312-Margulis-final.pdf

SYMBIOSIS (2009) 47, 51–58

ISSN 0334-5114

Position paper.

Spirochete round bodies

Syphilis, Lyme disease & AIDS: Resurgence of “the great imitator”?

Lynn Margulis 1* , Andrew Maniotis 2 , James MacAllister 1 , John Scythes 3 , Oystein Brorson 4 , John Hall 1 ,

Wolfgang E. Krumbein 5 , and Michael J. Chapman 1

1 Department of Geosciences, University of Massachusetts, Amherst, MA 01003, USA, Emails. celeste@geo.umass.edu,

jimmymac@geo.umass.edu, johnloomishall@gmail.com, oenothera1@yahoo.com, www.geo.umass.edu/margulislab;

2 Department of Pathology, University of Illinois at Chicago, College of Medicine, 840 South Wood Street,

Chicago, IL 60612, USA, Email. amanioti@uic.edu;

3 Research co-coordinator, Community Initiative for AIDS Research, proprietor and owner, Glad Day Bookshop,

Toronto, Ontario, Canada, Email. john@gladdaybookshop.com;

4 Sentralsykehuset i Vestfold, Department Microbiology, Tonsberg, Norway, Tel. +47-33-342475,

Email. abrorson@newmedia.no;

5 Geomicrobiology, ICBM, Carl von Ossietzky University, Oldenburg, Germany, Email. wek@uni-oldenburg.de

(Received August 18, 2008; Accepted August 31, 2008)

Abstract

We advocate investigation of spirochete cyclical symbioses (e.g., Borrelia sp ., Leptospira sp., Treponema sp.) given the

newly established verification of a developmental history in these gram-negative motile helical eubacteria, both in pure

culture and in mammals. Symbiotic spirochetes can be compared to free-living relatives for their levels of integration

(behavioral, metabolic, gene product or genetic levels). Detailed research that correlates life histories of symbiotic

spirochetes to changes in the immune system of associated vertebrates is sorely needed. Genome analyses show that in

necrotrophic symbioses ( Borrelia and Treponema sp.) of humans and other primates, integration of the bionts occurs at the

gene product and genetic level. Spirochete round bodies (also called cysts, L-forms and sphaeroplasts) can be induced by

many types of unfavorable conditions (e.g., threats of starvation, desiccation, oxidation, penicillin and other antibiotics).

Reversion to familiar helical, motile active swimmers by placement of pure cultures into favorable environments in some

cases can be controlled. These observations are supported by a European literature, especially Russian, apparently unknown

to American medicine and medical research.

Keywords: Spirochete cysts, Treponema pallidum , Borrelia burgdorferi , AIDS co-factor, immune suppression, STD,

spirochetoses, Spirosymplokos , fossil spirochetes, spirochete life histories, Mixotricha paradoxa , round

body reversion

1. Introduction

Powerful new techniques of microbiology, including

molecular ecology and evolution inspire us to urge

reinvestigation of the natural history of mammalian,

tick-borne, and venereal transmission of spirochetes in

relation to impairment of the human immune system.

At a small meeting, Spirochaete Co-evolution in the

Proterozoic Eon: Ecology, symbiosis, and pathogenesis (an

excursion into environmental immunology) organized by

Prof. Dr. Wolfgang E. Krumbein and Prof. Lynn Margulis

held in the Museum für Naturkunde (Berlin, May 1–2,

2008) we scientists, medical researchers, historians and

physicians endorsed this statement.

We, the signatories of this paper, limit ourselves to four

issues. First, that current medical discussions of two

spirochetoses (spirochete-associated infirmities, e.g., Lyme

disease, syphilis) omit mention of “round bodies” or state

that they have no clinical relevance (Feder et al.,

2007). Round bodies are viable, motile, slowly reproductive

morphologies assumed by spirochetes when they are

* The author to whom correspondence should be sent.

L. MARGULIS ET AL.

threatened by environmental insult such as changes in

solution chemistry: acidity-alkalinity, salts, gas composition

(i.e., oxygen, hydrogen sulfide); changes in chemical

concentrations (i.e., antibiotics, antibodies, carbohydrates,

amino acids, vitamins); or changes in viscosity or

temperature. Both starvation and threat of desiccation

induce round body formation. Some cultures of spirochetes

(e.g., Leptonema, Perfilievia , Dubinina et al., 2008) seem to

persist more as round bodies than as typical spirochetes.

