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piątek, 17 czerwca 2016

Cysty - dowody 1905-2005

pirochetal Cysts, L-Forms, and Blebs Observations from 1905

1905 Contribution à l'étude de la structure et du cycle évolutif du Spirochaete pallida de Bull Acad Sc Cracovie, 9:713. Rev prat Mal cutan,Siedlicki M. Schaudinn. 1906, 5:43.
[According to Campbell, 1950:
 
Described most of the forms ascribed to the evolution or involution of the spirochete of syphilis.]
 
253. Krzystalowicz F; 1905 Spostrzezenia nad budowa i rozwajem Spirochaeta pallida Schaudinn. Rozpr. wydz. mat, przyrold. Polska Akad., 5:414.Siedlicki M.
[According to Campbell, 1950:
 
Described most of the forms ascribed to the evolution or involution of the spirochete of syphilis.]
 
254. Schaudinn F; 1905 Über Spirochaeta pallida bei Syphilis und die Unterschiede dieser Form gegenuber Berlin. Klin. Wochschr., 42:673-675.Hoffman S. anderen Arten dieser Gattung.
 
Found that the classic spiral form is not the only form that spirochetes may assume.
[According to Novy & Knapp, Schaudinn believed that spirochetes were protozoa, not bacteria.]
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10 June 2005
Author Year Title Journal
Borrelia burgdorferi
1. Duray PH; 2005 Invasion of human tissues ex vivo by Borrelia burgdorferi. J Infect Dis, 191(10):1747-54.Yin SR; Ito Y;Bezrukov L; Cox C;
 
“In agreement with earlier studies by other groups, we also found spirochetes arranged in cystlike, or “encysted," structures. Similar structures of B. burgdorferi Cho MS; Firzgerald W; have been reported within insect and arthropod vectors. It has been speculated that these cystic structures could give rise to intact spirochetes, suggesting that
Dorward D; cystic forms represent dormant structures that subsequently cycle into spirochetes. In the present study, we did not examine how these cysts are formed nor did we
Zimmerberg J; notice any signs of these forms switching back to single spirochetes. These and other questions regarding cystic forms of this organism can now be addressed in
Margolis L. the ex vivo tissue system.” … “Most spirochetes visualized by electron microscopy were located extracellularly, but intracellular forms were occasionally observed in cultures in vitro, as described elsewhere.”
2. Zajkowska J; 2005 Atypical Forms of Borrelia burgdorferi—Clinical Consequences. Pol Merkuriusz Lek, 18(103):115-119.
Hermanowska-Szpakowicz T; [Abstract:]
 
“Borrelia burgdorferi utilizes a variety of mechanisms to counteract eradication by its host and establish chronic infection. We discuss several of these
Rubel J. mechanisms, including plasmid encoded genes, morphologic variants, cysts formation, colonies formation, antigenic variation, and resistance to iron deprivation. These mechanisms, as well as the possible survival of Borrelia burgdorferi in forms with low metabolic activity, may explain relapsing Lyme disease, and may also account for the difficulties with eradicating this pathogen.”
3. Murgia R; 2004 Induction of cystic forms by different stress conditions in Borrelia burgdorferi. APMIS, 112(1):57-62.Cinco M.
[Abstract:]
 
 “Cystic forms of Borrelia burgdorferi might represent a low metabolic activity state or phase of B. burgdorferi cells that allows the spirochete to survive in a hostile environment until conditions are favourable to multiply again. In this study we evaluated the rate of cyst formation induced by oxidative stress, pH variations, and heating, reconversion of cysts to vegetative forms, and some aspects of their metabolic activity. We observed cyst formation in the presence of extreme pH values, and at high temperature, but the best production of cystic forms was observed in the presence of H2O2. When transferred to BSK II medium, the cystic forms reconverted to spirochetes in relation to their age and type of induction treatment. Furthermore, we demonstrated a low metabolic activity of cystic forms by measuring amino acid incorporation. Overall, these data suggest that the phenomenon of conversion to cysts by B. burgdorferi provides a limited survival potential. This short-term survival, however, gives borreliae an additional chance to overcome unfavourable environmental conditions.”
 
4. Embers ME; 2004 Survival strategies of Borrelia burgdorferi, the etiologic agent of Lyme disease. Microbes and Infection, 6:312-318.RamamoorthyR;Philipp MT.
 
