scholarly journals Genotyping of Giardia duodenalis isolates in asymptomatic children attending daycare centre: evidence of high risk for anthroponotic transmission

2015 ◽  
Vol 144 (7) ◽  
pp. 1418-1428 ◽  
Author(s):  
A. P. OLIVEIRA-ARBEX ◽  
E. B. DAVID ◽  
T. C. G. OLIVEIRA-SEQUEIRA ◽  
G. N. BITTENCOURT ◽  
S. GUIMARÃES

SUMMARYGiardia duodenalis is a common intestinal parasite infecting children attending daycare centres. This study aimed to verify Giardia occurrence and the genotypes of isolates infecting children aged 0–6 years and workers at a daycare centre in the state of São Paulo, Brazil. The families of children who tested positive for Giardia, were asked to provide stool samples from household members and their dogs. Samples (123 children, 14 centre employees, 44 household members, 19 children after treatment, and 20 dogs) were examined for intestinal parasites using concentration methods. DNA extracted from all samples was submitted for polymerase chain reaction (PCR) testing and the amplicons generated were used for multilocus sequence typing of beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes. Giardia was detected in 15·9% and 28·6% of the 220 samples by microscopy and PCR, respectively. Analysis of sequences retrieved from 29 isolates revealed both assemblages A (31%) and B (69%). Sub-assemblages AII, BIII and BIV were identified and the alignment of the bg, gdh and tpi sequences revealed the presence of some single nucleotide polymorphisms, especially in assemblage B sequences. The higher predominance of assemblage B and the identification of the AII type support the view that anthroponotic transmission appears to be an important route of transmission in environments that concentrate children at an age when poor hygiene practices make them more vulnerable to such infection.

2020 ◽  
Vol 9 (1) ◽  
pp. 54
Author(s):  
Salem Belkessa ◽  
Daniel Thomas-Lopez ◽  
Karim Houali ◽  
Farida Ghalmi ◽  
Christen Rune Stensvold

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.


2014 ◽  
Vol 8 (06) ◽  
pp. 786-789 ◽  
Author(s):  
Ana Maria Fonseca ◽  
Natercia Fernandes ◽  
Filipa S Ferreira ◽  
Joana Gomes ◽  
Sónia Centeno-Lima

Introduction: Intestinal parasites are important contributors to the global disease burden, especially in children of low-income countries. The present study determined the frequency of intestinal parasites in children hospitalized at the diarrhea section of the Infectious-Contagious Diseases ward and at the Malnutrition ward of the Department of Pediatrics of the Maputo Central Hospital in Mozambique. Methodology: This pilot study conducted between February and March 2009 enrolled a total of 93 children between 1.5 and 48.2 months of age; 87.1% were younger than 24 months. Parasite detection in stool samples was achieved using direct microscopic observation and Ritchie’s concentration technique. Results: Infection with pathogenic intestinal parasites was detected in 16.1% (15/93) of the children. Giardia duodenalis and Trichuris trichiura were the most common parasites (6.5%, 6/93 each), followed by Ascaris lumbricoides (2.2%, 2/93). One case of mixed infection with A. lumbricoides plus T. trichiura was also detected. Conclusion: This study reinforces the importance of routinely examining stool samples for the diagnosis of intestinal parasites (including protozoa) in children hospitalized in endemic areas.


2019 ◽  
Vol 13 (10) ◽  
pp. 927-932
Author(s):  
Sedat Vezir ◽  
Filiz Kaya ◽  
Emine Vezir ◽  
Nermin Karaosmanoğlu ◽  
Ali Kudret Adiloğlu

Introduction: Chronic spontaneous urticaria (CSU) which develops without a known stimulation is defined as the occurrence of spontaneous wheals, angioedema or both for longer than six weeks. Infections, autoimmunity, food intolerance and internal parasitic infections are supposed to be underlying causes of CSU. The aim of this study was to evaluate the intestinal parasites in children and adult patients diagnosed as CSU, to determine the frequency of parasites in chronic urticaria, and to compare these patients with healthy demographic control groups. Methodology: Seventy six children and 38 adult patients with CSU were examined in terms of parasitic infections. The patients whom parasites were detected received anti-parasitic therapy and the improvements in CSU symptoms were evaluated. Stool samples were examined with direct microscopic examination (native-lugol), stool concentration and trichrome staining methods. Results: In pediatric patient group, 18.4% (n = 14) of the stool samples were positive for Blastocystis sp., 2.6% (n = 2), Dientamoeba fragilis and 1.3% (n = 1), Giardia duodenalis. In adult patient group, Blastocystis sp. was detected in 18.4% (n = 7) of the stool samples. Anti-parasitic therapy yielded substantial improvement in urticaria symptoms in 57.1% of pediatric and 60.0% of adult patients. Conclusions: Blastocystis sp. and D. fragilis may play a role in chronic urticaria which seriously disrupts the patient's quality of life. Parasitic infections should not be neglected in patients with cutaneous manifestations.