Round bodies, often called by other names such as "cysts",

granular bodies, L-forms, non-growing bodies, sphaero-

plasts, vesicles, etc. revert to the active helical swimmers

when conditions favorable to growth return (Fig. 1).

Second, that infections by spirochetes in humans, when

seen in their evolutionary and ecological context, are

examples of cyclical symbioses that have evolved over

geologic time. Certain symbioses have been shown to be

associated with viral-like particles capable of synthesis of

reverse transcriptases (Fig. 2). These are posited by Ryan

(2007) based significantly on the work of Luis Villareal to

be part of the integration process between the symbiont

partners (i.e., in this case human and spirochete).

Third, we caution that antibiotic treatment may be

effective only in the earliest stages of these spirochetoses.

Indeed antibiotics such as penicillin and its derivatives

induce round body formation and quiescence of symptoms

rather than cure. Suspension of round bodies in growth

media causes rapid, days to weeks, reversion to helical

swimmers as the Norwegian investigators have shown (Fig.

3; Brorson and Brorson, 2004).

Fourth, we question the accuracy of screening tests and

clinical diagnoses for either of these infections, Treponema

pallidum (the syphilis “germ”) or Borrelia burgdorferi (the

Lyme disease “germ”). Particularly vulnerable to mis-

interpretation are immunological tests in cases of re-

infection, later secondary or tertiary syphilis.

2. Spirochetes: Past and Present

Most spirochete species live freely, are unrelated to any

disease and therefore are unfamiliar to clinicians. We have

studied or taught cell biology, environmental science,

evolution, genetics, geobiology, natural history or

microbiology. Our interest, perhaps summarized around the

question “ What is the consequence of life's evolution on the

Earth as a planet?” has led us to scientific investigation of

Figure 2. Detail of Fig. 4. of Hoogenraad et al. (1967) captioned:

Electron micrograph of phage-infected spirochaetes, negatively

stained as in Fig. 1 and using the agar stripping technique (3). The

edge of the organism is lined with many small phages. The

fibrillar structure of the spirochaetes is clearly shown. Also

present in the lower section of the micrograph is a bacterial cell

wall typical of those found in large numbers in sheep tureen

contents. Scale bar = 0.5 µm.

Figure 1. Live spirochete reversible round bodies in mixed

culture. (The largest are Spirosymplokos deltaeiberi , from seaside

microbial mats, Alfacs peninsula, Catalunya, Spain). Nomarski

differential contrast light microscopy. A: Note several sizes of

round bodies come from different kinds of spirochetes. B: Higher

magnification shows the largest round bodies (rb) contain live,

motile Spirosysmplokos. Scale bars = 5 µm.

SPIROCHETE ROUND BODIES, SYPHILIS AND AIDS

Figure 3. Formation and reversion of round bodies (rb=cysts,

arrows) of Borrelia burgdorferi in pure culture. A, D: Conversion

to round bodies induced by "unfavorable conditions" most rapidly

by suspension in distilled water (starvation), spinal fluid or

antibiotics. B: Rbs formed after three weeks starvation, some

delayed, correspond to electron micrographs (by Sverre-Henning

Brorson). C, E, and H: Round bodies can reproduce as in C.

Fluorescein-conjugated polyclonal antibodies to B. burgdorferi

stains spirochete rb membranes. F: Reversion of rbs to mobile

spirochetes induced by resuspension in complete medium with

rabbit serum (for 2–6 weeks) shown in G. Scale bars: A, C, E, H =

2 µm; B, D = 3 µm; F, G = 10 µm.

and occasionally isolated spirochetes from ponds rich in

vegetation, digestive organs of marine mollusks (the

“crystalline style” of oysters and clams) and intestines of

wood-feeding termites and cockroaches. In these anoxic or

low-oxygen habitats spirochetes swim and proliferate.

Population densities often quickly reach over a thousand

million per milliliter.

