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PubMed1996

Phylogeny of not-yet-cultured spirochetes from termite guts.

Comparisons of 16S rDNA sequences were used to determine the phylogeny of not-yet-cultured spirochetes from hindguts of the African higher termite, Nasutitermes lujae (Wasmann). The 16S rRNA genes were amplified directly from spirochete-rich hindguts by using universal primers, and the amplified products were cloned into Escherichia coli. Clones were screened with a spirochete-specific DNA probe. Analysis of 1,410 base positions of the 16S rDNA insert from one spirochete clone, designated NL1, supported its assignment to the genus Treponema, with average interspecies similarities of ca. 85%. The sequence of NL1 was most closely related (ca. 87 to 88% similarity) to sequences of Spirochaeta stenostrepta and Spirochaeta caldaria and to a previously published sequence (ca. 87% similarity) of spirochetal clone MDS1 from the Australian lower termite, Mastotermes darwiniensis (Froggatt). On the basis of 16S rRNA sequence comparisons and individual base signatures, clones NL1 and MDS1 clearly represent two novel species of Treponema, although specific epithets have not yet been proposed. The gross morphology of NL1 was determined from in situ hybridization experiments with an NL1-specific, fluorescently labeled oligonucleotide probe. Cells were approximately 0.3 to 0.4 by 30 microns in size, with a wavelength and amplitude of about 10 microns and 0.8 to 1.6 micron, respectively. Moreover, electron microscopy of various undulate cells present in gut contents confirmed that they possessed ultrastructural features typical of spirochetes, i.e., a wavy protoplasmic cylinder, periplasmic flagella, and an outer sheath. The sequence data suggest that termite gut spirochetes may represent a separate line of descent from other treponemes and that they constitute a significant reservoir of previously unrecognized spirochetal biodiversity.

Animals↗

PubMed1981

Distribution of superoxide dismutase, catalase, and peroxidase activities among Treponema pallidum and other spirochetes.

Representative members of Spirochaetales were surveyed for their content of superoxide dismutase (SOD), catalase, and peroxidase activities. Only Leptospira exhibited peroxidase activity. Obligately anaerobic cultivable Treponema and Spirochaeta possessed no SOD or peroxidative capabilities. Upon polyacrylamide gel electrophoresis, Spirochaeta aurantia, Borrelia hermsi, and five Leptospira biflexa serovars showed SOD activity associated with one electrophoretic band which was inhibited by H2O2, suggesting that they were iron-containing dismutases. These spirochetes could be distinguished by differences in relative mobilities of their SODs. SOD activity, but not catalase activity, was induced aerobically in S. aurantia. All Leptospira interrogans serovars and two L. biflexa serovars lacked significant SOD activity. These SOD-deficient strains of Leptospira, with one exception, possessed high levels of catalase activity. The Nichols strain of virulent Treponema pallidum possessed SOD and catalase activities, but lacked peroxidase activity. The SOD in T. pallidum exhibited two electrophoretic bands containing copper and zinc, and its relative mobility was identical to that of purified rabbit SOD. Immunization of sheep with purified rabbit SOD resulted in antiserum which inhibited both rabbit SOD and T. pallidum SOD assayed by spectrophotometric analysis or activity staining following polyacrylamide gel electrophoresis. In agarose gel diffusion, precipitin lines of identity were observed between purified rabbit SOD and cell extracts of T. pallidum. These data indicated that the SOD activity detected in T. pallidum was host derived.

Borrelia↗

PubMed1973

Peptidoglycan of free-living anaerobic spirochetes.

Electron microscope examination of negatively stained or thin-sectioned cells of Spirochaeta stenostrepta treated with penicillin or lysozyme showed that the peptidoglycan was present as a thin, electron-dense layer adjacent and external to the cytoplasmic membrane. The peptidoglycan was isolated from cells of S. stenostrepta and Spirochaeta litoralis by a procedure including treatments with sodium lauryl sulfate and Pronase. Hydrolysates of the isolated S. stenostrepta and S. litoralis peptidoglycans contained glucosamine, muramic acid, glutamic acid, l-ornithine, and alanine in molar ratios of 0.90:0.85:1.00:1.00:1.40 and of 0.63:0.63:0.99:1.00:1.41, respectively. Determination of N-terminal residues suggested that nearly 50% of the ornithine in S. stenostrepta and S. litoralis peptidoglycans was involved in peptide cross-linkage. The peptidoglycan layer of S. stenostrepta was sensitive to lysozyme and myxobacter AL-1 protease.