Parasitology ◽  
2013 ◽  
Vol 141 (2) ◽  
pp. 206-215 ◽  
Author(s):  
ÁNGELA FERNÁNDEZ-ÁLVAREZ ◽  
AARÓN MARTÍN-ALONSO ◽  
NÉSTOR ABREU-ACOSTA ◽  
CARLOS FELIU ◽  
JEAN-PIERRE HUGOT ◽  
...  

SUMMARYThe flagellated parasite Giardia duodenalis is known as one of the most common causes of protozoal diarrhoea in both humans and animals worldwide. The aim of the present work was to perform the first study of G. duodenalis in rodents in the Canary Islands (Spain) and analyse the level of genetic variation and the potential zoonotic role of the isolates. Stool samples were collected from 284 wild rodents and Giardia cysts were detected by light microscopy. The overall prevalence of giardiasis was 25·4% and ranged from 19·4% in El Hierro to 34% in Gran Canaria. Positive samples were further characterized by PCR and nucleotide sequencing of the triose phosphate isomerase (TPI), β-giardin (BG) and glutamate dehydrogenase (GDH) genes. Our study revealed assemblage G as the most frequent genotype and identified two rodent-infecting G. duodenalis haplotypes of this assemblage, HI and HII. Phylogenetic analysis supported the monophyly of haplotype HI, which we suggest to be considered as a novel G. duodenalis sub-assemblage GII, due to the high genetic distances among this sub-genotype and assemblage G. Furthermore, G. duodenalis assemblage B was detected in an inhabited area in La Palma, a fact that may pose a potential risk of G. duodenalis transmission from rodents to humans.


2019 ◽  
Vol 24 (43) ◽  
Author(s):  
Lucia Reh ◽  
Aly Salimo Muadica ◽  
Pamela Carolina Köster ◽  
Sooria Balasegaram ◽  
Neville Q Verlander ◽  
...  

Background Protozoan enteroparasites Cryptosporidium species and Giardia duodenalis are major contributors to the burden of gastrointestinal illness in children globally, whereas the stramenopile Blastocystis species has been associated with irritable bowel syndrome and skin disorders. Aim To investigate the carriage of these parasites in voluntary asymptomatic schoolchildren (4‒14 years) in 12 different primary and secondary schools in Leganés (Madrid, Spain). Methods In a prospective cross-sectional study, stool samples and epidemiological questionnaires on demographics and potential risk factors were collected from participating schoolchildren. Detection of enteric parasites was conducted by PCR-based methods and confirmed by sequence analysis. We calculated prevalence and odds ratios (OR) with logistic regression. Results Stool samples and questionnaires were provided by 1,359 schoolchildren from 12 schools. The individual prevalence for any parasite was 28%; Blastocystis sp.: 13%; G. duodenalis: 18%; Cryptosporidium spp.: 1%. Two schoolchildren were infected with all three species and 53 with two species. Multivariable risk factor analysis using logistic regression models indicated that an existing infection with one parasite increased the odds for an additional infection with another parasite. The odds of Blastocystis sp. carriage increased up to the age of 10 years and being female increased the odds of Cryptosporidium spp. infection. Washing vegetables before preparing a meal was protective for Blastocystis sp. infection. Conclusion We detected a larger than expected proportion of asymptomatic cases in the participanting schoolchildren. Further investigation of asymptomatic children should be considered. Good hygiene measures should be encouraged for individuals of all ages to protect from protozoal infections.


Parasitologia ◽  
2021 ◽  
Vol 1 (2) ◽  
pp. 83-94
Author(s):  
Pamela Carolina Köster ◽  
Alejandro Dashti ◽  
Lucia Reh ◽  
Begoña Bailo ◽  
Marta Hernández-de-Mingo ◽  
...  