Margulis’s laboratory explores an evolutionary

hypothesis. She and her colleagues posit that a certain

spirochete genome provided an ancestral component to the

earliest nucleated cells (eukaryotes). Spirochete remnant

DNA hypothesized to be present in all nucleated organisms

should be detectable in the proteomes of fully sequenced

genomes. Simply stated, spirochete ancestors of Perfilievia

russae free-living spirochetes presented at the Berlin

meeting by Galina Dubinina (Institute of Microbiology of

the Russian Academy of Sciences, Moscow, Dubinina et

al., 2008) by hypothesis are the closest co-descendants of

the cytoskeleton of our nucleated cell lineage (Margulis et

al., 2006). We envision these (sulfide-oxidizing, 0.25 µm

diameter spirochetes) are related to ancestors of cilia, sperm

tails, haptonemes and myriad other organelles of motility in

nucleated organisms. If the evidence is correctly interpreted

spirochete remnants have dwelled in stable symbioses in

eukaryotes since their origin in the Proterozoic eon over

1000 million years ago (mya) (Hall, 2008). The most

ancient intestinal spirochete symbiont in the fossil record is

much younger (Miocene c. 20 mya). A Pillotina sp . large

spirochete was discovered inside Mastotermes

electrodominicus, a kalotermitid (dry wood-feeding termite)

embedded in amber (Wier et al., 2002; 2007). Intestinal

spirochetes lived as symbionts in insects long before the

appearance, fewer than 0.5 million years ago, of any human

animal on Earth.

Spirochetes are motile helical Gram-negative

eubacteria. As heterotrophs, at optimal temperatures for

growth, they require moisture and abundant food. Most

ferment sugar in the absence of oxygen. They form a

cohesive taxon detectable by the DNA sequence that

corresponds precisely to the 16 Svedberg-unit ribosomal

RNA (16S rRNA) component of the small 30S ribosomal

subunit. Spirochetes with their Gram-negative cell walls

and periplasmic (internal) flagella between their inner and

outer membranes are distinctive at the level of thin section-

electron microscopy (EM) (Fig. 2). The inner or plasma

membrane is universal in all prokaryotic (bacterial) and

eukaryotic cells. However the presence of an outer

lipoprotein membrane typifies Gram-negative bacteria. To

assign them to a lower taxon, a “species” or “genus”,

spirochete morphology is definitively discerned in EM thin

section and less well by negative stain images that permit

assessment of their flagella insertions and numbers.

Spirochetes share a distinguishable flagellar pattern

summarized as n: 2n:n where n=number of flagella at one

end, 2n (or zero for leptospiras) refers to the overlap of

spirochetes in nature. Our comments here are generated by

an abundant international literature and years of our own

work much of it on harmless spirochetes. We have observed

L. MARGULIS ET AL.

flagella in the middle and n=number of flagella at the

opposite end. The flagella pattern of Treponema pallidum

(1:2:1; 2:4:2), Borrelia burgdorferi (4:8:4/5:10:5), or any

other spirochete can be detected in three ways:

(1) morphology, especially active motility behavior in

tissue, (2) thin section transmission EM and (3) negative

stain whole mount EM. No reliable definitive tests for the

presence of T. pallidum in patients exists short of quality

high magnification observation of the spirochetes and/or

their round bodies in affected tissue by an experienced

expert microscopist. Reported cures of either infection, the

syphilis Treponema or the Lyme disease Borrelia , lack this

level of verification. Spirochetoses (e.g., leptospiroses,

yaws, syphilis, Lyme disease) are bacterial diseases

correlated with continued presence in the body of specific

spirochetes (i.e., obey Koch's postulates).

4. Mistaken for Dead

An extensive round body (=cyst) literature exists in

Russian, but has remained relatively unknown even to

spirochete experts elsewhere. In many spirochetes

formation and reversion of round bodies has been

documented by video microscopy techniques. The

approximate number of genes of the entire sequenced

genome of each genus in which round bodies are seen is

listed in parenthesis: Borrelia (950), Brachyspira (?),

Leptospira interrogans (4300), Perfilievia = aerotolerant

“ Thiodendron " spirochetes of Dubinina et al. (2008) (?),

Spirosymplokos deltaeiberi (?), Treponema pallidum

(1100), and the Treponema -like spirochetes epibiotic in

the cortex of the giant trichomonad Mixotricha paradoxa

(?). Both T. pallidum and Borrelia burgdorferi are

anaerobic heterotrophs that require complex organic food

under anoxic conditions. They die if exposed to ambient

oxygen. The round bodies, propagules that, until they revert

to swimming helices, seem incapable of at least rapid

growth by reproduction, form quickly. Within less than an

hour, under adverse conditions round bodies develop in

large population numbers when the spirochete’s needs are

not met. They survive for extended periods of time. They

revert to helical swimmer populations that grow vigorously

when food, salt, temperature, acidity, media viscosity and

other conditions become adequate. The controlled

formation and reversion of round bodies of Borrelia are

seen in many studies by the Brorsons. Negative stain EM

images show canonical treponemes with the 1-2-1 flagellar

pattern: “Borrelia”-type (4-8-4) or higher number flagella-

pattern-spirochetes recognizable in tissue preparations,

even in the same thin section from a rabbit syphiloma

(Ovcinnikov and Delektorskij, 1968; 1975).

