Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 29-1984. A 21-year-old man with headache, fever, and facial diplegia.
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OBJECTIVES: Recent molecular approaches have revealed that fastidious organisms such as Bacteroides forsythus and oral treponemes were frequently found in root canals with apical periodontitis. The purpose of this study was to identify the isolates of oral spirochetes at the species level in endodontic infections and to determine their association with B forsythus and Porphyromonas gingivalis. STUDY DESIGN: Seventy-nine teeth with apical periodontitis were selected for this study. After sampling from the root canals aseptically, polymerase chain reaction amplification for the 16S rRNA gene was performed with eubacterial universal primers. Subsequently, dot-blot hybridization was performed with 8 species-specific oligonucleotide probes. The microbial associations were analyzed by using the odds ratio. RESULTS: The most frequently found species was P gingivalis (27.4%), followed by Treponema maltophilum (26%), B forsythus (16.4%), and Treponema socranskii (2.7%). Other treponemes, including Treponema denticola, were not detected in our samples. Significant microbial associations were identified between T maltophilum, B forsythus, and P gingivalis by performing analysis with the odds ratio. CONCLUSION: Our results indicate that T maltophilum should be included in etiologic studies of endodontic diseases.
OBJECTIVE: Our objective was to determine the relationship between various types of perinatal infections and stillbirths. STUDY DESIGN: By use of various textbooks on perinatal infections, multiple MEDLINE searches, and the reference list of all appropriate manuscripts, the appropriate English language literature was reviewed to define the relationship between various perinatal infections and stillbirths. RESULTS: Infection may cause stillbirth by a number of mechanisms, including direct infection, placental damage, and severe maternal illness. A large variety of organisms have been associated with stillbirth, including many bacteria, viruses, and protozoa. In developed countries, between 10% and 25% of stillbirths may be caused by an infection, whereas in developing countries, which often have much higher stillbirth rates, the contribution of infection is much greater. Ascending bacterial infection, both before and after membrane rupture, with organisms such as Escherichia coli, group B streptococci, and Ureaplasma urealyticum is usually the most common infectious cause of stillbirth. However, in areas where syphilis is very prevalent, up to half of all stillbirths may be caused by this infection alone. Malaria may be an important cause of stillbirth in women infected for the first time in pregnancy. The two most important viral causes of stillbirth are parvovirus and Coxsackie virus, although a number of other viral infections appear to be causal. Toxoplasma gondii, leptospirosis, Listeria monocytogenes, and the organisms that cause leptospirosis, Q fever, and Lyme disease have all been implicated as etiologic for stillbirth. CONCLUSION: Because infection-related stillbirth is relatively rare in developed countries, and those that do occur are caused by a wide variety of organisms, reducing this etiologic component of stillbirth much further will be difficult. However, in certain developing countries, the stillbirth rate is so high and the infection-related component so great that achieving a substantial reduction in stillbirth should be possible simply by reducing maternal infections.
We develop a symbiogenetic concept of the origin of eukaryotic intracellular motility systems from anaerobic but aerotolerant spirochetes in sulfide-rich environments. The last eukaryotic common ancestors (LECAs) have extant archaeprotist descendants: motile nucleated cells with Embden-Meyerhof glycolysis and substrate-level phosphorylation that lack the alpha-proteobacterial symbiont that became the mitochondrion. Swimming and regulated O(2)-tolerance via sulfide oxidation already had been acquired by sulfidogenic wall-less archaebacteria (thermoplasmas) after aerotolerant cytoplasmic-tubule-containing spirochetes (eubacteria) attached to them. Increasing stability of sulfide-oxidizing/sulfur-reducing consortia analogous to extant sulfur syntrophies (Thiodendron) led to fusion. The eubacteria-archaebacteria symbiosis became permanent as the nucleus evolved by prokaryotic recombination with membrane hypertrophy, analogous to Gemmata obscuriglobus and other delta-proteobacteria with membrane-bounded nucleoids. Histone-coated DNA, protein-synthetic RNAs, amino-acylating, and other enzymes were contributed by the sulfidogen whereas most intracellular motility derives from the spirochete. From this redox syntrophy in anoxic and microoxic Proterozoic habitats LECA evolved. The nucleus originated by recombination of eu- and archaebacterial DNA that remained attached to eubacterial motility structures and became the microtubular cytoskeleton, including the mitotic apparatus. Direct LECA descendants include free-living archaeprotists in anoxic environments: archamoebae, metamonads, parabasalids, and some mammalian symbionts with mitosomes. LECA later acquired the fully aerobic Krebs cycle-oxidative phosphorylation-mitochondrial metabolism by integration of the protomitochondrion, a third alpha-proteobacterial symbiont from which the ancestors to most protoctists, all fungi, plants, and animals evolved. Secondarily anaerobic eukaryotes descended from LECA after integration of this oxygen-respiring eubacterium. Explanatory power and experimental predictions for molecular biology of the LECA concept are stated.
