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Parasites & Vectors Jun 2016Blastocystis is a common intestinal parasite with worldwide distribution but the distribution of Blastocystis and its subtypes in East Africa is largely unknown. In this...
BACKGROUND
Blastocystis is a common intestinal parasite with worldwide distribution but the distribution of Blastocystis and its subtypes in East Africa is largely unknown. In this study, we investigate the distribution of Blastocystis subtypes in Zanzibar, Tanzania and report the prevalence of intestinal parasites using both molecular methods and microscopy.
METHODS
Stool samples were collected from both diarrhoeic and non-diarrhoeic outpatients in Zanzibar. In addition to microscopy, real-time PCR for Blastocystis, Entamoeba histolytica and E. dispar, Giardia intestinalis, Cryptosporidium spp., and Dientamoeba fragilis was used. Blastocystis subtypes were determined by a conventional PCR followed by partial sequencing of the SSU-rRNA gene. Genetic assemblages of Giardia were determined by PCR with assemblage specific primers.
RESULTS
Intestinal parasites were detected in 85 % of the 174 participants, with two or more parasites present in 56 %. Blastocystis sp. and Giardia intestinalis were the most common parasites, identified by PCR in 61 and 53 % of the stool samples respectively, but no correlation between carriage of Blastocystis and Giardia was found. The Blastocystis subtype distribution was ST1 34.0 %, ST2 26.4 %, ST3 25.5 %, ST7 0.9 %, and 13.2 % were positive only by qPCR (non-typable). The Giardia genetic assemblages identified were A 6.5 %, B 85 %, A + B 4.3 %, and non-typable 4.3 %. The detection rate with microscopy was substantially lower than with PCR, 20 % for Blastocystis and 13.8 % for Giardia. The prevalence of Blastocystis increased significantly with age while Giardia was most prevalent in children two to five years old. No correlation between diarrhoea and the identification of Giardia, Blastocystis, or their respective genetic subtypes could be shown and, as a possible indication of parasite load, the mean cycle threshold values in the qPCR for Giardia were equal in diarrhoeic and non-diarrhoeic patients.
CONCLUSIONS
Carriage of intestinal parasites was very common in the studied population in Zanzibar. The most commonly detected parasites, Blastocystis and Giardia, had different age distributions, possibly indicating differences in transmission routes, immunity, and/or other host factors for these two species. In the Blastocystis subtype analysis ST1-3 were common, but ST4, a subtype quite common in Europe, was completely absent, corroborating the geographical differences in subtype distributions previously reported.
Topics: Adolescent; Adult; Aged; Blastocystis; Blastocystis Infections; Child; Child, Preschool; Feces; Female; Giardia lamblia; Giardiasis; Humans; Infant; Intestinal Diseases, Parasitic; Male; Middle Aged; Tanzania; Young Adult
PubMed: 27356981
DOI: 10.1186/s13071-016-1637-8 -
Antioxidants (Basel, Switzerland) Oct 2022The diagnosis of obesity comprises subjects with totally different phenotypes and metabolic profiles. Systemic inflammation and oxidative stress derived from the white...
The diagnosis of obesity comprises subjects with totally different phenotypes and metabolic profiles. Systemic inflammation and oxidative stress derived from the white adipose tissue are suggested as the link between this disease and the development of insulin resistance and metabolic comorbidities. The presence of unicellular eukaryotic parasites colonizing the human gut ecosystem is a common circumstance, and yet their influence on the inflammatory and redox status of the obese host has not been assessed. Herein, a set of inflammatory and redox biomarkers were assessed together with a parasitological analysis of 97 severely obese subjects. Information was also collected on insulin resistance and on the antioxidant composition of the diet. The global prevalence of intestinal unicellular parasites was 49.5%, with sp. the most prevalent protozoan found (42.3%). Colonized subjects displayed a higher total antioxidant capacity and a trend towards higher extracellular superoxide dismutase activity, regardless of their insulin resistance status, along with lower reduced glutathione/oxidized glutathione (GSH/GSSG) ratios in plasma in the insulin-resistant subgroup. No changes in malondialdehyde levels, or in inflammatory cytokines in plasma, were found in regard to the colonization status. In conclusion, enteric eukaryotic unicellular parasites may play an important role in modulating the antioxidant defenses of an obese host, thus could have beneficial effects with respect to the development of systemic metabolic disorders.
