study had positive Lyme ELISAs. Under the current recommendations for two-tier testing by the CDC/ASTPHLD, 91% of the patients in the study would have been misdiagnosed as not having Lyme borreliosis.

It is hoped that our work will help to end a medical controversy which has been going on for far too long. This study proves that chronic Lyme disease is of chronic infectious etiology, and that even antibiotic treatment well in excess of current recommendations is not necessarily curative. Given the flaws in currently accepted serologic diagnostic criteria, it is likely that Lyme borreliosis is vastly underdiagnosed. May this research help to focus the scientific community on effective curative therapies for patients with chronic Lyme disease.

It should also be noted that, in addition to its utility in growing B. burgdorferi, the MPM medium may be useful for culturing a variety of other spirochetes from patients."

[soijuv]

Sivulla on useita mikroskooppikuvia Bb:n eri muodoista yms. http://www.molecularalzheimer.org/

[soijuv]

Keskustelin jokin aika sitten borreliabakteerin kystamuodoista suomalaisen borrelioosilääkärin kanssa. Hänen mukaansa kystamuodoista ei ole riittävää näyttöä. Lähetin hänelle lukuisia tutkimuksia aiheesta mm. seuraavan Gruntarin tutkimuksen jossa bakteerin kystamuodot muuntuivat takaisin spirokeetoiksi in vivo (elimistössä). Useiden tutkijoiden mukaan kystamuotojen merkitystä ja bakteerin eri muotoja tulisi tutkia huomattavasti enemmän. Kystamuodot olivat seuraavan tutkimuksen mukaan erittäin selvitymiskykyisiä.

http://www.ncbi.nlm.nih.gov/sites/entre ... ool=Entrez
System2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum

APMIS. 2001 May;109(5):383-8. Links
Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo.
Gruntar I, Malovrh T, Murgia R, Cinco M.
Institute of Microbiology and Parasitology, Veterinary Faculty, Ljubljana, Slovenia. gruntaig@mail.vf.uni-lj.si

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 [PubMed - indexed for MEDLINE]
_________________________________________________________________


Lähettäjä: Soijuv Lähetetty: 22.10.2007 13:31

Italialainen tutkimus borreliabakteerin kystamuodosta. Tutkijoiden mukaan bakteerin muuntuminen kystamuotoon on todettu myös koeputkiolosuhteiden ulkopuolella eli ihmisen elimistössä.

"Viimeaikaisissa tutkimuksissa on osoitettu borreliabakteerin muuntuvan spirokeettamuodosta liikkumattomaksi kystamuodoksi silloin, kun bakteeri joutuu epäedullisiin olosuhteisiin. Kystamuodot kykenevät muuntumaan takaisin liikkuviksi spirokeetoiksi sekä in vitro että in vivo."

(Suom. huom. Useimmat lääkärit eivät vielä tunne kyseistä asiaa.)


Wien Klin Wochenschr. 2002 Jul 31;114(13-14):574-9
Cystic forms of Borrelia burgdorferi sensu lato: induction,
development, and the role of RpoS.

Murgia R, Piazzetta C, Cinco M.
Dipartimento di Scienze Biomediche, sez. Microbiologia, Università
degli Studi di Trieste, Trieste, Italy. rmurgia@dsbmail.units.it

It has been demonstrated recently that cells of Borrelia burgdorferi sensu lato, the etiological agent of Lyme disease, transform from mobile spirochetes into nonmotile cystic forms in the presence of certain unfavourable conditions, and that cystic forms are able to reconvert to vegetative spirochetes in vitro and in vivo. The purpose of this study was to investigate the kinetics of conversion of borreliae to cysts in different stress conditions such as starvation media or the presence of different antibiotics. Using the same experimental conditions we also investigated the possible role in cyst formation of RpoS, an alternative sigma factor that controls a regulon in response to starvation and transition to stationary phase. We observed that beta-lactams penicillin G and ceftriaxone, the antibiotics of choice in Lyme borreliosis treatment, favoured the production of cysts when used with serum-depleted BSK medium. In contrast, we observed a low level of cyst formation in the presence of macrolides and tetracyclines. In order to elucidate the role of the rpoS gene in cyst formation we analyzed the reaction of the rpoS mutant strain in comparison with its wild-type in different conditions. Under the same stimuli, both the wild-type borrelia and the rpoS knock-out isogenic strain produced cystic forms with similar kinetics, thus excluding the participation of the gene in this phenomenon.

Our findings suggest that cyst formation is mainly due to a physical-chemical rearrangement of the outer membrane of Borrelia burgdorferi sensu lato leading to membrane fusion and controlled by different regulation mechanisms.

PMID: 12422604 [PubMed - indexed for MEDLINE]

[soijuv]

Useat tutkijat, esim. norjalaiset Brorsonit, ovat jo vuosien ajan kertoneet borreliabakteerin kykenevän muuntumaan erilaisiin muotoihin. Spirokeettamuoto ei näin ollen ole ainoa muoto bakteerista. Suomalaisten taholta asiaa ei ole vielä tunnustettu. Pari viikkoa sitten yksi "borrelioosilääkäri" kertoi borrelioosia sairastavalle henkilölle, että sellaista asiaa kuin kystamuotoinen borreliabakteeri ei ole olemassakaan.

Spirokeettojen kyvystä muuntua esim. kystamuotoon on kuitenkin esitetty sivuillamme tietoa jo vuosien ajan. Kirjallisuudessa asian puolesta on esitetty tutkimuksia, toisinaan jopa kuvien kera, jo sadan vuoden ajan. Asia on todettu kupan aiheuttajan Treponema pallidumin kohdalla. Kyseinen spirokeetta on borreliabakteerin sukulainen.

Miklossyn ym. (2008) tutkimuksessa kolmelta neuroborrelioosia sairastavalta henkilöltä löytyi histokemiallisissa, immunohistokemiallisissa ja pimeäkenttätutkimuksissa borreliabakteerin kysta- ja granulamuotoja. In vivo -löydökset olivat identtisiä koeputkiolosuhteisissa (in vitro) tehtyihin löydöksiin.

Tulokset vahvistavat borreliabakteerin pleomorfisten ja kystamuotojen aiheuttavan keskushermostulehduksia borrelioosia sairastaville. Kyseiset muodot saattavat selittää taudissa esiintyvät pitkät oireettomat kaudet sekä taudin kroonistumisen. Tutkimuksen mukaan bakteeri saattaa aiheuttaa myös soluvaurioita ja solukuolemaa. Tutkimuslöydös on merkittävä taudin diagnosoinnin ja hoidon kannalta. Bakteerin epätyypillisiä muotoja saattaa esiintyä elimistössä myös ilman tyypillistä spirokeettamuotoa.

Suom.huom. tutkimuksesta on saatavissa koko PDF. Klikkaa tiivistelmän alapuolella olevaa PDF-linkkiä. Sivun vas.puolella olevasta valikosta löytyy useita valokuvia esim. kystamuodosta.
_________________________________________________________________


Lähettäjä: jukka61 Lähetetty: 30.9.2008 20:20

Osaatko Soijuv kertoa, miksi suomalaiset tutkijat ja lääkärit eivät myönnä (usko) kystamuotoisen Borrelia-bakteerin olemassaoloon, tyhnyys lienee liian helppo selitys, mutta toisaalta ei voi oikein muuhunkaan johtopäätöksen tulla, kun useat paljon kokeneemmat (mm. Brorsonit) ja pelkästään Borreliaa tutkivat ovat tehneet tuon johtopäätöksen jo vuosia sitten ja siitä lienee lisäksi kiistatonta näyttöä.

Eihän homma voi Suomessa mitenkään edetä, jos jo perusasioita ei tunnusteta, vaan intetään kuin kirkko aikanaan, että maapallo on tasainen ja reunalta voi pudota
_________________________________________________________________


Lähettäjä: Soijuv Lähetetty: 1.10.2008 13:10

Olen ollut asian tiimoilta jonkin verran yhteydessä joihinkin suomalaisiin lääkäreihin ja mediassakin olen maininnut asiasta(kansainvälisiä yhteyksiä 22:n maan tutkijan/lääkärin kanssa olen jo pitänyt usean vuoden ajan). Useimmat suomalaiset kyllä uskovat, mutta eivät tee asialle mitään- odottavat saavansa valmiin "reseptin" siitä miten toimia. Toiset sen sijaan selittävät, että asiasta ei ole ollut in vivo näyttöä (ihmiselimistössä), vaikka sitäkin on kyllä ollut jonkin verran jos tietoa vain on halunnut saada ja siitä kiinnostua. Miklossyn tutkimuksessa toisaalta todettiin in vitro ja in vivo muutosten olleen identtisiä. Olen laittanut tietoa in vivo näytöstä eteenpäin joten toiveissa on, että asia etenee jonkin verran Suomessakin.

Koska tietoa asiasta on joka tapauksessa ollut vuosien ajan runsaasti kuten sivujamme lukeneet ovat voineet todeta, voi todellakin ihmetellä miksi suomalaiset eivät ole paneutuneet asiaan. Jotta bakteerin voisi edes teoriassa kuvitella olevan tuhottavissa, on se ensin tunnettava. Jo vuosien ajan ja myös tällä hetkellä hoitoa annetaan tuntematta vihollisen käyttäytymistä ja erilaisia puolustautumiskeinoja tuhoamisyrityksiä (esim. antibiootteja) kohtaan. Tämän seurauksena hoito ei ole suuntautunut oikein.

