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PloS One 2022The increasing interest to perform and investigate the efficacy of fecal microbiota transplantation (FMT) has generated an urge for feasible donor screening. We report...
BACKGROUND
The increasing interest to perform and investigate the efficacy of fecal microbiota transplantation (FMT) has generated an urge for feasible donor screening. We report our experience with stool donor recruitment, screening, follow-up, and associated costs in the context of clinical FMT trials.
METHODS
Potential stool donors, aged between 18-65 years, underwent a stepwise screening process starting with an extensive questionnaire followed by feces and blood investigations. When eligible, donors were rescreened for MDROs and SARS-CoV-2 every 60-days, and full rescreening every 4-6 months. The costs to find and retain a stool donor were calculated.
RESULTS
From January 2018 to August 2021, 393 potential donors underwent prescreening, of which 202 (51.4%) did not proceed primarily due to loss to follow-up, medication use, or logistic reasons (e.g. COVID-19 measures). 191 potential donors filled in the questionnaire, of which 43 (22.5%) were excluded. The remaining 148 candidates underwent parasitology screening: 91 (61.5%) were excluded, mostly due to Dientamoeba fragilis and/or high amounts of Blastocystis spp. After additional feces investigations 18/57 (31.6%) potential donors were excluded (mainly for presence of Helicobacter Pylori and ESBL-producing organisms). One donor failed serum testing. Overall, 38 out of 393 (10%) potential donors were enrolled. The median participation time of active stool donors was 13 months. To recruit 38 stool donors, €64.112 was spent.
CONCLUSION
Recruitment of stool donors for FMT is challenging. In our Dutch cohort, failed eligibility of potential donors was often caused by the presence of the protozoa Dientamoeba fragilis and Blastocystis spp.. The exclusion of potential donors that carry these protozoa, especially Blastocystis spp., is questionable and deserves reconsideration. High-quality donor screening is associated with substantial costs.
Topics: Humans; Adolescent; Young Adult; Adult; Middle Aged; Aged; Fecal Microbiota Transplantation; Donor Selection; SARS-CoV-2; COVID-19; Feces; Clostridium Infections
PubMed: 36264933
DOI: 10.1371/journal.pone.0276323 -
Nutrients Aug 2022Obesity is an epidemic causing a metabolic health crisis. Herein, the interactions between the gut prokaryotic and eukaryotic communities, metabolic comorbidities and...
Obesity is an epidemic causing a metabolic health crisis. Herein, the interactions between the gut prokaryotic and eukaryotic communities, metabolic comorbidities and diet were studied. Stool samples from 56 subjects, 47 with type III obesity and 9 with type II obesity and cardiovascular risk or metabolic disease, were assessed for the richness, diversity and ecology of the bacterial gut community through metagenomics, together with the study of the presence of common unicellular eukaryote parasites ( sp., and ) by qPCR. Clinical information regarding metabolic comorbidities and non-alcoholic hepatic fatty liver disease was gathered. To assess the quality of the patients' diet, each participant filled in three dietary questionnaires. The most prevalent parasite sp. (46.4%), together with (8.9%), was found to be associated with higher mean diversity indexes regarding non-colonized subjects; the opposite of that which was observed in those with (16.1%). In terms of phyla relative abundance, with sp. and , very slight differences were observed; on the contrary, was related to an increase in Bacteroidetes and Proteobacteria, and a decrease in Firmicutes and Actinobacteria, presenting the lowest Firmicutes/Bacteroidetes ratio. At genus level, sp. and/or was accompanied with an increase in spp., and a decrease in spp., spp. and spp., while was associated with an increase in spp., and a decrease in spp., spp. and spp., and the highest spp./ spp. ratio. Participants with non-alcoholic hepatic fatty liver presented a higher Firmicutes/Bacteroidetes ratio, and those with type 2 diabetes displayed a significantly lower spp./ spp. ratio, due to an overrepresentation of the genus spp. The presence of parasites was associated with variations in the richness, diversity and distribution of taxa in bacterial communities, confirming a gain in diversity associated with sp. and providing different functioning of the microbiota with a potential positive effect on comorbidities such as type 2 diabetes, insulin resistance and metabolic syndrome. Future basic and clinical studies should assess the beneficial or pathogenic effect of these eukaryotes on obese subjects and focus on deciphering whether they may imply a healthier metabolic profile.