3. Chronic Infections as Symbioses

The likelihood that these two spirochete infections,

syphilis and Lyme disease, correlate with the establishment

of permanent human-spirochete symbioses soon after entry

of the bacteria into tissue has been insufficiently

investigated. It is reported that reverse transcriptases and

virus-like particles are generally abundant in cyclical

symbioses and it is suggested that they may facilitate the

integration of the association of the partners (Ryan, 2007).

Our intent is to improve and expand awareness of the

relationship between spirochetoses and symptoms

associated with immune suppression. We posit that the

spirochete disease syphilis persists in the human population

where its signs and symptoms may be overlooked or

misinterpreted for those of AIDS.

There may be many new drugs, but these two

spirochetoses, syphilis and Lyme disease, are not new.

Long-term association of symbiotic bacteria in animal

tissue tends toward massive gene loss when compared to

related bacteria that live freely in water, sand or mud. The

fact that Treponema pallidum and Borrelia burgdorferi are

no longer free-living and have lost many genes implies that

these spirochetes have long co-evolved with mammals (and

arthropods in the case of tick-borne Borrelia burgdorferi

Lyme disease). Contrast these integrated symbionts to

strains of Leptospira that live freely in rivers, streams and

coastal ocean waters that cause acute infection. Compared

to the fully genome-sequenced Leptospira interrogans

spirochete, over 80% of the genome of T. pallidum when

cultivated in vitro is absent. Dependency of T. pallidum on

the gene products of the human has rendered it incapable of

independent survival, growth or reproduction. Indeed

Borrelia burgdorferi has lost relatively even more of its

genophore (prokaryote “chromosome”) genes than

T. pallidum.

5. Spirochetoses and AIDS

Human tissue provides food and other conditions for

growth for both Treponema pallidum and Borrelia

burgdorferi spirochetes. Electron micrographic samples, in

principle, could verify the persistence of round bodies in

patients with symptoms, including Alzheimer's-type

dementia. Examination of biopsies from AIDS patients or

autopsies of brains from people who showed sudden

personality disturbance could test the hypothesis. Round

body formation in the test tube is induced by penicillin

especially in the presence of glycine (a protein amino acid,

read “food”). This discovery formed the major contribution

of the PhD dissertation (and accompanying patent

application) of Andrei Belichenko (2006). Dr. Belichenko

was a student of a well-known medical microbiologist Dr.

Igor Bazikov, Stavropol, Russia (Bazikov et al., 200).

Belichenko reports that decrease in penicillin concentration

induces reversion of round bodies to active hungry

SPIROCHETE ROUND BODIES, SYPHILIS AND AIDS

spirochetes. Russian research and that by Brorson and

Brorson (2004) on spirochetal life histories lead us to think

that both the presentation and the course of syphilis, Lyme

disease and other spirochetoses are altered by penicillin,

other antibiotics and possibly by “anti-retroviral” or

“protease inhibitor” drugs.

“Far from eradicating syphilis, antibiotics are driving

the disease underground and increasing the difficulty of

detection. Although the incidence of disease has more than

tripled since 1955, the chancre and secondary rash no

longer are commonly seen. Undoubtedly, some of these

lesions are being suppressed and the disease masked by the

indiscriminate use of antibiotics. The ominous prospect of a

widespread resurgence of the disease in its tertiary forms

looms ahead” (Pereyra and Voller, 1970).

We recommend studies to demonstrate or negate the

hypothesis that relapse, grave illness or death may ensue

from the reversion of round bodies to active spirochetes.