Tutkimuksissahan on esim. osoitettu nykyisin yleisimmin käytetyn suonensisäisen antibiootin Rocephalinin (keftriaksoni) aiheuttavan borreliabakteerin muuntumisen kystamuotoon erittäin nopeasti. Tällöin penisilliinityyppiset antibiootit, jotka vaikuttavat bakteerin solun seinämään ovat tehottomia. Tällaisissa tapauksissa on mahdollista, että hoidon teho on ollut vain väiaikainen - ainakin niissä tapauksissa joissa mitään muuta hoitoa ei ole ollut, sairastunut jatkaa entisiä elämäntapojaan eikä hänen immuunipuolustustaan ole tuettu millään tavoin.

Mielenkiintoista on, että kystamuodoista on todellakin tutkimuksia jo noin sadan vuoden ajalta ja useat tämänkin päivän tutkijat ovat kertoneet usean vuoden ajan havainnoistaan esim. siitä että he ovat nähneet nimenomaan kystamuotoisia borreliabakteereita, eivät spirokeettoja, eri keskushermostosairauksia kuten Parkinson-, Alzheimer- ja MS-oireita, sairastavien keskushermostossa.


Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis
Judith Miklossy , Sandor Kasas , Anne D Zurn , Shermann McCall , Sheng Yu and Patrick L McGeer

Journal of Neuroinflammation 2008, 5:40doi:10.1186/1742-2094-5-40


Published: 25 September 2008

Abstract (provisional)

Background
The long latent stage seen in syphilis, followed by chronic central nervous system infection and inflammation, can be explained by the persistence of atypical cystic and granular forms of Treponema pallidum. We investigated whether a similar situation may occur in Lyme neuroborreliosis. Method: Atypical forms of Borrelia burgdorferi spirochetes were induced exposing cultures of Borrelia burgdorferi (strains B31 and ADB1) to such unfavorable conditions as osmotic and heat shock, and exposure to the binding agents Thioflavin S and Congo red. We also analyzed whether these forms may be induced in vitro, following infection of primary chicken and rat neurons, as well as rat and human astrocytes. We further analyzed whether atypical forms similar to those induced in vitro may also occur in vivo, in brains of three patients with Lyme neuroborreliosis. We used immunohistochemical methods to detect evidence of neuroinflammation in the form of reactive microglia and astrocytes.

Results: Under these conditions we observed atypical cystic, rolled and granular forms of these spirochetes. We characterized these abnormal forms by histochemical, immunohistochemical, dark field and atomic force microscopy (AFM) methods.

The atypical and cystic forms found in the brains of three patients with neuropathologically confirmed Lyme neuroborreliosis were identical to those induced in vitro. We also observed nuclear fragmentation of the infected astrocytes using the TUNEL method. Abundant HLA-DR positive microglia and GFAP positive reactive astrocytes were present in the cerebral cortex.

Conclusion: The results indicate that atypical extra- and intracellular pleomorphic and cystic forms of Borrelia burgdorferi and local neuroinflammation occur in the brain in chronic Lyme neuroborreliosis. The persistence of these more resistant spirochete forms, and their intracellular location in neurons and glial cells, may explain the long latent stage and persistence of Borrelia infection. The results also suggest that Borrelia burgdorferi may induce cellular dysfunction and apoptosis. The detection and recognition of atypical, cystic and granular forms in infected tissues is essential for the diagnosis and the treatment as they can occur in the absence of the typical spiral Borrelia form.

...The persistence of more resistant atypical cystic and granular forms of Treponema pallidum, which are less sensitive to chemicals and antibiotics, are responsible for the long latent stage in chronic syphilis and for the infectivity of tissues devoid of the demonstrable vegetative form of spirochetes. The intracellular localization of Treponema pallidum is another way of evading from destruction by the host immune system [30, 39]. Virtually all types of mammalian cells can be invaded by Treponema pallidum resulting ultimately in functional cell damage and cell destruction.


Recently we reported evidence that Borrelia burgdorferi can also persist in the brain in chronic Lyme neuroborreliosis and, in analogy to Treponema pallidum, may cause dementia, cortical atrophy and amyloid deposition [3, 49, 51]. Only limited data have previously been available on the presence of atypical, cystic forms of spirochetes in the brain in chronic Lyme neuroborreliosis. Whether such forms may eventually cause functional damage and cell death is still not certain.

Here we analyzed atypical, cystic forms of Borrelia burgdorferi induced by unfavorable culture conditions and compared these with forms observed following 1 week of infection of primary chicken and rat neurons, as well as primary rat and human astrocytes. We also analyzed whether similar atypical and cystic forms may occur in vivo in brains of patients with pathologically and serologically confirmed Lyme neuroborreliosis and compared them to the atypical forms of Treponema pallidum in brains of patients with general paresis. The results show that under harmful culture conditions, the typical forms of Borrelia spirochetes are replaced by atypical forms varying from ring-shaped and cystic forms to fine single granules of almost submicroscopic size. These results are in harmony with previous observations [8, 55, 66]. The effect of osmotic shock induced with cold distilled water or heat shock was identical to those previously observed in other spirochetes [25, 67] Thioflavine S and Congo red had a similar effect. The mechanism of the harmful effect of these dyes is not known. They may act by binding to the outer sheath of Borrelia spirochetes [e.g. 56]. Thioflavin S and Congo red are widely used to detect amyloid deposits in affected tissues. Several observations suggested that Borrelia burgdorferi possesses amyloidogenic proteins [51, 68, 69]. Peptides derived from the OspA single-layer beta-sheet showed fibrillary amyloid formation, which may be an explanation of the binding of Thioflavin S and Congo red to the outer surface of Borrelia burgdorferi.

Atomic force microscopy (AFM) analysis showed rolled Borrelia spirochetes inside of a cyst covered by a thin outer membrane. This has also been observed in various types of spirochetes [e.g. 11, 38, 70] including Borrelia burgdorferi [8] by transmission electron microscopy analyses. Uni- or multi-spirochetal cysts may be formed. We illustrated by atomic force microscopy (AFM) rolling of two Borrelia spirochetes to form a cyst. The size of such cysts depends on the number of spirochetes packed inside of the cyst [8]. We observed bleb formation, connected to Borrelia spirochetes by a fine stalk, in both Borrelia strains. Thin newly formed spirochetes attached to spirochete cells, and to free minute granules were also observed.

Similar atypical, cystic and granular forms were observed in primary neuronal and astrocytic cell cultures exposed for 1 week to the Borrelia burgdorferi strains B31 and ADB1. ...
_________________________________________________________________


Lähettäjä: Soijuv Lähetetty: 30.9.2008 18:45

Edellisestä artikkelista jäi vahingossa pois linkki + artikkelin lopussa oleva linkki PDF tiedostoon josta löytyvät kystamuodon kuvat + tutkimus kokonaisuudessaan.


http://www.jneuroinflammation.com/content/5/1/40

Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis
Judith Miklossy , Sandor Kasas , Anne D Zurn , Shermann McCall , Sheng Yu and Patrick L McGeer

Journal of Neuroinflammation 2008, 5:40doi:10.1186/1742-2094-5-40


Published: 25 September 2008

Abstract (provisional)

Background
The long latent stage seen in syphilis, followed by chronic central nervous system infection and inflammation, can be explained by the persistence of atypical cystic and granular forms of Treponema pallidum. We investigated whether a similar situation may occur in Lyme neuroborreliosis. Method: Atypical forms of Borrelia burgdorferi spirochetes were induced exposing cultures of Borrelia burgdorferi (strains B31 and ADB1) to such unfavorable conditions as osmotic and heat shock, and exposure to the binding agents Thioflavin S and Congo red. We also analyzed whether these forms may be induced in vitro, following infection of primary chicken and rat neurons, as well as rat and human astrocytes. We further analyzed whether atypical forms similar to those induced in vitro may also occur in vivo, in brains of three patients with Lyme neuroborreliosis. We used immunohistochemical methods to detect evidence of neuroinflammation in the form of reactive microglia and astrocytes. Results: Under these conditions we observed atypical cystic, rolled and granular forms of these spirochetes. We characterized these abnormal forms by histochemical, immunohistochemical, dark field and atomic force microscopy (AFM) methods. The atypical and cystic forms found in the brains of three patients with neuropathologically confirmed Lyme neuroborreliosis were identical to those induced in vitro. We also observed nuclear fragmentation of the infected astrocytes using the TUNEL method. Abundant HLA-DR positive microglia and GFAP positive reactive astrocytes were present in the cerebral cortex. Conclusion: The results indicate that atypical extra- and intracellular pleomorphic and cystic forms of Borrelia burgdorferi and local neuroinflammation occur in the brain in chronic Lyme neuroborreliosis. The persistence of these more resistant spirochete forms, and their intracellular location in neurons and glial cells, may explain the long latent stage and persistence of Borrelia infection. The results also suggest that Borrelia burgdorferi may induce cellular dysfunction and apoptosis. The detection and recognition of atypical, cystic and granular forms in infected tissues is essential for the diagnosis and the treatment as they can occur in the absence of the typical spiral Borrelia form.

The complete article is available as a provisional PDF. The fully formatted PDF and HTML versions are in production.