“It has been suggested that B. burgdorferi may avoid immune surveillance mechanisms altogether by seeking a refuge within a host cell or by encapsulating itself inside a cyst membrane… Several studies in vitro have motivated the suggestion that B. burgdorferi may transform into cysts in vivo… cyst formation also has been shown to occur in body fluids such as the cerebrospinal fluid and in response to the addition of β-lactam antibiotics in vitro. The reasonable, yet unproven notion that an inability to resolve infection with antibiotics in some patients could result from such cyst formation may follow.”
 
5. Brorson O; 2004 An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole. Int Microbiol, 7(2):139-40.Brorson SH.
[Abstract:]
 
 “The susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole (TZ) was examined. The minimal bactericidal concentration (MBC) of TZ against the mobile spirochetes was >128 microg/ml at 37 degrees C in micro-oxic atmosphere when incubated for 14 days. TZ significantly reduced the conversion of mobile spirochetes to cystic forms during incubation. The MBC for older (10-months-old) cysts at 37 degrees C in a micro-oxic atmosphere was >0.5 microg/ml, but >0.125 microg/ml for young (1-day-old) cysts. Acridine orange staining, dark-field microscopy and transmission electron microscopy revealed that, when the concentration of TZ was > or = MBC, the contents of the cysts were partly degraded, core structures did not develop inside the young cysts, and the amount of RNA in these cysts decreased significantly. When cysts were exposed to TZ, both the spirochetal structures and core structures inside the cysts dissolved, and the production of blebs was significantly reduced. These observations may be valuable in the treatment of resistant infections caused by B. burgdorferi, and suggest that a combination of TZ and a macrolide antibiotic could eradicate both cystic and mobile forms of B. burgdorferi.”
 

Brorson O; 2002 An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi Int Microbiol, 5(1):25-31.Brorson SH. to hydroxychloroquine.
[Abstract:]
 
 “In this work the susceptibility of mobile and cystic forms of Borrelia burgdorferi to hydroxychloroquine (HCQ) was studied. The minimal bactericidal concentration (MBC) of HCQ against the mobile spirochetes was > 32 microg/ml at 37 degrees C, and > 128 microg/ml at 30 degrees C. Incubation with HCQ significantly reduced the conversion of mobile spirochetes to cystic forms. When incubated at 37 degrees C, the MBC for young biologically active cysts (1-day old) was > 8 microg/ml, but it was > 32 microg/ml for old cysts (1-week old). Acridine orange staining, dark-field microscopy and transmission electron microscopy revealed that the contents of the cysts were partly degraded when the concentration of HCQ was > or = MBC. At high concentrations of HCQ (256 microg/ml) about 95% of the cysts were ruptured. When the concentration of HCQ was > or = MBC, core structures did not develop inside the cysts, and the amount of RNA in these cysts decreased significantly. Spirochetal structures inside the cysts dissolved in the presence of high concentrations of HCQ. When the concentration of HCQ was > or = MBC, the core structures inside the cysts were eliminated. These observations may be valuable in the treatment of resistant infections caused by B. burgdorferi, and suggest that a combination of HCQ and a macrolide antibiotic could eradicate both cystic and mobile forms of B. burgdorferi.”
 
7. Murgia R; 2002 Cystic forms of Borrelia burgdorferi sensu lato: induction, development, and the role of RpoS. Wiener Klinische Wochenschrift, 114(13-14):574-9.Piazzetta C;Cinco M.
 [Abstract:]
 
 “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.”
 
8. Zajkowska JM; 2002 New aspects of the pathogenesis of Lyme disease. Przegl Epidemiol, 56 Suppl 1:57-67.Hermanowska-Szpakowicz T.
[From the abstract:]
 
“Morphological changes of B. burgdorferi as well as changes in expression of surface proteins caused by environmental determinants are essential in pathogenesis of Lyme disease. Cysts, spherical form (spheroplasts, L-form) and ‘blebs’ (gemmae) can be responsible for long lasting antigenic stimulation, signs of chronic borreliosis, and even probably connected with MS and Alzheimer disease.”
 
9. Gruntar I; Malovrh T; 2001 Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. APMIS,109(5):383-8.Murgia R; Cinco M.
[Abstract:]
 
“Cystic forms (also called spheroplasts or starvation forms) and their ability to reconvert into normal motile spirochetes have already been demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of this study was to determine whether motile B. garinii could develop from cystic forms, not only in vitro but also in vivo, in cyst-inoculated mice. The cysts prepared in distilled water were able to reconvert into normal motile spirochetes at any time during in vitro experiments, lasting one month, even after freeze-thawing of the cysts. Motile spirochetes were successfully isolated from 2 out of 15 mice inoculated intraperitoneally with cystic forms, showing the infectivity of the cysts. The demonstrated capacity of the cysts to reconvert into motile spirochetes in vivo and their surprising resistance to adverse environmental conditions should lead to further studies on the role and function of these forms in Lyme disease.”
 
