Spiral-shaped Organisms Research Articles

Batch-Mask: Automated Image Segmentation for Organisms with Limbless or Non-Standard Body Forms.

Efficient comparisons of biological color patterns are critical for understanding the mechanisms by which organisms evolve in nature, including sexual selection, predator-prey interactions, and thermoregulation. However, limbless, elongate, or spiral-shaped organisms do not conform to the standard orientation and photographic techniques required for many automated analyses. Currently, large-scale color analysis of elongate animals requires time-consuming manual landmarking, which reduces their representation in coloration research despite their ecological importance. We present Batch-Mask: an automated, customizable workflow to automatically analyze large photographic datasets to isolate non-standard biological organisms from the background. Batch-Maskis completely open-source and does not depend on any proprietary software. We also present a user guide for fine-tuning weights to a custom dataset and incorporating existing manual visual analysis tools (e.g., micaToolbox) into a single automated workflow for comparing color patterns across images. Batch-Maskwas 60x faster than manual landmarking and produced masks that correctly identified 96% of all snake pixels. To validate our approach, we used micaToolbox to compare pattern energy in a sample set of snake photographs segmented by Batch-Maskand humans and found no significant difference in the output results. The fine-tuned weights, user guide, and automated workflow substantially decrease the amount of time and attention required to quantitatively analyze non-standard biological subjects. With these tools, biologists can compare color, pattern, and shape differences in large datasets that include significant morphological variation in elongate body forms. This advance is especially valuable for comparative analyses of natural history collections across a broad range of morphologies. Through landmark-free automation, Batch-Maskcan greatly expand the scale of space, time, or taxonomic breadth across which color variation can be quantitatively examined.

Helicobacter spp. in the Stomach of Cats: Successful Colonization and Absence of Relevant Histopathological Alterations Reveals High Adaptation to the Host Gastric Niche.

In addition to Helicobacter pylori, many non-Helicobacter pylori Helicobacters (NHPH) are able to cause gastric disease in humans. Cats are a natural reservoir for many of these species. Accordingly, living in close and intimate contact with animals has been identified as a risk factor, and an important zoonotic significance has therefore been attributed to NHPH. To determine the prevalence and associated gastric histopathological changes of Helicobacter species, the gastric mucosa of 71 cats were evaluated. Only four presented normal histopathological mucosa with the absence of spiral-shaped organisms. Normal gastric mucosa and the presence of spiral-shaped bacteria were observed in 13 cats. The remaining animals presented histopathological changes representative of gastritis. Helicobacter species were detected in 53 cats (74.6%) by at least one detection method. None of the animals were positive for H. pylori or for H. ailurogastricus. Helicobacter heilmannii organisms were identified in 20 animals, predominantly in the body gastric region. Helicobacter salomonis was the second most prevalent species (57.1%), although it was mainly found in association with other NHPH. Helicobacter felis and H. bizzozeronii were less frequently detected. The great majority of the Helicobacter spp. PCR-positive animals presented normal features regarding fibrosis/mucosal atrophy, neutrophils, eosinophils, or other inflammatory cells and lymphofollicular hyperplasia. Given the controversy and the strong evidence of absence of significant histopathological alterations associated with the presence of Helicobacter spp. in cats, it is possible to hypothesize that these bacteria may be able to adapt to the feline gastric microenvironment or even to comprise part of the gastric microbiome of this animal species. Thus, prudency must be taken when prescribing an antibiotic therapy based solely on the presence of these bacteria in the feline stomach.

Mutations in the Borrelia burgdorferi Flagellar Type III Secretion System Genes fliH and fliI Profoundly Affect Spirochete Flagellar Assembly, Morphology, Motility, Structure, and Cell Division.