PubMed: 36358463
DOI: 10.3390/antiox11112090 -
Le Infezioni in Medicina Jun 2017The aim of this study was to evaluate the distribution of parasitic intestinal infections in patients attending Padua teaching hospital during a two-year period. Between...
The aim of this study was to evaluate the distribution of parasitic intestinal infections in patients attending Padua teaching hospital during a two-year period. Between 1st March 2011 and 28th February 2013, we examined stool specimens from 7341 patients (6127 Italians, 1214 non-Italians) for ova and parasites using microscopy, rapid enzyme immunoassays, culture techniques and molecular methods. Stools of 1080 patients (14.71%) were positive for parasites; a total of 1349 intestinal parasites were counted. Protozoa were detected in 1028/1080 patients (95.19%), while helminths were present in 80/1080 patients (7.41%). The protozoa most commonly detected were Blastocystis spp., Dientamoeba fragilis and Giardia duodenalis. Enterobius vermicularis was the helminth most frequently encountered. Of the 1080 infected patients, 227 (21.02%) had more than one parasite in their stool. To conclude, in Italy intestinal parasitoses must be unquestionably considered in differential diagnoses of gastrointestinal diseases. For this purpose, sound knowledge of epidemiology is essential.
Topics: Adolescent; Adult; Aged; Child; Diagnosis, Differential; Feces; Female; Helminthiasis; Hospitals, Teaching; Humans; Inpatients; Intestinal Diseases, Parasitic; Italy; Male; Middle Aged; Outpatients; Parasite Egg Count; Prevalence; Protozoan Infections; Retrospective Studies; Young Adult
PubMed: 28603232
DOI: No ID Found -
Parasitology Apr 2021This epidemiological study assesses the occurrence of enteric parasites in 4303 patients attended at two public hospitals in Ankara (Turkey) during 2018-2019. Microscopy...
This epidemiological study assesses the occurrence of enteric parasites in 4303 patients attended at two public hospitals in Ankara (Turkey) during 2018-2019. Microscopy was used as a screening test. Giardia duodenalis was also identified using a commercial ELISA for the detection of parasite-specific coproantigens. Giardia-positive samples by microscopy/ELISA were confirmed by real-time PCR and characterized using a multilocus genotyping scheme. Blastocystis sp. was genotyped in a sample subset. Blastocystis sp. (11.1%, 95% CI 11.4‒14.8%) and G. duodenalis (1.56%, 95% CI 1.22‒1.96) were the most prevalent pathogens found. Cryptosporidium spp., Entamoeba histolytica and intestinal helminths were only sporadically (<0.5%) found. For G. duodenalis, sequence (n = 30) analyses revealed the presence of sub-assemblages AII (23.3%), discordant AII/AIII (23.3%) and mixed AII + AIII (6.7%) within assemblage A, and BIII (10.0%), BIV (3.3%) and discordant BIII/BIV (23.3%) within assemblage B. Two additional sequences (6.7%) were assigned to the latter assemblage but sub-assemblage information was unknown. No associations between G. duodenalis assemblages/sub-assemblages and sociodemographic and clinical variables could be demonstrated. For Blastocystis sp., sequence (n = 6) analyses identified subtypes ST1, ST2 and ST3 at equal proportions. This is the first molecular characterization of G. duodenalis based on MLG conducted in Turkey to date.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blastocystis; Blastocystis Infections; Child; Child, Preschool; Feces; Female; Giardia lamblia; Giardiasis; Humans; Infant; Male; Middle Aged; Phylogeny; Turkey; Young Adult
PubMed: 32981546
DOI: 10.1017/S0031182020001821 -
Clinical Microbiology and Infection :... Jun 2015The introduction of molecular detection of infectious organisms has led to increased numbers of positive findings, as observed for pathogens causing gastroenteritis...
