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THE NATURE OF THE PHYLUM
OVERVIEW:
Spirochetes are their own phylum. The nature of the phylum
in persistence and extensive preservation mechanisms to
environmental stresses of temperature, gases, moisture, and
chemicals, is demonstrated across genera.
From the PBS "Life on Earth" Series interview with Eugene
Shapiro, Yale Pediatrician, "They would have you believe
that form of the disease, somehow this is, the bacteria
knows to act differently and it doesn't respond to
antibiotics in this sense. It really doesn't make any
sense."
In fact, spirochetes *do* behave differently from other
bacteria, as will be demonstrated below. One must study
nature to become familar with human disease. The answers
to the persistance of Lyme disease come from the
characteristics of the phylum. Simply put, the
spheroplast is not the "end stage". When that
morphological variant is observed in vitro, with
exposure to various antibiotics, it should not be
concluded that the organism has been killed.
The argument that the observation that spirochetes
persist, empirically, in patients' symptoms is
necessary to be understood per the politics of Lyme:
http://www.faim.org/Lyme%20hearing.htm
To understand the controversy, one must understand that
the CDC says "Lyme disease" is 5 of 10 bands on a Western
Blot, but they say this with no proof. A physician may
be paid to testify against a patients' right to treatment
by saying they don't have "Lyme disease" (5 of 10 bands)
and then in that sense, they are correct. Most Lyme-
infected patients do not have 5 of 10 bands. It has
not been published, how many people who have been
*treated* have 5 of 10 bands, although it is known, as
this data was collected by Mark Klempner, formerly of
Tufts, with his $4.7 million research grant. The
anecdotal evidence (Dr. Klempner's comment in a
public talk, among others) is that that range is 0.5 to
3.9% of all treated Lyme patients. Treatment abrogates
antibody response and many patients are completely
seronegative. Treatment does not always entirely wipe
out the infection *or* symptoms, as is the case in animal
and human spirochetal infections. It will be shown how
this is so, below.
THE NATURE OF THE PHYLUM- Defies Managed Care
What can Spirochetes do?
Morphological changes- the spheroplast or "cyst"
Dessication
Antimicrobial resistance genes sharing
Lateral gene tranfser
Intracellular persistence- immune and antibiotic evasion
Antigenic variation- immune evasion
Freezing- Lyme patients cannot donate blood, per the Red Cross
Porins- resistance to antimicrobials
Free-Living Spirochete, microbial mats, from dessication
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8346204&dopt=Abstract
Proc Natl Acad Sci U S A 1993 Aug 1;90(15):6966-6970,
Composite, large spirochetes from microbial mats:
spirochete structure review., Margulis L, Ashen JB,
Sole M, Guerrero R., Department of Biology, University
of Massachusetts, Amherst 01003. Full Text: GOOD ONE!
MORPHOMETRICS! SEE LAST PAGE, BOTTOM LEFT!
http://www.pnas.org/cgi/reprint/90/15/6966.pdf
Other, dessication
US army document (aerosolized)
http://www.google.com/search?q=cache:WpRbE1BgHKM:www.afpmb.org/pubs/dveps/haiti.pdf+US+Army,+Leptospira,+Haiti&hl=en
Haiti, 1994, Armed Forces Pest Management Board, Defense
Pest Management Analysis Center, Forest Glen Section,
WRAMC, Washington, DC, 20301-5001
"INFECTIOUS AGENT : Leptospira interrogans, over 200
serovars are known to exist. EPIDEMIOLOGY : This disease
occurs in both rural and urban environments, and it is
particularly common among people who work outdoors. The
case fatality rate is usually less than 20%. Reservoirs
include numerous wild and domestic animals, reptiles and
amphibians. Transmission is effected through pathogen
contact with skin and mucous mem branes (especially if
these are cut or abraded) via contaminated food or water,
moist soil, vegetation, the urine or tissues of infected
animals or, rarely, aerosols."
Intravector spheroplast
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=6414200&dopt=Abstract
Zh Mikrobiol Epidemiol Immunobiol 1983 Jul;7:47-51,
[Processes of bacterial L transformation and L form
reversion in the body of argasid ticks]. [Article in
Russian], Levina GA, Podboronov VM, Prozorovskii SV,
Grokhovskaia IM, Shlygina KN.
