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PubMed1998

Molecular phylogenetic identification of the intestinal anaerobic microbial community in the hindgut of the termite, Reticulitermes speratus, without cultivation.

A termite maintains an anaerobic microbial community in its hindgut, which seems to be the minimum size of an anaerobic habitat. This microbial community consists of bacteria and various anaerobic flagellates, and it is established that termites are totally dependent on the microbes for the utilization of their food. The molecular phylogenetic diversity of the intestinal microflora of a lower termite, Reticulitermes speratus, was examined by a strategy that does not rely on cultivation of the resident microorganisms. Small subunit ribosomal RNA (ssrRNA) genes were directly amplified from the mixed-population DNA of the termite gut by polymerase chain reaction (PCR) and clonally isolated. Most sequenced clones were phylogenetically affiliated with the four major groups of the domain Bacteria: the Proteobacteria group, the Spirochete group, the Bacteroides group, and the Low G + C gram-positive bacteria. The 16S rRNA sequence data show that the majority of the intestinal microflora of the termite consists of new species that are yet to be cultured. The phylogeny of a symbiotic methanogen inhabiting the gut of a lower termite (R. speratus) was analyzed without cultivation. The nucleotide sequence of the ssrDNA and the predicted amino acid sequence of the mcrA product were compared with those of the known methanogens. Both comparisons indicated that the termite symbiotic methanogen belonged to the order Methanobacteriales but was distinct from the known members of this order. The diversity of nitrogen-fixing organisms was also investigated without culturing the resident microorganisms. Fragments of the nifH gene, which encodes the dinitrogenase reductase, were directly amplified from the mixed-population DNA of the termite gut and were clonally isolated. The phylogenetic analysis of the nifH amino acid sequences showed that there was a remarkable diversity of nitrogenase genes in the termite gut. The molecular phylogeny of a symbiotic hypermastigote Trichonympha agilis (class Parabasalia; order Hypermastigida) in the hindgut of R. speratus was also examined by the same strategy. The whole-cell hybridization experiments indicated that the sequence originated from a large hypermastigote in the termite hindgut, Trichonympha agilis. According to the phylogenetic trees constructed, the hypermastigote represented one of the deepest branches of eukaryotes. The hypermastigote along with members of the order Trichomonadida formed a monophyletic lineage, indicating that the hypermastigote and trichomonads shared a recent common ancestry.

Animals↗
PubMed2001

Association between multiple sclerosis and cystic structures in cerebrospinal fluid.

BACKGROUND: The aim of the study was to search for infectious agents in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS). PATIENTS AND METHODS: CSF from ten patients with the diagnosis relapsing remitting MS and from five controls without MS were examined by transmission electron microscopy (TEM), dark field microscopy (DF), interference contrast microscopy (ICM) and UV-microscopic examination of acridine orange staining (AO). All CSF samples from patients and controls were cultured. RESULTS: 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 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 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. Our study may encourage other researchers to study larger patient groups.

Adult↗
PubMed2002

Brachyspira (Serpulina) pilosicoli spirochetemia in an immunocompromised patient.

The case of an elderly immunocompromised man with non-Hodgkin's lymphoma who presented with fever, abdominal pain and bloody diarrhea is described. Brachyspira pilosicoli was isolated from culture. The patient was treated with penicillin G i.v. and became afebrile. B. pilosicoli is a recently recognized enteric pathogen of humans and animals. Intestinal spirochetosis should be included in the differential diagnosis of any immunocompromised or critically ill patient with dysentery.

Aged↗
PubMed1985

Diagnosing early Lyme disease.

The diagnostic value of clinical, culture, and serologic findings was studied prospectively in 41 patients with early Lyme disease. Fifteen patients had erythema chronicum migrans alone, and 26 had clinical evidence of disseminated infection, most commonly affecting the brain or meninges, other skin sites, lymph nodes, or joints. Of 40 blood cultures, only one, from a patient with disseminated infection, yielded spirochetes. One of 10 patients tested with localized infection had an elevated IgM response to the Lyme spirochete (200 units or greater) during acute disease. Two to three weeks after beginning antibiotic therapy, four of the 10 patients had elevated specific IgM or IgG responses (200 units or greater). Of the 22 patients tested with disseminated disease, 10 initially had elevated levels of specific IgM or IgG, and 12 had such responses by convalescence. Because of the low yield of cultures and the delay in the specific antibody response, recognition of the clinical picture remains very important in diagnosing early Lyme disease.

