Intestinal Microbiota in Children With Symptomatic Dientamoeba fragilis Infection: A Case-control Study.

BACKGROUND: Dientamoeba fragilis in children has been associated with gastrointestinal symptoms, like abdominal pain and diarrhea. The mechanism underlying these symptoms in children with D. fragilis remains unclear. We hypothesized that concomitant microbial alterations, which have been described in other parasitic infections, may be associated with gastrointestinal symptoms in D. fragilis. METHODS: In this case-control study performed in 2 centers, 19 children referred to a pediatrician because of gastrointestinal symptoms and with a positive fecal PCR for D. fragilis were included as cases. We included 19 healthy children as controls and matched for age and gender, selected from an existing cohort of 63 children. A PCR for D. fragilis was performed on fecal samples of the 19 controls to assess D. fragilis carriership in this asymptomatic group. Microbiota was analyzed with the IS-pro technique, and the intestinal microbiota composition and diversity were compared between the 2 groups. RESULTS: Microbiota of children with D. fragilis and gastrointestinal symptoms did not significantly differ in terms of composition and diversity compared with controls, both on phylum and species level. In the asymptomatic controls, a positive fecal PCR for D. fragilis was found in 16 of 19 (84.2%). CONCLUSION: Intestinal microbiota does not seem to play a key role in the presence of clinical symptoms in children with D. fragilis. The pathogenicity of D. fragilis and pathophysiologic pathways underlying the development of gastrointestinal symptoms remains yet to be clarified.

A clinical guideline on Dientamoeba fragilis infections.

Dientamoeba fragilis (D. fragilis) is an intestinal parasite frequently detected in humans with abdominal pain and diarrhoea, but it is also commonly found in asymptomatic subjects. Hence its clinical relevance is often disputed. The introduction of polymerase chain reaction (PCR) is a versatile and sensitive diagnostic technique for the detection of intestinal parasites, and in some Western world countries PCR has almost completely replaced microscopic diagnostics. PCR has however resulted in an increase in the number of D. fragilis-positive patients. The disputed pathogenic nature of this intestinal parasite and an apparent increase in the incidence of patients with positive PCR results have renewed the discussions between clinicians and microbiologists on how to deal with an infected patient. Moreover, treatment guidelines differ throughout the world which makes it difficult for clinicians to choose an optimal therapeutic regimen.AimTo summarize and discuss the current knowledge on the pathogenicity, best diagnostic approach, treatment and follow-up of children and adults infected with D. fragilis.

Clinical and Epidemiological Characteristics of Patients with Dientamoeba fragilis Infection.

Dientamoeba fragilis is an intestinal protozoan, usually considered nonpathogenic. However, in the last years, there has been an attempt to clarify its possible pathogenic role. We aim to evaluate the clinical and epidemiological characteristics of D. fragilis-infected patients. Adults with D. fragilis detection in feces who attended the Vall d'Hebron University Hospital (Barcelona, Spain) were evaluated retrospectively from April 2009 to March 2014. We classified the patients in asymptomatic, symptomatic without other causes except infection of D. fragilis, and symptomatic with another cause. Among symptomatic patients, treatment response was evaluated. One hundred eight patients were included. Sixty-three percent of the patients were immigrants, 29.6% were autochthonous, and 7.4% were travelers. Forty-nine (45.3%) patients presented symptoms, and eosinophilia was observed in 26 (24.1%) patients. Overall, 59 (54.7%) patients were asymptomatic, 15 (13.8%) presented symptoms which were attributable to other causes, and 34 (31.5%) patients presented symptoms with no other causes. In this last group, 29 patients received specific treatment and 65.5% of them presented a complete resolution of the symptoms. The group of symptomatic patients with no other cause had more proportion of women, more proportion of autochthonous people, and were older compared with the group of asymptomatic patients. Dientamoeba fragilis infection should be considered as pathogenic when other causes are ruled out.

The role of Blastocystis sp. and Dientamoeba fragilis in irritable bowel syndrome: a systematic review and meta-analysis.