Alanine↗

PubMed1977

Relationship between cell coiling and motility of spirochetes in viscous environments.

The lowest viscosity that stops translational motility of cells (minimum immobilizing viscosity [MIV] was determined for various spirochetes. The viscous agent used was polyvinylpyrrolidone, The MIV for either Spirochaeta halophila P1 or Spirochaeta aurantia J4T was approximately 1,000 centipoise (cp), and for Leptospira interrogans (biflexa) B16 the MIV was greater than 500 cp. In comparison, the MIV for the flagellated bacteria Escherichia coli and Spirillum serpens was 60 cp. MIV values for two S. halophila mutant strains lacking the characteristic cell coiling (Hel-mutants) were 70 and 120 cp, approximately one-tenth the MIV for the wild-type strain. MIV values for cells of S. aurantia strains with fewer coils than comparably long cells of S. aurantia J4T were 300 to 600 cp. The average velocity of strains of S. aurantia and S. halophila decreased at viscosities higher than 2 to 3 cp. At 2 cp the average velocity of S. halophila P1 was 16 micron/s, whereas the average velocities of Hel-mutant strains were 7 to 9 micron/s. This study indicates that the coiling of spirochetes plays a role in their ability to move through environments of realtively high viscosity. Among the spirochetes we investigated, this ability is greater in the more extensively coiled strains.

Culture Media↗

PubMed1980

Involvement of periplasmic fibrils in motility of spirochetes.

Nonmotile (Mot-) strains of Spirochaeta aurantia and Spirochaeta halophila were isolated with a procedure involving mutagenesis of motile wild-type cells. Electron microscopy showed that a Mot- mutant strain of S. halophia possessed incomplete periplasmic fibrils, inasmuch as most or all of the filamentous portion of the periplasmic fibrils was absent. Some of the cells of this Mot-, fibril-defective mutant strain lacked the filamentous portion of the periplasmic fibrils and formed proximal hooks, whereas other cells appeared to have a very small segment of the filamentous portion of the periplasmic fibrils attached to the proximal hooks. Motile revertants were isolated repeatedly from cultures of the Mot-, fibril-defective mutant and from S. halophila Mot- mutants that completely lacked periplasmic fibrils. The motile revertants possessed periplasmic fibrils ultrastructurally indistinguishable from wild-type periplasmic fibrils. This study indicates that periplasmic fibrils play an essential role in the motility of spirochetes.

Cytoplasm↗

PubMed1982

Enzymatic activities for interconversion of purines in spirochetes.

Enzymatic activities that catalyze the interconversion of purines and purine derivatives were detected in cell extracts of Spirochaeta aurantia, Spirochaeta stenostrepta, Treponema succinifaciens, and Treponema denticola. Phosphoribosyltransferase activities present in cell extracts of each of the four spirochete species functioned in the conversion of adenine, hypoxanthine, and guanine to AMP, IMP, and GMP, respectively. Nucleotidase activities in the extracts mediated the formation of nucleosides from nucleotides. The conversion of adenosine, inosine, and guanosine to the respective purine bases was catalyzed by nucleoside phosphorylase and, in some instances, by nucleoside hydrolase activities. Guanine deaminase activity was found in both S. aurantia and S. stenostrepta, whereas adenosine deaminase activity was detected only in S. aurantia. Adenine deaminase activity in T. succinifaciens extracts was sensitive to O2 and was relatively resistant to heating. Our results indicate that the four species of spirochetes studied possess a broad spectrum of purine interconversion enzymes. It is suggested that these enzymes may function in metabolic processes important for the survival of spirochetes in nutrient-poor natural environments.

Adenosine Deaminase↗

PubMed1994

16S rDNA sequence and phylogenetic position of an uncultivated spirochete from the hindgut of the termite Mastotermes darwiniensis Froggatt.

