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Emerging from obscurity: biological, clinical, and diagnostic aspects of Dientamoeba fragilis.

Ever since its first description in 1918, Dientamoeba fragilis has struggled to gain recognition as a significant pathogen. There is little justification for this neglect, however, since there exists a growing body of case reports from numerous countries around the world that have linked this protozoal parasite to clinical manifestations such as diarrhea, abdominal pain, flatulence, and anorexia. A number of studies have even incriminated D. fragilis as a cause of irritable bowel syndrome, allergic colitis, and diarrhea in human immunodeficiency virus patients. Although D. fragilis is most commonly identified using permanently stained fecal smears, recent advances in culturing techniques are simplifying as well as improving the ability of investigators to detect this organism. However, there are limitations in the use of cultures since they cannot be performed on fecal samples that have been fixed. Significant progress has been made in the biological classification of this organism, which originally was described as an ameba. Analyses of small-subunit rRNA gene sequences have clearly demonstrated its close relationship to Histomonas, and it is now known to be a trichomonad. How the organism is transmitted remains a mystery, although there is some evidence that D. fragilis might be transmitted via the ova of the pinworm, Enterobius vermicularis. Also, it remains to be answered whether the two distinct genotypes of D. fragilis recently identified represent organisms with differing virulence.

Adult↗

Application of indirect immunofluorescence to detection of Dientamoeba fragilis trophozoites in fecal specimens.

An indirect fluorescent-antibody (IFA) assay was carried out to examine for the presence of Dientamoeba fragilis trophozoites in preserved fecal specimens. Antiserum to D. fragilis trophozoites was raised in a rabbit with a dixenic culture of D. fragilis (ATCC 30948) from the American Type Culture Collection. After absorption with Klebsiella pneumoniae and Bacteroides vulgatus, the immune rabbit serum was used for examination by the IFA assay. A total of 155 clinical samples were tested; 42 with no parasites, 9 with D. fragilis, and 104 with other parasites. The IFA assay identified seven D. fragilis organisms. Two specimens with doubtful IFA assay readings showed very scanty amounts of D. fragilis trophozoites on stained smears. There were no false-positive IFA assay readings. The IFA assay appeared to be a promising method because of its speed in screening. The specificity of the IFA assay indicates that other diagnostic tests such as an enzyme-linked immunosorbent assay could be developed to identify D. fragilis antigens in fecal specimens.

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Cryptic genetic diversity in Dientamoeba fragilis.

Uncertainty surrounding the role of Dientamoeba fragilis in human disease could be due in part to the existence of pathogenic and nonpathogenic variants. Evidence for two genetically distinct forms was obtained using PCR-restriction fragment length polymorphism analysis of ribosomal genes. Future studies in humans will need to take D. fragilis diversity into account.

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Direct amplification and genotyping of Dientamoeba fragilis from human stool specimens.

Dientamoeba fragilis is a globally occurring parasite that has been recognized as a causative agent of gastrointestinal symptoms. A single-round PCR was developed to detect D. fragilis DNA directly from human stool samples. The genetic diversity of D. fragilis from 93 patients and 6 asymptomatic carriers was examined by PCR followed by restriction fragment length polymorphism and sequencing of part of the small-subunit rRNA gene. The data show that D. fragilis sequences can be studied directly from fecal specimens despite the absence of a cyst stage and without the need for prior culturing. In addition, the results suggest strongly that D. fragilis shows remarkably little variation in its small-subunit rRNA gene.

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Prospective study of the prevalence, genotyping, and clinical relevance of Dientamoeba fragilis infections in an Australian population.

