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Sample records for giardia duodenalis purified

  1. Infections by Intestinal Coccidia and Giardia duodenalis.

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    Cama, Vitaliano A; Mathison, Blaine A

    2015-06-01

    The coccidians Cryptosporidium spp, Cyclospora cayetanensis, and Cystoisospora belli and the flagellate Giardia duodenalis are pathogenic protozoa associated with gastrointestinal manifestations. Diagnosis relies heavily on microscopy, and although ova-and-parasite examinations can detect Giardia and Cystoisospora, Cryptosporidium and Cyclospora often require specific diagnostic requests. Approved non-microscopy methods are available for Giardia and Cryptosporidium, although negative results are frequently followed by microscopic assays. Polymerase chain reaction-based methods are not frequently used for diagnosis of Giardia and Cryptosporidium and have been used primarily for epidemiologic or outbreak investigations of Giardia and Cryptosporidium. Published by Elsevier Inc.

  2. Giardia duodenalis genetic assemblages and hosts

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    Heyworth Martin F.

    2016-01-01

    Full Text Available Techniques for sub-classifying morphologically identical Giardia duodenalis trophozoites have included comparisons of the electrophoretic mobility of enzymes and of chromosomes, and sequencing of genes encoding β-giardin, triose phosphate isomerase, the small subunit of ribosomal RNA and glutamate dehydrogenase. To date, G. duodenalis organisms have been sub-classified into eight genetic assemblages (designated A–H. Genotyping of G. duodenalis organisms isolated from various hosts has shown that assemblages A and B infect the largest range of host species, and appear to be the main (or possibly only G. duodenalis assemblages that undeniably infect human subjects. In at least some cases of assemblage A or B infection in wild mammals, there is suggestive evidence that the infection had resulted from environmental contamination by G. duodenalis cysts of human origin.

  3. Molecular characterization of Giardia duodenalis in Yemen.

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    Alyousefi, Naela A; Mahdy, Mohammed A K; Xiao, Lihua; Mahmud, Rohela; Lim, Yvonne A L

    2013-06-01

    Giardia duodenalis is an important intestinal protozoan in Yemen with infection rates ranging from 18% to 27%. To date, there has been no genotyping study to provide a better understanding of the transmission dynamic. This study was conducted to genotype and subtype G. duodenalis in Yemen. Stool samples were collected from 503 Yemeni outpatients between 1 and 80 years old, including 219 males and 284 females. Giardia cysts were detected via microscopy after the formal-ether concentration. Genotyping of Giardia was carried out using PCR and sequence analysis of the 16s rRNA and b-giardin genes. Of the 89 microscopy-positive Giardia samples, 65 were successfully sequenced, of which 66% (43 of 65) were identified as G. duodenalis assemblage A and 34% (22 of 65) as assemblage B. Further subtyping analysis based on b-giardin gene identified the presence of subtypes A2 and A3, which belong to the anthroponotic sub-assemblage AII. Data of the study suggest that anthroponotic transmission played a potential role in the transmission of giardiasis in the community. However, further genotyping and subtyping studies of specimens from humans and animals living in the same households are needed for a more definitive understanding of giardiasis transmission in Yemen. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Multilocus genotyping of Giardia duodenalis in Malaysia.

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    Huey, Choy Seow; Mahdy, Mohammed A K; Al-Mekhlafi, Hesham M; Nasr, Nabil A; Lim, Yvonne A L; Mahmud, Rohela; Surin, Johari

    2013-07-01

    Giardia duodenalis is considered the most common intestinal parasite in humans worldwide. In Malaysia, many studies have been conducted on the epidemiology of giardiasis. However, there is a scarcity of information on the genetic diversity and the dynamics of transmission of G. duodenalis. The present study was conducted to identify G. duodenalis assemblages and sub-assemblages based on multilocus analysis of the glutamate dehydrogenase (gdh), beta-giardin (bg) and triose phosphate isomerase (tpi) genes. Faecal specimens were collected from 484 Orang Asli children with a mean age of 7 years and examined using light microscopy. Specimens positive for Giardia were subjected to PCR analysis of the three genes and subsequent sequencing in both directions. Sequences were edited and analysed by phylogenetic analysis. G. duodenalis was detected in 17% (84 of 484) of the examined specimens. Among them, 71 were successfully sequenced using at least one locus. Genotyping results showed that 30 (42%) of the isolates belonged to assemblage A, 32 (45%) belonged to assemblage B, while discordant genotype results were observed in 9 specimens. Mixed infections were detected in 43 specimens using a tpi-based assemblage specific protocol. At the sub-assemblages level, isolates belonged to assemblage A were AII. High nucleotide variation found in isolates of assemblage B made subtyping difficult to achieve. The finding of assemblage B and the anthroponotic genotype AII implicates human-to-human transmission as the most possible mode of transmission among Malaysian aborigines. The high polymorphism found in isolates of assemblage B warrants a more defining tool to discriminate assemblage B at the sub-assemblage level. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Identification of zoonotic genotypes of Giardia duodenalis.

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    Hein Sprong

    Full Text Available Giardia duodenalis, originally regarded as a commensal organism, is the etiologic agent of giardiasis, a gastrointestinal disease of humans and animals. Giardiasis causes major public and veterinary health concerns worldwide. Transmission is either direct, through the faecal-oral route, or indirect, through ingestion of contaminated water or food. Genetic characterization of G. duodenalis isolates has revealed the existence of seven groups (assemblages A to G which differ in their host distribution. Assemblages A and B are found in humans and in many other mammals, but the role of animals in the epidemiology of human infection is still unclear, despite the fact that the zoonotic potential of Giardia was recognised by the WHO some 30 years ago. Here, we performed an extensive genetic characterization of 978 human and 1440 animal isolates, which together comprise 3886 sequences from 4 genetic loci. The data were assembled into a molecular epidemiological database developed by a European network of public and veterinary health Institutions. Genotyping was performed at different levels of resolution (single and multiple loci on the same dataset. The zoonotic potential of both assemblages A and B is evident when studied at the level of assemblages, sub-assemblages, and even at each single locus. However, when genotypes are defined using a multi-locus sequence typing scheme, only 2 multi-locus genotypes (MLG of assemblage A and none of assemblage B appear to have a zoonotic potential. Surprisingly, mixtures of genotypes in individual isolates were repeatedly observed. Possible explanations are the uptake of genetically different Giardia cysts by a host, or subsequent infection of an already infected host, likely without overt symptoms, with a different Giardia species, which may cause disease. Other explanations for mixed genotypes, particularly for assemblage B, are substantial allelic sequence heterogeneity and/or genetic recombination. Although the

  6. Giardia duodenalis Infections in Humans and Other Animals in China

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    Li, Junqiang; Wang, Haiyan; Wang, Rongjun; Zhang, Longxian

    2017-01-01

    Giardia duodenalis is an important zoonotic pathogen in both public and veterinary health, and has been genotyped into at least eight assemblages (A–H), each with a distinct host range. In recent years, this intestinal protozoan parasite has been identified widely in humans and various other animals, and has even been recorded in environmental contaminants. Along with whole genome sequencing of G. duodenalis, multilocus sequence typing is increasingly being used to characterize G. duodenalis isolates. Here, we review the epidemiology, genotyping, and subtyping of G. duodenalis from humans and a wide range of other animals, as well as from wastewater, in China. PMID:29081771

  7. Giardia duodenalis Infections in Humans and Other Animals in China

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    Junqiang Li

    2017-10-01

    Full Text Available Giardia duodenalis is an important zoonotic pathogen in both public and veterinary health, and has been genotyped into at least eight assemblages (A–H, each with a distinct host range. In recent years, this intestinal protozoan parasite has been identified widely in humans and various other animals, and has even been recorded in environmental contaminants. Along with whole genome sequencing of G. duodenalis, multilocus sequence typing is increasingly being used to characterize G. duodenalis isolates. Here, we review the epidemiology, genotyping, and subtyping of G. duodenalis from humans and a wide range of other animals, as well as from wastewater, in China.