We have analyzed the 16S rDNA sequence and the phylogenetic position of an uncultivated spirochete from the hindgut contents of the Australian termite Mastotermes darwiniensis Froggatt. The 16S rRNA genes of bacteria from the hindgut contents of Mastotermes darwiniensis were amplified by polymerase chain reaction. The amplification products were cloned and sequenced. The sequences were compared to known homologous primary structures. Two of the clones (MDS1 and MDS3) had an insert of 1498 nucleotides showing typical signatures of spirochete 16S rRNA sequences. The sequences of the two clones were most similar to the 16S rRNA sequence of Spirochaeta stenostrepta (89.8%) and Treponema sp. strain H1 (90.7%). Phylogenetical analysis positioned the hindgut spirochete sequence with that of the free-living anaerobic Spirochaeta stenostrepta and Treponema sp. strain H1 as its nearest relatives within the cluster of the spirochetes. We conclude that analyzed SSU rDNA sequences originate from a spirochete related to the genus Treponema. It is possibly one of the uncultivated unique spirochetes symbiotic in termite hindguts.

Animals↗

PubMed1982

Anaerobic spirochete from a deep-sea hydrothermal vent.

An obligately anaerobic spirochete, designated strain GS-2, was selectively isolated from samples collected at a deep-sea (2,550 m) hydrothermal vent of the Galapagos Rift ocean floor spreading center. The morphological and physiological characteristics of strain GS-2 resembled those of Spirochaeta strains. However, strain GS-2 failed to grow consistently in any liquid medium tested. In addition, strain GS-2 grew more slowly and to lower yields than other Spirochaeta species. The occurrence of obligately anaerobic bacteria in hydrothermal vents indicates that the water in at least some of the vent areas is anoxic. The presence of strain GS-2 shows that these areas are favorable for anaerobic marine spirochetes.

Journal Article↗

PubMed1992

Polymerase chain reaction primers and probes derived from flagellin gene sequences for specific detection of the agents of Lyme disease and North American relapsing fever.

By cloning and sequencing the flagellin gene of Borrelia hermsii and comparing this sequence with that of the corresponding gene from B. burgdorferi, I identified a central region within the two genes which showed a reduced level of sequence similarity. Oligonucleotide sequences selected from this region produced species-specific amplimers when used in polymerase chain reaction experiments. Thus, primers derived from the B. burgdorferi sequence amplified a 276-bp fragment from 22 strains of B. burgdorferi of diverse geographic origin but not from 5 strains of B. hermsii, 5 other Borrelia species, 16 Treponema, Leptospira, and Spirochaeta species, or representatives of 10 other bacterial genera. However, when the amplified fragments were tested for hybridization with an oligonucleotide probe derived from the nonhomologous region, seven strains from either Germany or Switzerland did not hybridize. Cloning and sequencing of the amplified fragments from these strains revealed that the 22 strains of B. burgdorferi tested could be divided into three groups based on the nucleic acid sequence of the central region of the flagellin gene. With this information, oligonucleotide probes that hybridized to the amplified fragments and were able to differentiate the three groups of B. burgdorferi were designed. The corresponding primers, derived from the B. hermsii gene sequence, were tested for their ability to amplify DNA from this collection of strains. Although no amplification was obtained with representatives of the three groups of B. burgdorferi or various Treponema, Leptospira, and Spirochaeta species, amplification was obtained with the five other Borrelia species (B. parkeri, B. turicatae, B. crocidurae, B. anserina, and B. coriaceae) in addition to the five strains of B. hermsii. Sequencing of the amplified fragments from one strain of B. hermsii as well as B. parkeri and B. turicatae allowed the design of oligonucleotide probes that were able to differentiate the three species of North American relapsing fever spirochetes into two separate groups. These studies suggest that there is sufficient diversity within the flagellin gene sequences of closely related Borrelia species to differentiate them into groups and to pursue taxonomic studies both within and between species.

Amino Acid Sequence↗

PubMedDate not supplied

[Clinical contribution to the knowledge of Lyme disease. Case reports].