Irritable bowel syndrome (IBS) is globally one of the most prevalent gastrointestinal disorders with a negative impact on quality of life and socio-economic status of patients. Recently, controversial evidences suggest that Blastocystis sp. and Dientamoeba fragilis infections may be implicated in the development of IBS. We performed a systematic review and meta-analysis to examine the possible association regarding this issue. PubMed, ScienceDirect, Scopus, Web of Science, and Cochrane electronic databases were searched (up to February 2017) to identify the relevant studies. Pooled odds ratio (OR) and 95% confidence intervals were estimated using a random effects meta-analysis model on data from included studies. A total of 17 studies including 5882 participants (2527 patients and 3310 controls) met the eligibility criteria. Individuals with Blastocystis infection were found to have a positive association with IBS (OR, 2.19; 95% CI, 1.54-3.13), while this association was not observed for D. fragilis infection (OR, 1.13; 95% CI, 0.22-5.72). In subgroup analysis for Blastocystis infection, the pooled ORs were OR 2.29, 95% CI 1.55-3.41; OR 1.70, 95% CI 0.83-3.44; and OR 3.83, 95% CI 2.34-6.27 for hospital-based, healthy volunteers, and combined controls, respectively. Considering the subtypes, meta-analysis result demonstrated significant positive ORs for ST1 (OR, 4.40; 95% CI, 2.81-6.90) and ST3 (OR, 1.94; 95% CI, 1.36-2.77) to be potential risk factors for IBS. Our results support the existence of a positive association between Blastocystis sp. and IBS. Further studies with more sample size should be performed to better investigate the real impact of these parasites on the occurrence of IBS.

The Transcriptome Sequence of Dientamoeba fragilis Offers New Biological Insights on its Metabolism, Kinome, Degradome and Potential Mechanisms of Pathogenicity.

Dientamoeba fragilis is a human bowel parasite with a worldwide distribution. Dientamoeba was once described as a rare and harmless commensal though recent reports suggest it is common and potentially pathogenic. Molecular data on Dientamoeba is scarce which limits our understanding of this parasite. To address this, sequencing of the Dientamoeba transcriptome was performed. Messenger RNA was extracted from cultured Dientamoeba trophozoites originating from clinical stool specimens, and sequenced using Roche GS FLX and Illumina HiSeq technologies. In total 6,595 Dientamoeba transcripts were identified. These sequences were analysed using the BLAST2GO software suite and via BLAST comparisons to sequences available from TrichDB, GenBank, MEROPS and kinase.com. Several novel KEGG pathway maps were generated and gene ontology analysis was also performed. These results are thoroughly discussed guided by knowledge available for other related protozoa. Attention is paid to the novel biological insights afforded by this data including peptidases and kinases of Dientamoeba, as well as its metabolism, novel chemotherapeutics and possible mechanisms of pathogenicity. Currently, this work represents the largest contribution to our understanding of Dientamoeba molecular biology and also represents a major contribution to our understanding of the trichomonads generally, many of which are important pathogens of humans and animals.

A review of Dientamoeba fragilis carriage in humans: several reasons why this organism should be considered in the diagnosis of gastrointestinal illness.

Dientamoeba fragilis is a protozoan that inhabits the human gut. It is approximately 100 years since Dientamoeba's discovery and first description when it was described as a rare and harmless commensal. Since then it has struggled to gain recognition as a pathogen despite the evidence supporting its pathogenicity. Dientamoeba remains neglected, probably due to the misconceptions that it is uncommon and non-pathogenic. Usually, carriage of Dientamoeba is associated with symptoms such as abdominal pain and diarrhea. Moreover, antimicrobial therapy followed by resolution of symptoms coincides with the eradication of Dientamoeba. This manuscript reviews the scientific literature relating to Dientamoeba's prevalence and pathogenicity. While much of the evidence supporting its pathogenicity is only circumstantial, it is apparent that most researchers agree that Dientamoeba is pathogenic. Therefore, in symptomatic patients who harbor Dientamoeba and no other pathogen, Dientamoeba should be considered as the etiological agent and treated as such.

The ambiguous life of Dientamoeba fragilis: the need to investigate current hypotheses on transmission.

Dientamoeba fragilis is an inhabitant of the human bowel and is associated with gastrointestinal illness. Despite its discovery over a century ago, the details of Dientamoeba's life cycle are unclear and its mode of transmission is unknown. Several theories exist which attempt to explain how Dientamoeba may be transmitted. One theory suggests that animals are responsible for the transmission of Dientamoeba. However, reports of Dientamoeba in animals are sporadic and most are not supported by molecular evidence. Another theory suggests that Dientamoeba may be transmitted via the ova of a helminth. Given that the closest relative of Dientamoeba is transmitted via the ova of a helminth, this theory seems plausible. It has also been suggested that Dientamoeba could be transmitted directly between humans. This theory also seems plausible given that other relatives of Dientamoeba are transmitted in this way. Despite numerous investigations, Dientamoeba's mode of transmission remains unknown. This review discusses the strengths and weaknesses of theories relating to Dientamoeba's mode of transmission and, by doing so, indicates where gaps in current knowledge exist. Where information is lacking, suggestions are made as to how future research could improve our knowledge on the life cycle of Dientamoeba.