Asymptomatic carriage of diarrhoea-causing enteric protist parasites in the general population is poorly understood, particularly in medium- to high-income countries. This molecular epidemiological survey investigates the presence, molecular diversity, and household transmission of Giardia duodenalis, Cryptosporidium spp., Blastocystis sp., and Enterocystozoon bieneusi in schoolchildren aged 2–13 years (n = 74) and their legal guardians (n = 6) in Madrid, Spain. Enteroparasite detection and genotyping was conducted in stool samples by molecular (PCR and Sanger sequencing) methods. Potential associations linked to infections were investigated through epidemiological questionnaires. Giardia duodenalis was the most prevalent enteric parasite found (14%, 95% CI: 7.1–23), followed by Blastocystis sp. (10%, 95% CI: 6.2–22) and Cryptosporidium spp. (3.8%, 95% CI: 0.78–11). None of the participants tested positive for E. bieneusi. Sequence analyses revealed the presence of G. duodenalis assemblage B, sub-assemblage BIV in a single child. The three Cryptosporidium isolates obtained were assigned to C. hominis, two of them belonging to the gp60 subtype IbA10G2. Four Blastocystis subtypes were identified including ST2 (38%, 3/8), ST3 (25%, 2/8), ST4 (25%, 2/8), and ST8 (12%, 1/8). All G. duodenalis and Cryptosporidium isolates were detected in children only. Blastocystis ST3 and ST4 were circulating in members of the same household. Blastocystis carriage rates increased with the age of the participants. Presence of diarrhoea-causing enteric protists was common in apparently healthy children.


2019 ◽  
Author(s):  
Aly Salimo Muadica ◽  
Pamela C Köster ◽  
Alejandro Dashti ◽  
Begoña Bailo ◽  
Marta Heernández de Mingo ◽  
...  

Abstract Background: The protozoa Giardia duodenalis, Cryptosporidium spp., the stramenopile Blastocystis sp. and the microsporidia Enterocytozoon bieneusi are among the most frequent diarrheal pathogens affecting humans globally. This molecular epidemiological study assesses the frequency and molecular diversity of these intestinal parasites in schoolchildren in central Spain, complementing previously published data on risk and protective factors associated with parasite infection (Reh et al., Euro Surveill. 2019;24).Methods: Stool samples were collected from voluntary asymptomatic schoolchildren (4‒14 years) and their siblings (1‒16 years) attending 12 primary and secondary schools in Leganés (Madrid). Initial detection of pathogens was conducted by PCR-based methods targeting the small subunit (ssu) ribosomal RNA or the internal transcribed spacer (ITS) genes of these parasite species. Genotyping of G. duodenalis-positive samples was carried out by PCR and Sanger sequencing of appropriate markers including the glutamate dehydrogenase (gdh), the ß-giardin (bg), and triose phosphate isomerase (tpi) loci. For C. hominis/C. parvum-positive samples the 60-kDa glycoprotein (gp60) locus was used.Results: A total of 1,512 stool samples were analysed. Giardia duodenalis was the most prevalent pathogen (17.4%, 95% CI: 15.5‒19.4%), followed by Blastocystis sp. (13.0%, 95% CI: 11.4‒14.8%), and Cryptosporidium spp. (0.9%, 95% CI: 0.5%‒1.5%). Enterocytozoon bieneusi was not detected. Sequence analyses of the 24 G. duodenalis isolates genotyped at the gdh, bg, and/or tpi loci revealed the presence of sub-assemblages AII (16.6%, 4/24) and BIV (79.2%, 19/24). An additional sequence (4.2%, 1/24) represented an ambiguous BIII/BIV result. Analyses of the 14 Cryptosporidium sequences generated at the ssu rRNA allowed the identification of C. hominis (71.4%; 10/14) and C. parvum (21.4%; 3/14). An additional sequence (7.2%, 1/14) was only identified at the genus level. A total of 162 Blastocystis sp. isolates were successfully genotyped, revealing the presence of five subtypes including ST1 (22.8%; 37/162), ST2 (36.4%; 59/162), ST3 (21.6%; 35/162), ST4 (18.6%; 30/162), and ST8 (0.6%; 1/162).Conclusions: Giardia duodenalis sub-assemblage BIV, Cryptosporidium hominis and Blastocystis ST2 were the genetic variants of these parasite species more prevalent in the asymptomatic schoolchildren population investigated. These findings are very similar to those previously reported in clinical, symptomatic populations in Spain. Enterocytozoon bieneusi was absent in apparently healthy schoolchildren.


2021 ◽  
Vol 15 (1) ◽  
pp. e0008987 ◽  
Author(s):  
Augusto Messa ◽  
Pamela C. Köster ◽  
Marcelino Garrine ◽  
Carol Gilchrist ◽  
Luther A. Bartelt ◽  
...  