The introduction of molecular detection of infectious organisms has led to increased numbers of positive findings, as observed for pathogens causing gastroenteritis (GE). However, because little is known about the prevalence of these pathogens in the healthy asymptomatic population, the clinical value of these additional findings is unclear. A case-control study was carried out in a population of patients served by general practitioners in the Netherlands. A total of 2710 fecal samples from case and matched control subjects were subjected to multiplex real-time PCR for the 11 most common bacterial and four protozoal causes of GE. Of 1515 case samples, 818 (54%) were positive for one or more target organisms. A total of 49% of the controls were positive. Higher positivity rates in cases compared to controls were observed for Campylobacter spp., Salmonella spp., Clostridium difficile, enteroinvasive Escherichia coli/Shigella spp., enterotoxigenic E. coli, enteroaggregative E. coli, atypical enteropathogenic E. coli (EPEC), Cryptosporidium parvum/hominis, and Giardia lamblia. However, Dientamoeba fragilis and Shiga-like toxigenic E. coli were detected significantly less frequent in cases than in controls, while no difference in prevalence was found for typical EPEC and enterohemorrhagic E. coli. The association between the presence of microorganisms and GE was the weakest in children aged 0 to 5 years. Higher relative loads in cases further support causality. This was seen for Campylobacter spp., Salmonella spp., enterotoxigenic E. coli, and C. parvum/hominis, and for certain age categories of those infected with C. difficile, enteroaggregative E. coli, and atypical EPEC. For D. fragilis and Shiga-like toxigenic E. coli/enterohemorrhagic E. coli, pathogen loads were lower in cases. Application of molecular diagnostics in GE is rapid, sensitive and specific, but results should be interpreted with care, using clinical and additional background information.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Bacteria; Bacterial Infections; Case-Control Studies; Child; Child, Preschool; Feces; Female; Gastroenteritis; Humans; Infant; Infant, Newborn; Male; Middle Aged; Molecular Diagnostic Techniques; Multiplex Polymerase Chain Reaction; Netherlands; Parasites; Protozoan Infections; Real-Time Polymerase Chain Reaction; Young Adult
PubMed: 25700890
DOI: 10.1016/j.cmi.2015.02.007 -
Clinical Microbiology and Infection :... Dec 2018Multiplex PCR assays offer highly sensitive and specific tools for the detection of enteric pathogens. This prospective study aimed at comparing the novel Roche LightMix... (Comparative Study)
Comparative Study
Evaluation of the Roche LightMix Gastro parasites multiplex PCR assay detecting Giardia duodenalis, Entamoeba histolytica, cryptosporidia, Dientamoeba fragilis, and Blastocystis hominis.
OBJECTIVES
Multiplex PCR assays offer highly sensitive and specific tools for the detection of enteric pathogens. This prospective study aimed at comparing the novel Roche LightMix Modular Assay Gastro Parasites (LMAGP) detecting Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Blastocystis hominis, and Dientamoeba fragilis with routine laboratory procedures.
METHODS
Stool specimens (n = 1062 from 1009 patients) were consecutively examined by LMAGP, R-Biopharm Ridascreen enzyme immunoassays (EIAs) detecting G. duodenalis or E. histolytica/dispar, and microscopy of wet mounts. Discrepant results were analysed by in-house PCR.
RESULTS
D. fragilis or B. hominis were detected by LMAGP in 131 (14.4%) and 179 (19.9%; 16 samples positive by microscopy; p < 0.0001) of 909 samples, respectively. Of 918 samples analysed for Cryptosporidium spp., six were positive by LMAGP (three could be confirmed by Kinyoun staining and one by in-house PCR). G. duodenalis was detected by LMAGP, EIA, or microscopy in 20, 16, or 9 of 1039 stool samples, respectively; all four samples missed by EIA were confirmed by in-house PCR. In total, 938 stool samples were analysed for E. histolytica/dispar. Nine of ten EIA-positive samples were negative by LMAGP but positive by in-house PCR for E. dispar. One E. histolytica infection (positive by both LMAGP and in-house PCR) was missed by EIA and microscopy. Parasites only detected by microscopy included Enterobius vermicularis eggs (n = 3) and apathogenic amoebae (n = 27).