Lyme disease (LD) is a well-recognized multi-system disorder, caused by the spirochaeta Borrelia burgdorferi. It involves the skin, nervous system, joints and heart. Our group reported the first case of Lyme Borreliosis in Italy in 1985. Italian Group for Lyme Borreliosis has recently reported the epidemiology of Lyme disease in Italy. The epidemiological data seem to underestimate the prevalence of Lyme arthritis in Italy. The pattern of articular involvement is most often mono- or oligoarthritis with polyarticular arthralgias and frequently needs laboratory confirmation. In this study, some clinical observations of Lyme Borreliosis occurred in the last year are reported. Analysis of the B. burgdorferi immune response by ELISA and Western Blot techniques has the potential to support or exclude a diagnosis of early stage as well as active LD infection was associated with Polymerase Chain Reaction assay based on amplification of the DNA of B. burgdorferi. When the direct evidence of spirochaeta in tissues is lacking, this approach permits the best evaluation of Lyme borreliosis.

Adolescent↗

PubMed2005

[Genetic variants of Borrelia afzelii, a pathogen of the ixodes tick borrelioses].

Borrelia afzelii is one of two most important pathogens of the ixodes tick borrelioses (ITB) in Russia and neighboring countries. This pathogen circulates in various ecosystems and has a wide range of reservoir hosts and transmitters. The results of studies of genetic heterogeneity of the spirochaetae B. afzelii are considered. A total of 139 primary isolates were studied. The isolates were isolated from three species of Ixodes ticks at different stages of development and obtained from the laboratory of infection transmitters, Gamaleya Scientific Research Institute of Epidemiology and Microbiology, Russran Academy of Medical Sciences, Moscow. The transmitters and reservoir hosts of borrelias were caught in natural foci of Russia (from Kaliningrad region irk the west to south Sakhalin in the east), Czechia, Lithuania, Estonia, Ukraine, and Moldavia. Analysis of genotype sequence similarity obtained by sequencing of the rrf(SS)-rrl(23S) spacer demonstrated that the B. afzelii genospecies incorporated no less than 10 genetic variants of spirochaetae, most variants being geographically widespread.

Animals↗

PubMed1990

[Syphilitic arthro-osteitis].

We report a black female with past history of sexual promiscuity who developed arthroosteitis of clavicula and sternoclavicular joint and cranial osteitis during secondary syphilis. Spirochaeta were identified in the material from osteoarticular biopsy. The patient was treated with benzathine penicillin, 2,400,000 U weekly for three weeks. There was a complete clinical recovery. We emphasize the uncommon occurrence of osteoarticular involvement in secondary syphilis and the exceptional character, according to the literature, of the finding of spirochaeta in the biopsy material. We discuss the differential diagnosis and we review the literature on secondary syphilis. We suggest that syphilis should be considered in the differential diagnosis of any acute arthritis or osteitis, particularly when the involved joint is the sternoclavicular and the patient is sexually promiscuous.

Adult↗

PubMed1996

Archaeal-eubacterial mergers in the origin of Eukarya: phylogenetic classification of life.

A symbiosis-based phylogeny leads to a consistent, useful classification system for all life. "Kingdoms" and "Domains" are replaced by biological names for the most inclusive taxa: Prokarya (bacteria) and Eukarya (symbiosis-derived nucleated organisms). The earliest Eukarya, anaerobic mastigotes, hypothetically originated from permanent whole-cell fusion between members of Archaea (e.g., Thermoplasma-like organisms) and of Eubacteria (e.g., Spirochaeta-like organisms). Molecular biology, life-history, and fossil record evidence support the reunification of bacteria as Prokarya while subdividing Eukarya into uniquely defined subtaxa: Protoctista, Animalia, Fungi, and Plantae.

Animals↗

PubMed2001

Nitrogen fixation by symbiotic and free-living spirochetes.

Spirochetes from termite hindguts and freshwater sediments possessed homologs of a nitrogenase gene (nifH) and exhibited nitrogenase activity, a previously unrecognized metabolic capability in spirochetes. Fixation of 15-dinitrogen was demonstrated with termite gut Treponema ZAS-9 and free-living Spirochaeta aurantia. Homologs of nifH were also present in human oral and bovine ruminal treponemes. Results implicate spirochetes in the nitrogen nutrition of termites, whose food is typically low in nitrogen, and in global nitrogen cycling. These results also proffer spirochetes as a likely origin of certain nifHs observed in termite guts and other environments that were not previously attributable to known microbes.