Topics: Animals; Bacteria; Blastocystis; Diabetes Mellitus, Type 2; Feces; Humans; Obesity, Morbid; Parasites
PubMed: 35956387
DOI: 10.3390/nu14153211 -
Revista Do Instituto de Medicina... 2021In order to provide additional data on the prevalence and genetic diversity of Dientamoeba fragilis in human populations, we conducted a study in children from...
In order to provide additional data on the prevalence and genetic diversity of Dientamoeba fragilis in human populations, we conducted a study in children from low-income communities in Sao Paulo State, Brazil. Fecal samples from daycare center attendees up to 6 years old (n=156) and staff members (n=18) were submitted to PCR and sequencing of D. fragilis as well as to microscopic examination for the presence of other intestinal parasites. All children assessed were asymptomatic and 10.3% (16/156) were positive for D. fragilis. No worker was found to be positive. An association between Dientamoeba and coinfection with other intestinal parasites was observed. Concerning the genetic diversity, 14 and only two isolates were genotype 1 and genotype 2, respectively. Our findings outline interesting aspects: (1) asymptomatic children as carriers of Dientamoeba in communities in which environmental conditions ensure parasite transmission and, (2) association between Dientamoeba infection in young children and coinfection with other enteric parasites, reinforcing its transmission via the fecal-oral route.
Topics: Brazil; Child; Child, Preschool; Dientamoeba; Dientamoebiasis; Feces; Humans; Intestinal Diseases, Parasitic; Prevalence
PubMed: 33978095
DOI: 10.1590/S1678-9946202163039 -
Archives of Disease in Childhood Jul 2019To study the association between colonisation and faecal calprotectin to see whether the parasite is a harmless commensal or a gut pathogen.
OBJECTIVE
To study the association between colonisation and faecal calprotectin to see whether the parasite is a harmless commensal or a gut pathogen.
DESIGN
Cross-sectional study of previously collected stool samples.
SETTING AND PATIENTS
Two hundred stool samples originated from children aged 5-19 years with chronic abdominal pain and diarrhoea, who were seen in paediatric clinics in the Netherlands and Belgium and in whom somatic gastrointestinal disorders were excluded. Another 122 samples came from a healthy community-based reference population of the same age. All stool samples were analysed with real-time PCR for the detection of and with an ELISA for calprotectin-a biomarker of gastrointestinal inflammation.
MAIN OUTCOME MEASURES
Prevalence of colonisation and results of stool calprotectin testing.
RESULTS
was detected in 45% (95% CI 38% to 51%) of patients and in 71% (95% CI 63% to 79%) of healthy children. Median (IQR) concentrations of calprotectin in patients and healthy children with a positive PCR result were not different from those with a negative PCR result (40 (40-55) μg/g vs 40 (40-75) μg/g, respectively).
CONCLUSION
Since colonisation is most prevalent in healthy children and is not associated with an increase in faecal calprotectin concentration, our data do not support the inference that is a pathogenic parasite. Routinely testing for in children with chronic abdominal pain should therefore be discouraged.