  8. Genetic characterisation of Giardia duodenalis in dairy cattle in Brazil.

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    Paz e Silva, Flávio Medeiros; Lopes, Raimundo Souza; Araújo, João Pessoa

    2012-02-01

    The intestinal protozoan parasite Giardia duodenalis (Lambl, 1859) Kofoid & Christiansen, 1915 [syn. Giardia intestinalis and Giardia lamblia] has emerged as a widespread enteric pathogen in humans and domestic animals. In recent years, G. duodenalis has been found in cattle worldwide and longitudinal studies have reported cumulative prevalence of 100% in some herds. In the present study, we determined the prevalence and genetic characterisation of G. duodenalis in 200 dairy cattle from 10 dairy farms in São Paulo state, Brazil. All faecal specimens were screened for the presence of G. duodenalis using microscopy examination, enzyme immunoassay (EIA) and polymerase chain reaction (PCR). DNA was extracted from faecal samples and G. duodenalis were identified by amplification of the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes followed by restriction fragment length polymorphism (RFLP) or sequencing analysis. Giardia was identified in eight farm locations (80% prevalence). Overall, 15/200 (7.5%) animals were positive for infection, only one of which was a cow. Giardia duodenalis genotype E was present in 14 of the animals tested. Zoonotic genotype AI was present in one positive sample. Genotype E and genotype A represented 93% and 7% of G. duodenalis infections, respectively. This study demonstrates that G. duodenalis infection was prevalent in dairy calves in São Paulo state and that the non-zoonotic genotype E predominates in cattle in this region. Nevertheless, calves naturally infected in Brazil can shed Giardia cysts that can potentially infect humans, and thus, they may represent a public health risk.

  9. First studies on Giardia duodenalis in the water buffalo

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    G. Cringoli

    2010-02-01

    Full Text Available A cross-sectional survey of Giardia duodenalis infection in the water buffalo was carried out in Central Italy. The survey was conducted on a sample of 90 farms, selected using a grid approach within a Geographical Information System, followed by proportional allocation. On each farm, faecal samples were collected from three to five asymptomatic buffalo calves, aged from 1 to 9 weeks (total number = 347. Each faecal sample was tested for the presence of copro-antigens of G. duodenalis using a commercially available ELISA. Out of the 90 farms, 27 (30.0% resulted positive. With respect to animals, out of the 347 faecal samples, 63 (18.1% were found to have antigens of G. duodenalis. The results of the logistic regression model showed a positive association between the positivity to G. duodenalis and the presence of sheep on farm.

  10. Molecular Characterization of Giardia duodenalis in Children in Kenya.

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    Mbae, C; Mulinge, E; Guleid, F; Wainaina, J; Waruru, A; Njiru, Z K; Kariuki, S

    2016-03-22

    Giardia duodenalis is an important intestinal protozoan in humans worldwide with high infection rates occurring in densely populated and low resource settings. The parasite has been recorded to cause diarrhea in children. This study was carried out to identify G. duodenalis assemblages and sub-assemblages in children presenting with diarrhea in Kenya. A total of 2112 faecal samples were collected from children aged ≤ 5 years and screened for the presence of Giardia cysts using microscopy. A total of 96 (4.5%) samples were identified as Giardia positive samples and were genotyped using glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and β-giardin loci. The three markers successfully genotyped 72 isolates and grouped 2 (1.4) isolates as Assemblage A, 64 (88.9) as Assemblage B and 7 (9.7%) consisted of mixed infections with assemblage A and B. A further analysis of 50 isolates using GDH Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) categorized 2 assemblage A isolates as sub-assemblage AII while 6 and 14 assemblage B isolates were categorized into sub-assemblage BIII and BIV respectively. A mixed infection with sub-assemblage BIII and BIV was recorded in 28 isolates. Over half (55.6%) of Giardia infections were recorded among the children between 13 to 48 months old. This paper reports the first data on the assemblages and sub-assemblages of Giardia duodenalis in children representing with diarrhea in Kenya.

  11. Molecular Identification of Giardia duodenalis Isolates from Fars Province, Iran.

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    Mohammad Rayani

    2014-03-01

    Full Text Available Giardia duodenalis is one of the most common human intestinal protozoan parasites worldwide and is endemic throughout the world with a vast range of mammalian hosts. The present study aimed to identify the prevalence of G. duodenalis isolates and determine the most common of its assemblages in the patients referring to health centers and hospitals in Fars province, Iran that will be subjected to further molecular investigation.We collected 1000 human fecal samples from health centers and hospitals in Shiraz, Iran in a one year period from September 2009 to August 2010. Microscopic examination for the presence of G. duodenalis cysts and trophozoites was performed by direct wet mount before and after the concentration techniques. Extraction of DNA was performed by Phenol-Chloroform-Isoamylalcohol (PCI. G. duodenalis-positive specimens were analyzed by PCR. A fragment of the SSU-rDNA (292 bp gene was amplified by PCR using the forward primer RH11 and the reverse primer RH4. Genotyping was performed using sequence analysis of G. duodenalis glutamate dehydrogenase gene using primers GDHeF, GDHiF, and GDHiR.The prevalence of Giardia infection was 10.7% (107/1000 examined based on microscopic examination. PCR identified 80% (40/50 of the samples as positive for G. duodenalis based on SSU-rDNA amplification on sucrose gradient samples. Besides, genotyping results indicated 32 isolates (80% as assemblage AII and 8 isolates (20% as assemblage BIII and BIV based on the DNA sequence analysis of the glutamate dehydrogenase locus of G. duodenalis.The findings of this study emphasize that Iran (Fars Province is a favorable area for giardiasis with an anthroponotic infection route.

  12. A report on a case of Giardia duodenalis

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    Sujata Pudasaini

    2017-09-01

    Full Text Available ABSTRACTGiardia is caused by protozoa, Giardia lamblia. It is one of the most common causes of water born or food born diarrhea and it has a worldwide distribution. The prevalence is approximately 20-30% in developing countries. Giardia duodenalis is the most commonly reported intestinal protozoan in humans mainly affecting children. The infection is usually associated with malabsorption, weight loss and growth retardation.We report a case of four years old boy who presented with diarrhea and malabsorption for last 2 years. Its clinical presentation and histopathological diagnosis on duodenal biopsy is discussed.

  13. Identification of Zoonotic Genotypes of Giardia duodenalis

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    Sprong, H.; Cacciò, S.M.; van der Giessen, J.W.B

    2009-01-01

    , through ingestion of contaminated water or food. Genetic characterization of G. duodenalis isolates has revealed the existence of seven groups (assemblages A to G) which differ in their host distribution. Assemblages A and B are found in humans and in many other mammals, but the role of animals...... the potential to tackle intricate epidemiological questions concerning protozoan diseases....

  14. Diagnosis of Giardia duodenalis Infection using Dot Blot in Comparison with Microscopy.

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    Yazdani, Hajar; Sharafi, Seyedeh Maryam; Yousefi, Hoseynali; Hadipur, Mahbobeh; Sepahvand, Akram; Darani, Hossein Yousofi

    2016-01-01

    Giardia duodenalis is an intestinal flagellate parasite which spreads all over the world and is considered as a health problem in the most rural and low sanitation areas. Many diagnostic tests have been developed for the detection of Giardia parasite in stool samples but all of them have some disadvantages such as lack of sensitivity and specificity. In search for a simple and accurate test, diagnosis of Giardia infection using dot blot method has been investigated in this work. In this descriptive study, 30 stool samples which their infection with Giardia were confirmed by direct examination and formalin ether considered as case group. Thirty stool samples without Giardia infection according to formalin ether examination were also considered as a control group. Giardia cysts were isolated from the stool samples using sucrose method. In order to raise antiserum against Giardia cysts, the purified cysts were then sonicated and injected to a rabbit. A mono specific antiserum against the 66KDa band of Giardia cyst antigen was also prepared. The two antisera were used in the dot blot test. Finally, the sensitivity and specificity of the dot-blot method were estimated by considering formalin ether as the gold standard. When Poly specific antiserum was used, the sensitivity and specificity of the dot blot for detection of Giardia infection were 77% and 64% respectively. However the sensitivity and specificity of this assay were 97% and 64% respectively when monospecific antiserum was used. It seems that dot blot is an easy method for the diagnosis of Giardia especially in the rural areas. However more work is recommended for further development of this test.

  15. Pathophysiology of enteric infections with Giardia duodenalis

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    Buret A.G.

    2008-09-01

    Full Text Available Giardia is the most prevalent human intestinal parasitic protist in the world, and one of the most common parasite of companion animals and young livestock. Giardia is a major cause of diarrhea in children and in travelers. The host-microbial interactions that govern the outcome of infection remain incompletely understood. Findings available to date indicate that the infection causes diarrhea via a combination of intestinal malabsorption and hypersecretion. Malabsorption and maldigestion mainly result from a diffuse shortening of epithelial microvilli. This enterocytic injury is mediated by activated host T lymphocytes. Pathophysiological activation of lymphocytes is secondary to Giardia-induced disruption of epithelial tight junctions, which in turn increases intestinal permeability. Loss of epithelial barrier function is a result of Giardia-induced enterocyte apoptosis. Recent findings suggest that these effects may facilitate the development of chronic enteric disorders, including inflammatory bowel disease, irritable bowel syndrome, and allergies, via mechanisms that remain poorly understood. A newly discovered SGLT-1 glucose uptake-mediated host cytoprotective mechanism may represent an effective modulator of the epithelial apoptosis induced by this parasite, and, possibly, by other enteropathogens. A better understanding of the pathogenesis of giardiasis will shed light on new potential therapeutic targets.