10. Brorson O; 2001 Susceptibility of motile and cystic forms of Borrelia burgdorferi to ranitidine bismuth citrate. Int Microbiol, 4(4):209-15.Brorson SH.
[Abstract:]
 
“Gastrointestinal symptoms accompanying Lyme disease have not been considered in the treatment of Lyme patients yet. Here we examine the effect of ranitidine bismuth citrate (RBC) on motile and cystic forms of Borrelia burgdorferi in vitro, to determine whether it could cure this bacterial infection in the gastrointestinal tract. When motile forms of B. burgdorferi were exposed to RBC for 1 week at 37 degrees C, the minimal bactericidal concentration (MBC) was > 64 mg/ml. At 30 degrees C, the MBC was > 256 mg/ml. When the incubation lasted for 2 weeks at 37 degrees C, the MBC dropped to > 2 mg/ml. Bismuth aggregates were present on the surface of B. burgdorferi when RBC > or = MBC, as shown by transmission electron microscopy (TEM). Cystic forms of B. burgdorferi, exposed to RBC for 2 weeks at 37 degrees C, were examined by cultivation in BSK-H medium (Sigma B3528). They were stained with acridine orange (pH 6.4, pH 7.4) and studied by TEM. The MBC for RBC for young cystic forms (1 day old) and old cysts (8 months old) was estimated to be > 0.125 mg/ml and > 2 mg/ml, respectively. Bismuth aggregates were attached to the cysts and, in some, the pin-shaped aggregates penetrated the cyst wall. The bismuth aggregates also bound strongly to blebs and granules of B. burgdorferi when RBC > or = MBC. When B. burgdorferi is responsible for gastrointestinal symptoms, bismuth compounds may be candidates for eradication of the bacterium from the gastrointestinal tract.”
 

Brorson O; 2001 Association between multiple sclerosis and cystic structures in cerebrospinal fluid. Infection, 29(6):315-9.Brorson SH; Henriksen TH;
 
“Cystic structures were observed in CSF of all ten patients by AO and TEM. DF revealed eight cyst-positive patients out of nine. One of five control persons Skogen PR; had such structures in the CSF; this person had suffered from erythema migrans. Spirochete or rod-like structures emerged after culturing two of the MS patient CSF Schoyen R. samples and these structures could be propagated. CONCLUSION: A significant association of CSF cysts and MS was identified in this small study among residents in a coastal area of southern Norway. The cysts could be of spirochetal origin.”
 
12. Alban PS; Johnson 2000 Serum-starvation-induced changes in protein synthesis and morphology of Borrelia Microbiology, 146 (Pt 1):119-27.PW; Nelson DR. burgdorferi.
 
"In a recent study, Brorson & Brorson (1997) demonstrated that B. burgdorferi cells transform from vegetative spirochaetes into spherical ‘cyst-forms’ when
incubated in BSKII medium lacking rabbit serum (BSKII-S). We confirmed these observations. ...Within 24 h, cells started of serum were completely non-motile and 30-40% had begun to encyst. After 48 h incubation in RPMI, ~90% of serum-starved cells had formed cysts (Fig. 1). ...In contrast to typical helical vegetative cells, most 48 h serum-starved cells were coiled within a membrane. ...
When rabbit serum or BSK was added to RPMI containing 48 h serum-starved cells, the cysts opened within 10 s to yield intact, but non-motile spirochaete cells (Fig. 2). ...Cells begain to regain motility 12-15 h after emerging from the cysts.
...the Western blots displayed consistent differences between the protein antigens recognized in vegetative cells and cysts. ...By forming cysts, it is also
conceivable that B. burgdorferi cells evade detection by the immune system.
Cyst formation is an active cellular response to serum starvation. The addition of tetracycline inhibits cyst formation, demonstrating that cyst formation requires protein synthesis and that cysts are not merely degenerative forms."
 
13. Amosova LI. 2000 An electron microscopic study of Borrelia in the body of the female ixodid tick Ixodes Parazitologiia, May-Jun;34(3):234-40.
persulcatus.
[From the abstract:]
 
 "Borrelia burgdorfery s. lato in naturally infected females of tick Ixodes persulcatus were examined by transmission electron microscopy.
The Borreliae were found in midgut and ovary. ...Two morphological types of borreliae were observed."
 
14. Beermann C; 2000 Lipoproteins from Borrelia burgdorferi applied in liposomes and presented by Cell Immunology, May 1;201(2):124-131.
Wunderli-Allenspach dendritic cells induce CD8(+) T-lymphocytes in vitro.
H; Groscurth P;Filgueira L.
 