A prospective study was conducted over a 30-month period, in which fecal specimens from 6,750 patients were submitted to the Department of Microbiology at St. Vincent's Hospital, Sydney, Australia. Trophozoites of Dientamoeba fragilis were detected in 60 (0.9%) patients by permanent staining, and confirmation was performed by PCR. Gastrointestinal symptoms were present in all patients, with diarrhea and abdominal pain the most common symptoms. Thirty-two percent of patients presented with chronic symptoms. The average age of infected patients was 39.8 years. No correlation was found between D. fragilis and Enterobius vermicularis, a proposed vector of transmission for D. fragilis. The genetic diversity of 50 D. fragilis isolates was examined by PCR, and the PCR products were analyzed for the presence of restriction fragment length polymorphisms. These results showed no variation in the small-subunit rRNA gene and demonstrated a single genotype for all Australian isolates. This study shows the potential pathogenic properties of D. fragilis and the need for all laboratories to routinely test for this organism.

Abdominal Pain↗

Evaluation of three diagnostic methods, including real-time PCR, for detection of Dientamoeba fragilis in stool specimens.

Dientamoeba fragilis is a protozoan parasite of humans that infects the mucosa of the large intestine and is associated with gastrointestinal disease. We developed a 5' nuclease (TaqMan)-based real-time PCR assay, targeting the small subunit rRNA gene, for the detection of D. fragilis in human stool specimens and compared its sensitivity and specificity to conventional PCR and microscopic examination by a traditional modified iron-hematoxylin staining procedure. Real-time PCR exhibited 100% sensitivity and specificity.

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Dientamoeba fragilis: a review with notes on its epidemiology, pathogenicity, mode of transmission, and diagnosis.

Dientamoeba fragilis was found in 4.2% of approximately 43,000 individuals who submitted stools for parasitological examination during 1970 to 1974. The parasite was more frequently found in the younger age group (less than 20 years) than in older age groups, and more often in females than in males. Symptoms in 255 of patients in whom D. fragilis was the only parasite found and for whom detailed symptoms had been supplied, included: diarrhea, abdominal pains, anal pruritus, and loose stools. Analysis of mixed infections of D. fragilis with intestinal helminths suggests that such infections are random except for the combination of D. fragilis and Enterobius vermicularis. This combination occurred 9 times more often than theoretically expected. Daily periodicity and distribution of D. fragilis within stools of one patient were studied over a period of 6 months. More than twice as many organisms per ml of stool were present in the last than in the first portion evacuated. The total number of organisms excreted fluctuated markedly from day to day.

Adolescent↗

Incidence of Dientamoeba fragilis in faecal samples submitted for routine microbiological analysis.

Over a six-month period, 857 faecal samples were submitted to the Department of Microbiology and Immunology at Sultan Qaboos University Hospital in Oman, for routine microbiological examination. All samples were stained using the Gomori trichrome method. Trophozoites of Dientamoeba fragilis were detected in 41 (5.1%) patients, making it the most common enteropathogen found in the study. Of the patients with pure D. fragilis infection, 83% had abdominal pain, the duration of which varied from one month to two years. The use of permanently stained smears allowed detection of D. fragilis for the first time in the Sultanate of Oman.

Adolescent↗

Dientamoeba fragilis: the unflagellated human flagellate.

Dientamoeba fragilis is a pathogenic protozoan parasite with a world-wide distribution. Although originally described as an amoeboid organism, it has been reclassified as a flagellate, on the basis of a number of electron microscopic and immunological findings. Except for its lack of a flagellum, D. fragilis closely resembles Histomonas and Trichomonas. Interestingly, a resistant cyst stage has not been demonstrated and it is unlikely that its trophozoites can survive successfully outside the human host. As a consequence of its higher than anticipated coincidence of infection with Enterobius vermicularis, transmission may occur via ova of this pinworm. D. fragilis infection may be acute or chronic, and has been reported in both children and adults. The most common clinical symptoms include abdominal pain, persistent diarrhoea, loss of appetite, weight loss and flatulence. Occasionally, eosinophilia, urticaria and pruritus have been described. Demonstration of the characteristic nuclear structure of D. fragilis, needed for a definitive diagnosis, cannot be achieved in unstained faecal material; therefore, permanently stained smears are essential. Treatment is recommended in symptomatic cases, and iodoquinol, tetracycline and metronidazole have been used successfully.