ABSTRACTThe Lyme disease spirochete Borrelia burgdorferi migrates to distant sites in the tick vectors and mammalian hosts through robust motility and chemotaxis activities. FliH and FliI are two cytoplasmic proteins that play important roles in the type III secretion system (T3SS)-mediated export and assembly of flagellar structural proteins. However, detailed analyses of the roles of FliH and FliI in B. burgdorferi have not been reported. In this study, fliH and fliI transposon mutants were utilized to dissect the mechanism of the Borrelia type III secretion system. The fliH and fliI mutants exhibited rod-shaped or string-like morphology, greatly reduced motility, division defects (resulting in elongated organisms with incomplete division points), and noninfectivity in mice by needle inoculation. Mutants in fliH and fliI were incapable of translational motion in 1% methylcellulose or soft agar. Inactivation of either fliH or fliI resulted in the loss of the FliH-FliI complex from otherwise intact flagellar motors, as determined by cryo-electron tomography (cryo-ET). Flagellar assemblies were still present in the mutant cells, albeit in lower numbers than in wild-type cells and with truncated flagella. Genetic complementation of fliH and fliI mutants in trans restored their wild-type morphology, motility, and flagellar motor structure; however, full-length flagella and infectivity were not recovered in these complemented mutants. Based on these results, disruption of either fliH or fliI in B. burgdorferi results in a severe defect in flagellar structure and function and cell division but does not completely block the export and assembly of flagellar hook and filament proteins.

Presence and significance of Helicobacter spp. in the gastric mucosa of Portuguese dogs.

BackgroundNon-Helicobacter pylori Helicobacters (NHPH) are also able to cause disease in humans. Dogs are a natural reservoir for many of these species. Close and intense human contact with animals has been identified as a risk factor and therefore, an important zoonotic significance has been attributed to NHPH.MethodsTo determine the prevalence of Helicobacter species and the gastric histopathological changes associated, gastric mucosa samples of 69 dogs were evaluated.ResultsOnly one dog presented a normal histopathological mucosa with absence of spiral-shaped organisms. A normal gastric mucosa and the presence of spiral-shaped bacteria was observed in two dogs. All remaining animals presented histopathological changes representative of gastritis. Helicobacter species were detected in 60 dogs (87.0%) by at least one detection method. Histological, histochemical and immunohistochemical evaluations revealed that Helicobacter spp. were present in 45 (65.2%), 52 (75.4%) and 57 (82.6%) dogs, respectively. Spiral-shaped bacteria were detected by qPCR analysis in 33 (47.8%) dogs. H. heilmannii-like organisms were identified in 22 animals (66.7%) and predominantly in the antral gastric region. H. salomonis was the second most prevalent species (51.5%) although it was mainly found in association with other Helicobacter spp. and in the body gastric region. H. bizzozeronii and H. felis were less frequently detected.ConclusionsIt was concluded that, despite the high incidence and worldwide distribution of gastric NHPH in dogs, the presence of specific Helicobacter species may vary between geographic regions. NHPH infections were significantly accompanied by mild to moderate intraepithelial lymphocyte infiltration and mild to moderate gastric epithelial injury, but a clear relationship between gastritis and Helicobacter infection could not be established.

Species of the Family Helicobacteraceae Detected in an Australian Sea Lion ( Neophoca cinerea ) with Chronic Gastritis

We describe the first case of gastritis in a male Australian sea lion (Neophoca cinerea) in which members of the family Helicobacteraceae, particularly the genus Wolinella, were detected. The sea lion exhibited clinical signs of gastrointestinal disease, including abdominal pain, lack of appetite, and lethargy. Examination of one ileal and five gastric biopsy specimens collected over a 10-year period revealed persistent fibrosis and/or superficial focal erosion and ulceration of the lamina propria. Spiral-shaped organisms 5 to 12 microm long were observed in two of the gut biopsy specimens. While Helicobacter species were detected by PCR in one of the gastric biopsy specimens, Wolinella species were detected in four of the five gastric specimens, including those in which spiral-shaped organisms were observed. Comparisons of biopsy specimen ribosomal DNA sequences with those obtained from the feces of this animal, the gastric tissue of a clinically healthy individual, and the feces of several other cohoused sea lions and fur seals revealed a separate and possibly novel gastric Helicobacter species. A possibly novel Wolinella species, along with Wolinella succinogenes, was also identified. These findings highlight the pathogenic potential of other members of this family in the etiopathogenesis of gastric disease in these animals.

Helicobacter heilmannii gastroduodenal disease and clinical aspects in children with dyspeptic symptoms

To evaluate the occurrence and clinical characteristics of Helicobacter heilmannii infection among children presenting with dyspeptic symptoms. Prospective cohort study of 580 patients. Of all examined dyspeptic children, 26.4% were infected with spiral-shaped organisms, and 0.9% of patients were found to be infected with spiral H. heilmannii-like organisms. In children with dyspeptic symptoms, the possible presence of gastroduodenal disease due to H. heilmannii should be considered. Further studies are needed to clarify H. heilmannii-related gastroduodenal pathology in the paediatric population.