The prevalence of Dientamoeba fragilis in patients with suspected enteroparasitic disease in a metropolitan area in Denmark.

The prevalence of Dientamoeba fragilis in patients from a metropolitan area in Denmark was determined by examination of paired stool samples using two techniques: a formol ethyl-acetate concentration technique with unpreserved faeces and a permanent staining technique on faeces preserved with sodium acetate-acetic acid-formalin (SAF). Using the SAF permanent staining technique and the formol ethyl-acetate concentration technique, 25% and 15% of the specimens, respectively, were parasite-positive. D. fragilis was detected in 12 of the 103 patients, only two of whom harboured other recognised pathogenic parasites. Overall, D. fragilis had a remarkably high prevalence in the metropolitan area of Denmark investigated.

[Role of Dientamoeba fragilis in human bowel infections]. / Dientamoeba fragilis: suo ruolo nelle infezioni intestinali umane.

During 2004, we identified 39 intestinal infections caused by Dientamoeba fragilis and 23 caused by Giardia duodenalis, on 1141 home subjects (3.4% and 2.0% respectively): D. fragilis was observed in 4.5% of O and P (29/644 subjects suffering from aspecific bowel disorders) and in 2.0% of diarrhoeas (10/497); G. duodenalis was observed in 3.1% (20/644) and in 0.6% (3/497) of cases respectively. No other pathogenic parasites were identified (only 1 case of enterobiasis in an asymptomatic child). Commensal protozoa were observed, among O and P, in 4.3% of cases (28/644). The non-pathogenic B. hominis, often associated with other protozoa, was observed in 4.1% of all 1141 cases. D. fragilis is undoubtedly more frequent in adults (36/39 cases, 92.3%) than in children (3/39, 7.7%), and is prevalent among females (24/39, 61.5%) in respect of males (15/39, 38.5%). G. duodenalis is more prevalent among adults (16/23, 69.5%) than children (7/23, 30.5%), but is more frequent among males (13/23, 56.5%) than females (10/23, 43.5%). Clinical correlations of dientamoebiasis and giardiasis are reported, seasonal and epidemiological features of these protozooses are outlined, and the authors emphasize the importance and need of Giemsa stain, among O and P and acute or prolonged diarrhoeas, on the basis of previous good direct microscopic observations of faecal specimens, for correct and complete diagnosis of intestinal infections.

Dientamoebiasis: clinical importance and recent advances.

Dientamoeba fragilis, an unusual single-celled parasite that was described first in 1918, is found worldwide in the gastrointestinal tract of humans. D. fragilis has emerged from obscurity recently because it is now recognized as a common cause of chronic diarrhoea and is treatable with drugs. Recent molecular studies have described D. fragilis as having two genotypes. Diagnostic tests, based on conventional and real-time PCR, have been developed that will provide a rapid, sensitive and specific diagnosis of D. fragilis. These tests will also aid the elucidation of the host distribution and the life cycle of this pathogen.

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.

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.

Dientamoeba fragilis, a neglected cause of diarrhea, successfully treated with secnidazole.

OBJECTIVE: To evaluate the pathogenicity of Dientamoeba fragilis by comparing it with Giardia lamblia and to investigate the effect of a single dose of secnidazole in dientamoebiasis. METHODS: Stool samples of 400 patients, admitted to the Department of Parasitology, Celal Bayar University, were examined by direct wet mount, formalin-ethyl acetate concentration and trichrome staining methods on three consecutive days. All cases positive for D. fragilis were treated with a single dose of secnidazole, 30 mg/kg for children, and 2 g for adults. On the seventh and fourteenth days, at the end of the treatment, stool samples were examined by the same methods, and clinical symptoms were again evaluated. RESULTS: D. fragilis and G. lamblia were detected in 35 (8.8%) and 34 (8.5%) cases, respectively. The most frequent symptoms were found to be abdominal pain and diarrhea in both infections. D. fragilis was eradicated in 34 (97.1%) patients with a single dose of secnidazole, but a second dose was necessary in one patient. Clinical symptoms disappeared in 27 (77.1%) and decreased in eight (22.9%) cases, after eradication. CONCLUSIONS: These data suggested that D. fragilis is as prevalent and pathogenic as G. lamblia, and secnidazole seems to be highly effective in achieving parasitologic and clinical cure. To our knowledge, this is the first report of secnidazole being used in the treatment of dientamoebiasis.

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.