Topics: Abdominal Pain; Adolescent; Belgium; Child; Child, Preschool; Cohort Studies; Cross-Sectional Studies; Dientamoeba; Dientamoebiasis; Feces; Female; Humans; Male; Netherlands; Prevalence; Prospective Studies; Real-Time Polymerase Chain Reaction; Retrospective Studies; Young Adult
PubMed: 30798256
DOI: 10.1136/archdischild-2018-316383 -
Clinical Microbiology Reviews Jul 2016Dientamoeba fragilis is a protozoan parasite of the human bowel, commonly reported throughout the world in association with gastrointestinal symptoms. Despite its... (Review)
Review
Dientamoeba fragilis is a protozoan parasite of the human bowel, commonly reported throughout the world in association with gastrointestinal symptoms. Despite its initial discovery over 100 years ago, arguably, we know less about this peculiar organism than any other pathogenic or potentially pathogenic protozoan that infects humans. The details of its life cycle and mode of transmission are not completely known, and its potential as a human pathogen is debated within the scientific community. Recently, several major advances have been made with respect to this organism's life cycle and molecular biology. While many questions remain unanswered, these and other recent advances have given rise to some intriguing new leads, which will pave the way for future research. This review encompasses a large body of knowledge generated on various aspects of D. fragilis over the last century, together with an update on the most recent developments. This includes an update on the latest diagnostic techniques and treatments, the clinical aspects of dientamoebiasis, the development of an animal model, the description of a D. fragilis cyst stage, and the sequencing of the first D. fragilis transcriptome.
Topics: Animals; Dientamoeba; Dientamoebiasis; Disease Models, Animal; Humans; Intestines; Life Cycle Stages; Phylogeny
PubMed: 27170141
DOI: 10.1128/CMR.00076-15 -
Trends in Parasitology Jul 2014Trichomonads are common parasites of many vertebrate and invertebrate species, with four species classically recognized as human parasites: Dientamoeba fragilis,... (Review)
Review
Trichomonads are common parasites of many vertebrate and invertebrate species, with four species classically recognized as human parasites: Dientamoeba fragilis, Pentatrichomonas hominis, Trichomonas vaginalis, and Trichomonas tenax. The latter two species are considered human-specific; by contrast, D. fragilis and P. hominis have been isolated from domestic and farm mammals, demonstrating a wide host range and potential zoonotic origin. Several new studies have highlighted the zoonotic dimension of trichomonads. First, species typically known to infect birds and domestic mammals have been identified in human clinical samples. Second, several phylogenetic analyses have identified animal-derived trichomonads as close sister taxa of the two human-specific species. It is our opinion, therefore, that these observations prompt further investigation into the importance of zoonotic trichomonads for human health.
Topics: Animals; Bird Diseases; Birds; Humans; Phylogeny; Trichomonas Infections; Trichomonas vaginalis; Zoonoses
PubMed: 24951156
DOI: 10.1016/j.pt.2014.05.005 -
Tropical Parasitology 2022() diagnosis is an intestinal protozoan parasite globally found in rural and urban areas and is attracting a growing interest. Its prevalence in stool varies from 0.2%...
INTRODUCTION
() diagnosis is an intestinal protozoan parasite globally found in rural and urban areas and is attracting a growing interest. Its prevalence in stool varies from 0.2% to more than 19% depending upon the population studied.
MATERIALS AND METHODS
This study was based on the examination of 100 stool samples of randomly referred cases in a rural area in Motobus district, Kafr El-Sheikh governorate, Egypt. Our aim was to investigate the presence of in stool of the examined individuals using conventional polymerase chain reaction (PCR) compared to wet mount and trichrome stain with confirmation of infection by transmission electron microscopy.
RESULTS
was detected in 13/100 of the stool samples examined using wet mount smears, while trichrome stain detected 17/100. Conventional PCR diagnosed 41 cases of in the studied group. A very good agreement was found between wet mount and trichrome stain for diagnosing , while there was fair agreement between conventional PCR and both microscopy methods. Transmission electron microscope was performed on pooled positive samples that revealed the internal structures of trophozoite with its characteristic nucleus.
CONCLUSIONS
PCR technique was superior to microscopy for the detection of . Trichrome stain remains vital for microscopic diagnosis.
PubMed: 36643979
DOI: 10.4103/tp.tp_93_21 -
PLoS Neglected Tropical Diseases Oct 2021The clinical significance of Blastocystis sp. and Dientamoeba fragilis in patients with gastrointestinal symptoms is a controversial issue. Since the pathogenicity of...
Investigation of neglected protists Blastocystis sp. and Dientamoeba fragilis in immunocompetent and immunodeficient diarrheal patients using both conventional and molecular methods.