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[Dientamoeba fragilis--an old parasite sparks new interest].

Dientamoeba fragilis is a protozoan parasite of the human colon with only the trophozoite stage identified. The exact mode of transmission is still unknown. Formerly considered to be apathogenic, several studies have suggested that D. fragilis has pathogenic properties. Common symptoms of the infection include diarrhea and abdominal pain. After reviewing the literature we suggest that treatment can be considered for patients with prolonged symptoms, provided no other causative agent has been found.

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[Dientamoeba fragilis and dientamoebiasis: aspects of clinical parasitology and laboratory diagnosis].

The flagellate Dientamoeba fragilis is a potential intestinal pathogen of humans. It is probably undervalued overall because its identification relies on permanent stains directly on fresh fecal specimens (Giemsa) or preserved in PVA or SAF (Trichrome or Iron-hematoxylin). During 1999 we processed stoll samples of 151 subjects and we searched also D. fragilis. We processed 105 cases of enteritis (82 children and 23 adults) and 46 samples of formed stools to investigate only for intestinal parasites (12 children and 34 adults). The microscopical observations were made on smears directly in saline solution, and after Dobell solution, MIF and Giemsa stained. Iron hematoxylin and trichrome stains were performed in some circumstances only. D. fragilis was observed in 11.3% of cases, 7.6% among the group of enteritis and 13.8% among the other subjects. These positive 17 cases were observed more often among adults (76.5% of cases) than among children (23.5%). In our study area D. fragilis seems more frequent than Giardia intestinalis and represents the third important intestinal parasite, following Enterobius vermicularis (children) and Strongyloides stercoralis (elders).

Adolescent↗

[Dientamoeba fragilis: possibly an important cause of persistent abdominal pain in children].

OBJECTIVE: To determine the effect of an antiprotozoic treatment on children with persistent abdominal pain and infection with Dientamoeba fragilis. DESIGN: Retrospective. METHOD: A total of 43 children with D. fragilis infection and persistent gastrointestinal complaints were included in the study. Of these 27 were treated with clioquinol and 16 with a nitroimidazole drug: metronidazole or tinidazole. The parasitological and clinical effects of the treatment were assessed. RESULTS: In 33 of the 43 (77%) children, no parasites were detected during follow-up with a triple faeces test: 22/27 following treatment with clioquinol and 11/16 following treatment with a nitroimidazole drug. In 27 of the 33 (82%) children with a negative follow-up result, gastrointestinal complaints were considerably less or had completely disappeared. In 2 of the 10 (20%) children in which D. fragilis had not disappeared in the follow-up period, the complaints were less or had disappeared. CONCLUSION: Effective treatment of D. fragilis infection in children with longstanding gastrointestinal complaints often resulted in a reduction or disappearance of the complaints.

Abdominal Pain↗

[Dientamoeba fragilis: is it really fragile? Approach to specimen handling and rapid microscopic diagnosis].

Dientamoeba fragilis is a pathogenic protozoan parasite with a world-wide distribution. Interestingly, a resistant cyst stage has not been demonstrated and it is still an unsolved problem how this parasite can survive successfully outside the human host. D. fragilis was found in 2% of approximately 2500 individuals unselected who submitted stools for parasitological examination during 2001 in Padua (Italy). The goal of this study was to detect the protozoan stages and the duration of persistence of this protozoa in faeces stored in different environmental conditions. The trophozoites of D. fragilis were detected up to 60 days after the collection of the faeces stored at 4 degrees C and Giemsa stained. The laboratory detection rate of the organism is greatly enhanced by use of preservative to fix stool specimens immediately after passage. Alternatively, a microscopic observation of the collected stool has to be performed immediately after passage followed by examination of permanently-stained smears. Demonstration of the charateristic "golf-club" and "acanthopodia-like" structures in unstained fixed faecal material by direct microscopy (400x) are suitable for a rapid identification of D. fragilis.

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Dientamoeba fragilis detection methods and prevalence: a survey of state public health laboratories.