Acetylene↗

PubMed1985

Isolation of a spirochete from the soft tick, Ornithodoros coriaceus: a possible agent of epizootic bovine abortion.

A Borrelia-like spirochete was detected in all three parasitic stages of Ornithodoros coriaceus, the soft tick implicated in the epizoology of epizootic bovine abortion. After the spirochete had been isolated, its distinctness from other North American tick-borne borreliae as well as from Spirochaeta aurantia, Treponema pallidum, and Leptospira interrogans serovar pomona was established on the basis of its morphology, protein components, and inability to infect mice. The spirochete is passed trans-stadially and via eggs by ticks, and it is also excreted in coxal fluid after ticks have fed and detached. Circumstantial evidence suggests that the spirochete may be causally related to epizootic bovine abortion.

Abortion, Veterinary↗

PubMed1975

Carotenoid pigments of facultatively anaerobic spirochetes.

Carotenoid pigments were purified from a previously undescribed, red, halophilic spirochete (spirochete RS1), and from Spirochaeta aurantia strain J1. Both spirochetes are facultative anaerobes and produce pigments when growing aerobically. The major pigments of the two spirochetes were identified by means of chromatographic analysis, absorption spectroscopy, hydride reduction, acetylation and silylation experiments, and mass spectrometry. It was concluded that the major pigment from spirochete RS1 was 4-keto-1',2'-dihydro-1'-hydroxytorulene. This conclusion was further supported by infrared spectroscopy and additional analytical data. The evidence showed that the major pigment from S. aurantia was 1',2'-dihydro-1'-hydroxytorulene. Chromatographic and spectrophotometric evidence indicated that this pigment was also present, as a minor carotenoid component, in spirochete RS1. These pigments have been previously detected almost exclusively in gliding bacteria, such as species of Flexibacter, Stigmatella, and Myxococcus. The occurrence of 4-keto-1',2'-dihydro-1'-hydroxytorulene and 1',2'-dihydro-1'-hydroxytorulene in both spirochetes and gliding bacteria may have significance with respect to the evolutionary development of these organisms.

Acetylation↗

PubMed1985

Chemotactic transducer proteins of Escherichia coli exhibit homology with methyl-accepting proteins from distantly related bacteria.

Transducers are transmembrane, methyl-accepting proteins central to the chemotactic systems of the enteric bacteria Escherichia coli and Salmonella typhimurium. Methyl-accepting proteins have been reported in a number of species in addition to these enteric bacteria. Those species include Bacillus subtilis and Spirochaeta aurantia, representatives of groups that diverged from ancestral enteric bacteria and from each other very early in bacterial evolution. An antiserum that reacts with all transducers of E. coli precipitated specifically methyl-accepting proteins from B. subtilis and S. aurantia, indicating that these proteins share antigenic determinants with transducers of E. coli. In addition, analysis of tryptic peptides by high-pressure liquid chromatography revealed similarities in the regions of methyl-accepting sites for proteins from all three species. These observations imply that structural features have been preserved in the three species from transducers contained in a common ancestor of eubacteria. It is thus reasonable to predict that other flagellated, chemotactic bacteria will be found to contain methyl-accepting proteins homologous to transducers of enteric bacteria.

Bacillus subtilis↗

PubMed2000

Phylogenetic foundation of spirochetes.

The spirochetes are free-living or host-associated, helical bacteria, some of which are pathogenic to man and animal. Comparisons of 16S rRNA sequences demonstrate that the spirochetes represent a monophyletic phylum within the bacteria. The spirochetes are presently classified in the Class Spirochaetes in the order Spirochetales and are divided into three major phylogenetic groupings, or families. The first family Spirochaetaceae contains species of the genera Borrelia, Brevinema, Cristispira, Spirochaeta, Spironema, and Treponema. The second family Brachyspiraceae contains the genus Brachyspira (Serpulina). The third family Leptospiraceae contains species of the genera Leptonema and Leptospira. Novel spirochetal species, or phylotypes, that can not be presently cultivated in vitro, have been identified from the human oral cavity, the termite gut, and other host-associated or free-living sources. There are now over 200 spirochetal species or phylotypes, of which more than half is presently not cultivable. It is likely that there is still a significant unrecognized spirochetal diversity that should be evaluated.

Borrelia↗