Adolescent↗
PubMed1986

Lyme carditis. Electrophysiologic and histopathologic study.

To further define the nature of Lyme carditis, electrophysiologic study and endomyocardial biopsy were performed in a patient with Lyme disease, whose principal cardiac manifestation was high-degree atrioventricular block. Intracardiac recording demonstrated supra-Hisian block and complete absence of an escape mechanism. Gallium 67 scanning demonstrated myocardial uptake, and right ventricular endomyocardial biopsy revealed active lymphocytic myocarditis. A structure compatible with a spirochetal organism was demonstrated in one biopsy specimen. It is concluded that Lyme disease can produce active myocarditis, as suggested by gallium 67 imaging and confirmed by endomyocardial biopsy. Furthermore, the presence of high-grade atrioventricular block in this disease requires aggressive management with temporary pacemaker and corticosteroid therapy.

Adult↗
PubMed1987

Intestinal spirochetosis in homosexual men.

Previous studies have demonstrated intestinal spirochetosis in rectal biopsy specimens from 2 to 7 percent of heterosexual and 36 percent of homosexual patients, but the role of intestinal spirochetosis in the pathogenesis of intestinal disease remains unclear. To assess the clinical, histologic, and microbiologic correlates of intestinal spirochetosis in a high-risk group, rectal biopsy specimens from 130 homosexual men, 92 percent of whom had intestinal symptoms, were evaluated. All men were extensively evaluated for rectal and enteric pathogens. Intestinal spirochetosis was identified in rectal biopsy specimens from 39 (30 percent) men; 15 percent of biopsy specimens revealed intestinal spirochetosis on hematoxylin and eosin plus alcian blue staining, and positive results were found in 30 percent on silver staining. No rectal biopsy specimens from 79 control patients with a variety of gastrointestinal symptoms demonstrated evidence of spirochetosis on hematoxylin and eosin, alcian blue, or silver staining (p less than 0.0001). Fifty-six percent of rectal biopsy specimens from men with intestinal spirochetosis were normal, and no specific histologic abnormality was correlated with intestinal spirochetosis. There were no differences in the presence of or type of intestinal symptoms, sigmoidoscopic appearance of the mucosa, type of sexual practice, or prior antibiotic use in men with and without spirochetosis. Other intestinal pathogens were frequent in both groups, and only rectal gonorrhea was significantly associated with intestinal spirochetosis. It is concluded that homosexual men with intestinal symptoms have an increased prevalence of spirochetosis, often in association with Neisseria gonorrhoeae. Independent association of spirochetosis with clinical or histologic findings could not be demonstrated.

Biopsy↗
PubMed1985

Orthodontic therapy in patients with juvenile periodontitis: clinical and microbiologic effects.

The correction of malocclusions in juvenile periodontitis (JP) patients completing periodontal therapy is a problem of increasing clinical concern to orthodontists, since many teeth with severe alveolar bone loss in these patients can now be successfully treated without extraction. In this report, fixed edgewise orthodontic therapy was carried out after the completion of periodontal therapy on four JP patients. The orthodontic therapy included extensive intrusion of teeth severely affected by JP. Phase-contrast microscopic analysis of subgingival plaque from orthodontically treated teeth was used to monitor longitudinally the effects of fixed orthodontic bands on the subgingival flora and also to monitor the efficacy of topical and systemic antimicrobial therapy aimed at suppression of suspected periodontopathic bacteria. Orthodontic movement was completed on most periodontally compromised teeth without significant evidence of additional deterioration in periodontal status. However, within the first 6 months of orthodontic band placement, all patients had significant increases in the number of spirochetes and motile rods in their subgingival flora. Three of the patients also developed high levels of crevicular polymorphonuclear leukocytes around orthodontically treated teeth, indicating significant subgingival inflammation. Intensive antimicrobial measures, including topical inorganic salt applications and systemic tetracycline, were helpful in limiting clinical inflammation and subgingival colonization by periodontopathogens during orthodontic therapy. The results demonstrate that successful orthodontic repositioning can be carried out in treated JP patients. In addition, bacteriologic monitoring and chemotherapeutic suppression of periodontal pathogens may be valuable in the prevention of further destructive periodontal disease activity in periodontitis patients undergoing orthodontic therapy.