  16. Presence of Cryptosporidium spp. and Giardia duodenalis through drinking water.

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    Castro-Hermida, José Antonio; García-Presedo, Ignacio; Almeida, André; González-Warleta, Marta; Correia Da Costa, José Manuel; Mezo, Mercedes

    2008-11-01

    To evaluate the presence of Cryptosporidium spp. and Giardia duodenalis in the influent and final effluent of sixteen drinking water treatment plants located in a hydrographic basin in Galicia (NW Spain) - in which the principal river is recognised as a Site of Community Importance (SCI) - estimate the efficiency of treatment plants in removing these protozoans and determine the species and genotypes of the parasites by means of a molecular assay. All plant samples of influent and final effluent (50-100 l) were examined in the spring, summer, autumn and winter of 2007. A total of 128 samples were analysed by method 1623, developed by US Environmental Protection Agency for isolation and detection of both parasites. To identify the genotypes present the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and b-giardina (G. duodenalis). The mean concentrations of parasites in the influent were 0.0-10.5 Cryptosporidium spp. oocysts per litre and 1.0-12.8 of G. duodenalis cysts per litre. In the final treated effluent, the mean concentration of parasites ranged from 0.0-3.0 oocysts per litre and 0.5-4.0 cysts per litre. The distribution of results by season revealed that in all plants, the highest numbers of (oo)cysts were recorded in spring and summer. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. Cryptosporidium spp. and G. duodenalis were consistently found at high concentrations in drinking water destined for human and animal consumption in the hydrographic basin under study, in Galicia (NW Spain). It is important that drinking water treatment authorities rethink the relevance of contamination levels of both parasites in drinking water and develop adequate countermeasures.

  17. Giardia duodenalis induces pathogenic dysbiosis of human intestinal microbiota biofilms.

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    Beatty, Jennifer K; Akierman, Sarah V; Motta, Jean-Paul; Muise, Stacy; Workentine, Matthew L; Harrison, Joe J; Bhargava, Amol; Beck, Paul L; Rioux, Kevin P; McKnight, Gordon Webb; Wallace, John L; Buret, Andre G

    2017-05-01

    Giardia duodenalis is a prevalent cause of acute diarrheal disease worldwide. However, recent outbreaks in Italy and Norway have revealed a link between giardiasis and the subsequent development of chronic post-infectious irritable bowel syndrome. While the mechanisms underlying the causation of post-infectious irritable bowel syndrome remain obscure, recent findings suggest that alterations in gut microbiota communities are linked to the pathophysiology of irritable bowel syndrome. In the present study, we use a laboratory biofilm system to culture and enrich mucosal microbiota from human intestinal biopsies. Subsequently, we show that co-culture with Giardia induces disturbances in biofilm species composition and biofilm structure resulting in microbiota communities that are intrinsically dysbiotic - even after the clearance of Giardia. These microbiota abnormalities were mediated in part by secretory-excretory Giardia cysteine proteases. Using in vitro cell culture and germ-free murine infection models, we show that Giardia-induced disruptions of microbiota promote bacterial invasion, resulting in epithelial apoptosis, tight junctional disruption, and bacterial translocation across an intestinal epithelial barrier. Additionally, these dysbiotic microbiota communities resulted in increased activation of the Toll-like receptor 4 signalling pathway, and overproduction of the pro-inflammatory cytokine IL-1beta in humanized germ-free mice. Previous studies that have sought explanations and risk factors for the development of post-infectious irritable bowel syndrome have focused on features of enteropathogens and attributes of the infected host. We propose that polymicrobial interactions involving Giardia and gut microbiota may cause persistent dysbiosis, offering a new interpretation of the reasons why those afflicted with giardiasis are predisposed to gastrointestinal disorders post-infection. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights

  18. High sensitivity of Giardia duodenalis to tetrahydrolipstatin (orlistat in vitro.

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    Juliane Hahn

    Full Text Available Giardiasis, a gastrointestinal disease caused by Giardia duodenalis, is currently treated mainly with nitroimidazoles, primarily metronidazole (MTZ. Treatment failure rates of up to 20 percent reflect the compelling need for alternative treatment options. Here, we investigated whether orlistat, a drug approved to treat obesity, represents a potential therapeutic agent against giardiasis. We compared the growth inhibitory effects of orlistat and MTZ on a long-term in vitro culture adapted G. duodenalis strain, WB-C6, and on a new isolate, 14-03/F7, from a patient refractory to MTZ treatment using a resazurin assay. The giardiacidal concentration of the drugs and their combined in vitro efficacy was determined by median-effect analysis. Morphological changes after treatment were analysed by light and electron microscopy. Orlistat inhibited the in vitro growth of G. duodenalis at low micromolar concentrations, with isolate 14-03/F7 (IC50(24h = 2.8 µM being more sensitive than WB-C6 (IC50(24h = 6.2 µM. The effect was significantly more potent compared to MTZ (IC50(24h = 4.3 µM and 11.0 µM, respectively and led to specific undulated morphological alterations on the parasite surface. The giardiacidal concentration of orlistat was >14 µM for 14-03/F7 and >43 µM for WB-C6, respectively. Importantly, the combination of both drugs revealed no interaction on their inhibitory effects. We demonstrate that orlistat is a potent inhibitor of G. duodenalis growth in vitro and kills parasites at concentrations achievable in the gut by approved treatment regimens for obesity. We therefore propose to investigate orlistat in controlled clinical studies as a new drug in giardiasis.

  19. Prevalence and genotyping of Giardia duodenalis among Arabian horses in Ahvaz, southwest of Iran

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    H. Jafari

    2016-09-01

    Full Text Available Giardia duodenalis is globally recognized as an important zoonotic intestinal protozoan parasite. So far, eight assemblages of G. duodenalis (A-H have been identified. Substantial evidence suggests the zoonotic potential of assemblages A, B, and E in livestock. In this study, the genotype of Giardia duodenalis isolates was genetically identified by determining the sequence of ssu-rRNA gene and performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP on glutamate dehydrogenase gene of the parasite in Arabian horses from Ahvaz, located in southwest of Iran. The results revealed that assemblages AI and E (livestock-associated G. duodenalis were present in horse isolates. Also, based on the findings, prevalence of G. duodenalis infection among horses was estimated at 35.7%. The results indicated that G. duodenalis is highly prevalent among Arabian horses, posing a zoonotic risk for giardiasis in Ahvaz, Iran.

  20. Giardia duodenalis in Alpine (Rupicapra rupicapra rupicapra) and Apennine (Rupicapra pyrenaica ornata) chamois

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    De Liberato, Claudio; Berrilli, Federica; Marangi, Marianna; Santoro, Maristella; Trogu, Tiziana; Putignani, Lorenza; Lanfranchi, Paolo; Ferretti, Francesco; D?Amelio, Stefano; Giangaspero, Annunziata

    2015-01-01

    Background Although chamois Rupicapra spp. are the most abundant mountain ungulates in Europe, no data are available on the presence of Giardia duodenalis infecting these species. Methods A total of 157 fecal samples from Alpine Rupicapra rupicapra rupicapra and Apennine Rupicapra pyrenaica ornata chamois were tested for the presence of G. duodenalis by immunofluorescence test, quantitative Real Time PCR and end-point PCR?for genotype characterization. Results G. duodenalis was detected in R....

  1. Molecular characterization of Giardia duodenalis and Cryptosporidium parvum in fecal samples of individuals in Mongolia.

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    Hong, Sung-Hee; Anu, Davaasuren; Jeong, Young-Il; Abmed, Davaajav; Cho, Shin-Hyeong; Lee, Won-Ja; Lee, Sang-Eun

    2014-01-01

    The Giardia and Cryptosporidium species are widespread and frequent diarrhea-related parasites affecting humans and other mammalian species. The prevalence of these parasites in Mongolia is currently unknown. Therefore, we performed molecular analyses of G. duodenalis and C. parvum in stool samples from 138 patients hospitalized with diarrhea in Mongolia using nested polymerase chain reaction (PCR). A total of 5 (3.62%) and 7 (5.07%) fecal samples were positive for G. duodenalis and C. parvum, respectively. Giardia duodenalis and C. parvum infections were prevalent in children Mongolia.