INTRODUCTION
The clinical significance of Blastocystis sp. and Dientamoeba fragilis in patients with gastrointestinal symptoms is a controversial issue. Since the pathogenicity of these protists has not been fully elucidated, testing for these organisms is not routinely pursued by most laboratories and clinicians. Thus, the prevalence of these organisms and the subtypes of Blastocystis sp. in human patients in Turkey are not well characterized. This study aimed to determine the prevalence of Blastocystis sp. and D. fragilis in the diarrheic stool samples of immunodeficient and immunocompetent patients using conventional and molecular methods and to identify Blastocystis sp. subtypes using next generation sequencing.
MATERIAL AND METHODS
Individual stool specimens were collected from 245 immunodeficient and 193 immunocompetent diarrheic patients between March 2017 and December 2019 at the Gazi University Training and Research Hospital in Ankara, Turkey. Samples were screened for Blastocystis sp. and D. fragilis by conventional and molecular methods. Molecular detection of both protists was achieved by separate qPCRs targeting a partial fragment of the SSU rRNA gene. Next generation sequencing was used to identify Blastocystis sp. subtypes.
RESULTS
The prevalence of Blastocystis sp. and D. fragilis was 16.7% and 11.9%, respectively as measured by qPCR. The prevalence of Blastocystis sp. and D. fragilis was lower in immunodeficient patients (12.7% and 10.6%, respectively) compared to immunocompetent patients (21.8% and 13.5%, respectively). Five Blastocystis sp. subtypes were identified and the following subtype distribution was observed: ST3 54.4% (n = 37), ST2 16.2% (n = 11), ST1 4.4% (n = 3), ST6 2.9% (n = 2), ST4 1.5% (n = 1), ST2/ST3 11.8% (n = 8) and ST1/ST3 8.8% (n = 6). There was no statistically significant difference in the distribution of Blastocystis sp. subtypes between immunocompetent and immunodeficient patients.
CONCLUSION AND RECOMMENDATION
Our findings demonstrated that Blastocystis sp. and D. fragilis are commonly present in immunocompetent and immunodeficient patients with diarrhea. This study is the first to use next generation sequencing to address the presence of Blastocystis sp. mixed subtypes and intra-subtype variability in clinical samples in Turkey.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blastocystis; Diarrhea; Dientamoeba; Feces; Female; High-Throughput Nucleotide Sequencing; Humans; Immunocompetence; Male; Middle Aged; Primary Immunodeficiency Diseases; Turkey; Young Adult
PubMed: 34613993
DOI: 10.1371/journal.pntd.0009779 -
Pathogens (Basel, Switzerland) Oct 2022The enteric protozoan parasites spp., and are-to various extents-contributors to the burden of gastrointestinal illness in high-income countries. Detection of these...
The enteric protozoan parasites spp., and are-to various extents-contributors to the burden of gastrointestinal illness in high-income countries. Detection of these pathogens by microscopy examination is challenging because of the limited sensitivity and need for specific staining procedures. We developed and optimised a new multiplex real-time PCR assay for the simultaneous detection of spp., and in clinical (stool) samples. The diagnostic performance of the assay was evaluated against a large panel of well-characterised DNA samples positive for spp. ( = 126), ( = 132) and ( = 49). The specificity of the test was assessed against a DNA panel from other intestinal or phylogenetically related parasites ( = 105) and faecal DNA from individuals without clinical manifestations ( = 12). The assay exhibited a diagnostic sensitivity of 0.90-0.97 and a diagnostic specificity of 1. The limit of detection was estimated for (1 oocyst) and (5 × 10 cysts). The method allowed the detection of four species (, , and ) and five assemblages (A-E) without cross-reacting with other parasites belonging to the phyla Amoebozoa, Apicomplexa, Euglenozoa, Microsporidia, Nematoda and Platyhelminthes. This newly developed multiplex real-time PCR assay represents a novel alternative for the rapid and accurate detection of , and in clinical settings.
PubMed: 36365028
DOI: 10.3390/pathogens11111277