Treatment of Helicobacter pylori gastritis improves dyspeptic symptoms in children.

In adults, the treatment of Helicobacter pylori infection is only recommended for patients with active gastric or duodenal ulcers. It is not known whether similar guidelines can be applied to children because the prevalence of peptic ulcer disease in childhood is estimated to be much lower than in adults. The purpose of this study was to determine whether treatment of H. pylori gastritis would improve symptoms of dyspepsia in children. Sixteen patients (5 boys, 11 girls) aged 14 +/- 1.2 years who had symptoms of dyspepsia were evaluated using upper gastrointestinal endoscopy with biopsies to establish the diagnosis of H. pylori gastritis. They were treated for 2 weeks with clarithromycin, amoxicillin, and a proton pump inhibitor. Dyspepsia symptoms were evaluated by a questionnaire before and after treatment of the infection. The effect of H. pylori treatment on the total symptom score was analyzed with use of the Student t test. Values are presented as mean +/- SEM. All patients had antral nodularity and chronic active gastritis with spiral-shaped organisms but no evidence of peptic ulcer disease. Mean total symptom score decreased significantly at 2 to 4 weeks after treatment (12.6 +/- 0.9 vs. 2.1 +/- 0.5 P < 0.001), and it remained low (2.9 +/- 0.7) at follow-up 9.7 +/- 1.4 months (range, 2-24 months later). These results suggest that the treatment of H. pylori gastritis can improve dyspeptic symptoms in children.

Helicobacter pylori in endoscopy patients in Zimbabwe: value of enzyme-linked immunosorbent assay and a rapid urease test

Biopsy and serum specimens were obtained from 95 patients undergoing endoscopy at the University of Zimbabwe Medical School. Common presenting features were epigastric pain, bleeding and dyspepsia. Ulcers were detected in 16 patients (17%), and were more common in men (24%) than in women (7%). Histological examination of biopsies showed that all 95 patients had spiral-shaped organisms that were indistinguishable microscopically from Helicobacter pylori, though the numbers of organisms varied considerably. There was evidence that the degree of inflammation in the mucosa was related to the numbers of H. pylori-like organisms (HPLO) present. Fifty-one biopsy specimens (55%) gave a positive rapid urease test (RUT), with colour change occurring within 4 h. In all but one case, the gastric mucosa from these patients contained moderate to numerous HPLO. We defined the ‘gold standard’ of H. pyori-associated gastritis as the presence of both moderate to numerous HPLO and moderate to severe inflammation in the gastric mucosa. Using these criteria, RUT had a sensitivity of 67% and a specificity of 68%. Sera from 92 patients were tested for immunoglobulin G antibodies reactive with a glycine-extract antigen of H. pylori, using an enzyme-linked immunosorbent assay ( elisa). Sera giving an indeterminate reaction in the elisa were also tested by Western blotting. In all, 36 sera (39%) gave a positive elisa or Western blot reaction. There was poor correlation between serology and RUT results, with only 57% of biopsy specimens from seropositive patients giving a positive RUT, compared with 45% from seronegative patients. Positive serology was found in only 35 patients (61%) with histological evidence of H. pylori-associated gastritis, and the specificity of the test was only 54%. When used in combination with the RUT result, however, 79% of patients with a positive RUT and positive serology had histological evidence of H. pylori-associated gastritis. There was a general trend for increased seropreyalence in patients with mild to moderate atypia. These findings indicate that serology, using an antigen derived from the type strain of H. pylori, is unreliable in detecting H. pylori infection in Zimbabwe. Current studies are aimed at characterizing antigens from organisms isolated from Zimbabwean patients.