Adolescent↗
PubMed1989

Relationship of human gingival crevicular fluid polyamine concentration to the percentage of spirochaetes in subgingival dental plaque.

This relationship was examined at 45 inflamed periodontitis sites in 28 patients. A statistically significant positive correlation between polyamine concentration and spirochaetes was observed overall (r = 0.551, p = 0.0001). Within subgroups of this population (n = 15 each), there was no such relationship among midly inflamed sites with 5-6 mm probing depths (r = 0.0003, p = 0.999), but significant relationships among moderately inflamed sites with 5-6 mm depths (r = 0.567, p = 0.027) and 7-11 mm depths (r = 0.639, p = 0.010). Thus polyamines may be involved in subgingival plaque homeostasis at moderately inflamed periodontal sites.

Dental Plaque↗
PubMed1990

Spirochaetes and granulocytes at sites involved in periodontal disease.

Eighty-six men and 63 women with periodontitis participated in this investigation, at the start of which clinical indices were recorded and the pocket contents from one periodontally involved site for each patient were analysed. The numbers of spirochaetes, other motile microorganisms, non-motile filaments, rods or cocci and of granulocytes, monocytes and epithelial cells were determined. After non-surgical treatment 98 patients (P-group) were unsuccessfully and 51 (C-group) were successfully treated. At baseline the P-group had significantly higher numbers of spirochaetes (p = 0.0001) and polymorphonuclear leucocytes (p = 0.0256) than the C-group. The number of rods was statistically higher in the C-group (p = 0.0254). There was no significant difference between the groups with respect to the number of remaining teeth, plaque or calculus scores. Significantly higher values were found in the P-group for bleeding on probing (p = 0.0434), number of pockets greater than or equal to 5 mm (p = 0.0001), mean pocket depths (p = 0.0001), percentage bone loss per site (p = 0.0001) and the number of sites with greater than or equal to 20% bone loss on radiographs (p = 0.0001).

Bacteria↗
PubMed1985

Electron microscopy of the intestine and rectum in acquired immunodeficiency syndrome.

To provide a better understanding of the morphologic changes that take place in the intestine and colon in acquired immunodeficiency syndrome (AIDS), electron microscopy was performed on intestinal or colonic biopsy specimens obtained from 6 patients with AIDS and from 2 patients with AIDS-related lymphadenopathy syndrome. Cryptosporidia were attached to the plasma membrane of epithelial cells in 2 patients and were noninvasive. An invasive protozoan organism identified as Microsporidia was found in 1 patient. Evidence for epithelial cell injury was limited. Unusually prominent secretory granules in colonic epithelial cells (a morphologic counterpart of secretion) was found in 2 patients. Tubuloreticular structures were observed in 7 patients. The structures were found in endothelial cells, lymphocytes, monocytes, intraepithelial lymphocytes, and free in the capillary lumen. Tube- and ring-shaped forms were observed in 2 patients, prominent intraepithelial mast cells in 4 patients, rectal spirochetosis in 1 patient, and pseudomembranous colitis in 1 patient with intestinal and systemic shigellosis. Vesicular rosettes, retroviruses, other viruses, and Mycobacterium avium-intracellulare were not observed. These observations expand our knowledge of morphologic changes in the colonic and intestinal mucosa in patients with AIDS. Tubuloreticular structures are so prominent, in contrast to our previous electron-microscopic observations in other disease and normal states of the intestine and colon, that their finding (though clearly nonspecific) may be a clue to the diagnosis of AIDS in an otherwise equivocal situation.

Acquired Immunodeficiency Syndrome↗