CONCLUSIONS
The data call for routine use of multiplex PCR assays for the detection of enteric protozoan parasites in laboratory diagnostics.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Blastocystis hominis; Child; Child, Preschool; Clinical Laboratory Techniques; Cryptosporidium; Dientamoeba; Entamoeba histolytica; Feces; Female; Giardia lamblia; Humans; Immunoenzyme Techniques; Infant; Infant, Newborn; Intestinal Diseases, Parasitic; Male; Microscopy; Middle Aged; Multiplex Polymerase Chain Reaction; Prospective Studies; Reagent Kits, Diagnostic; Sensitivity and Specificity; Young Adult
PubMed: 29581055
DOI: 10.1016/j.cmi.2018.03.025 -
Parasitology Research Aug 2020Shotgun metagenomics with high-throughput sequencing (HTS) techniques is increasingly used for pathogen identification and characterization. While many studies apply...
Shotgun metagenomics with high-throughput sequencing (HTS) techniques is increasingly used for pathogen identification and characterization. While many studies apply targeted amplicon sequencing, here we used untargeted metagenomics to simultaneously identify protists and helminths in pre-diagnosed faecal and tissue samples. The approach starts from RNA and operates without an amplification step, therefore allowing the detection of all eukaryotes, including pathogens, since it circumvents the bias typically observed in amplicon-based HTS approaches. The generated metagenomics datasets were analysed using the RIEMS tool for initial taxonomic read assignment. Mapping analyses against ribosomal reference sequences were subsequently applied to extract 18S rRNA sequences abundantly present in the sequence datasets. The original diagnosis, which was based on microscopy and/or PCR, could be confirmed in nearly all cases using ribosomal RNA metagenomics. In addition to the pre-diagnosed taxa, we detected other intestinal eukaryotic parasites of uncertain pathogenicity (of the genera Dientamoeba, Entamoeba, Endolimax, Hymenolepis) that are often excluded from routine diagnostic protocols. The study clearly demonstrates the applicability of untargeted RNA metagenomics for the parallel detection of parasites.
Topics: Animals; Feces; High-Throughput Nucleotide Sequencing; Humans; Intestinal Diseases, Parasitic; Metagenomics; Molecular Diagnostic Techniques; Parasites; RNA, Ribosomal, 18S; Sequence Analysis, DNA
PubMed: 32591865
DOI: 10.1007/s00436-020-06754-9 -
Annals of Parasitology 2015This cross-sectional study was carried to determine the prevalence of intestinal parasitic infections among mentally handicapped individuals in Alexandria, Egypt, in the...
This cross-sectional study was carried to determine the prevalence of intestinal parasitic infections among mentally handicapped individuals in Alexandria, Egypt, in the period from December 2012 till November 2013. The study was conducted on 200 institutionalized and non-institutionalized mentally handicapped individuals. Fresh stool samples were subjected to different stains including; trichrome for detecting intestinal protozoa, modified acid fast stain for intestinal coccidia and quick hot gram chromotrope stain for Microsporidia. Also they were processed by Kato-Katz and formol ethyl acetate techniques for intestinal helminths. Additionally, blood samples were collected for measuring hemoglobin levels. Out of 200 mentally handicapped individuals, 87 (43.5%) were infected. The infection rates were 44.6% and 42.6% for non-institutionalized and institutionalized people, respectively. Regarding gender, 46.7% and 38.5% were reported for the males and females respectively. The most common parasites detected were: Cryptosporidium sp. (23.5%), microsporidia (15%), Giardia lamblia (8.5%), Dientamoeba fragilis (8%), Cyclospora cyatanensis (7.5%), Blastocystis hominis (6.5%), Entamoeba histolytica (5.5%) and Entamoeba coli (2.5%). Rates for Isospora belli and Enterobius vermicularis were estimated to be 1.5% for each, while lower rate was reported for Iodamoeba butschlii (1.0%). Prevalence of infections among mentally handicapped individuals are indications for several risk factors, including improper sanitary hygiene and illiteracy about personal hygiene. Therefore, frequent investigations, health care and medical intervention are needed.