Acute gastritis associated with spiral organisms from cats

Numerous studies implicated Helicobacter pylori as one causative agent producing gastritis and dyspepsia. Recent reports focus on another bacterium, Gastrospirillum hominis, as a possible pathogen producing gastritis. We report a 30-year-old researcher who became acutely ill with epigastric pain indicative of esophagitis or peptic ulcer disease. Gastritis and a gastric ulcer were observed endoscopically. Histological examination of the gastric mucosa revealed an acute gastritis and large spiral-shaped organisms. The spiral forms were present in large quantities in the gastric mucosa of experimental animals (cats) handled by the patient in his research. Electron microscopy confirmed that the organisms from the cat and patient were morphologically identical. The patient was successfully treated with bismuth subsalicylate. His symptoms resolved and the organisms were cleared from his stomach. This study provides evidence that another bacterium, a Gastrospirillum, may cause gastritis in man and may be transmitted from animal to man.

Chronic gastroenterocolitis in nine cats.

Lesions of a chronic gastroenterocolitis were found in 9 cats of the Persian breed that were euthanized after a prolonged period of bloody diarrhea. Gross lesions consisted of gastrointestinal edema with prominent Payer's patches, multiple grayish nodules, and a few irregular erosions within the colonic mucosa. Microscopically, the changes were composed of degeneration, necrosis, and proliferation of gastric glandular epithelium, dilated intestinal crypts with lymphoplasmacytic cell infiltration of the lamina propria, and in 1 cat, severe transmural necrosis of the colon. With the Giemsa stain, spiral-shaped organisms in the gastrointestinal lumen and intracellularly in the gastric and the colonic epithelium were observed. These organisms could not be cultured. Although the role of these spiral-shaped organisms was not determined, other agents that could explain the disorder were not found.

Bacterial Populations Associated with Different Regions of the Human Colon Wall

The microorganisms associated with the undiseased human colon wall were examined in material obtained from four sudden-death victims. In traffic accident subjects (aged 45 and 16 years) the anaerobe-aerobe ratio was about 10(4):1 in all areas of the colon examined, whereas in acute heart failure subjects (aged 74 and 46 years) the ratio was as low as 1.2:1. The flora of each individual was distinct and complex. Although the predominant anaerobes isolated were Bacteroides and Fusobacterium spp., which composed over 50% of the flora in some samples, the species isolated (indicated by morphology and glucose fermentation products) varied between individuals. Other major types observed were gram-positive nonsporing rods, including Bifidobacterium spp., and anaerobic cocci (between 8 and 20% of isolates). Clostridia were only isolated in significant numbers from one individual. Scanning electron microscopy showed that most of the organisms were present below the top surface of the mucin layer overlying the mucosa. The use of several different preparative procedures for microscopy showed a complex microbial structure within the mucus, but major variations in the bacterial populations in different areas of the colon were not found. Spiral-shaped organisms up to 60 mum long in the form of double helices were found in two subjects by scanning electron microscopy but were not isolated during the parallel bacteriological investigation. The differences between this and previous studies are discussed in relation to experimental procedures and also in contrast to results with animals that showed a particularly specialized flora associated with the colon wall.

Infection of the Colon of the Rhesus Monkey by Spiral-Shaped Organisms

Intestinal spirochetosis, an infection of the mucosa by spiral-shaped organisms, was studied in clinically normal rhesus monkeys (Macaca mulatta) by histology, transmission and scanning electron microscopy. The incidence of intestinal spirochetosis was 42% in 221 monkeys. Spiral organisms stained with hematoxylin and eosin (HE) appeared as a broad basophilic haze on the colonic surface and were strongly positive by the Warthin-Starry stain. Spiral-shaped bacteria include two structurally different organisms: spirochetes and flagellated microbes. They intimately populated the brush border of the surface of the epithelium of the large intestine. They were absent in the crypts and in the small intestine. Infection by spirochetes produced no alteration of cytocomponents of the underlying host structures. Spirochetes and flagellates infrequently penetrated beyond the brush border into the epithelial cytoplasm and also into the lamina propria. Even in cases where invasion was documented, no inflammatory response was found.

Intestinal Microflora in the Guinea Pig as Observed by Scanning Electron Microscopy. Effect of Fibrous Dietary Supplements

The gastrointestinal ecosystems of weanling guinea pigs fed a purified diet with and without supplements of cabbage, alfalfa and holocellulose were examined by scanning electron microscopy. The diets, which were based on soybean protein, sucrose and wood cellulose, were fed for 28 days and the mucosa and associated microbes were fixed in situ. Bacteria, rods and cocci being the predominant morphological types, were commonly observed on the epithelial surface of the jejunum and ileum and much less frequently in the duodenum. In the cecum, fusiform- and spiral-shaped organisms were the most common types. The fusiforms, which produced unique indentations in the intestinal mucosa, were found in the intestines of the cabbage and alfalfa-supplemented guinea pigs but not in those fed the basal diet. There was considerably less mucus on the intestinal epithelium when the cabbage supplement was supplied. Whether promotion of the fusiform population or elimination of the mucus secretion are causally related to the growth stimulating properties of the supplements is not clear, but the results suggest a relationship.