  2. Population-based analyses of Giardia duodenalis is consistent with the clonal assemblage structure

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    Takumi Katsuhisa

    2012-08-01

    Full Text Available Abstract Background Giardia duodenalis is a common protozoan parasite of humans and animals. Genetic characterization of single loci indicates the existence of eight groups called assemblages, which differ in their host distribution. Molecular analyses challenged the idea that G. duodenalis is a strictly clonal diplomonad by providing evidence of recombination within and between assemblages. Particularly, inter-assemblage recombination events would complicate the interpretation of multi-locus genotyping data from field isolates: where is a host infected with multiple Giardia genotypes or with a single, recombined Giardia genotype. Methods Population genetic analyses on the single and multiple-locus level on an extensive dataset of G. duodenalis isolates from humans and animals were performed. Results Our analyses indicate that recombination between isolates from different assemblages are apparently very rare or absent in the natural population of Giardia duodenalis. At the multi-locus level, our statistical analyses are more congruent with clonal reproduction and can equally well be explained with the presence of multiple G. duodenalis genotypes within one field isolate. Conclusions We conclude that recombination between G. duodenalis assemblages is either very rare or absent. Recombination between genotypes from the same assemblage and genetic exchange between the nuclei of a single cyst needs further investigation.

  3. Cysteine Protease-Dependent Mucous Disruptions and Differential Mucin Gene Expression in Giardia duodenalis Infection.

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    Amat, Christina B; Motta, Jean-Paul; Fekete, Elena; Moreau, France; Chadee, Kris; Buret, Andre G

    2017-11-01

    The intestinal mucous layer provides a critical host defense against pathogen exposure and epithelial injury, yet little is known about how enteropathogens may circumvent this physiologic barrier. Giardia duodenalis is a small intestinal parasite responsible for diarrheal disease and chronic postinfectious illness. This study reveals a complex interaction at the surface of epithelial cells, between G. duodenalis and the intestinal mucous layer. Here, we reveal mechanisms whereby G. duodenalis evades and disrupts the first line of host defense by degrading human mucin-2 (MUC2), depleting mucin stores and inducing differential gene expression in the mouse small and large intestines. Human colonic biopsy specimens exposed to G. duodenalis were depleted of mucus, and in vivo mice infected with G. duodenalis had a thinner mucous layer and demonstrated differential Muc2 and Muc5ac mucin gene expression. Infection in Muc2 -/- mice elevated trophozoite colonization in the small intestine and impaired weight gain. In vitro, human LS174T goblet-like cells were depleted of mucus and had elevated levels of MUC2 mRNA expression after G. duodenalis exposure. Importantly, the cysteine protease inhibitor E64 prevented mucous degradation, mucin depletion, and the increase in MUC2 expression. This article describes a novel role for Giardia's cysteine proteases in pathogenesis and how Giardia's disruptions of the mucous barrier facilitate bacterial translocation that may contribute to the onset and propagation of disease. Copyright © 2017 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  4. Population genetic analysis of Giardia duodenalis: genetic diversity and haplotype sharing between clinical and environmental sources.

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    Durigan, Mauricio; Ciampi-Guillardi, Maisa; Rodrigues, Ricardo C A; Greinert-Goulart, Juliane A; Siqueira-Castro, Isabel C V; Leal, Diego A G; Yamashiro, Sandra; Bonatti, Taís R; Zucchi, Maria I; Franco, Regina M B; de Souza, Anete P

    2017-04-01

    Giardia duodenalis is a flagellated intestinal protozoan responsible for infections in various hosts including humans and several wild and domestic animals. Few studies have correlated environmental contamination and clinical infections in the same region. The aim of this study was to compare groups of Giardia duodenalis from clinical and environmental sources through population genetic analyses to verify haplotype sharing and the degree of genetic similarity among populations from clinical and environmental sources in the metropolitan region of Campinas. The results showed high diversity of haplotypes and substantial genetic similarity between clinical and environmental groups of G. duodenalis. We demonstrated sharing of Giardia genotypes among the different populations studied. The comparison between veterinary and human sequences led us to identify new zoonotic genotypes, including human isolates from genetic assemblage C. The application of a population genetic analysis in epidemiological studies allows quantification of the degree of genetic similarity among populations of Giardia duodenalis from different sources of contamination. The genetic similarity of Giardia isolates among human, veterinary, and environmental groups reinforced the correlation between clinical and environmental isolates in this region, which is of great importance for public health. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  5. Multilocus genotyping of Giardia duodenalis in lambs from Spain reveals a high hetrogeneity

    Science.gov (United States)

    Fecal specimens from 120 lambs in Valencia (Spain) were analyzed for Giardia duodenalis by IFA and nested-PCR using the beta giardin, glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and small subunit ribosomal RNA (ssurRNA) genes. The highest prevalence was obtained using the ssurRN...

  6. A large scale molecular study of Giardia duodenalis in horses in Colombia

    Science.gov (United States)

    The prevalence of Giardia duodenalis genotypes in horses is poorly known. The present study examined feces from 195 horses, 1 month to 17 years of age, in 4 locations in Colombia. Prevalence of infection was determined by PCR and all positives were sequenced to determine the genotypes. Thirty four (...

  7. Prevalence and genotype of Giardia duodenalis in dairy cattle from Northern and Northeastern part of Thailand.

    Science.gov (United States)

    Inpankaew, Tawin; Jiyipong, Tawisa; Thadtapong, Nalumon; Kengradomkij, Chanya; Pinyopanuwat, Nongnuch; Chimnoi, Wissanuwat; Jittapalapong, Sathaporn

    2015-09-01

    The aims of this study were to determine prevalence and genotype of Giardia duodenalis in feces of dairy cattle from the northern part and the northeastern part of Thailand. A total of 900 fecal samples were collected directly from rectum and examined by using zinc sulphate centrifugal flotation technique and polymerase chain reaction (PCR). The overall prevalence of G. duodenalis in dairy cows was 5.0 % (45/900) by zinc sulphate centrifugal flotation and 6.0 % (54/900) by PCR. Genotypes of G. duodenalis found in this study were Assemblage AI and E. The results indicated that dairy cattle may act as a potential risk of Giardia transmission among animals and humans (especially Assemblage AI).

  8. Molecular characterization of Giardia duodenalis isolated from Semai Pahang Orang Asli (Peninsular Malaysia aborigines).

    Science.gov (United States)

    Mahdy, A K Mohammed; Surin, Johari; Mohd-Adnan, A; Wan, K-L; Lim, Y A L

    2009-09-01

    This study was conducted to determine the genotypes of Giardia duodenalis isolated from human faecal samples at Pos Betau, Pahang, Malaysia. Faecal specimens were collected and examined for G. duodenalis cysts using Trichrome staining techniques. Molecular identification was carried out by the amplification of a region of the small subunit of the nuclear ribosomal RNA (SSU rRNA) gene using nested PCR and subsequent sequencing. The sequences from 15 isolates from G. duodenalis were subjected to phylogenetic analysis (including appropriate outgroups) using the neighbor-joining and maximum parsimony methods. The trees identified G. duodenalis assemblages A and B, with a predominance of assemblage B. The predominance of anthroponotic genotypes indicates the possibility of anthroponotic transmission of these protozoa in this Semai Pahang Orang Asli community.

  9. Molecular identification of Giardia duodenalis isolates from domestic dogs and cats in Wroclaw, Poland.

    Science.gov (United States)

    Piekarska, Jolanta; Bajzert, Joanna; Gorczykowski, Michał; Kantyka, Magdalena; Podkowik, Magdalena

    2016-09-01

    Giardia duodenalis (G. intestinalis) is a common protozoan causing gastrointestinal disorders in many species of mammals. The genus of Giardia has high molecular diversity. Dogs and cats, in addition to their typical infection with assemblages C, D and F, may be a reservoir of zoonotic assemblages (A and B). The aim of this study was a genetic characteristic of Giardia isolates of dogs and cats from the area of Wroclaw (Poland). A total of 128 and 33 faecal samples from dogs and cats, respectively, were analyzed by routine coprological methods. The animals were diagnosed on the presence of G. duodenalis antigens in faeces soluble with the use of SNAP Giardia (IDEXX Laboratories) immunosorbent assay. 27 DNA isolates of Giardia were subjected to molecular identification (PCR-RFLP). The prevalence of G. duodenalis was 21.1% (27/128) in dogs and 15.1% (5/33) in cats. In dogs, C assemblage was present in 18 (81%) positive stool samples, D assemblage in 2 (9%) samples, B assemblage present in one (4.5%), and mixed assemblages (C and D) occurred in one (4.5%) sample. F assemblage was found in 4 (80%) cats' positive stool samples and A assemblage occurred in one case (20%). Confirmation of the presence of A and B zoonotic assemblages suggests that infected pets can be a threat to human health. This study describes for the first time the presence of mixed infections within host-specific C and D assemblages in dogs in Poland.