Phenomena previously unknown in free-living spirochetes are reported: large-sized cells with variable diameter (length to 100 microns, width between 0.4 and 3.0 microns), composite structure (smaller spirochetes inside larger ones), and positive phototropic behavior. These bacteria, Spirosymplokos, are compared with all other spirochete genera. The large spirochete, grown in mixed culture, was studied live and by transmission EM. The protoplasmic cylinder was replete with spherical granules 20-32 nm in diameter, and three to six periplasmic 26-nm flagella were inserted subterminally. Comparably granulated and flagellated small spirochetes were located inside the protoplasmic cylinder and in the periplasm of the large ones. When exposed to air, movement became erratic, protoplasmic cylinders retracted to lie folded inside the outer membrane, and refractile membranous structures formed. From one to four structures per still-moving spirochete were seen. Spirosymplokos was enriched from laboratory samples exposed to oxygen-rich and desiccating, but not dry, conditions for at least 4 mo after removal of microbial mat from the field.
BBK32 is a fibronectin-binding protein from the Lyme disease-causing spirochete Borrelia burgdorferi. In this study, we show that BBK32 shares sequence similarity with fibronectin module-binding motifs previously identified in proteins from Streptococcus pyogenes and Staphylococcus aureus. Nuclear magnetic resonance spectroscopy and isothermal titration calorimetry are used to confirm the binding sites of BBK32 peptides within the N-terminal domain of fibronectin and to measure the affinities of the interactions. Comparison of chemical shift perturbations in fibronectin F1 modules on binding of peptides from BBK32, FnBPA from S. aureus, and SfbI from S. pyogenes provides further evidence for a shared mechanism of binding. Despite the different locations of the bacterial attachment sites in BBK32 compared with SfbI from S. pyogenes and FnBPA from S. aureus, an antiparallel orientation is observed for binding of the N-terminal domain of fibronectin to each of the pathogens. Thus, these phylogenetically and morphologically distinct bacterial pathogens have similar mechanisms for binding to human fibronectin.
The present study was designed to evaluate the effect of increased viscosity of the intestinal digesta on proliferation of enterotoxigenic Escherichia coli and the intestinal spirochaete Brachyspira pilosicoli in weaned pigs. Pigs were fed an experimental diet based on cooked white rice (R), which was supplemented with carboxymethylcellulose (CMC; 40 g/kg diet) to increase digesta viscosity. Thirty-six piglets weaned at 21 d of age were divided into six groups, three of which were fed R and three Addition of CMC increased digesta viscosity in the ileum (P=0.01), caecum (P=0.0007) and colon (P=0.0035), without increasing indices of large intestinal fermentation. Pigs fed developed a natural infection with enterotoxigenic E. coli after weaning and had more (P<0.0001) diarrhoea than pigs fed R. Subsequent experimental infection of two groups of pigs with B. pilosicoli resulted in more (P<0.0001) colonisation in pigs fed than R. At this time, all pigs fed had wetter (P<0.0001) faeces than those fed R, irrespective of whether they were infected with B. pilosicoli, but infected pigs also had an increased (P=0.025) number of days with diarrhoea post-infection irrespective of diet. In pigs fed it was not clear to what extent the increased viscosity associated with CMC, or the concurrent infection with enterotoxigenic E. coli, was responsible for the increased proliferation of B. pilosicoli. In a second experiment, five pigs that were weaned onto an R diet were transferred onto 3 weeks later. These pigs did not develop a natural infection with enterotoxigenic E. coli after the diet change, confirming the particular susceptibility of pigs to enterotoxigenic E. coli proliferation immediately post-weaning.