Purulent Rectal Discharge Associated With a Nontreponemal Spirochete

A nontreponemal spirochete was isolated from the rectum of a homosexual man with a chronic purulent discharge. Known infectious causes of the disease were excluded. Although the pathogenicity of the organism was not proved, the patient's symptoms rapidly resolved following penicillin G benzathine therapy. When culture of a nonhemorrhagic, purulent rectal discharge fails to verify Neisseria gonorrhoeae, health care personnel are encouraged to carefully examine a Gram's-stained smear of the discharge for spiral-shaped organisms.

Erratum: Microbial degradation of [14C]polystyrene and 1,3-diphenylbutane

guinea pig specimens were made. All of the isolates were identified as Torlllol~sis pit7tolope~ii. The isolation of T. pirltolopesii from the intestinal tract of guinea pigs has been reported previously (Parle 1957). Tor~rlopsis pir7tolopesii is known to associate with tissue surfaces in the stomach of mice and rats (Savage 1969), but we do not believe that such an association has been previously reported for guinea pigs. Microbial associations with epithelial sulfaces in the alimentary canal of mice and rats (Savage 1969), man and monkeys (Takeuchi et 01. 1974), chickens (Fuller and Turvey 197 l) , swine (Tannock and Smith 1970), nematodes (Tannock and Savage 1974), insects (Breznak and Pankratz 1977), and sheep (Bauchop et 01. 1975) have been described. We have demonstrated that microbial associations with tissue surfaces also occur in the alimentary tract of guinea pigs. ,P~\)ipe.s (Kol1:ir) and Coptot~rr~re.s,forttio.~c~~~~~.s Shilxki]. Appl. Environ. Microbiol. 33: 406-426. DAVIS. C. P., and D. C. SAVAGE. 1974. Habitat. succession, attachment and morphology of segmented, filamentous microbes indigenous to the murine gastrointestinal tract. Infect. Immun. 10: 948-956. FULLER. R.. and A . TLJRVEY. 1971. Bacteria associated with the intestinal wall of the fowl. (Gallris rlott~esticrrs). J . Appl. Bacteriol. 34: 617-622. LODDER, J . . and N. J. W. KREGER-VAN RIJ. 1970. The Yeasts. North Holland Publishing Company, Amsterdam. pp. 1235-1292. PARLE, J. N. 1957. Yeasts isolated from the mammalian ;dimentary tract. J. Gen. Microbiol. 17: 363-367. SAVAGE. D. C. 1969. Microbial interference between indigenous yeast and lactobacilli in the rodent stomach. J . Bacteriol. 98: 12781283. SAVAGE, D. C.. J . S. MCALLISTER, and C. P. DAVIS. 1971. Anaerobic bacteria on the mucosal epithelium of the murine large bowel. Infect. Immun. 4: 492-502. TAKEUCHI. A,, H. R. JERVIS, H. NAKAZAWA, and D. M. ROBINSON. 1974. Spiral-shaped organisms on the surface colonicepitheliumofthe monkey and man. Am. J. Clin. Nutr. 27: 12871296. ~ ~

Spiral-shaped organisms on the surface colonic epithelium of the monkey and man

Spiral-shaped organisms on the surface colonic epithelium of the monkey and man

Microbial colonization of the intestinal epithelium in suckling mice.