  10. Are species-specifi antigen detection tests needed in the diagnosis of Giardia duodenalis infection?

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    Flávia Fernandes de Mendonça Uchôa

    2017-08-01

    Full Text Available Objective: To assess the diagnostic performance in human stool samples of a rapid, qualitative, solid-phase immunochromatographic test (Alere® originally developed to detect Giardia duodenalis antigens in fecal samples of dogs. Methods: Samples from 54 patients with a previous diagnosis of giardiasis were tested by the microscopic examination to assess the performance of an immunochromatographic kit developed to detect Giardia duodenalis coproantigen in dog feces. Results: The agreement between the microscopic and the immunological methods was 83.3%. These findings are consistent with those of other studies using human specific kits. Conclusions: It is suggested that the same immunochromatographic test could be used for Giardia diagnosis in both species.

  11. Low risk for transmission of zoonotic Giardia duodenalis from dogs to humans in rural Cambodia

    DEFF Research Database (Denmark)

    Inpankaew, Tawin; Schär, Fabian; Odermatt, Peter

    2014-01-01

    BACKGROUND: A number of epidemiological studies have demonstrated Giardia as prevalent in both humans and dogs worldwide and have postulated the occurrence of anthroponotic, zoonotic and animal-specific cycles of transmission, which may be geographically and regionally unique in its epidemiology....... The aim of this study was to utilise molecular tools to determine the prevalence and compare genotypes of Giardia duodenalis infecting humans and dogs living in a previously identified Giardia-endemic village in rural Cambodia in order to ascertain zoonotic transmission risk. FINDINGS: The prevalence of G....... duodenalis in humans and dogs was 18.3% (40/218) and 10.6% (10/94) by PCR, respectively. Molecular characterisation of the small subunit of ribosomal RNA (SSU rRNA) gene, triose phosphate isomerase (TPI) gene and sub-assemblage characterisation of the glutamate dehydrogenase (gdh) gene placed 27.5% (11...

  12. Cryptosporidium cuniculus and Giardia duodenalis in rabbits: genetic diversity and possible zoonotic transmission.

    Science.gov (United States)

    Zhang, Weizhe; Shen, Yujuan; Wang, Rongjun; Liu, Aiqin; Ling, Hong; Li, Yihong; Cao, Jianping; Zhang, Xiaoyun; Shu, Jing; Zhang, Longxian

    2012-01-01

    Cryptosporidium and Giardia are the two important zoonotic pathogens causing diarrhea of humans and animals worldwide. Considering the human cryptosporidiosis outbreak and sporadic cases caused by C. cuniculus, the important public health significance of G. duodenalis and little obtained information regarding rabbit infected with Cryptosporidium and Giardia in China, the aim of this study is to determine the prevalence and molecularly characterize Cryptosporidium and Giardia in rabbits in Heilongjiang Province, China. 378 fecal samples were obtained from rabbits in Heilongjiang Province. Cryptosporidium oocysts and Giardia cysts were detected using Sheather's sugar flotation technique and Lugol's iodine stain method, respectively. The infection rates of Cryptosporidium and Giardia were 2.38% (9/378) and 7.41% (28/378), respectively. Genotyping of Cryptosporidium spp. was done by DNA sequencing of the small subunit rRNA (SSU rRNA) gene and all the nine isolates were identified as Cryptosporidium cuniculus. The nine isolates were further subtyped using the 60-kDa glycoprotein (gp60) gene and two subtypes were detected, including VbA32 (n = 3) and a new subtype VbA21 (n = 6). G. duodenalis genotypes and subtypes were identified by sequence analysis of the triosephosphate isomerase (TPI) gene. The assemblage B (belonging to eight different subtypes B-I to B-VIII) was found in 28 G. duodenalis-positive samples. The rabbits have been infected with Cryptosporidium and Giardia in Heilongjiang Province. The results show that the rabbits pose a threat to human health in the studied areas. Genotypes and subgenotypes of C. cuniculus and G. duodenalis in this study might present the endemic genetic characterization of population structure of the two parasites.

  13. Cryptosporidium cuniculus and Giardia duodenalis in rabbits: genetic diversity and possible zoonotic transmission.

    Directory of Open Access Journals (Sweden)

    Weizhe Zhang

    Full Text Available BACKGROUND: Cryptosporidium and Giardia are the two important zoonotic pathogens causing diarrhea of humans and animals worldwide. Considering the human cryptosporidiosis outbreak and sporadic cases caused by C. cuniculus, the important public health significance of G. duodenalis and little obtained information regarding rabbit infected with Cryptosporidium and Giardia in China, the aim of this study is to determine the prevalence and molecularly characterize Cryptosporidium and Giardia in rabbits in Heilongjiang Province, China. METHODOLOGY/PRINCIPAL FINDINGS: 378 fecal samples were obtained from rabbits in Heilongjiang Province. Cryptosporidium oocysts and Giardia cysts were detected using Sheather's sugar flotation technique and Lugol's iodine stain method, respectively. The infection rates of Cryptosporidium and Giardia were 2.38% (9/378 and 7.41% (28/378, respectively. Genotyping of Cryptosporidium spp. was done by DNA sequencing of the small subunit rRNA (SSU rRNA gene and all the nine isolates were identified as Cryptosporidium cuniculus. The nine isolates were further subtyped using the 60-kDa glycoprotein (gp60 gene and two subtypes were detected, including VbA32 (n = 3 and a new subtype VbA21 (n = 6. G. duodenalis genotypes and subtypes were identified by sequence analysis of the triosephosphate isomerase (TPI gene. The assemblage B (belonging to eight different subtypes B-I to B-VIII was found in 28 G. duodenalis-positive samples. CONCLUSIONS/SIGNIFICANCE: The rabbits have been infected with Cryptosporidium and Giardia in Heilongjiang Province. The results show that the rabbits pose a threat to human health in the studied areas. Genotypes and subgenotypes of C. cuniculus and G. duodenalis in this study might present the endemic genetic characterization of population structure of the two parasites.

  14. Evidence of heterozygosity and recombinant alleles in single cysts of Giardia duodenalis

    OpenAIRE

    Aguiar, Juliana Martins; Silva, Sheila Oliveira; Santos, Valdir Azevedo dos; Taniwaki, Sueli Akemi; Oliveira, Tricia Maria Ferreira de Sousa; Ferreira, Helena Lage; Keid, Lara Borges; Gregori, Fábio; Soares, Rodrigo Martins

    2016-01-01

    Abstract Giardia duodenalis is divided into eight assemblages (named A to H). Isolates of assemblage A are divided into four sub-assemblages (AI, AII, AIII and AIV). While isolates of sub-assemblage AII are almost exclusively detected in human hosts, isolates of assemblage B are encountered in a multitude of animal hosts and humans. Here, we isolated single cysts of G. duodenalis from a human stool sample and found that one of them had overlaps of assemblage AII and B alleles and an unexpecte...

  15. Performance of microscopy and ELISA for diagnosing Giardia duodenalis infection in different pediatric groups.

    Science.gov (United States)

    Silva, Renata K N R; Pacheco, Flávia T F; Martins, Adson S; Menezes, Joelma F; Costa-Ribeiro, Hugo; Ribeiro, Tereza C M; Mattos, Ângela P; Oliveira, Ricardo R; Soares, Neci M; Teixeira, Márcia C A

    2016-12-01

    Techniques for Giardia diagnosis based on microscopy are usually applied as routine laboratory testing; however, they typically exhibit low sensitivity. This study aimed to evaluate Giardia duodenalis and other intestinal parasitic infections in different pediatric groups, with an emphasis on the comparison of Giardia diagnostic techniques. Feces from 824 children from different groups (diarrheic, malnourished, with cancer and from day care) were examined by microscopy and ELISA for Giardia, Cryptosporidium sp. and Entamoeba histolytica coproantigen detection. Giardia-positive samples from day-care children, identified by either microscopy or ELISA, were further tested by PCR targeting of the β-giardin and Gdh genes. Statistically significant differences (PGiardia duodenalis was more frequent in day-care children and Cryptosporidium sp. in diarrheic and malnourished groups; infections by Entamoeba histolytica were found only in children with diarrhea. Considering positivity for Giardia by at least one method, ELISA was found to be more sensitive than microscopy (97% versus 55%). To examine discrepancies among the diagnostic methods, 71 Giardia-positive stool samples from day-care children were tested by PCR; of these, DNA was amplified from 51 samples (77.4%). Concordance of positivity between microscopy and ELISA was found for 48 samples, with 43 confirmed by PCR. Parasite DNA was amplified from eleven of the 20 Giardia samples (55%) identified only by ELISA. This study shows the higher sensitivity of ELISA over microscopy for Giardia diagnosis when a single sample is analyzed and emphasizes the need for methods based on coproantigen detection to identify this parasite in diarrheic fecal samples. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  16. Giardia duodenalis infection: risk factors for children living in sub-standard settlements in Brazil

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    Júlio César Teixeira

    2007-06-01

    Full Text Available The objective of the present study was to characterize Giardia duodenalis infection among children living in sub-standard settlement areas in the municipality of Juiz de Fora, Minas Gerais State, Brazil. The cross-sectional epidemiological study included 590 children from 1 to 5 years of age. Data were collected from one child per selected family through home interviews with the parent or guardian and parasitological examination of stool samples. Thirty-one putative risk factors concerning family structure, socioeconomic status, and environmental factors were analyzed by multivariate logistic regression. Prevalence of G. duodenalis infection was 18% (106 children. Four potential risk factors were heavily associated with G. duodenalis infection: number of under-five children in the same household, index child's birth order, existence of a bathroom in the home, and drinking water source.