"Spherical bodies"- Oral Spirochetes
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10895696&dopt=Abstract
Oral Microbiol Immunol 1999 Dec;14(6):384-6, Factors
affecting the formation of spherical bodies in the
spirochete Treponema denticola., De Ciccio A, McLaughlin
R, Chan EC., Faculty of Dentistry, McGill University,
Montreal, Quebec, Canada.
"The oral spirochete Treponema denticola typically is a
helically shaped, motile bacterial cell. However,
morphological variations of T. denticola cells in the
form of "spherical bodies" are sometimes seen. Little
is known about the environmental factors that cause their
formation. The effects of oxygen, growth temperature,
nutrient depletion and the addition of metabolic end-
products were tested to determine their role in the
morphogenesis of the spherical bodies. It was found
that the age of the culture, the omission of individual
components (yeast extract, rabbit serum, volatile fatty
acids or thiamine pyrophosphate) from the medium and
the addition of the metabolic end product lactic acid
enhanced the formation of these bodies. However, their
formation was decreased upon omission of the medium
components asparagine and sodium bicarbonate."
PMID: 10895696
Borrelial spheroplasts
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9266264&dopt=Abstract
Infection 1997 Jul;25(4):240-246, Transformation of
cystic forms of Borrelia burgdorferi to normal, mobile
spirochetes., Brorson O, Brorson SH., Dept. of
Microbiology, Ulleval University Hospital, Oslo, Norway.
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9646104&dopt=Abstract
Infection 1998 May;26(3):144-150, 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 SH., Dept. of
Microbiology, Vestfold Sentralsykehus, Tonsberg.
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10052721&dopt=Abstract
APMIS 1998 Dec;106(12):1131-1141, A rapid method for
generating cystic forms of Borrelia burgdorferi, and
their reversal to mobile spirochetes., Brorson O, Brorson
SH., Department of Microbiology, Vestfold Sentralsykehus,
Tonsberg, Norway.
Antimicrobial resistance in spheroplast form of B burgdorferi
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10379684&dopt=Abstract
APMIS 1999 Jun;107(6):566-576, An in vitro study of the
susceptibility of mobile and cystic forms of Borrelia
burgdorferi to metronidazole., Brorson O, Brorson SH.,
Department of Microbiology, Vestfold Sentralsykehus,
Tonsberg, Norway.
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11478686&dopt=Abstract
APMIS 2001 May;109(5):383-388, 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
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11081332&dopt=Abstract
Pol Merkuriusz Lek 2000 Aug;9(50):584-588, [Neurologic
syndromes in Lyme disease]. Zajkowska JM, Hermanowska-
Szpakowicz T, Kondrusik M, Pancewicz SA. Kliniki Chorob
Pasozytniczych i Neuroinfekcji AM w Bialymstoku
PHAGE TRANSDUCTION; TRANSFER of ANTIBIOTIC RESISTENCE GENES
Bacteriophage transduction of Brachyspira (swine dysentery)
http://www.nps.ars.usda.gov/publications/publications.htm?lognum=0000091627
Online publication of the United States Department of
Agricultural Research Service, Identification And
Detection Of Genes Of Vsh-1, An Unusual Bacteriophage
(host Genome Packaging Agent) Of Serpulina Hyodysenteriae
"Interpretive Summary: We recently reported the transfer
of antibiotic resistance genes between cells of Serpulina
hyodysenteriae. This spiral-shaped bacterium causes swine
dysentery, a worldwide disease estimated to cost the U.S.
pork industry $100 million per year. This was the first
report of gene transfer between S. hyodysenteriae cells.
We discovered that the genes were transferred by a
previously unknown gene transfer agent that we named
VSH-1. In the present studies, we identified several
VSH-1 genes. We found VSH-1 genes in every known species
of Serpulina which cause intestinal disease in chickens,
pigs, and possibly in humans. These pioneering studies
suggest the gene transfer agent, VSH-1, is common among
Serpulina species. As a result, VSH-1 and similar agents
should be evaluated for bacteria in the intestinal tract
to trade genes. ***The importance of these results is
that we have described a new method by which genes such
as antibiotic resistance genes can be spread among
bacteria.*** The resistance of bacterial pathogens to
antibiotics is anemerging and serious problem in human
and veterinary medicine."