[Sar_ani]

BIOFILMIT

Jo 80-luvulla on alettu puhua bakteerien muodostamista biofilmeistä. Ne ovat jonkinlaisia `koteloita`, joihin bakteeri suojautuu hyökkäyksiltä, niin elimistön kuin antibioottienkin.

Bakteereita ,joilla tällainen kyyky on, kutsutaan persister bakteereiksi. Biofilmejä muodostavia bakteereita ovat mm. mycobakteeri ja yllätys, yllätys: treponema (l. syfiliksen aiheuttava spirokeette-bakteeri).

Ja niinkuin on ollut puhetta siitä treponemasta; jo 40-luvulla mikrobiologit havaitsivat sen eri olomuodot. Jossain vaiheessa on sitten ilmeisesti tullut ilmeisesti täydellinen blackout, kun tiede on kieltänyt vanhat löydökset.

Mielenkiintoista alla olevassa tiivistelmässä on nuo neljä strategiaa, jotka on nostettu esiin, miten ongelmaa voitaisiin lähestyä. Nimittäin kohdassa 1 otetaan esille bakteerien ´herättäminen´, että ne voitaisiin sen jälkeen eliminoida. Tätähän se Viljasen ryhmä nimenomaan äskettäin tutki.

Muuallakin ollaan kiinnostuneita TNF:n ja gamma interferon:n vaikutuksista borrelioositartunnassa. javascript:emoticon(':roll:')




: Pathol Biol (Paris). 1990 Apr;38(4):276-80.Links




Mycobacteria, cytokines and antibiotics.

Rook GA.
Department of Medical Microbiology, University College and Middlesex

School of Medicine, School of Pathology, London.

We still do not understand the mechanism of immunity ty mycobacteria in

man, and convincing reproducible kill of M. tuberculosis by human

macrophages has not been achieved. The pathways so far elucidated,

involving gamma interferon, 1,25(OH)2 vitamin D3, and TNF release

seem more likely to lead to immunopathology than to protection.

Meanwhile the major problem for the clinician is the existence of

"persister" bacteria, which are not eliminated by the immune response,

even when therapy has greatly reduced the bacterial load. It seems

unlikely that it will be possible to design antibiotics which will rapidly kill

dormant persister bacilli, so new strategies for therapy may need to

concentrate on modulation of the host response. The objectives of such

therapies would be: 1) "Reawakening" of dormant persisters. 2) Rapid

immune recognition of persisters. 3) Suppression of the tissue-damaging

pathway. 4) Enhancement of the optimally protective mechanism, but this

has not yet been defined.


PMID: 2198523 [PubMed - indexed for MEDLINE

::

[soijuv]

Bakteerien muuntuminen L-muotoon on osoitettu jälleen kerran.

Molekyylimikrobiologian ja immunologian professori Y. Zhang ym. onnistuivat viljelemään L-muotoisia E.coli -bakteereja (2009). Jotkut bakteerit, kuten e.coli ja borreliabakteeri, muuntuvat L-muotoon stressitilanteissa esim. antibioottihoidon aikana. Muutamat muutkin bakteerit kuten mahahaavan ja tuberkuloosin aiheuttajat pystyvät muuntumaan tähän vaikeasti hoidettavaan muotoon. L-muotoisilla bakteereilla ei ole soluseinää. Tällöin ne kerääntyvät yhteen "paistetun kananmunan" kaltaisiksi ryhmiksi. Tässä muodossa bakteeria on vaikea esim. viljellä koeolosuhteissa. Ne ovat myös vastustuskykyisiä esim. tavallisille penisilliiniryhmän antibiooteille.

Bakteerit kykenevät muuntumaan useisiin eri muotoihin, L-muoto on vain yksi esimerkki muodonmuutoksesta joka tapahtuu antibioottihoidon aiheuttaman stressin vaikutuksesta. L-muodot saattavat olla perussyynä erilaisiin kroonisiin sairauksiin kuten reuma, sarkoidoosi, erilaiset tulehdukselliset ruuansulatuskanavan sairaudet jne. Zhangin ym. tekemän löydön pohjalta on mahdollista löytää uusia, nykyistä tehokkaampia antibiootteja. Niiden myötä immuunipuolustus saattaa havaita ja tuhota "paljastuneet" bakteerit entistä paremmin.

Bacterial antibiotic resistance genes discovered

Artikkeli: http://media.www.jhunewsletter.com/medi ... age2.shtml

Tutkimus: http://www.plosone.org/article/info:doi ... -Domingue1

By Aleena Lakhanpal
Issue date: 11/5/09

Antibacterial soap, hand sanitizer and antibiotics are all substances that we use in an attempt to kill bacteria that might make us sick.Whether we are concerned about getting strep throat, bacterial meningitis or something else, these prevention methods can offer protection.

However, some bacteria, such as those that cause Staph and MRSA infections, are becoming increasingly resistant to antibiotics. Since the 1930s, researchers have been aware that bacteria may be able to resist treatment because they can morph into the L-form, or bacteria lacking cell walls.

Until the 1980s, not much else could be known about the L-form, but now, researchers at the Bloomberg School of Public Health have used a wide variety of modern molecular tools to learn more about the origin and biological functions of the L-form bacteria.

Ying Zhang, a professor of molecular microbiology and immunology at Bloomberg, is the senior author of the study, which was published in PLoS ONE last month.

Not all bacteria can transform into the L-form, but those that can include Bacillus anthracis (anthrax), Treponema pallidum (syphilis), Mycobacterium tuberculosis (tuberculosis), Heliobacter pylori (stomach ulcers and cancer), Borrelia burgdorferi (Lyme disease) and Escherichia coli (food poisoning). Zhang's team used E. coli to create a culture of L-form bacteria.

Although it had been difficult to culture L-form bacteria before, Zhang and his team created a new method that more closely simulated the in vivo conditions in which these bacteria form.

"The presence of antibiotic stress is cell wall inhibiting, like penicillin," Zhang said. To prevent the cells from bursting because of this increased stress, Zhang's team added sucrose to the cell media.
This culture represented the mechanism that occurs in the body. "L forms are formed in response to stress," Zhang said. "They have a different mode of survival and replication from classical bacteria." The cell wall-deficient bacteria cluster together in the shape of a fried egg rather than the smooth, homogeneous appearance of wild-type bacteria cultures.

Not only are L-form bacteria difficult to culture and therefore study, but this "fried egg" cluster is part of what makes the L-form bacteria resistant to antibiotics, in addition to the fact that they do not have cell walls for commonly used antibiotics to disintegrate.

Once Zhang and his team were able to successfully culture L-form E. coli, they screened for and identified mutants that fail to grow at the L-form. From these mutants, they were able to discover a series of genes that were linked with the inability to grow in the L-form.

"These fall into four to five different categories involving extracellular matrix synthesis, membrane proteins, membrane biogenesis, DNA repair as well as iron metabolism and energy metabolism," Zhang said.

Their identification of these genes and their effect on L-form bacterial expression is a resounding discovery because it was impossible to do before, what with the difficulty of culturing the L-forms of various bacteria. Zhang noted, however, that although his team managed to create and study a culture of L-form bacteria, their study cannot be universal.

"What we can culture is only a small percentage - probably less than 1 percent - of all bacteria on earth," Zhang said.
"They exist in nature and grow easily, but we're limited to what we can grow and the form of bacteria that can grow. Bacteria can grow a variety of different forms even for the same species, and can change forms under different conditions. L-forms are one example of changing under antibiotic stress."

These L-forms of various bacteria may be the underlying reason for chronic resistant and recurring diseases, such as sarcoidosis, various forms of inflammatory bowel diseases and rheumatoid arthritis. Zhang is confident that there will be many practical applications of this discovery.

"It is possible, with our discovery of the L-form genes to develop new antibiotics and more effective ones that can be used with current ones as well as new vaccines to . . . allow these forms to be eliminated by the immune system," he said.

[soijuv]

Borreliabakteeri muuntuu stressin esim. antibioottien vaikutuksesta vaikeasti hoidettavaan L-muotoon. (Johns Hopkins 2009)

The article below (by Steve Nery) was published in at least three Maryland newspapers last week- the Star Democrat (front page), the Kent County News and the Record Observer.

The Kent County News

Lyme disease victims eye beneficial study.

By Steve Nery

BALTIMORE Lyme disease patient advocates are hopeful that new studies on especially resistant forms of bacteria at Johns Hopkins could help lead to less restrictive treatment guidelines.

The November issue of the Johns Hopkins newsletter contains an article about research at the Bloomberg School of Public Health on bacteria that can morph into the "L-form" bacteria that lack cell walls that are resistant to antibiotics. They include Borrelia burgdorferi, the bacteria that causes Lyme disease, as well as the bacteria responsible for anthrax, syphilis, tuberculosis, food poisoning and stomach ulcers.

The Infectious Diseases Society of America recommends the use of an antibiotic for a maximum of four weeks to treat Lyme disease, but many patients, especially those not diagnosed swiftly following a tick bite, report effects that can continue to linger for life. Droves of Lyme disease patients spend thousands in out-of-pocket dollars to receive treatment from the few doctors that don't follow the IDSA guidelines.

Ying Zhang, Yanking Yang and William A. Glover authored a study published last month in PloS ONE. The researchers used modern molecular tools to learn more about the origin and biological functions of the L-form bacteria. Researchers have known about the existence of this form of bacteria since the 1930's, but little beyond that.