  17. Giardia duodenalis in Alpine (Rupicapra rupicapra rupicapra) and Apennine (Rupicapra pyrenaica ornata) chamois.

    Science.gov (United States)

    De Liberato, Claudio; Berrilli, Federica; Marangi, Marianna; Santoro, Maristella; Trogu, Tiziana; Putignani, Lorenza; Lanfranchi, Paolo; Ferretti, Francesco; D'Amelio, Stefano; Giangaspero, Annunziata

    2015-12-21

    Although chamois Rupicapra spp. are the most abundant mountain ungulates in Europe, no data are available on the presence of Giardia duodenalis infecting these species. A total of 157 fecal samples from Alpine Rupicapra rupicapra rupicapra and Apennine Rupicapra pyrenaica ornata chamois were tested for the presence of G. duodenalis by immunofluorescence test, quantitative Real Time PCR and end-point PCR for genotype characterization. G. duodenalis was detected in R. r. rupicapra and R. p. ornata, with a percentage value of 4.45 (5.82 and 1.85 %, respectively), and a cyst burden of up to 31,800 cysts/g of feces. Assemblages A/AI and E were identified in R. r. rupicapra and assemblage A/AIII in R. p. ornata. The present study represents the first record of Giardia duodenalis in Rupicapra spp., suggesting that these wild bovids can play an epidemiological role in environmental contamination and transmission of both zoonotic and non-zoonotic genotypes.

  18. Morphological and physiological characteristics of a virulent and zoonotic assemblage A Giardia duodenalis canine strain.

    Science.gov (United States)

    Coelho, Camila Henriques; Silva, Ana Carolina Carvalho; Costa, Adriana Oliveira; Fernandes, Ana Paula

    2017-10-01

    Giardiasis is an intestinal parasitosis that affects millions of people worldwide and is considered a zoonotic disease. Frequently in contact with humans, dogs are the main host involved in this zoonotic transmission. Here, we compared some aspects of Giardia duodenalis biology between two strains: a recently isolated dog strain (BHFC1) and a human reference strain (Portland-1). Growth curve analysis revealed that BHFC1 trophozoites multiply faster than the human isolate Portland-1 in axenic culture, but has a lower rate of cysts formation. Scanning electron microscopy revealed that BHFC1 trophozoites have the same conventional shape and morphological structures expected for G. duodenalis trophozoites, but presented a more prominent flange. For the best of our knowledge, this work is the first description of morphological aspects and encystation process of a G. duodenalis strain isolated from a dog. Since BHFC1 and Portland-1 have been maintained in axenic cultures for different periods of time, differences observed in growth, encystation rates and flange size may be attributed to adaptation of Portland-1 to axenic culture and lack of the environmental pressures. BHFC1 can be useful as tool for better understanding of Giardia duodenalis biology. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Characterization of Giardia duodenalis infections in dogs in Trinidad and Tobago.

    Science.gov (United States)

    Mark-Carew, Miguella P; Adesiyun, Abiodun A; Basu, Asoke; Georges, Karla A; Pierre, Theresa; Tilitz, Sophie; Wade, Susan E; Mohammed, Hussni O

    2013-09-01

    To our knowledge, the zoonotic potential of Giardia duodenalis has not been assessed in companion animals in Trinidad and Tobago. This report details the first attempt to evaluate the potential zoonotic risk of G. duodenalis in dogs and identify assemblages of G. duodenalis found in dog populations on both islands. Fecal samples were collected from free-roaming dogs and dogs at the Trinidad and Tobago Society for the Prevention of Cruelty to Animals from October 2010 to June 2011. A total of 168 samples were collected of which 104 samples were analyzed for the presence of G. duodenalis by PCR amplification of the ssu-rRNA gene with subsequent assemblage-typing. A subset of samples was also analyzed by ELISA. Twenty-six samples were positive for G. duodenalis by PCR for an overall prevalence of 25%. Four samples were identified as assemblage C (15.4%), 21 as assemblage D (80.8%), and one as assemblage E (3.8%). Puppies were four-times more likely to be infected with G. duodenalis than adult dogs (OR 4.61, 95% CI 1.73-12.2). There was a significant agreement between ELISA and PCR in the detection of the protozoa (κ=0.67). We infer from our results that while the prevalence of G. duodenalis is relatively high in Trinidad and Tobago, the zoonotic risk of infection in humans is low since neither assemblage A nor B was identified in the study population. Published by Elsevier B.V.

  20. Occurrence and diversity of Giardia duodenalis assemblages in livestock in the UK.

    Science.gov (United States)

    Minetti, C; Taweenan, W; Hogg, R; Featherstone, C; Randle, N; Latham, S M; Wastling, J M

    2014-12-01

    Giardia duodenalis is a common intestinal parasite in humans and a wide range of livestock species. It is a genetically heterogeneous parasite that has been characterized in seven distinct genetic assemblages or cryptic species, and molecular markers can be used to differentiate both animal-specific and potentially zoonotic genotypes. Little is known about G. duodenalis and the range of assemblages occurring in domestic livestock species in the UK. Here, we present data on the occurrence and molecular diversity of G. duodenalis detected in the faeces or large intestinal contents of cattle, sheep, pigs, goats and camelids from farms in the north-west of England. Both healthy and clinically diseased animals were included in the survey. The presence of Giardia spp. and assemblages was determined by sequencing of the small-subunit ribosomal RNA gene. The potential association of infection with various clinical and epidemiological parameters was studied in cattle using both univariate and multivariate analyses. Giardia spp. were detected in 127 (34.3%) of the 370 animals tested. G. duodenalis assemblage E was found to be predominant in cattle and sheep, followed by assemblage A. Mixed infections with assemblages A and E were also detected. Interestingly, some cattle, sheep and pigs were found to be infected with more unexpected assemblages (C, D, F). Pre-weaned calves were more likely to test positive than adult animals, but no association between the occurrence of overt intestinal disease and G. duodenalis infection was detected. The common occurrence of assemblage A and the finding of unusual assemblages in atypical hosts suggest that in future, a multilocus analysis should be used to confirm the actual diversity of G. duodenalis in livestock and the presence of potentially zoonotic genotypes. These data also suggest that there is a need to re-evaluate the clinical significance of G. duodenalis infection in livestock. © 2014 The Authors. Transboundary and

  1. Burden of Giardia duodenalis infection and its adverse effects on growth of schoolchildren in rural Malaysia.

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    Hesham M Al-Mekhlafi

    Full Text Available BACKGROUND: Giardia duodenalis infection and malnutrition are still considered as public health problems in many developing countries especially among children in rural communities. This study was carried out among Aboriginal (Orang Asli primary schoolchildren in rural peninsular Malaysia to investigate the burden and the effects of Giardia infection on growth (weight and height of the children. METHODS/FINDINGS: Weight and height of 374 children aged 7-12 years were assessed before and after treatment of Giardia infection. The children were screened for Giardia parasite using trichrome staining technique. Demographic and socioeconomic data were collected via face-to-face interviews using a pre-tested questionnaire. Overall, 22.2% (83/374 of the children were found to be infected with Giardia. Nutritional status of children was assessed and the results showed that the mean weight and height were 23.9 kg (95% CI = 23.3, 24.5 and 126.6 cm (95% CI = 125.6, 127.5, respectively. Overall, the prevalence of severe underweight, stunting and wasting were 28.3%, 23.8% and 21.0%, respectively. Multiple linear regression analyses showed sex, Giardia infection and household monthly income as the significant determinants of weight while sex and level of mother's education were the significant determinants of height. Weight and height were assessed at 3 and 6 months after treatment of Giardia infection. It was found that Giardia infection has a significant association with the weight of children but not with height. CONCLUSIONS/SIGNIFICANCE: This study reveals high prevalence of Giardia infection and malnutrition among Aboriginal children in rural Malaysia and clearly highlights an urgent need to identify integrated measures to control these health problems in the rural communities. Essentially, proper attention should be given to the control of Giardia infection in Aboriginal communities as this constitutes one of the strategies to improve the

  2. Burden of Giardia duodenalis Infection and Its Adverse Effects on Growth of Schoolchildren in Rural Malaysia

    Science.gov (United States)