Same-MEDLINE
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11319078&dopt=Abstract
Appl Environ Microbiol 2001 May;67(5):2037-2043
Brachyspira (Serpulina) hyodysenteriae gyrB mutants
and interstrain transfer of coumermycin (1) resistance.,
Stanton TB, Matson EG, Humphrey SB., Pre-Harvest Food
Safety and Enteric Diseases Research, National Animal
Disease Center, USDA Agricultural Research Service,
Ames, IA 50010, USA., tstanton@nadc.ars.usda.gov
LEPTOSPIRA BACTERIOPHAGE REPLICATES AS A PLASMID
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11004167&dopt=Abstract
J Bacteriol 2000 Oct;182(20):5700-5705, The LE1
bacteriophage replicates as a plasmid within Leptospira
biflexa: construction of an L. biflexa-Escherichia coli
shuttle vector., Girons IS, Bourhy P, Ottone C, Picardeau
M, Yelton D, Hendrix RW, Glaser P, Charon N., Unite de
Bacteriologie Moleculaire et Medicale, Institut Pasteur,
75724 Paris Cedex 15, France. isgirons@pasteur.fr
LEPTOSPIRAL LATERAL GENE TRANSFER
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11075918&dopt=Abstract
J Mol Microbiol Biotechnol 2000 Oct;2(4):455-62,
Technological advances in the molecular biology of
Leptospira., Zuerner R, Haake D, Adler B, Segers R.,
Bacterial Diseases of Livestock, National Animal
Disease Center, Agricultural Research Service, US
Department of Agriculture, Ames, Iowa50010, USA.
rzuerner@nadc.ars.usda.gov
SWINE LAGOONS AND RESISTANCE TO THE TETRACYCLINES
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11282596&dopt=Abstract
Appl Environ Microbiol 2001, Apr;67(4):1494-1502,
Occurrence and diversity of tetracycline resistance
genes in lagoons and groundwater underlying two swine
production facilities., Chee-Sanford JC, Aminov RI,
Krapac IJ, Garrigues-Jeanjean N, Mackie RI., Department
of Animal Sciences, University of Illinois at
Urbana-Champaign,Urbana, IL 61801, USA.
PHAGE TRANSDUCTION/BACTERIOPHAGES OF BORRELIA burgdorferi
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11466280&dopt=Abstract
J Bacteriol 2001 Aug;183(16):4771-4778, Transduction
by phiBB-1, a Bacteriophage of Borrelia burgdorferi.,
Eggers CH, Kimmel BJ, Bono JL, Elias AF, Rosa P, Samuels
DS., Division of Biological Sciences, The University of
Montana, Missoula, Montana 59812. "The kan cassette
recombined into a resident cp32 and was stably maintained.
The cp32 containing the kan cassette was packaged by
phiBB-1 released from this B. burgdorferi strain. phiBB-1
has been used to transduce this antibiotic resistance
marker into naive CA-11.2A cells, as well as two other
strains of B. burgdorferi. This is the first direct
evidence of a mechanism for lateral gene transfer in B.
burgdorferi."
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=6853449&dopt=Abstract
J Bacteriol 1983 Jun;154(3):1436-1439, Bacteriophage
in the Ixodes dammini spirochete, etiological agent
of Lyme disease., Hayes SF, Burgdorfer W, Barbour AG.
"A bacteriophage with a B-3 morphology was detected by
electron microscopy in a spirochete isolated from the
tick Ixodes dammini. It has a 40- to 50-nm elongated
head and a tail 50 to 70 nm in length. It appears
devoid of collars or kite-tail structure. The
spirochete has been identified as the causative agent
of Lyme disease."
ANTIGENIC VARIATION IN BRACHYSPIRA
Brachyspira Antigenic variation
http://www.nps.ars.usda.gov/publications/publications.htm?lognum=0000114138
The Search For Brachyspira Outer Membrane Proteins
That Interact With The Host "Technical Abstract:
Little is known about the outer membrane structure
of Brachyspira hyodysenteriae and Brachyspira
pilosicoli or the role of outer membrane proteins
(OMPs) in host colonization and the development of
disease. The isolation of outer membrane vesicles
from B. hyodysenteriae has confirmed that cholesterol
is a significant outer membrane constituent and that
it may impart unique characteristics to the lipid
bilayer structure including a reduced density. Unique
proteins that have been identified in the B.
hyodysenteriae outer membrane include the variable
surface (Vsp) proteins and lipoproteins such as SmpA
and BmpB. While the function of these proteins
remains to be determined, there is evidence to
suggest that they may be involved in immune evasion.