Please see bibliography, 254 references on 49 pages

compiled by Joanna Rubel “Spirochetal Cysts, L-forms, and

Blebs, Observations from 1905 to 2005”, at

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

A three-decade-long gap ushered in by the touted "cure

of penicillin" separates physicians today from the bulk of

medical literature on “the great imitator”. T. pallidum

symbiosis may help explain the high correlation of the

presence of viruses, pneumonias, other opportunistic

infections and the general symptoms of immune

suppression so well described in the "old syphilology"

medical literature (Colman Jones

www.cbc.ca/ideas/features/Aids/aidsspin.html). We suspect

that many patients carry the latent disease that has become

invisible because of the "syphilization effect" and

misdiagnosis.

T. pallidum spirochetes that cover themselves with

human proteins to which people make antibodies (Radolf

and Lukehart, 2006) cause "autoimmune diseases".

The vigorous antigenic response of early infection fades

to the classical secondary-to-tertiary symptoms of paresis.

“PARESIS”, a mnemonic, refers to a coherent and varying

set of symptoms, a syndrome. “Pe rsonality disturbances,

A ffect abnormalities, R eflex hyperactivity, E ye

abnormalities, S ensorium changes, I ntellectual impairment

and Sl urred speech. PARESIS may begin with a dramatic

delusional episode (e.g., Nietzsche January 1889 in the

Turin Plaza; Margulis, 2004 and 2007). However, over the

years, dementia may alternate with periods of such clarity

that there seems to have been a cure” (Hayden, 2003).

Since the research group of Luc Montagnier first

described LAV “virus-like particles” (later called “HIV-1”)

from “Patient 1”, a close connection has been shown

between AIDS and a history of syphilis in multi-partner

men (Barre-Sinoussi et al., 1983). “Patient 1” sought

medical consultation for swollen lymph nodes, muscle

weakness without fever or weight loss, and for episodes of

ubiquitous mycobacterium avium intracellulare (MAI

group) diarrhea, and emaciation associated with refractory

bowel infections in emaciated homosexuals and in immune

compromised patients generally. TB and other myco-

bacteria correlate with amoebic dysentery. Death

records report causes as Pneumocystis carini pneumonia,

Entamoeba histolytica , Candida albicans or other

“opportunistic infection” (Coulter, 1987). In sub-Saharan

Africa, the historic overuse of antibiotics and malnutrition

also contribute to immune suppression. One of us (John

Scythes) reports that he has not found a single documented

case of an immune suppressed patient, whether HIV-

positive or -negative, who has died of complications of

syphilis since HIV records began being maintained in the

early 1980s. Is it possible that the narrow focus on "HIV as

the cause of AIDS", an example of scientific "misplaced

concreteness" typical in explanation of evolution (Cobb,

2008), has facilitated missed diagnosis of syphilis?

Indeed, investigators in Toronto and San Francisco

found an inverse relationship between treponemal antibody

and AIDS symptoms that could be interpreted as the

immune deficiency typical of disseminated syphilis

(MacFadden et al., 1989; Haas et al., 1990; Fralick et al.,

1994).

Contrary to the statements on many official government

and medical websites that "syphilis is easily curable by

antibiotics", the disease is often refractory to antibiotic and

other treatments except perhaps in very early immuno-

responsive stages (Musher et al., 1990). It has not been

adequately shown that T. pallidum infection in its

secondary and later stages is curable after any therapy.

Because the lesions of secondary and tertiary syphilis are

autoimmune, there is often an inability to react to a skin test

of the delayed type. A loss of specificity against syphilis

antigens is noted. Chronicity, or changes in immune

response with time well established in spirochetoses is

common to other infections: herpes viruses, tuberculosis

and symptoms attributed to HIV. Syphilis, both early ulcers

and the later immunoregulation problems, seems to

facilitate acquisition of opportunistic bacteria, viruses,

fungi and the progression to full-blown collapse of the

immune system (Scythes and Jones, 2006).

gonorrhea. He did not have AIDS. He had been previously

treated for syphilis, but was he cured? Patient 1 tested

positive for antibodies to three viruses: cytomegalovirus

(CMV), Epstein-Barr virus and Herpes simplex. The first

“HIV isolate” reported by Montagnier's group was from

Patient 1. Since Montagnier's work, many centers that used

immunological tests not sensitive for all stages of syphilis

have documented a close relationship between a history of

treponematoses and HIV/AIDS (Veugelers et al., 1992;

Renzullo et al., 1991; Blocker et al., 2000). Chronic

syphilitics and AIDS patients, those unmistakably ill and

immune suppressed, do not succumb to HIV or syphilis

directly. They die of reactivation tuberculosis (TB) and

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