    Al-Mekhlafi, Hesham M.; Al-Maktari, Mohamed T.; Jani, Rohana; Ahmed, Abdulhamid; Anuar, Tengku Shahrul; Moktar, Norhayati; Mahdy, Mohammed A. K.; Lim, Yvonne A. L.; Mahmud, Rohela; Surin, Johari

    2013-01-01

    Background Giardia duodenalis infection and malnutrition are still considered as public health problems in many developing countries especially among children in rural communities. This study was carried out among Aboriginal (Orang Asli) primary schoolchildren in rural peninsular Malaysia to investigate the burden and the effects of Giardia infection on growth (weight and height) of the children. Methods/Findings Weight and height of 374 children aged 7–12 years were assessed before and after treatment of Giardia infection. The children were screened for Giardia parasite using trichrome staining technique. Demographic and socioeconomic data were collected via face-to-face interviews using a pre-tested questionnaire. Overall, 22.2% (83/374) of the children were found to be infected with Giardia. Nutritional status of children was assessed and the results showed that the mean weight and height were 23.9 kg (95% CI = 23.3, 24.5) and 126.6 cm (95% CI = 125.6, 127.5), respectively. Overall, the prevalence of severe underweight, stunting and wasting were 28.3%, 23.8% and 21.0%, respectively. Multiple linear regression analyses showed sex, Giardia infection and household monthly income as the significant determinants of weight while sex and level of mother's education were the significant determinants of height. Weight and height were assessed at 3 and 6 months after treatment of Giardia infection. It was found that Giardia infection has a significant association with the weight of children but not with height. Conclusions/Significance This study reveals high prevalence of Giardia infection and malnutrition among Aboriginal children in rural Malaysia and clearly highlights an urgent need to identify integrated measures to control these health problems in the rural communities. Essentially, proper attention should be given to the control of Giardia infection in Aboriginal communities as this constitutes one of the strategies to improve the nutritional status of

  3. Burden of Giardia duodenalis infection and its adverse effects on growth of schoolchildren in rural Malaysia.

    Science.gov (United States)

    Al-Mekhlafi, Hesham M; Al-Maktari, Mohamed T; Jani, Rohana; Ahmed, Abdulhamid; Anuar, Tengku Shahrul; Moktar, Norhayati; Mahdy, Mohammed A K; Lim, Yvonne A L; Mahmud, Rohela; Surin, Johari

    2013-01-01

    Giardia duodenalis infection and malnutrition are still considered as public health problems in many developing countries especially among children in rural communities. This study was carried out among Aboriginal (Orang Asli) primary schoolchildren in rural peninsular Malaysia to investigate the burden and the effects of Giardia infection on growth (weight and height) of the children. Weight and height of 374 children aged 7-12 years were assessed before and after treatment of Giardia infection. The children were screened for Giardia parasite using trichrome staining technique. Demographic and socioeconomic data were collected via face-to-face interviews using a pre-tested questionnaire. Overall, 22.2% (83/374) of the children were found to be infected with Giardia. Nutritional status of children was assessed and the results showed that the mean weight and height were 23.9 kg (95% CI = 23.3, 24.5) and 126.6 cm (95% CI = 125.6, 127.5), respectively. Overall, the prevalence of severe underweight, stunting and wasting were 28.3%, 23.8% and 21.0%, respectively. Multiple linear regression analyses showed sex, Giardia infection and household monthly income as the significant determinants of weight while sex and level of mother's education were the significant determinants of height. Weight and height were assessed at 3 and 6 months after treatment of Giardia infection. It was found that Giardia infection has a significant association with the weight of children but not with height. This study reveals high prevalence of Giardia infection and malnutrition among Aboriginal children in rural Malaysia and clearly highlights an urgent need to identify integrated measures to control these health problems in the rural communities. Essentially, proper attention should be given to the control of Giardia infection in Aboriginal communities as this constitutes one of the strategies to improve the nutritional status of Aboriginal children.

  4. Giardia duodenalis Surface Cysteine Proteases Induce Cleavage of the Intestinal Epithelial Cytoskeletal Protein Villin via Myosin Light Chain Kinase.

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    Amol Bhargava

    Full Text Available Giardia duodenalis infections are among the most common causes of waterborne diarrhoeal disease worldwide. At the height of infection, G. duodenalis trophozoites induce multiple pathophysiological processes within intestinal epithelial cells that contribute to the development of diarrhoeal disease. To date, our understanding of pathophysiological processes in giardiasis remains incompletely understood. The present study reveals a previously unappreciated role for G. duodenalis cathepsin cysteine proteases in intestinal epithelial pathophysiological processes that occur during giardiasis. Experiments first established that Giardia trophozoites indeed produce cathepsin B and L in strain-dependent fashion. Co-incubation of G. duodenalis with human enterocytes enhanced cathepsin production by Assemblage A (NF and S2 isolates trophozoites, but not when epithelial cells were exposed to Assemblage B (GSM isolate trophozoites. Direct contact between G. duodenalis parasites and human intestinal epithelial monolayers resulted in the degradation and redistribution of the intestinal epithelial cytoskeletal protein villin; these effects were abolished when parasite cathepsin cysteine proteases were inhibited. Interestingly, inhibition of parasite proteases did not prevent degradation of the intestinal tight junction-associated protein zonula occludens 1 (ZO-1, suggesting that G. duodenalis induces multiple pathophysiological processes within intestinal epithelial cells. Finally, this study demonstrates that G. duodenalis-mediated disruption of villin is, at least, in part dependent on activation of myosin light chain kinase (MLCK. Taken together, this study indicates a novel role for parasite cathepsin cysteine proteases in the pathophysiology of G. duodenalis infections.

  5. Giardia duodenalis Surface Cysteine Proteases Induce Cleavage of the Intestinal Epithelial Cytoskeletal Protein Villin via Myosin Light Chain Kinase.

    Science.gov (United States)

    Bhargava, Amol; Cotton, James A; Dixon, Brent R; Gedamu, Lashitew; Yates, Robin M; Buret, Andre G

    2015-01-01

    Giardia duodenalis infections are among the most common causes of waterborne diarrhoeal disease worldwide. At the height of infection, G. duodenalis trophozoites induce multiple pathophysiological processes within intestinal epithelial cells that contribute to the development of diarrhoeal disease. To date, our understanding of pathophysiological processes in giardiasis remains incompletely understood. The present study reveals a previously unappreciated role for G. duodenalis cathepsin cysteine proteases in intestinal epithelial pathophysiological processes that occur during giardiasis. Experiments first established that Giardia trophozoites indeed produce cathepsin B and L in strain-dependent fashion. Co-incubation of G. duodenalis with human enterocytes enhanced cathepsin production by Assemblage A (NF and S2 isolates) trophozoites, but not when epithelial cells were exposed to Assemblage B (GSM isolate) trophozoites. Direct contact between G. duodenalis parasites and human intestinal epithelial monolayers resulted in the degradation and redistribution of the intestinal epithelial cytoskeletal protein villin; these effects were abolished when parasite cathepsin cysteine proteases were inhibited. Interestingly, inhibition of parasite proteases did not prevent degradation of the intestinal tight junction-associated protein zonula occludens 1 (ZO-1), suggesting that G. duodenalis induces multiple pathophysiological processes within intestinal epithelial cells. Finally, this study demonstrates that G. duodenalis-mediated disruption of villin is, at least, in part dependent on activation of myosin light chain kinase (MLCK). Taken together, this study indicates a novel role for parasite cathepsin cysteine proteases in the pathophysiology of G. duodenalis infections.

  6. Prevalence of Cryptosporidium spp. and Giardia duodenalis in pigs in Lusaka, Zambia

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    Joyce Siwila

    2012-02-01

    Full Text Available This study was aimed at determining the prevalence of Cryptosporidium spp. and Giardia duodenalis in pigs which were being raised in intensive management systems. Faecal samples were collected from pigs of all age groups from three different piggery units. Samples were collected directly from the rectum for piglets and weaners and from the floor within 2 min – 5 min of excretion for sows and boars. At the time of collection, faecal consistency was noted as being normal, pasty or diarrhoeic. Samples were analysed further using the Merifluor® Cryptosporidium/Giardia immunofluorescence assay. All piggeries had at least one pig infected with either parasite. From a total 217 samples collected, 96 (44.2%; confidence interval [CI] = 37.6% – 50.9% were positive for Cryptosporidium spp., whilst 26 (12%; CI = 7.6% – 16.3% had G. duodenalis parasites. Of all the pigs, 6.9% (15/217 harboured both parasites. With regard to Cryptosporidium spp. infection, statistically significant differences were observed amongst the three units (p = 0.001, whereas no significant differences were observed for G. duodenalis infection (p = 0.13. Prevalence was higher in weaners as compared to other pig classes for both parasites, with significant differences being observed for G. duodenalis infection (p = 0.013. There was, however, no difference in infection between male and female pigs for both parasites. Furthermore, most infections were asymptomatic. From the study results it was clear that Cryptosporidium spp. and G. duodenalis infections were prevalent amongst pigs in the piggeries evaluated and, as such, may act as a source of infection for persons who come into contact with them.