These data may explain the ability of the organism
to initiate chronic infection."
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10510046&dopt=Abstract
Vet Microbiol 1999 Aug 31;68(3-4):273-283, Sequence
characterization of two new members of a multi-gene
family in Serpulina hyodysenteriae (B204) with
homology to a 39 kDa surface exposed protein: vspC and
D., McCaman MT, Auer K, Foley W, Gabe JD., Berlex
Biosciences, Process Development Department, Richmond,
CA, USA.
"Previous cloning and sequencing of clones from a
genomic library constructed from Serpulina hyodysenteriae
B204 had identified a tandem pair of open reading frames,
identified as vspA and vspB (variable surface protein)
expected to encode proteins with homology to (but not
identical with) a 39 kDa surface exposed membrane protein
from this animal pathogen. Additional screening of the
genomic library was performed to retrieve the remainder of
the vspB gene using new oligonucleotide probes based
upon the cloned gene sequences. Not only was this goal
met but we also discovered two more adjacent and related
vsp genes (vspC and vspD) and have completely sequenced
them.."
ANTIGENIC VARIATION IN THE OTHER BORRELIA
http://www.cdc.gov/ncidod/eid/vol6no5/barbour.htm
Emerging Infectious Diseases, Vol. 6, No. 5, Sep-Oct
2000, Synopsis Antigenic Variation in Vector-Borne
Pathogens, Alan G. Barbour* and Blanca I. Restrepo,
*University of California Irvine, Irvine, California;
and Corporación para Investigaciones Biológicas,
Medellín, Colombia
"Several pathogens of humans and domestic animals
depend on hematophagous arthropods to transmit them
from one vertebrate reservoir host to another and
maintain them in an environment. These pathogens
use antigenic variation to prolong their circulation
in the blood and thus increase the likelihood of
transmission. By convergent evolution, bacterial
and protozoal vector-borne pathogens have acquired
similar genetic mechanisms for successful antigenic
variation. Borrelia spp. and Anaplasma marginale
(among bacteria) and African trypanosomes, Plasmodium
falciparum, and Babesia bovis (among parasites) are
examples of pathogens using these mechanisms.
Antigenic variation poses a challenge in the development
of vaccines against vector-borne pathogens.
Antigenic Variation and Borrelia [~80 citations 26 October 01]
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=PureSearch&db=PubMed&details_term=%28%28%22antigenic%20variation%22%5BMeSH%20Terms%5D%20OR%20antigenic%20variation%5BText%20Word%5D%29%20AND%20%28%22borrelia%22%5BMeSH%20Terms%5D%20OR%20borrelia%5BText%20Word%5D%29%29
INTACELLULARITY AND ANTIMICROBIAL RESISTANCE:
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=1634816&dopt=Abstract
J Infect Dis 1992 Aug;166(2):440-444, Fibroblasts
protect the Lyme disease spirochete, Borrelia burgdorferi,
from ceftriaxone in vitro., Georgilis K, Peacocke M,
Klempner MS., Department of Medicine, New England Medical
Center, Boston, Massachusetts.
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8486939&dopt=Abstract
J Infect Dis 1993 May;167(5):1074-1081, Invasion of
human skin fibroblasts by the Lyme disease spirochete,
Borrelia burgdorferi., Klempner MS, Noring R, Rogers RA.,
Division of Geographic Medicine and Infectious Diseases,
New England Medical Center, Tufts University School of
Medicine, Boston, Massachusetts 02111.
Romanian Fibroblast, Intracellular study
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9558977&dopt=Abstract
Roum Arch Microbiol Immunol 1997 Jan;56(1-2):77-96,
Interactions between Borrelia burgdorferi and
eukaryote cells: comparative ultrastructural aspects.,
Ionescu MD, Ionescu AD, Hristescu S, Ionescu M,
Orasanu M, Coman N. Cantacuzino Institute, Bucharest,
Romania.