Colonization by indigenous microorganisms of the mucosal epithelia of the large bowels of suckling mice was followed by microbial culture techniques and by light, fluorescence, and electron microscopy. Certain microbes colonize in distinctive patterns the cecal and colonic epithelia in these mice. Coliforms and enterococci colonize the large bowel 7 to 9 days after birth and reach high population levels during the second week. During that period, these facultative anaerobes can be detected by immunofluorescence techniques in microcolonies in the mucin on the epithelium. During the third week, however, after their populations decline to the low levels characteristic of adult mice, coliforms and enteroccoci can be observed only infrequently in the mucous layer. Anaerobic fusiform-shaped bacteria appear in the mucous layers along with the microcolonies of enterococci and coliforms during the second week after birth. These anaerobes increase in numbers in the mucin until they form thick layers on the mucosal epithelium by the end of the third week. They remain in the mucous layer throughout the life of the normal mouse. Anaerobic spiral-shaped microbes also colonize the mucous layer on the cecal and colonic epithelium. But these organisms can be detected by immunofluorescence in 1-week-old mice, well in advance of the time the fusiform-shaped bacteria can be found. In the second week, the latter microbes co-inhabit the mucous layer with the spiral-shaped organisms. The fusiform- and spiral-shaped microbes remain associated in the mucin on the cecal and colonic mucosal epithelia into the adult life of mice.

Ultrastructural identification of spirochetes and flagellated microbes at the brush border of the large intestinal epithelium of the rhesus monkey.

Spiral-shaped organisms exclusively and intimately populate the brush border of the cecal and colonic epithelium of healthy monkeys (Macaca mulatta). These organisms replace the glycocalyx, destroy most microvilli, and attenuate the terminal web of the brush border. Despite these remarkable alterations, the remaining host cellular structure is unchanged. Two structurally distinct microbes, a spirochete and a flagellate, were recognized by electron microscopy. These spirochetes share the general characteristics of other known spirochetes: they are 3 to 6 mum long and 0.2 to 0.4 mum wide, spiral 2 to 6 times, and have axial fibrils of 6-12-6 and 4-8-4 arrangements. Flagellated microbes are 4 to 6 mum long and 0.2 to 0.4 mum wide, spiral 2 to 4 times, and are characterized by a polar flagellum which originates from the terminal button at each end of the cytoplasmic body. This in vivo ultrastructural study of anaerobic spiral-shaped organisms bypasses the difficulties of in vitro culture techniques and provides a detailed description and identification of these organisms in their host environment.

Anaerobic bacteria on the mucosal epithelium of the murine large bowel.

Anaerobic bacteria can be detected at population levels of 10(11) organisms per g of cecum or colon in adult mice from four different colonies widely spaced in the United States. Most of these microorganisms are oxygen-intolerant fusiform-shaped bacteria. At least one type of these tapered, rod-shaped bacteria can be seen in layers in the epithelial mucin in frozen-section histological preparations of the large bowels of mice. In addition, such microorganisms can be seen within 0.5 mum of the epithelium in ultrathin sections of colon or cecum examined in an electron microscope. These fusiform-shaped bacteria predominate in the mucin layers. However, spiral-shaped microorganisms can be found as well near the mucosal epithelia in ultrathin sections of colon. Also, such organisms can be seen in negatively-stained preparations of washings of the colonic mucosal epithelia examined in an electron microscope. At least three types of spiral-shaped organisms, including both spiral-shaped bacteria and spirochetes, can be found in preparations from mice from three of the four colonies. Such spiral-shaped microorganisms can be detected at population levels as great as 10(9) organisms per g of cecum or colon in anaerobic cultures of the large bowels of mice from all four colonies. One anaerobic spiral bacterium was isolated in pure culture. This particular organism was found by immunofluorescence to be intermingled with the fusiform-shaped bacteria in the mucin on the mucosal epithelium in the mouse large bowel.

VINCENT'S SPIRILLUM AND BACILLUS FUSIFORMIS IN PSEUDOMEMBRANOUS ANGINAS

Since Plaut, in 1894, described a spirillum and an associated spindle-shaped bacillus in cases of membranous tonsillitis, and Vincent, in 1896, described similar organisms in cases of hospital gangrene, a large number of observers have reported similar lesions in which the spirillum and the Bacillus fusiformis have been constantly associated. These lesions have been described as acute, or subacute, ulcerative or membranous inflammations in which were found the characteristic spirilla and bacilli. Morphologically, the fusiform bacilli have been described as spindle-shaped rods with tapering ends, slightly bulging in the center, usually straight, and from 6 to 12 microns in length. The spirilla, usually associated with the bacilli, are delicate spiral-shaped organisms, staining somewhat faintly, and varying considerably in length and in the number of twists or curves. Both organisms take the ordinary stains but are Gram-negative. They are always found associated