 "We could document invasion of Bb into the dermis and shedding of Bb-blebs into the tissue under in vitro conditions (Fig. 1b). ...We show with electron microscopy that shedding of blebs by Bb also takes place in the tissue which confirms earlier observations (34).
Bb-liposomes were used as a model for Bb-blebs to study uptake by cells. "...we studied the uptake of Bb-liposomes by human DC, fibroblasts, and B- and T-lymphocytes. All tested cells incorporated Bb-liposomes, as visualized by immunofluorescence microscopy. ...we could document that Bb-liposomes were incorporated within seconds."
 
15. Zajkowska JM; 2000 [No title available]. Pol Merkuriusz Lek, 9(50):584-8.
Hermanowska-Szpakowicz T et al. [From the abstract:]
 
 “Spheroplast L-form of borrelia could be responsible for difficulties with their eradication.”
 
16. Zajkowska JM; 2000 [Selected aspects of immunopathogenesis in Lyme disease]. Pol Merkuriusz Lek, 9(50):579-83.
Hermanowska-Szpakowicz T; [From the abstract:]
 
“In pathogenesis of chronic and recurrent cases difficult to treat is essential is survive [sic] of metabolic inactive bacteria,
Pancewicz SA; antigens B. burgdorferi "blebs", cystic L-form or insoluble complexes antigen-antibody or possibility of intracellular survive [sic] of B. burgdorferi.”
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http://www.wple.net/plek/numery_2015/numer-11-2015/656-661.pdf.

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Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis  2008 Sep 25

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.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2564911/

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Marty Ross MD

Treating borrelia, the Lyme germ, with antibiotics is complicated. There is limited research showing which treatment regimens work best. So many Lyme Literate Medical Doctors (including me) prescribe antibiotics based on a theory and our collective observation that combinations of antimicrobials work better than single agents alone. What follows is not definitive or comprehensive guide. Rather it reflects how I organize and put together my effective Lyme disease antibiotic treatments

http://www.treatlyme.net/treat-lyme-book/lyme-disease-antibiotic-guide/

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Cystevorming door de spirocheet Borrelia burgdorferi sensu lato onder invloed van stressfactoren