Topics: Adolescent; Child; Egypt; Female; Hospitals, Psychiatric; Humans; Intestinal Diseases, Parasitic; Male; Persons with Mental Disabilities; Young Adult
PubMed: 26878626
DOI: 10.17420/ap6104.19 -
PloS One 2019In this study we characterized the presence and subtype (ST1-ST4) of Blastocystis in patients attended at a referral center for tropical diseases in Northern Italy. We...
In this study we characterized the presence and subtype (ST1-ST4) of Blastocystis in patients attended at a referral center for tropical diseases in Northern Italy. We also, evaluated the organism's association with other intestinal parasites. Parasite screening was performed on 756 patients, from different geographical origins (namely, Italians, Africans, South Americans, Asian and non-Italian Europeans) in which Italians represented the largest group. Blastocystis was seen to be the most prevalent parasite in the study. Subtype 3 and 1 were the most frequently found in the Italians and Africans. Our data confirmed previous studies performed in Italy, in which ST3 proved to be the most prevalent subtype, but we highlighted also a high frequency of mixed subtypes, which were probably underestimated in former analyses. Interestingly, the mixed subtypes group was the most prevalent in all the analysed geographical areas. About half of our cases showed other co-infecting parasites and the most frequent was Dientamoeba fragilis. Our study confirms that, in Blastocystis infection, multiple subtypes and co-infecting parasites are very frequently present, in particular Dientamoeba fragilis.
Topics: Adolescent; Adult; Aged; Blastocystis; Blastocystis Infections; Child; Coinfection; Dientamoeba; Dientamoebiasis; Emigrants and Immigrants; Feces; Female; Humans; Italy; Male; Middle Aged; Prevalence; Retrospective Studies; Tertiary Care Centers
PubMed: 30615638
DOI: 10.1371/journal.pone.0210171 -
Mikrobiyoloji Bulteni Oct 2019Aim of the present study was to identify protozoones which are difficult to define through wet slide in fresh fecal samples by using different fixatives with modified...
Aim of the present study was to identify protozoones which are difficult to define through wet slide in fresh fecal samples by using different fixatives with modified Trichrome stain within five minutes. Two different fixatives prepared for the alternative approach. The slides were fixed by two different fixatives, one of them (fixative-1) was based ethylalcohol, formalin, acetic acid, distilled water and the other one (fixative-2) based ethylalcohol, formalin, citric acid, distilled water included a mordant [divalent or polyvalent metals which make coordination complex with some dyes] consisted copper sulphate pentahydrate (CuSO4 .5H2 O). Slides prepared by the two different fixatives were stained by a different modification of Gomori's trichrome stain that we made. Samples fixed by Schaudinn fixative including mercury chloride were stained by Wheatley modification of Gomori's trichrome stain as a gold standard for control and comparison. We worked with 50 fecal samples which we thought included human intestinal protozoones after the wet slide examination. Comparing the methods, slides prepared with the method including citric acid gave almost similar results with the classical method excluded Entamoeba coli cystes. Slides prepared with the methode including acetic acid gave low performance compared with the classical method especially E.coli cystes and Blastocystis spp., Endolimax nana, Iodamoeba bütschlii, E.hartmanni. Both new fixatives gave superior performance at the slides included Dientamoeba fragilis and approximately shorten the procedure process ten times than the classical method. When the both alternative methods compared in each other, the slides prepared with fixative-2 exposed better performance for the protozoones Blastocystis spp., E.nana, I.bütschlii and E.hartmanni while the fixative-1 displayed minimal superiority for D.fragilis including criterias that we based. The fixative-2 and modified stain methode that we used in our study, makes available the diagnostic phase ten times faster than the classical method in human stool parasitological tests excluding the E.coli cystes at parasitology and microbiolgy laboratories. It seems to be a good option to the classical method for routine usage.
Topics: Eukaryota; Feces; Fixatives; Formaldehyde; Humans; Microscopy; Parasitic Diseases; Parasitology
PubMed: 31709939
DOI: 10.5578/mb.68633