Giardia duodenalis is an enteric parasite commonly detected in children. Exposure to this organism may lead to asymptomatic or symptomatic infection. Additionally, early-life infections by this protozoan have been associated with impaired growth and cognitive function in poor resource settings. The Global Enteric Multicenter Study (GEMS) in Mozambique demonstrated that G. duodenalis was more frequent among controls than in diarrhoeal cases (≥3 loosing stools in the previous 24 hours). However, no molecular investigation was conducted to ascertain the molecular variability of the parasite. Therefore, we describe here the frequency and genetic diversity of G. duodenalis infections in children younger than five years of age with and without diarrhoea from the Manhiça district in southern Mozambique enrolled in the context of GEMS. Genomic DNA from 757 G. duodenalis-positive stool samples by immunoassay collected between 2007–2012, were reanalysed by multiplex PCR targeting the E1-HP and C1-P21 genes for the differentiation of assemblages A and B. Overall, 47% (353) of the samples were successfully amplified in at least one locus. Assemblage B accounted for 90% (319/353) of all positives, followed by assemblage A (8%, 29/353) and mixed A+B infections (1%, 5/353). No association between the presence of a given assemblage and the occurrence of diarrhoea could be demonstrated. A total of 351 samples were further analysed by a multi-locus sequence genotyping (MLSG) approach at the glutamate dehydrogenase (gdh), ß-giardin (bg) and triose phosphate isomerase (tpi) genes. Overall, 63% (222/351) of samples were genotyped and/or sub-genotyped in at least one of the three markers. Sequence analysis revealed the presence of assemblages A (10%; 23/222) and B (90%; 199/222) with high molecular diversity at the nucleotide level within the latter; no mixed infections were identified under the MLSG scheme. Assemblage A sequences were assigned to sub-assemblages AI (0.5%, 1/222), AII (7%, 15/222) or ambiguous AII/AIII (3%, 7/222). Within assemblage B, sequences were assigned to sub-assemblages BIII (13%, 28/222), BIV (14%, 31/222) and ambiguous BIII/BIV (59%, 132/222). BIII/BIV sequences accumulated the majority of the single nucleotide polymorphisms detected, particularly in the form of double peaks at chromatogram inspection. This study demonstrated that the occurrence of gastrointestinal illness (diarrhoea) was not associated to a given genotype of G. duodenalis in Mozambican children younger than five years of age. The assemblage B of the parasite was responsible for nine out of ten infections detected in this paediatric population. The extremely high genetic diversity observed within assemblage B isolates was compatible with an hyperendemic epidemiological scenario where infections and reinfections were common. The obtained molecular data may be indicative of high coinfection rates by different G. duodenalis assemblages/sub-assemblages and/or genetic recombination events, although the exact contribution of both mechanisms to the genetic diversity of the parasite remains unknown.


2021 ◽  
Vol 26 (35) ◽  
Author(s):  
Davide Resi ◽  
Stefania Varani ◽  
Anna Rosa Sannella ◽  
Alessandra M De Pascali ◽  
Margherita Ortalli ◽  
...  

Giardiasis, the disease caused by the flagellate Giardia duodenalis (syn. G.lamblia, G. intestinalis), is the most commonly reported among the five food- and waterborne parasitic diseases under mandatory surveillance in 24 EU countries. From November 2018 to April 2019, an outbreak of giardiasis occurred in a municipality of the Bologna province, in north-eastern Italy. Microscopy and immunochromatography identified cysts and antigens, respectively, of the parasite in stool samples of 228 individuals. Molecular typing of 136 stool samples revealed a vast predominance (95%) of G. duodenalis assemblage B. Investigations into potential sources indicated tap water as the most likely vehicle of infection, although cysts were not detected in water samples. Control measures mostly aimed at preventing secondary transmission by informing citizens about the outbreak, and by treatment of patients with anti-parasitic drugs. This is the first documented human outbreak of giardiasis in Italy; its investigation has highlighted the difficulties in the timely detection and management of this parasite, which is often overlooked as a cause of human gastroenteritis. The long and variable incubation time, absence of specific symptoms and a general lack of awareness about this pathogen contributed to delay in diagnosis.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Corinne Légeret ◽  
Céline Rüttimann ◽  
Hans Fankhauser ◽  
Henrik Köhler

Abstract Background A wide variation of causes can lead to gastrointestinal symptoms in children- an infection with parasites is one of them. The expansion of international travel might lead to an increase in testing children for a correspondent infection. Currently there are no guidelines available, which patients should be tested for a possible parasitical infection. The aim of the study was to characterize Swiss children suffering from intestinal parasites, in order to provide more knowledge for the clinician who should be tested. Methods This is a retrospective study of Swiss pediatric patients, whose stools have been tested for parasites and helminths. Results A total of 1855 stool samples, belonging to 572 different children with an average age of 7.9 years, were tested within a 10-year period. The prevalence of a positive result was 4.2%, of which all were positive for Blastocystis, and 12.5% had a co-infection with Endolimax nana. Conclusion Immigrants, immune compromised children with diarrhea and pediatric patients with bloody or protracted diarrhea should have 2 different stool specimens examined for a possible parasitical infection.


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