Samenvatting - Lyme-borreliose of de ziekte van Lyme wordt momenteel beschouwd als de snelst groeiende infectieziekte van de wereld en het aantal patiënten met deze door Ixodes-teken overgebrachte bacteriële ziekte neemt wereldwijd dan ook steeds verder toe. De veroorzaker is de spirocheet Borrelia burgdorferi sensu lato, een spiraalvormige bacterie waatoe drie pathogene soorten behoren: Borrelia burgdorferi sensu stricto, Borrelia garinii en Borrelia afzelii. Wat betreft de tekenbeetconsultaties bij de huisartsen in Nederland, nam de incidentie toe van 191/100.000 in 1994, naar 564 per 100.000 inwoners in 2009; een verdrievoudiging sinds 1994. In 1994 was de incidentie van erythema migrans 39/100.000 en in 2009 was deze incidentie gestegen tot 134/100.000. Indien men het aantal erythema migrans gevallen en tekenbeetconsulten omrekent naar de totale Nederlandse bevolking betekent dit dat in 2009 de huisartsen ongeveer 22.000 patiënten zagen met erythema migrans en 93.000 patiënten met een tekenbeet. Net als de geslachtsziekte syfilis is ook Lyme-borreliose een multisysteem ziekte waarbij verschillende stadia zijn te onderscheiden. Het bekendste stadium bij Lyme-borreliose is de vroege lokale huidinfectie erythema migrans of Bull’s eye op de plaats van de tekenbeet, die slechts in 50% van de gevallen optreedt. Door een speciaal bewegingsmechanisme dat bestaat uit een axiaalfibril en endoflagellen is de spirocheet Borrelia burgdorferi sensu lato in staat om actief de infectiehaard te verlaten om zich elders in het lichaam van zijn gastheer te vestigen. Via endotheelcellen van de bloedvaten kan B. burgdorferi s.l. de bloedbaan verlaten, om zich diep in de weefsels te vestigen, met een voorkeur voor zuurstofarme milieu’s zoals de gewrichten en het centrale zenuwstelsel. Het gevolg is de vroege gedissemineerde Lyme-borreliose die tot uiting kan komen als Lyme-artritis en neuroborreliose. Verder is er het stadium van de late of chronische Lyme-borreliose met chronische Lyme-artritis en chronische neuroborreliose als veelvoorkomende aandoeningen. De patiënten met deze aandoeningen vormen het grote probleem. Ondanks “adequaat” veronderstelde antibioticabehandelingen in de lokale- en vroege gedissemineerde Lyme-borreliose is de spirocheet Borrelia burgdorferi sensu lato, regelmatig in staat tot overleving in de weefsels. Korte antibioticumbehandelingen laten een percentage zien van 30-62% aan recidieven binnen 3 jaar na de behandeling van de vroege Lyme-borreliose. Als men dit percentage betrekt op de jaarlijkse 22.000 patiënten die in Nederland behandeld worden voor erythema migrans, dan betekent dit dat 6600 tot 13.640 patiënten te maken kunnen krijgen met een recidiverende Lyme-borreliose binnen 3 jaar na een “adequaat” veronderstelde antibioticumbehandeling. Dat er jaarlijks enkele duizenden chronische Lyme-borreliose patiënten bij komen, komt overeen met de gegevens van de Nederlandse vereniging van Lyme patiënten (NVLP). Volgens de NVLP zouden er in 2009 in Nederland ruim 70.000 personen zijn die min of meer te lijden zouden hebben van de neurologische gevolgen van een besmette tekenbeet. Het totale aantal chronische Lyme patiënten zou volgens de NVLP enige honderdduizenden groot zijn. Het exacte aantal is echter niet bekend. Wat wel bekend is, is dat het chronische Lyme-borreliose legioen met persisterende klachten alleen maar groeiende is, ondanks “adequate” antibioticumbehandelingen. De volgende vraag staat dan ook centraal in dit literatuuronderzoek: “Ontbreken er fundamentele schakels in de antibacteriële behandeling bij Lyme-borreliose?” Het mislukken van “adequaat” veronderstelde antibioticumbehandelingen bij Lyme-borreliose werd voor het eerst gemeld door onderzoeker Preac-Mursic in 1989, al meer dan 20 jaar geleden. Bij patiënten die met antibiotica waren behandeld, werd de persistentie van de spirocheet bevestigd met positieve culturen van B. burgdorferi s.l. van huidbiopsies, synoviaalvocht, liquor en bloed. Falende antibioticumbehandelingen bij Lyme-borreliose vormde de aanzet in Europa om uitgebreid in vitro onderzoek te doen naar het gedrag van de bacterie onder invloed van diverse stressfactoren waaronder “effectief ” veronderstelde antibiotica. Een nadere beschouwing van het in vitro onderzoek toont aan dat de spirocheet B. burgdorferi s.l. onder ongunstige omstandigheden als het ware (tijdelijk) onderduikt en kan.transformeren van een normale mobiele spirocheet in een inerte cyste, een afgegrensd ruststadium waarin de spirocheet zich verbergt en waarbij de buitenste membraan zich als een “cocon” om de spirocheet heeft gelegd. De cyste biedt zodoende bescherming tegen de meeste antibiotica en andere stressfactoren en ook tegen eventuele antilichamen van de gastheer. Men dient zich te realiseren dat de spirocheet razendsnel kan overgaan in de cystevorm zodra de stressfactor aanwezig is. Een in vitro onderzoek laat zien dat na één minuut 95% van de spirocheten van B. burgdorferi s.l. was overgegaan in cysten onder invloed van hypotone stress (aquadest) en na 4 uur werd er geen mobiele intakte spirocheet meer waargenomen. In vitro onderzoeken hebben ook laten zien dat na het verdwijnen van de stressfactor, dus als de omstandigheden weer gunstig worden voor de spirocheet B. burgdorferi s.l., deze reconversie kan vertonen van de cystevorm naar een normale spiraalvormige spirocheet. De reconversie kan afhankelijk zijn van de concentratie van een stressfactor en/of de tijdsduur van de beïnvloeding hiervan. Reconversie kan ook razendsnel gaan zoals een onderzoek heeft aangetoond bij cysten van 48 uur oud, die waren gevormd onder invloed van serum-starvation. Reconversie vond binnen 10 seconden plaats na toevoeging van konijnenserum aan het medium van de cysten. Uit de cysten kwamen onbeweeglijke maar intakte spirocheten. De spirocheten herkregen hun beweeglijkheid na 12 tot 15 uur na het uitkomen van de cysten.

http://dspace.ou.nl/handle/1820/4556

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http://boreliozaabc.blogspot.nl/2016/09/293-prac-na-temat-ze-borrelia-tworzy.html

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jak Borrelia zmienia się z formy cysty w forme podstawowa.Film - mikroscop

 
 
 
 

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