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We here report on three patients with gastrointestinal symptoms in whom spirochetes were found in colonic biopsies. The patients, heterosexual adults, were not immunocompromised. Electron microscopy was performed on colonic biopsies from each of the three patients. Apart from the basophilic band consisting of spirochetes, the mucosa was normal in two patients on light microscopy and showed mild inflammation in the other one. However on electron microscopy there was invasion of the colonic epithelial cells, macrophages, goblet cells and Schwann cells by spirochetes, and stunting of the microvilli. The spirochetes conformed to the morphology of Brachyspira aalborgi, and no other infective etiology or pathology could be identified in these patients to account for their symptoms. Since the clinical significance of intestinal spirochetosis is uncertain, antibiotics were not administered to any of the three patients and all three improved symptomatically with non-specific treatment. Intestinal spirochetosis, previously thought to be non-invasive and non-pathogenic in humans, may be invasive and may be the cause of gastrointestinal symptoms in some patients.
Three SPF-laying hens were inoculated into the crop with avian intestinal spirochaetes which previously had been passaged in broiler chicks by oral inoculation (isolate 1380). Mild persisting gastrointestinal disorder developed; at nine months post inoculation spirochaetes were readily demonstrated in caecal faeces. Histologic examination of the caecal mucosa revealed many spirochaetes covering the mucosal surface and filling up the crypts lumina. Spirochaetes were found in intra- and subepithelial locations and in gaps running through the epithelium. These gaps often opened into subepithelial cavities crowded with spirochaetes ('gaplike lesions'). These lesions were seen mostly on the tips of the villi and in the deeper parts of the crypts. Massive erosion or desquamation of epithelium heavily infested by spirochaetes occurred. These findings indicate colonisation of the mucosal surface and of the crypts, penetration of the mucosa and colonisation of subepithelial compartments with spirochasetes in poultry suffering from intestinal spirochaetosis.
The pathogenic potential of the anaerobic intestinal spirochaetes Brachyspira (Serpulina) pilosicoli and Brachyspira innocens was evaluated in adult chickens. Thirty 17-week-old Cobb broiler breeder hens were individually caged in three groups of 10 birds. Control birds (group A) were sham inoculated with sterile broth medium. Birds in the other two groups (groups B and C) were inoculated, respectively, with an isolate of B. innocens or of B. pilosicoli. Birds were monitored daily, and killed at 41 weeks of age. Infection had no consistent effect on body weight gain, but inoculation with B. pilosicoli resulted in a transient increase in faecal water content. B. innocens infection had no effect on egg production, but B. pilosicoli infection caused a delayed onset of laying, and a highly significant reduction in egg production over the first 11 weeks of lay. This study confirms that B. pilosicoli can cause serious egg production losses in adult chickens, while B. innocens is not obviously pathogenic.
This study aimed to evaluate the efficacy of some commonly used disinfectants in inactivating the pathogenic avian intestinal spirochaetes Brachyspira intermedia and Brachyspira pilosicoli, and to examine spirochaete survival in chicken caecal faeces held at 4 degrees C, 25 degrees C or 37 degrees C. Six disinfectants were evaluated at their recommended working concentrations: alkaline salts, quaternary ammonium, iodine as an iodophor, chlorine from a chlorine-release agent, glutaraldehyde and hydrogen peroxide. All but alkaline salts inactivated two different concentrations of both spirochaete species in less than 1 min in the presence of organic matter. Both spirochaete species at three different cell concentrations survived in caecal faeces at 37 degrees C for between 2 and 17 h. B. intermedia tended to survive for longer than B. pilosicoli, but the maximum survival time for both species at 4 degrees C was only 72 to 84 h. Hence, avian intestinal spirochaetes are rapidly inactivated by several common disinfectants, and their survival time in chicken caecal faeces is much less than has been reported for porcine intestinal spirochaetes in porcine faeces. It should be relatively easy to break the cycle of infection between batches of laying birds by resting sheds for a few days, and by using disinfectants on any residual faecal matter.