Zhang, a professor of molecular microbiology and immunology at Bloomberg and the senior author of the study, said in the newsletter that the bacteria take on the L form in response to stress caused by antibiotics. The wall-less cells cluster together in the shape of a fried egg, which makes them ever more resistant to antibiotics.

The team successfully cultured E. coli L-form bacteria and discovered genes that were linked with the inability to grow in the L-form.

These L-forms of various bacteria may be the underlying reason for chronic diseases such as rheumatoid arthritis, according to the newsletter.

"It is possible, with our discovery of the L-form genes, to develop new antibiotics and more effective ones that can be used with current ones as well as new vaccines to ? allow these forms to be eliminated by the immune system", Zhang said in the publication.

Patients with chronic Lyme disease hope new research will change the Infectious Diseases Society of America's Lyme disease treatment guidelines, which are under review as ordered by Connecticut Attorney General Richard Blumenthal.

"Hopkins proving L-form bacteria is not killed by antibiotics while in this form shows that the chronic and debilitating symptoms, disability and death can all be related to ongoing infection in those that have been exposed", said Lucy Barnes, director of the Lyme Disease Education and Support Groups of Maryland

[soijuv]

Borreliabakteerit voivat selvitä elimistössä vuosikausia kystamuodossa (pyöreä muoto). Siinä muodossa ne selviävät paremmin erilaisista uhista kuten antibiooteista. Uusi antibiootti "Tigesykliini" saattaa olla aiempia antibiootteja tehokkaampaa.


http://www.umass.edu/loop/talkingpoints ... /99534.php


Margulis and colleagues spirochete research may point to new Lyme treatment


[] In Proceedings of the National Academy of Sciences, Distinguished Professor Lynn Margulis, Geosciences, UMass Amherst advanced and former students James MacAllister and Andrew Wier and their colleagues in Norway and Canada, report new details about the life history of the spirochete bacterium, Borelia burgdorferi, which causes Lyme disease. Their study leads them to suggest that clinical trials of the antibiotic Tigecycline are warranted. This drug potentially provides a treatment regimen for people with Lyme disease that has worked, so far only in the laboratory, to destroy spirochetes, they point out.

The study represents a lifetime of work by medical microbiologist Oystein Brorson and his pathologist cousin, Sverre-Henning Brorson of Oslo, Norway, says Margulis, a winner of the Darwin and Wallace Medal from the Linnean Society of London. She recently returned to campus from a year as Eastman Professor at Balliol College, Oxford University.

B. burgdorferi spirochetes take up residence in tissues of people bitten by ticks that carry the bacteria in their guts and inject them via saliva. The spirochetes most familiar form is an agile, spiral-shaped swimmer associated with acute symptoms of Lyme disease and rapid reproduction of the bacteria. But as Margulis explains, these bacteria can go underground and persist for years by entering a self-protective, quiescent stage known as a spirochete round-body (RB) propagule. In this state, they better resist what scientists call unfavorable environmental conditions such as starvation, desiccation and exposure to antibiotics such as penicillin and deoxycillin. Chronic Lyme disease symptoms correlate with the continuous presence of reversible RB propagules in patients moist tissues.

What the Brorsons work shows is that, unlike other antibiotics, Tygecycline administered at the correct dosage and timing destroys the bacterium even when it has protected itself in this quiescent stage. Other antibiotics, if they do anything at all, simply cause B. burgdorferi to enter its RB propagule state and wait out the treatment. â??Tigecycline is, so far, the only known antibiotic that destroys the Lyme disease spirochete in both the growing and the quiescent RB stages of its life historyâ? Margulis notes.

She and her students are interested in spirochetes, in particular two individuals found embedded in ancient (15 million-year-old) amber and their RB propagules, because of the Serial Endosymbiosis Theory of the evolution of nucleated cells. It posits that animal, plant, fungal and other cells with nuclei evolved by symbiotic merger of two types of bacteria, at least: A spirochete (belonging to the eubacterium group) that reversibly develops RB propagules in response to changing conditions, and Thermoplasma, from the archaebacterium group, a sulfide-gas-making microbe that lives in nearly boiling and very acidic waters (more acid than our stomachs). In their PNAS paper, the authors state that chronic spirochete infections in humans when seen in their ecological-evolutionary context are examples of symbioses that have evolved over geologic time.

Image: Persistent quiescent Lyme disease spirochete (Borrelia burgdorferi) round bodies (RBs) may endure for years. They stain red as seen with here with a fluorescence microscope (scale bar = 8 micrometers). One RB is seen at very high magnification with an electron microscope (EM) thin section in the lower left inset). After incubation in a favorable supportive blood serum growth medium for over one month the RBs develop into normal helical swimmer spirochetes as seen in the EM photo of the upper right inset (bar = 1 micrometer). (Photo courtesy of Oystein and Sverre-Henning Brorson)
February 17, 2010.

[soijuv]

"Borrelia-bakteereita (B.garinii) kasvatettiin koeputkiolosuhteissa erilaisissa liuoksissa. Bakteeri muuntui eri kokoisiksi kystamuodoiksi (yksittäisiä ja ryhmiä) erityisesti epäedullisissa olosuhteissa. Brasiliassa esiintyvä Borrelioosin kaltainen sairaus, nimeltään Baggio-Yoshinari, on borrelia-bakteerin kystamuodon aiheuttama."

Growth, cysts and kinetics of Borrelia garinii (Spirochaetales: Spirochaetacea) in different culture media.

Angela de OliveiraI; Adivaldo Henrique FonsecaI, +; Catia Marques da CostaI; Elenice MantovaniII; Natalino Hajime YoshinariII

IPrograma de Pós-Graduação em Ciências Veterinárias, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, BR 465 Km 7, 23890-000 Seropédica, RJ, Brasil
IIDepartamento de Reumatologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil


ABSTRACT

The aim of the present paper was to evaluate cyst formation and growth parameters of Borrelia garinii in a range of media differing in formulation and cost. A qualitative assessment of morphology and motility of B. garinii was conducted. All media were prepared aseptically and used in test tubes or Petri dishes. For each medium, the initial spirochete concentration was standardized to 103 spirochets/mL. The following culture media were suitable to grow B. garinii: Barbour-Stoenner-Kelly, brain heart infusion and PMR. Growth was minimal at six weeks post-inoculation and maximum spirochete density was observed between 9-12 weeks.

Often, the cultures developed cysts of different sizes, isolated or in groups, with a spiraled portion of variable sizes, mainly in unfavorable culture media.

Brazilian Lyme disease-like illness, also known as Baggio-Yoshinari syndrome (BYS), is a new and interesting emerging tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, only during its cystic forms.

It has been assumed that the peculiar clinical and laboratory features of BYS are consequential to the absence of a human sucker Ixodes ricinus complex tick at risk areas in Brazil, supporting the concept that the borrelia phenotypic expression pattern is modified as it is transmitted through the host.

Key words: culture media - Borrelia garinii - spirochaetaceae - kinetic growth - cysts



full text pdf:

http://www.scielo.br/pdf/mioc/v105n5/20.pdf

[soijuv]

Borreliabakteerin spirokeettamuoto muuntuu epäedullisissa olosuhteissa pyöreään muotoon. Se saattaa selittää bakteerin pitkäaikaisen selviämisen elimistössä. (Saksa 10/2010)

Metamorphosis of Borrelia burgdorferi organisms ? RNA, lipid and protein composition in context with the spirochetes' shape

1. Samiya Al-Robaiy1,2,?,
2. Hassan Dihazi3,?,
3. Johannes Kacza4,
4. Johannes Seeger4,
5. Jürgen Schiller5,
6. Daniel Huster5,
7. Jens Knauer1,6,
8. Prof. Dr. Reinhard K. Straubinger1,7,*

Article first published online: 21 OCT 2010

Author Information

1. 1

Institute of Immunology, College of Veterinary Medicine, and Center for Biotechnology and Biomedicine, University of Leipzig, Germany
2. 2

Cardiothoracic Surgery, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany
3. 3

Department of Nephrology and Rheumatology, University Hospital Goettingen, Germany
4. 4

Institute of Veterinary Anatomy, University of Leipzig, Germany
5. 5

Institute of Medical Physics and Biophysics, Medical Department, University of Leipzig, Germany
6. 6

Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
7. 7

Institute for Infectious Diseases and Zoonoses, Department for Veterinary Sciences, Faculty of Veterinary Medicine, LMU Munich, Germany

Email: Prof. Dr. Reinhard K. Straubinger (R.Straubinger@lmu.de)

*Correspondence: Prof. Dr. Reinhard K. Straubinger, Phone: +49 (0)89 2180 2528, Fax: +49 (0)89 2180 99 2527

1. ?

Authors contributed equally to this work.


Abstract

Borrelia burgdorferi, the agent of Lyme borreliosis, has the ability to undergo morphological transformation from a motile spirochetal to non-motile spherical shape when it encounters unfavorable conditions. However, little information is available on the mechanism that enables the bacterium to change its shape and whether major components of the cells ? nucleic acids, proteins, lipids ? are possibly modified during the process.