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8240439&dopt=Abstract
Arthritis Rheum 1993 Nov;36(11):1621-1626, Persistence
of Borrelia burgdorferi in ligamentous tissue from a
patient with chronic Lyme borreliosis., Haupl T, Hahn
G, Rittig M, Krause A, Schoerner C, Schonherr U, Kalden
JR, Burmester GR., Department of Medicine III, University
of Erlangen-Nuremberg, Germany. [In this study, the
patient was treated for 6 weeks with oral doxy 200 mgs/
day and then 2 weeks IV ceftriaxone, and then 3 weeks
of roxithromycin, sulfatmethoxazole and trimethoprim,
and then surgery was performed.-KMD]
PERSISTENCE OF LEPTOSPIRA (Immune evasion)
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9009336&dopt=Abstract
Infect Immun 1997 Feb;65(2):729-38, Invasion of Vero
cells and induction of apoptosis in macrophages by
pathogenic Leptospira interrogans are correlated
with virulence., Merien F, Baranton G, Perolat P.,
Laboratoire des Leptospires, Institut Pasteur, Noumea,
New Caledonia, France. "...Invasion of epithelial
cells and induction of apoptosis in macrophages may
be related to the pathogenicity of Leptospira, and
both could contribute to its ability to survive in
the host and to escape from the immune response."
PERSISTENCE OF BORRELIA burgfdorferi PAST TREATMENT IN ANIMALS
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10048169&dopt=Abstract
Wien Klin Wochenschr 1998 Dec, 23;110(24):874-81,
Clinical manifestations, pathogenesis, and effect
of antibiotic treatment on Lyme borreliosis in dogs.
Straubinger RK, Straubinger AF, Summers BA, Jacobson
RH, Erb HN., James A. Baker Institute for Animal
Health, Ithaca, New York, USA. rks4@cornell.edu
"CONCLUSIONS: B. burgdorferi disseminates through
tissue by migration following tick inoculation,
produces episodes of acute arthritis, and establishes
persistent infection. The spirochete survives antibiotic
treatment and disease can be reactivated in
immunosuppressed animals."
VIABILITY AFTER FREEZING
LEPTOSPIRA
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=3804861&dopt=Abstract
J Appl Bacteriol 1986 Nov;61(5):407-11, Storage of
pathogenic leptospires in liquid nitrogen., Palit A,
Haylock LM, Cox JC. [to minus 70 C-KMD]
"The viability of nine strains has so far been
observed over a period of 8-22 months storage in
liquid nitrogen and full viability of all strains
has been preserved over this period. Virulence of
strains of serovars pomona and hardjo was well
preserved, as demonstrated by challenge tests in
guinea pigs and domestic pigs."
FREEZING AND SPIROCHETES [~30 citations]
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=PureSearch&db=PubMed&details_term=%28%28%22freezing%22%5BMeSH%20Terms%5D%20OR%20freezing%5BText%20Word%5D%29%20AND%20%28%22spirochaetales%22%5BMeSH%20Terms%5D%20OR%20spirochetes%5BText%20Word%5D%29%29
B. burgdorferi AND FREEZING
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=2773025&dopt=Abstract
Transfusion 1989 Sep;29(7):581-3, Survival of
Borrelia burgdorferi in blood products., Badon SJ,
Fister RD, Cable RG. American Red Cross Blood Services,
Farmington, Connecticut. "The organism was shown to
survive ***in RBCs*** (4 degrees C) and FFP (below -18
degrees C) for 45 days and in PCs (20-24 degrees C)
for 6 days. The results of this study do not exclude
the possibility of transmission of Lyme disease through
blood transfusion."
PORINS and Antimicrobial Resistance [~40 citations on MEDLINE]
http://www.ncbi.nlm.nih.gov:80/entrez/query.fcgi?cmd=PureSearch&db=PubMed&details_term=%28%28%22porins%22%5BMeSH%20Terms%5D%20OR%20PORIN%5BText%20Word%5D%29%20AND%20antimicrobial%5BAll%20Fields%5D%29
So, in conclusion, spirochetes do not behave like
other bacteria. They behave like spirochetes. They
are persistent, environmentally resistant, undergo
morphological changes and revert, can be intracellular,
resistant to antibiotics, can acquire virulence via
phage-vector.
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