Dientamoeba fragilis is a pathogenic protozoan parasite that has no cyst stage. Because of the lack of a cyst stage, the laboratory detection of D. fragilis in stool specimens is dependent on the stool processing and examination methods employed. Failure to use recommended stool fixation and permanent staining techniques almost precludes identification of D. fragilis, which is associated with gastrointestinal illness in humans. In this survey, questionnaires were mailed to all State and territorial public health laboratories requesting information on the number of ova and parasite examinations, methods of processing and examining stools, and the number of D. fragilis positive stools for 1985. Forty-three of 54 (80 percent) laboratories responded. Results showed that those laboratories which reported D. fragilis detection examined more stools using recommended stool fixation methods and were more likely to stain permanently all stools examined. Permanent staining of all stools, as compared to loose and watery stools only, resulted in a fivefold greater detection of D. fragilis. More State and territorial public health laboratories reported finding D. fragilis infections in 1985 than in a 1978 survey performed by the Centers for Disease Control. However, in 1985 only six laboratories reported 82 percent of all D. fragilis detections. To increase the probability of detecting D. fragilis in stool specimens, the findings suggest that all stools should be submitted fixed in polyvinyl alcohol fixative, sodium acetate-acetic acid-formalin fixative, or Schaudinn's fixative. Further, all specimens, regardless of consistency, should be permanently stained prior to microscopic examination.

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Dientamoeba fragilis: a bowel pathogen?

Although many patients infected with Dientamoeba fragilis suffer gastrointestinal symptoms, controversy still surrounds the pathogenic role of this intestinal protozoan. We describe three patients infected with D fragilis who had gastrointestinal symptoms. In the first patient symptoms resolved with therapy before the protozoan was eradicated from the stool, however in the second, symptoms persisted unabated despite clearing of the organism. In the third patient, symptoms resolution was associated with effective treatment. We discuss the life cycle, diagnosis and treatment of this organism and its role as a pathogen.

Adult↗

Colitis due to Dientamoeba fragilis.

Clinical reports have suggested that the parasite Dientamoeba fragilis may be a cause of acute and chronic colitis in children and adults. The mechanism by which this parasite produces colitis has not been determined. The clinical findings in this case report suggest that D. fragilis causes colitis through an invasive ulcerating process.

Adult↗

Dientamoeba fragilis: a gastrointestinal protozoan infection in adults.

Dientamoeba fragilis is a protozoan parasite of the large intestine of man. Individuals with infection may be asymptomatic or have gastrointestinal and systemic symptoms. We report a patient with symptomatic D. fragilis infection and negative extensive laboratory and radiological workup, with resolution of symptoms after diiodohydroxyquin therapy. No parasites were detected in three follow-up stool examinations. We then undertook retrospective study to define and describe further clinical symptoms in adults with this infection by analysis of data from medical records of 50 subjects with this parasite.

Abdomen↗

[3 emerging protozoal infections in The Netherlands: Cyclospora, Dientamoeba, and Microspora infections].

Increased international travelling, an increased number of patients with immunosuppression caused by HIV infection, and renewed interest in known but little studied microorganisms, resulted in a more frequent finding of certain medically important parasites. Three emerging protozoal infection, Cyclospora cayetanensis, Dientamoeba fragilis and Microspora (Enterocytozoon bieneusi and Encephalitizoon) are causative agents of diarrhoea. Encephalitozoon infections are also associated with hepatitis, hepatoconjunctivitis and nephritis. C. cayetanensis infection was diagnosed in 28 patients in the years 1992-1995 in the Academic Medical Centre in Amsterdam, half of these patients returning from a visit to Indonesia. D. fragilis has a prevalence in the Netherlands of 8% among patients with diarrhoea lasting longer than one week referred for parasitological investigation. The prevalence of E. bieneusi in HIV positive patients with diarrhoea was 8-10% in the Academic Medical Centre in the last five years. Infection with Encephalitozoon appears to be endemic in the Netherlands.

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