Deducing from investigations utilizing electron microscopy, it seems that shape alteration begins with membrane budding followed by folding of the protoplasmatic cylinder inside the outer surface membrane. Scanning electron microscopy confirmed that a deficiency in producing functioning periplasmic flagella did not hinder sphere formation. Further, it was shown that the spirochetes' and spheres' lipid compositions were indistinguishable. Neither phosphatidylcholine nor phosphatidylglycerol were altered by the structural transformation. In addition, no changes in differential protein expression were detected during this process. However, minimal degradation of RNA and a reduced antigen-antibody binding activity were observed with advanced age of the spheres.

The results of our comparisons and the failure to generate mutants lacking the ability to convert to spheres suggest that the metamorphosis of B. burgdorferi results in a conditional reconstruction of the outer membrane. The spheres, which appear to be more resistant to unfavorable conditions and exhibit reduced immune reactivity when compared to spirochetes, might allow the B. burgdorferi to escape complete clearance and possibly ensure long-term survival in the host. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

[soijuv]

http://www.lymenet.de/literatur/cystsl.htm


Historical Observations of Spirochetal Cysts
and L-Forms
Summary of historical observations of spirochetal cyst (and L-forms) prepared by an anonymous volunteer of actionlyme.

(P)= interesting photos, (R)= Highly recommended reading, (F)=foreign language
Borrelia - burgdorferi
Microbiology, 2000 Jan;146 (Pt 1):119-27

Serum-starvation-induced changes in protein synthesis and morphology of Borrelia burgdorferi.
Alban PS, Johnson PW, Nelson DR

B. burgdorferi spirochetes converted to a cystic form when placed in a deficient culture media. These cysts reverted to motile spirochetes when transferred to growth media.
[From the abstract:] "Cyst opening and recovery of spiral-shaped non-motile organisms was induced within 1 min by the addition of either BSKrs+ or rabbit serum (6%v/v, final concentration). Cells regained motility with additional incubation in BSKrs+."

APMIS, 1999;106(12):1131-1141

A rapid method for generating cystic forms of Borrelia burgdorferi, and their reversal to mobile spirochetes.
Brorson, O., & Brorson, S.
Motile B. burgdorferi spirochetes converted to cysts within 1 minute when placed in distilled water. The cysts reverted to spirochetes after transfer to a growth medium.

APMIS, 1999;107(6):566-576.

(R) Brorson, O., & Brorson
An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to metronidazole.
B. burgdorferi cysts were degraded upon incubation with metronidazole (in vitro).

12th International Conference on Lyme Disease and Other Spirochetal and Tick-Borne Disorders., April 8-9, 1999
Keynote Address - The Complexity of Vector-borne Spirochetes.
Burgdorfer, W.
"This relatively large Borrelia [Borrelia burgdorferi] is not readily detectable in blood smears or thick drops of Lyme disease patients and susceptible host animals, yet engorgement on infected hosts results in up to 100% infected ticks.... RML [NIH's Rocky Mountain Lab] scientists Dave Dorward and Claude Garon using silver staining, transmission and scanning electron microscopy investigated the nature of naturally elaborated membrane blebs on the surface of cultured B. burgdorferi or free in the medium, and found both linear and circular DNA (Fig.13)... These most recent findings [of RML researchers and others] do confirm the development of membrane-derived cysts, blebs, spherules, vesicles and the potential transformation to motile, helical spirochetes...as a "survival mechanism" of spirochetes to overcome or escape unfavorable conditions." [Willy Burgdorfer, Ph.D., of the National Institutes of Health, is the discoverer of Borrelia burgdorferi.]

Infection, 1998;26(3):144-50

(R) In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium
Brorson, O., & Brorson S.
. B. burgdorferi transformed into cysts (spheroplast L-forms) within 1-24h of inoculation into spinal fluid. When transferred to a growth medium, the cysts converted back to normal spirochetes after 9-17 days of incubation. The authors suggest that encystment of Bb may explain why cultivation of spinal fluid may be negative in patients with neuroborreliosis. The cysts may be recognized by microscopy.

American Journal of Dermatopathology,1996;18(6):571-9.

(R) Heterogeneity of Borrelia burgdorferi in the skin.
Aberer, E., Kersten, et al
Encysted B. burgdorferi were found in a skin biopsy. The authors conclude that these are in vivo variant forms of Bb and not staining artifacts. Includes remarkable color videomicroscopy photographs as supporting evidence.

[Diagnosis:] "Borrelia may escape immune surveillance by colony formation and masking within collagen, resulting in seronegativity."

Infection, 1996;24(3):218-26.

Formation and cultivation of Borrelia burgdorferi spheroplast L-form variants.
Mursic, V.P., Wanner, et al
[Cysts:] In vitro investiation of morphological variants of B. burdorferi, in an effort to explain the clinical persistence of active Lyme borreliosis despite antibiotic therapy. The authors suggest that these atypical forms may allow Borrelia to survive antibiotic treatment. [Persistence:] "...clinical persistence of Borrelia burgdorferi in patients with active Lyme borreliosis occurs despite obviously adequate

Antimicrobial Agents & Chemotherapy, 1995;39(5):1127-33.
(R) Effects of penicillin, ceftriaxone, and doxycycline on the morphology of Borrelia
Kersten, A, Poitschek C, Rauch S, Aberer, E
B. burgdorferi cultures gradually developed cysts and blebs [granules] when incubated in antibiotics. The degree of alteration was strongly correlated with dose and duration.The alterations occured most rapidly with ceftriaxone, then with penicillin; doxycycline caused a smaller percentage of organisms to undergo morphologic changes. Those organisms that remained in typical spirochetal form in the doxycycline cultures showed decreased motility. The authors suggest that these morphologic changes may shed light on the ability of B. burgdorferi to survive

Infection, 1994 ;22(6):401-406.

Ultrastructure of Borrelia burgdorferi after exposure to benzylpenicillin
Schaller, M. et al&
B. burgdorferi were observed to form cysts and blebs when treated with penicillin G. "These structures were not found under optimal culture conditions. One may speculate that the borreliae could escape the action of the antibiotic by developing such spherical bodies." (p. 404)

Abstracts of the V International Conference of Lyme Borreliosis, Arlington, VA.1992

Borrelia burgdorferi in the skin: a morphological and immunohistochemical study of the heterogenous appearance of this microorganism
Aberer, E., Kersten, et al
B. burgdorferi granules were detected in skin biopsies of erythema migrans and acrodermatitis chronica atrophicans lesions.

Annals New York Academy of Sciences, 1988;539:468-470.

Concurrent neocortical borreliosis and Alzheimer's disease: Demonstration of a spirochetal cyst form
MacDonald, A.B.
In vivo finding of Borrelia burgdoreri cysts in an autopsy of a human brain. "An unexpected observation was the identification of cystic forms of the Borrelia spirochete in dark-field preparations of cultured hippocampus, and in imprints of hippocampus... A cystic form of the Borrelia spirochete would explain the ability of the microbe to persist in the host during a prolonged period of asymptomatic clinical latency, which spans the period between primary infection and the expression of tertiary manifestations of neuroborreliosis."

Borrelia - duttoni
Lancet, 1920 ;2:1237-1244.

(R) The Horace Dobell lecture on an experimental investigation of Spirochaeta duttoni, the parasite of tick fever
Leishman, W.B.
The author argues that S. duttoni, when inside a tick, reproduce by a process of budding and extrusion of granules; the granules grow into young spirochetes. The granules are also themselves capable of multiplication. Their development into spirochetal form within a vertibrate host is an exceptional occurence, brought about by certain environmental circumstances. This theory rests on an accumulation of observations by the author and by other researchers cited, including a correlation of the temporary disappearance of spirochetes from the tick's stomach with the appearance of large numbers of granules. Temperature was found to influence the development of classic-shaped spirochetes.

Annales de l'Institut Pasteur, 1918;32:49-59.

A note on the "granule clumps" found in Ornithodorus moubata and their relation to the spirochaetes of African relapsing fever (tick fever).
Leishman, W.B.
Granules developed into spirochetes. Innoculation of tissues containing only granules produced spirochaetosis in mice. Observed periods of several days where few, if any, spirochetes could be found inside a tick, followed by sudden re-invasion of tissues with mostly young and vigorously motile spirochetes; then this sequence would repeat in a regular pattern. Concludes that this has to do with the reproductive habits

Parasitology, 1912;Vol. IV, p.133.

The transmission of Spirochaeta duttoni.
Hindle, E.
[According to W. Burgdorferi, 1999:] Hindle reported that in infected ticks kept at 21SC, the spirochetes disappeared from the midgut by the 10th day and could no longer be detected either in the gut or in the tissues. However, triturates of such ticks were able to infect mice, and an increase in temperature to 35SC led to the reappearance of typical spirochetes.

Annals of Tropical Medicine and Parasitology, 1907;No. 3

The morphology and life-history of Spirochaeta Duttoni
Breinl
[According to Dutton, 1907:] Observed encysted forms of S. duttoni in the spleen. The cysts broke into granular bodies from which new generations of spirochetes emerged.

Lancet, 1907;ii:1523.

(R) A note on the morphology of Spirochaeta Duttoni.
Dutton, J.S. & Todd,
Spirochetes within sporocyst-like bodies were found in the blood even when other forms had disappeared. The authors suggest that reproduction may occur by direct division, but possibly also by a process involving extrusion of granules which subsequently develop into new spirochetes.