This study investigated whether feeding different wheat varieties to laying hens could influence colonization with the intestinal spirochaete Brachyspira intermedia. Fifty ISA-Brown laying hens were divided into two groups. One group were fed a laying-hen diet formulated with wheat variety Westonia, and one were fed the diet incorporating variety Stilleto. Each group was divided into 15 hens experimentally infected with B. intermedia and 10 uninfected controls. The 30 infected hens were housed in individual cages in one room, and the controls were similarly housed in another room. Following administration of cultures of B. intermedia strain HB60 by crop-tube over 3 days, cloacal swabs were taken for spirochaete culture every 3 to 4 days. The water content of caecal faeces, and egg production and body weight were measured weekly. The hens were killed after 4 weeks, the caeca cultured for spirochaetes and the viscosity of the ileal contents measured. A total of 48/120 (40%) of the excreta samples from infected hens fed Westonia contained B. intermedia, compared with 21/120 (17.5%) for Stiletto (P = 0.0002). The ileal viscosity of hens fed Westonia also was higher (P = 0.048), but viscosity was not clearly related to the non-starch polysaccharide (NSP) content of the wheats. Westonia had a slightly higher total NSP content than Stiletto, but the ratio of soluble to insoluble NSP was lower. Infected hens developed wetter excreta, but neither infection nor diet altered egg production. In conclusion, the wheat variety can influence colonization with B. intermedia, apparently through diet-related alterations in the intestinal microenvironment.
There has been much confusion over the significance of spirochaetes found in the caeca of laying hens and the impact they may have on egg production. In recent years, the situation has been made clearer and the presence of such species as Brachyspira pilosicoli have been shown to cause a mild, chronic disease in both layers and breeders and to reduce egg production by reportedly 5%. In the United Kingdom, a multi-age caged laying site with three separate flocks of approximately 12 000 birds each was chronically infected with B. pilosicoli but displayed few clinical signs except for a noticeable reduction in egg production and an increased mortality. The flocks were treated for 3 days in the drinking water with tiamulin at 12.5 mg/kg bodyweight, and a steady improvement in performance was recorded. The production results were compared with a flock that had been untreated with tiamulin previously, as a control, and one that had been treated at 25 and 45 weeks of age. A 9.8% improvement in egg production/hen housed up to 72 weeks of age and 9.7% in total egg weight was recorded, as well as an 8.6% reduction in actual hen mortality, in the tiamulin-treated flock in comparison with the untreated control. After taking into account the difference in breeds used, there was only a 6% reduction in egg production but an 8.84% increase in mortality in the untreated flock compared with the individual breed's standard production data. The cost of the disease was estimated at 14 million pound in the United Kingdom, based on a national laying flock of 30 million or 1.5% of production. Faecal examination for potentially pathogenic spirochaetes should be part of the differential diagnosis of under-performing laying flocks.
Recently, a number of diseases that had been thought previously to be caused by something other than an infectious agent are now known to be caused by bacteria. It now appears that it is not uncommon that bacteria, viruses, or fungi can cause diseases even when these organisms have not been detected or cultured. The most recent, well-publicized case is that of stomach ulcers; these are largely due to Helicobacter pylori infections. Here, the possibility is explored that amylotrophic lateral sclerosis (ALS) is caused by a cell wall-deficient microorganism.
The amount of penicillin which suffices to abort pneumococcal and streptococcal infections in white mice and rabbits, as well as syphilitic infection in rabbits, has been shown to increase markedly with the number of organisms inoculated. The curative dose increases also with the age of the infection, presumably owing to the interim increase in the number of organisms in the nfected host. The necessity for the larger doses of penicillin is attributed to the longer itime for which effectively bactericidal concentrations of penicillin must then be provided in order to kill the larger number of organisms.
A series of observations with the phase contrast microscope on the occurrence of a complex life cycle in the pathogenic Treponema pallidum as it occurs in the syphilitic rabbit testis has been presented and it seems likely from these observations that there are two means of vegetative reproduction, consisting of (1) transverse division (the most important under usual conditions); and (2) the production of gemmae or buds which eventuate into unispirochetal cysts comparable to those described for saprophytic forms, within each of which single spirochetes develop and differentiate, and from which they subsequently emerge. In addition preliminary evidence is presented which suggests that a more complex process is involved in which multispirochetal cysts develop following aggregation of two or more organisms. Within each of these larger cysts numerous organisms develop and subsequently emerge as tangled ropes. Following emergence, they subsequently undergo transverse division and gemmae formation, and so reproduce vegetatively. Subsequent papers will elaborate upon these processes.
Further evidence for the occurrence of a complex life cycle in the pathogenic Treponema pallidum as visualized by means of stained preparations is presented.