Borrelia - recurrentis
Compt. rend. acad. sci., clix, 1914;pp. 119-122.
(F) De la periode de latence du Spirille chez le Pou infecte de fievre recurrent.
Sergent, E. & Foley,
[As described by Leishman, 1920): After ingestion into ticks, the Leptospira studied disappeared after 24 hours. After the 6th day, new, actively mobile spirochetes reappeared suddenly in great numbers. Infectivity was highest on the 6th day prior to this reappearance, despite of the absence of demonstrable spirochetes. Transverse fission of the spirochetes in the louse was only rarely observed. When classic-shapedspirochetes were present, their numbers did not increase.

Leptospira
Journal of Bacteriology 1954;67:619-627.

(R) Formation of granular structures by Leptospirae as revealed by the electron microscope.
Czekalowki, J.W., et al
Leptospira began to show granulation after 2 weeks in a culture. The granules were spaced regularly within the bodies of the spirochetes. After four weeks a larger type of granule appeared which was broader than the body of the spirochetes. These were later "shed free." By the 5th to 7th month, there were no spirochetes observed; the culture contained only granules. The granules consisted of "what appears to be short segments of leptospiral body embedded in homogeneous substance." The authors conclude that the "formation of granules represents a rhythmic and constant process and hence these granules must play a role in the life-cycle of leptospirae."

J. Hygiene, 1949;47:390-392.

The morphology of the genus Leptospira as shown by the electron microscopy.
Babudieri, B.

Acta Biologica Belge, 1943;3-4:245.

(F)Coloration des granules leptospiriens.
Herreweghe, E.

Bull. ass. diploms microbiol. fac. pharm.,1942;61:72-80.

(F) Le micro-manipulateur et les granules d'une souche de Leptospire aquicole non-pathogene.
Bessemans, A.,
[According to Czekalowski, 1954]: Granules from the culture of a leptospira were isolated using a micromanipulator and grown from single cell

Compt. rend. soc. biol., 1942;136:141-144.

(F) Sur l'evolution du L. ieterohaemorrhagiae, granule leptospirogene.
Gastinel, P. et al

Quoted by Van Thiel, P.H., 1948.
The leptospiroses.
Timmerman, H. 1927

Universitaire Pers, Leiden.

Granules develop in response to physical and chemical changes.

Journal of Experimental Medicine, 1916;23:377-402.

The etiology and mode of infection and specific therapy of Weil's disease.
Inada, R., Ido, Y., et al,
Interprets atypical spirochetal forms as "degenerative." Spirochetes were seen inside phagocytic cells and epithelial cells. Classic-shaped spirochetes were found in the blood in small numbers, and only in the early stages of the disease.

Non-Spirochetal
Journal of Bacteriology, 1942;44:37-70.
The significant of the large bodies and the development of L-type colonies in bacterial cultures.
Dienes, L.
"If they [L type colonies] represent a variant form [of bacterial morphology], the observations indicate that the variability of bacteria extends much further than is commonly supposed. The bacteria apparently are able to grow in the form of soft granules and to multiply by the development and germination of large bodies." (p.60)

Other/Mixed/Unknown

CRC Press, 1993

Cell wall deficient forms: Stealth Pathogens
Mattman, L.H.

Eur. J. Clin. Microbiol. Infect. Dis., 1989.

Unusual in vitro formation of cyst-like structures associated with human intestinal spirochaetosis
Gebbers, J.O. et al
In vitro findings suggest that spirochetes may develop in cysts, contrary to the traditional view that transverse fission is their main mode of reproduction. As cysts were found in centrifugates of cultures but not in biopsy speciments, the authors speculate that this mode of reproduction may occur only when in sub-optimal environments outside the host. Electron micrographs of maturation of spirochetes within the

Microbiol. Immunology, 1980;24:321-334.

Electron microscopy of the spherical bodies of oral spirochetes in vitro.
Umemoto, T., et al

J. Gifu Dent. Soc. 2:1-15.

Spherical body formation of oral spirochetes following addition of sucrose
Umemoto, T. et al 1974

Am J. MS, 1958;236:677-91.

Preliminary observations of the cultivation and morphology of a microorganism from the cerebrospinal fluid of patients with MS.
Myerson, R.M.

Journal of Neuropathology, 1954;13:221-29.

Morphology of Spirochaeta myelophthora in multiple sclerosis
Steiner, G.

Journal of Neuropathology, 11:343-72.

(R) Acute plaques in multiple sclerosis, their pathogenic significance and the role of spirochaetes as etiological factor.
Steiner, G. 1952

Journal of Bacteriology, 1951;62:347-349.

(P) Further studies on the significance of spirochetal granules.
Hampp, E.G.

American Journal of Syphilis, 1950;34:122-125.

Studies on the life cycles of spirochaetes: I. The use of phase contrast microscopy.
Delamater, E.D., et al
Includes several small photos of spirochetes emerging from "gemma," which the authors interpret as reproductive forms.

J Am Dental Assoc, 1950;40:1-11.

Morphologic characteristics of the smaller oral treponemes and Borrelia vincentii as revealed by stained smear, darkfield and electron microscopic technics.
Hampp, E.G. et al

Archives of Neurology and Psychiatry , 1939.

(R) "Silver cells" and "spirochete-like formations" in MS and other diseases of the central nervous system
Hassin, G.B. et al
Review confirms the findings of G. Steiner and other researchers who found "silver cells" [spirochetal granules that take a silver stain] in brain autopsies of MS cases. (G. Steiner contended that MS is an infectious disease caused by a spirochete that changes from a classic spirochetal form to a granular form, and which destroys myelin.) While the authors found granules in the CNS of all 8 MS patients they studied, they dispute Steiner's belief that they are the causative agent of the disease.

Deutsche Ztschr. f. Nervenheilkunde, 1928;107:112.

(F) Demonstration von Spirochäten im menschlichen Gehirn bei Multipler Sklerose.
Steiner, G.
(Demonstration of spirochetes in the human brain in multiple sclerosis)

Das Handbuch der pathogenen Protozoen, Leipzig, Johann Ambrosius Barth, Band 3, 1925

(F) Die Spirochäten
Zuelzer, M.

Münchner med. Wochenschrift, Med. Klin., 1919;66:1245

(F) (R) Forschungen über die Ätiologie der multiplen Sklerose.
Kuhn, P. & Steiner,
(Research on the etiology of multiple sclerosis)

Münchner med. Wochenschrift, Med Klin, 1917;13:1007

(F) (R) Über die Ursache der multiple Sklerose.
[The cause of multiple sclerosis.]
Kuhn, P., & Steiner,

British Medical Journal, 1916;i:409-411.

(R) Spirochaetes and their granular phase.
Fantham, H.B.
Observed the process of spirochetal formation of granules, and the emergence of small spirochetes from these granules. "It must also be borne in mind that coccoid bodies may be present when spirochaetes as such cannot be detected." (p.410)

Julius Springer, Berlin, 1914.
(F) Studien über die Fortpflanzung von Bakterien, Spirillen and Spirochaeten.
Meirowsky, S.E.

British Medical Journal, 1911;1:752.

(R) The infective granule in certain protozoal infections, as illustrated by the spirochaetosis of Sudanese fowl.
Balfour, A.
Spirochetes were observed to discharge large numbers of granules. "...the spirochaetes undergo an astonishing change. They discharge from their periplastic sheaths spherical granules, and it is apparently these granules which enter the red cells, develop in them and complete a cycle of schizogony...In process of time the spirochaete loses its activity, becomes difficult to see, and eventually all that is left of it is the limp and lifeless... [that the granules] do not appear to take on the Romanowsky stain may explain why they have not previously been noticed... I have found these granules to be resistant forms and their presence in countless numbers in the tissues might explain part of the mechanism of relapse and the difficulty of curing completely some of the more chronic spirochaetal infections, as, for example, syphilis and

Journal of Infectious Diseases, 1906;3:291-293.

Studies on the Spirillum obermeieri and related organisms.
Novy, F.G., & Knapp,
Found that the classic spiral form is not the only form that spirochetes may assume.

Treponema ...
Zbl. Bakt., 1994;280:297-303.

Formation of multiple treponemes.
Wolf, V. & Wecke, J.
[From the abstract:] "It was calculated that the formation of spherical bodies may reduce their surface by up to 75% as compared to the single form. Thus, the reaction surface for antibodies or other compounds produced by the host is considerably diminished. Therefore, such spherical structures being at resting states may represent a survival strategy of spirochetes. ...the spherical bodies may be the starting point of the new inflammatory episode. This wavelike process is typical of many spirochetal diseases."

Vestn Dermatol Venerol, 199?;4:32-6.

(F)The cytoarchitectonics of hard chancre in rabbits with experimental syphilis exposed to oliusulfon and cefamezine
[In Russian; English abstract available]
Delektorskii VV.,
Describes T. pallidum ultrastructure, and the process of formation of a granule. Cefamezin did not effect spirochetal cysts in the treatment of (snip)

Microbiol. Immunolo.,;1984;28:11-22.

Colonial morphology of treponemes observed by electron microscopy.
Umemoto, T., et al,
[From the abstract:] "Scanning and transmission electron microscopy revealed that the colonies of Reiter treponemes contained spherical forms almost up to 5 am in diameter, each consisting of an outer membrane and a treponemal main body."

Journal of Applied Bacteriology, 1983;55:417-428.

(R)A proposed life cycle for the Reiter treponeme.
Al-Qudah, A.A.
Demonstrates the viability of cysts and the existence of a complex manner of reproduction. "Although transverse fission may be the main mode of reproduction of Reiter treponemes in optimal growth conditions, the spontaneous formation of cysts increases in aging cultures to the extent that it is rare to find a typical treponeme in old cultures. We conclude that such cysts... [serve to] by-pass adverse environmental conditions and to ensure the propagation of the organism. ...the existence of the causative agent of syphilis in a nonspirochetal form has long been hypothesized to explain the latency of syphilis and the infectivity of tissues devoid of demonstrable treponemes...This agrees with what usually happens in protozoa in nature; ...the majority of cysts in protozoa are a means of protecting their contents against unfavorable conditions... Later, depending on conditions when the harmful exposure is past, protective cysts may become multiplication cysts. They are not

Microbiol. Immunolo. ,1982;26(3):191-198.

An internal view of the spherical body of Treponema macrodentium as revealed by scanning electron microscopy.
Umemoto, T., et al,
"External observation of a spherical body by scanning electron microscopy clearly revealed the main bodies [spirochetes] running beneath the inner surface of the spherical body membrane [cyst]. " Includes a freeze fracture photograph of a cross-section of a multispirochetal cyst,

Acta Pathol Microbiol Scand [A],1977;pertenue (sic KMD) Electron microscopy of lymph nodes of hamsters experimentally infected with Treponema
Blom J.
Treponemes were found intracellularly in macrophages. These treponemes did not show their typically helical shape, but were present as spherical forms or cysts.

J. Am. Vener. Dis. Assoc. , 1976;3(2):109-127.

Biopharmacology of syphilotherapy.
Rein, M.F.

British Journal of Venereal Diseases, 1971

(P) Current concepts of the morphology and biology of Treponema pallidum (syphilis) based on electron microscopy
Ovcinnikov, N.M., et al
[Granules:] "Another mode of reproduction resorted to in adverse circumstances consists in the formation of spores which subsequently develop into new treponemes. The breakdown into granules is especially pronounced under the action of penicillin and immune sera." [Cysts:] "Under stressful conditions, the treponeme 'packs' itself into a compact roll (Fig. 8) and becomes covered with a transparent mucoid capsule, which resists the pentration of drugs and antibodies." "Encystment as a mechanism of survival and mode of reproduction is widespread in nature, especially among protozoa." [Intracellular:] T. pallidum were found inside a cell taken from the site of a chancre; and L-forms were found

New England Journal of Medicine, 1971; 284: 642-653.

Diagnosis and treatment of syphilis.
Sparling, P.F.
Includes a review of recent [as of 1971] evidence indicating that penicillin treatment is not always curative in patients with late syphilis. "Penicillin therapy of neurosyphilis has not been as effective [as in early syphilis]. Several studies have reported relapses... Clinical progression of symptomatic neurosyphilis is relatively common despite antibiotics." (p.650) [Diagnostic issues:] Some infected patients also

British Journal of Venereal Diseases, 1968;44:1-34.

(P) Further study of ultrathin sections of Treponema pallidum under the electron microscrope.
Ovcinnikov, N.M., et al,
Observations of T.pallidum cystic and granular formations under the electron microscope. "...under unfavourable conditions of existence, treponemes form real cysts as a method of persistent survival and multiplication, as occurs not infrequently among protozoa." "As the treponeme moves, the thickness changes. This indicates that the body possesses a capacity for contraction... The sharply-marked structural elements of the treponeme and its complex and characteristic structure indicate that cysts are not a product of degeneration. In addition, in cultures where there are many cysts, they are very mobile, which is another argument against degeneration... When transfers are made from cultures containing cysts and almost no ordinary spiral forms, growth of ordinary spiral forms occurs." Includes photo of a treponeme packed into a cyst

British Journal of Venereal Diseases, 1968

Further observations on the persistence of Treponema pallidum after treatment in rabbits and humans.
Yobs, A.R.
Results of a 4-year study of rabbits treated with penicillin for late latent syphilis. Confirmed persistence of syphilis in numerous subjects after antibiotic treatment. Cortisone treatment can reactivate clinical disease. Offers various theories to explain the persistence of T. pallidum, including morphologic changes in the organism. However, the author believes that the existence of a complex life cycle with differing

Vestn Akad Med Nauk SSSR. 1965;20(8):46-50.

[L-forms of Treponema pallidum].
Ustimenko LM
in Russian. No abstract available.
PMID: 5328461 UI: 66154896

British Journal of Venereal Diseases , 1964

Significance of spiral organisms found after treatment in late human and experimental syphilis.
Collart, P., Borel, et al
Persistence of T. Pallidum after treatment. Organisms are still present but have lost their virulence. Cortisone reactivates clinical disease.

Journal of Bacteriology, 1963;85:932-939.

Morphology of Treponema microdentium as revealed by electron microscopy of ultrathin sections
Listgarten, M.A., et al,
[Findings:] Spirochetal granules were found in cultures of T. microdentium. They were more numerous in older cultures. [Observations pertaining to the classification of spirochetes:] "The [cell] envelope had an irregular contour, was easily disrupted during processing, and did not appear esstial in maintaining the shape of the protoplasmic cylinder. It is therefore probable that this envelope is quite distinct from bacterial cells walls, which in ultrathin sections appear as regular, well-defined, electron-dense structures." (p.938)

Journal of Bacteriology, 1961;82:967-978.

Influence of osmotic pressure on the morphology of the Reiter treponeme.
Hardy, P.H. & Nell
"[Reiter] Treponemes in saline solution were observed while distilled water was pulled into the preparation by capillary action, and it was found that although all treponemes in a field were not changed to spheres simultaneously, the conversion of any single one took place

American Journal of Syphilis, 1953;37:29-36.

Treponema pallidum buds, granules and cysts as found in human syphilitic chancres and seen in fixed unstained smears under darkground illumination.
Coutts, W.E. et al
"Spirochetogenic granules are by far more numerous than the cysts."

American Journal of Syphilis, 1951;35:164-179.

Studies on the life cycles of spirochaetes: V. The life cycle of the Nichols non-pathogenic Treponema in culture.
Delamater, E.D., et al
Formation of reproductive cysts.

American Journal of Syphilis, 1951;35:216-224.

Studies on the life cycles of spirochaetes: VII. The life cycle of the Kazan non-pathogenic Treponema pallidum in culture.
Delamater, E.D., et al

Yale Journal of Biology and Medicine, 1950.

The morphology and staining characteristics of Treponema pallidum. Review of the literature and description of a new technique for staining the organisms in tissues.
Campbell, R.E. et al

Journal of Experimental Medicine, 1950;92:247-250.

(P) Studies on the life cycles of spirochetes: IV. The life-cycle of the Nichols pathogenic Treponema pallidum in the rabbit testis as visualized by means of stained smears.
Delamater, E.D., et al

Experimental Medicine, 1950;92:239-246.

Studies on the life cycles of spirochaetes: III. The life cycle of the Nichols pathogenic Journal of Treponema pallidum in the rabbit testis as seen by phase contrast microscopy.
Delamater, E.D., et al
"...it seems likely from these observations that there are two means of vegetative reproduction, consisting of (1) transverse division (the most important under usual conditions); and (2) the production of gemmae or buds which eventuate into unispirochetal cysts comparable to those described for saprophytic forms, within each of which single spirochetes develop and differentiate, and from which they subsequently emerge."

American Journal of Syphilis, 1949;33:101-113.

Morphology, cultural characteristics and a method for mass cultivation of the Reiter spirochaetes.
Gelperin, A.

Journal of Bacteriology, 1948;56:755-769.

(P) Morphologic characteristics of certain cultured strains of oral spirochetes and Treponema pallidum as revealed by the electron microscope.
Hampp, E.G., Scott
"Typical free granules, the end products of granule "shedding," ... consist for the most part of what appear to be short sections of spirochetes closely packed together...Although it is not possible to determine from these micrographs that the granules are germinative units, their constant rhythmic occurrence in living cultures suggests this possibility. Further support of this hypothesis is provided by the fact that cultures up to 31 months old, showing only refractile granules by dark-field examination, have invariably given normal growths on transfer to fresh medium (Hampp, 1946)." (p.768). Also of interest: the authors did not find cell membranes on the spirochetes they examined.

Bulletin of Hygiene, 1947;23:548.

Study by means of micromanipulation of the virulence of one or several spirochaetes as well as viability of spirochaetes or granular forms of culture of supposed Treponema
Bessemans, A.,

American Journal of Syphilis, 1947;31:109-114.

Transmission of experimental syphilis from mouse to mouse in absence of S. pallida and pathologic changes in presence of successful innoculation
Wile, U.J.
Showed that syphilis can be transmitted by tissues from infected hosts in the absence of spirochetes, suggesting that the infectious agent is present in another form. Note: this study does not specifically mention cysts or granules.

J Am Dental Assoc, 1946;33:201-206.

Morphologic alteration of smaller oral treponemas during aging of cultures; Effect of age on viability of spirochetal cultures.
Hampp, E.G.

Journal of Bacteriology, 1943;46:15-24.

(P) Bacterial morphology as shown by the electron microscope; V. Treponema pallidum, Treponema macrodentium and Treponema microdentium.
Mudd, S., Polevitsky, et al
"Irregularly spheroidal, dense bodies... are often found attached to the spirochetal cell, frequently near the end; such a dense body may be in close apposition to the outside of the spirochetal cell-wall or may be connected to it by a short stalk. The evidence concerning these bodies seems to support the interpretation that they are asexual reproductive bodies." (p.23)

American Journal of Syphilis, 1942;26:565-573.

Some morphologic features of the Nichols strain of Treponema pallidum as revealed by the electron microscope
Morton, H.E. et al,

JAMA, 1942;199:880-881.

(P) The morphology of spirochaeta pallida in the electron microscope.
Wile, U.J., Picard, et al
"...in many specimens a curious knoblike structure was seen at the end of many organisms. Their almost uniform shape and density suggest that these are not extraneous particles of the preparation but a part of the organism itself."

Annales de l'Institut Pasteur, 1940;64:439-455.

(F) Etude morphologique du Spirochaeta pallida. Modes de devision. Spirochetogene syphilitique.
Manouelian, Y.

American Journal of Syphilis, 1938;22:294.

Morphologic variations of the syphilitic germ.
Bessemans, A.
Discusses pleomorphism in T. pallidum.

Annales de l'Institut Pasteur, 1935;55:698-708.

(F) Syphilis tardiva. Forms minuscules du Spirochaeta pallida. Spirochetogene syphilitique.
Manouelian, Y.

American Journal of Syphilis, 1932;16:155-190.

(R) The life history of Treponema pallidum. A Critical review of literature.
Ingraham, N.R., Jr.
Excellent, "must-read" review of the findings and various interpretations of T. pallidum prior to 1932. States that there have been 18 separate experiments in which tissues from infected hosts transmitted infection in the absence of spirochetes, suggesting that the organism is present in another form. Of particular interest to the reader: Roukavischnikilff's belief that the cause of syphilis circulates in the blood of an infected animal in an avisual (submicroscopic) stage; and McDonaugh's theory that Treponema pallidum is the adult male phase of a coccidial

Compt. rend. soc. biol., 1930;104:477-480.

(F) Gommes syphilitiques et formes anormales du treponemes, ultravirus syphilitiques.
Levaditi, C.
Compt. rend. soc. biol., 1930;104:736-740.

(F) Cycle evolutif du Treponema pallidum du Spirochaeta pertenuis et du Spirchaeta cunicola.
Levaditi, C. et al,

Compt. rend. soc. biol., 1930;104:72-75.

(F) Relation entre le cycle evolutif du "Treponema pallidum" et la genese des lesions syphilitiques.
Levaditi, C., et al

American Journal of Syphilis, 1930;14:433-437.

(P) Granular transformation of Spirochaeta pallida in aortic focal lesions.
Warthin, A.S. et al
Findings of T. pallidum spirochetes in atypical forms in aortic focal lesions suggested that the possibility that the spirochete may transform itself into a minute granule by a series of contractions. Atypical forms were found even when typical spirochetes were absent. Includes an interesting drawing of the transitional stages observed as a spirochete transforms itself into a minute granular form. The authors raise the question as to whether this progression represents evolution or involution, but seem to emphasize the possibility of involution.

British Medical Journal of Dermatology & Syphilis, 1913;25:1-14.

(R) The complete life history of the organism of syphilis
McDonagh, J.E.R.

Lancet, 1912;2:1011.

The life cycle of the organism of syphilis.
McDonagh, J.E.R.
Poses several questions: (1) Why is the incubation period of syphilis so long? (2) Why do 1-2 injections of salvarsan [used to treat syphilis prior to the discovery of penicillin] not cure every case? (3) In tertiary syphilis, fewer organisms are found; why then is this stage the hardest to treat? The author argues that the Treponema pallidum is the adult male phase of a coccidial protozoan, and that the spores that result from the conjugation of the two sexual phases are the actual infectious agent of syphilis. The spores were observed to develop inside of

Journal of Experimental Medicine, 1912;16:194-198.

Treponema mucosum (new species) a mucin producing spirochaeta from pyorrhea.
Noguchi, H.
Irregular spirochetal forms and many granules appeared when conditions were unfavorable to the organism. Small spirochaeta were seen attached to round bodies as if they had just sprouted from them. Journal of Experimental Medicine, 1911;XIV:99-112.

A method for the pure cultivation of pathogenic Treponema pallidum.
Noguchi, H.
Observed "spore-like round bodies" connected with young pallida. Also of interest: the author reports that he was unable to cultivate T. pallidum in any medium without the addition of tissue.

Journal of the Royal Army Medical Corps., 1911;Vol. XViI, p.225.

Granule-shedding in Treponema pallidum and associated Spirochaetae.
O'Farrel, W.R. Balfour, A

Münchner med. Wochenschr., 1906;53:310-312.

(F) Weitere Mitteilungen über die Spirochaeta Pallida.
Herxheimer, K.
Found that the classic spiral form is not the only form that spirochetes may assume.

Münchner med. Wochenschr., 1905;53:310-312.

(F) Zur Kenntnis der Spirochaeta Pallida.
Herxheimer, K.

Berlin. Klin. Wochschr., 1905;42:673-675.

(F) Über Spirochaeta pallida bei Syphilis und die Unterschiede dieser Form gegenüber anderen Arten dieser Gattung.
Schaudinn, F., & Hoffman, S.

Note: the American Journal of Syphilis was originally called the American Journal of Syphilis, Gonorrhea, and Venereal Diseases.

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Address of this page is http://www.lymenet.de/literatur/cystsl.htm Homepage of this Server is http://www.lymenet.de

[soijuv]

Tri Alan Mc Donaldin näkemys borreliabakteerin eri muodoista ja niiden koosta:

Gemmae -size from 7 microns to 30 microns { These are the largest round bodies of spirochetes - also called Cystic borrelia}

Granules - size from0.5 microns to 2 microns {these are the agents of Granulovacuolar Degeneration of neural cells
in Alzheimer's Disease. Granular spirochetal forms are very prominent inside of Biofilms of Borrelia
where they co-exist with other non-spiral specialized borrelia forms. Eisendle's images of
ACA lesions with FFM technique nicely demonstrate aggregates of pure granular borrelia [ which I now reclassify as
biofilm communities of Bb in ACA skin.
As to the maximum number of Granules which a single spirochete can produce, I would place an
upper limit of between 30 to 50 granules Maximum per single spirochete.

Liposomes - size from 0.01 microns to 0.03microns. Liposomes are continually shed from the surface of the spiral motile
borrelia as they move through the body. There seems to be no limit to the number of liposomes which can be shed
from a single borrelia spirochete. [Liposomes = "Blebs" = Microvescles]
The continuous shedding of liposomes [ without numerical upper limits on the number of liposomes emerging
from a single spirochete is a biological Multiplier of pathogenicity, vastly expanding the damage
that a single spirochete can cause in its lifetime.

All contain DNA ,But
Gemmae - DNa content is the entire chromosome and all 21 plasmids
granules - DNa content is the entire chromosome and all 21 plasmids
Liposomes - DNa content is probably limited to Plasmids only ( linear and circular - Garon's studies)

the Beermann paper from year 2000 clearly shows the profile of liposomes (blebs)
under electron microscopy.
Radolf's paper in year 2012 shows blebs to be much smaller than gemmae ( cystic borrelias)

liposomes can easily invade nonphagocytic mammalian cells [ Beermann 2000]
liposomes [blebs] are photographed inside the host mammalian cell nucleus.
Gemmae are incapable of doing this , due to their size.

Granules can also form inside mammalian cells, either as a a result of
penetration of the cell by the motile spiral form and then subsequent
segmentation breakup into granular forms
or
perhaps granular forms can directly penetrate mammalian cells
to reach the cytoplasm, [ but granular forms are probably incapable of
penetrating the nucleus of the mammalian cell- at least based on image
data now available]

[soijuv]

Tri A.McDonaldin mukaan (2013) Bb:n kystamuodot on mahdollista nähdä suoraan verinäytteistä. Piilossa (silmät, aivot jne) olevia Bb bakteereita ei löydetä ennenkuin näytteet otetaan kuoleman jälkeen kudoksista. Niitä ei löydetä millään nykyisellä testillä.

1. is there eve a circulating cystic borrelia form i human blood? answer: Yes - I can send to you a 200 slide lecture on Borrelia Cystic forms
if you wish to see it. It is also freely availabe on my website
www.alzheimerborreliosis.net
2. Can cysts of borrelia be demonstrated in direct analysis of human blood? answer --Yes --
see link to Round body (Cystic borrelia lecture-200 slides) - - DR Morten-Laane
3. Is the content of Cystic borrelia equal to the DNA content of spiral borrelia? answer --yes
4. Is a culture negative result ( of blood ) the end of the story as far as the possibility that cystic forms
might be present in tissue sites outside of blood? answer -- Negative in blood does not exclude the existence of cystic
forms elsewhere.
5. Does any lab testing method reflect the Status of :SANCTUARY SITES [ BRAIN,EYE,GONAD,FASCIA ETC] ?? Answer -- No Lab test other than the AUTOPSY
Is informative about the status of SANCTUARY SITES in the human body for the possible presence of any of the forms of borrelia
( Spiral, Straightened, Cystic,Granular, Cell Wall deficient, Biofilm communities,Liposomal forms of Borrelia)