  7. Encystation commitment in Giardia duodenalis: a long and winding road

    Directory of Open Access Journals (Sweden)

    Argüello-Garciá R.

    2009-12-01

    Full Text Available Cholesterol and bile salts are relevant modulators of Giardia encystation. Although several molecules within signaling cascades have been identified, and changes in their expression observed during giardial encystation, their underlying interactions leading to expression of cyst wall markers (CWPs and precursors of the GalNAc homopolymer are not well defined. Recent experimental data and the completion of the Giardia Genome Project Database (GiardiaDB allow us now to consider the role of bile salts as “natural stimuli” and the potential involvement of a Raf/MEK/ERK pathway mediating cholesterol-regulated expression of cyst-specific genes. These new findings may provide promising targets for diagnostics, drug design and prophylactic intervention against giardiasis.

  8. Fate of Cryptosporidium parvum and Cryptosporidium hominis oocysts and Giardia duodenalis cysts during secondary wastewater treatments.

    Science.gov (United States)

    Cheng, Hui-Wen A; Lucy, Frances E; Graczyk, Thaddeus K; Broaders, Michael A; Tamang, Leena; Connolly, Michelle

    2009-09-01

    This study investigates the fate of Cryptosporidium parvum and C. hominis oocysts and Giardia duodenalis cysts at four Irish municipal wastewater treatment plants (i.e., Plant A, B, C, and D) that utilize sludge activation or biofilm-coated percolating filter systems for secondary wastewater treatment. The fate of these pathogens through the sewage treatment processes was determined based on their viable transmissive stages, i.e., oocysts for Cryptosporidium and cysts for Giardia. Analysis of final effluent indicated that over 97% of viable oocysts and cysts were eliminated, except at Plant C, which achieved only 64% of oocyst removal. A significant correlation between the removal of oocysts and cysts was found at Plants A, B, and D (R = 0.98, P hominis, and G. duodenalis, with maximum concentrations of 20 oocysts and eight cysts per gram in primary sludge indicating the need for further sludge sanitization treatments. This study provides evidence that C. parvum and C. hominis oocysts and G. duodenalis cysts are present throughout the wastewater processes and in end-products, and can enter the aquatic environment with consequent negative implications for public health.

  9. Occurency of Giardia duodenalis assemblages in river water sources of Black Sea, Turkey.

    Science.gov (United States)

    Koloren, Zeynep; Seferoğlu, Onuralp; Karanis, Panagiotis

    2016-12-01

    A total of 420 environmental water samples and 120 drinking water samples from 45 different sampling sites of the Black Sea in Turkey were collected between 2012 and 2014. Genomic DNA was isolated from all the investigated water samples and comparativelly analyzed by Loop-mediated isothermal amplification (LAMP) of the elongation factor 1 Alfa (EF1α) gene, and by nested Polymerase Chain Reaction (nPCR) of the small subunit (SSU) rRNA and semi-nested PCR (snPCR) of the glutamate dehydrogenase gene (GDH). 141 (58.7%), 125 (52.1%) and 120 (50%) samples respectivelly were positive by each method. Out of 240 environmental samples collected from 25 sites of Samsun Province have been found positive for G. duodenalis by LAMP, nPCR and snPCR, respectively. 55 (30.5%), 50 (27.8%) and 47 (26.1%) of 180 environmental samples collected from 20 other sampling sites of Giresun Province were positive for Giardia by LAMP, nPCR and snPCR, respectively. Five PCR products from different samples of the Giresun Province and 10 other samples from the Samsun Province were found positive for G. duodenalis assemblage B. Five PCR products from Giresun Province and 5 samples from Samsun Province were found positive for G. duodenalis assemblage A. This is the first report about G. duodenalis assemblages A and B from water samples investigations in Black Sea of Turkey. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Differentiation of Giardia duodenalis from other Giardia spp. by using polymerase chain reaction and gene probes.

    OpenAIRE

    Mahbubani, M H; Bej, A K; Perlin, M H; Schaefer, F W; Jakubowski, W; Atlas, R M

    1992-01-01

    Giardia spp. are waterborne organisms that are the most commonly identified pathogenic intestinal protozoans in the United States. Current detection techniques for Giardia species in water include microscopy and immunofluorescence techniques. Species of the genus Giardia are classified on the basis of taxonomic criteria, such as cell morphology, and on host specificity. We have developed a polymerase chain reaction- and gene probe-based detection system specific for Giardia spp., which can di...

  11. Detection of zoonotic and livestock-specific assemblages of Giardia duodenalis in free-living wild lizards

    Directory of Open Access Journals (Sweden)

    Aurora Reboredo-Fernández

    Full Text Available Abstract Giardia duodenalis is a zoonotic parasite that infects the gut of a wide range of vertebrates, including numerous wildlife species. However, little is known about this protozoan parasite in reptiles. Fecal samples from 31 wild lizards were collected in Galicia (northwest Spain and screened for the presence of Giardia by PCR amplification and sequencing of the ITS1-5.8S-ITS2 region in the ribosomal unit. This allowed detection of the parasite in 5 samples (16.1%, and enabled identification of G. duodenalis assemblage A2 in two samples of Iberian rock lizard (Iberolacerta monticola, G. duodenalis assemblage B in other two samples of I. monticola, and G. duodenalis assemblage E in one sample of Bocage’s wall lizard (Podarcis bocagei. The results obtained after PCR amplification and sequencing of the SSU-rDNA gene confirmed the presence of G. duodenalis assemblage A in two samples of I. monticola. This is the first report of G. duodenalis in free-living lizards, although further studies are needed to distinguish between actual infection and mechanical dissemination of cysts. The detection of zoonotic and livestock-specific assemblages of G. duodenalis demonstrates the wide environmental contamination by this parasite, possibly due to human activities.

  12. Giardia duodenalis induces paracellular bacterial translocation and causes postinfectious visceral hypersensitivity

    Science.gov (United States)

    Halliez, Marie C. M.; Motta, Jean-Paul; Feener, Troy D.; Guérin, Gaetan; LeGoff, Laetitia; François, Arnaud; Colasse, Elodie; Favennec, Loic; Gargala, Gilles; Lapointe, Tamia K.; Altier, Christophe

    2016-01-01

    Irritable bowel syndrome (IBS) is the most frequent functional gastrointestinal disorder. It is characterized by abdominal hypersensitivity, leading to discomfort and pain, as well as altered bowel habits. While it is common for IBS to develop following the resolution of infectious gastroenteritis [then termed postinfectious IBS (PI-IBS)], the mechanisms remain incompletely understood. Giardia duodenalis is a cosmopolitan water-borne enteropathogen that causes intestinal malabsorption, diarrhea, and postinfectious complications. Cause-and-effect studies using a human enteropathogen to help investigate the mechanisms of PI-IBS are sorely lacking. In an attempt to establish causality between giardiasis and postinfectious visceral hypersensitivity, this study describes a new model of PI-IBS in neonatal rats infected with G. duodenalis. At 50 days postinfection with G. duodenalis (assemblage A or B), long after the parasite was cleared, rats developed visceral hypersensitivity to luminal balloon distension in the jejunum and rectum, activation of the nociceptive signaling pathway (increased c-fos expression), histological modifications (villus atrophy and crypt hyperplasia), and proliferation of mucosal intraepithelial lymphocytes and mast cells in the jejunum, but not in the rectum. G. duodenalis infection also disrupted the intestinal barrier, in vivo and in vitro, which in turn promoted the translocation of commensal bacteria. Giardia-induced bacterial paracellular translocation in vitro correlated with degradation of the tight junction proteins occludin and claudin-4. The extensive observations associated with gut hypersensitivity described here demonstrate that, indeed, in this new model of postgiardiasis IBS, alterations to the gut mucosa and c-fos are consistent with those associated with PI-IBS and, hence, offer avenues for new mechanistic research in the field. PMID:26744469

  13. Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children.

    Directory of Open Access Journals (Sweden)

    Catrin E Moore

    2016-07-01

    Full Text Available In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays.Cryptosporidium oocysts were detected in 2.2% (11/498 of samples using microscopy and in 7.7% (38/498 with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498 by microscopy and 27.7% (138/498 by molecular tests; 82% of the positive samples (by either method were from children aged 1-10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2% samples, followed by Cryptosporidium meleagridis in 9 (23.7%, Cryptosporidium parvum in 8 (21.1%, Cryptosporidium canis in 5 (13.2%, and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples was most common followed by A (12.3% and mixed infections (5.1%. Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67-55.46, chronic medical diagnoses (AOR 4.51, 95% CI 1.79-11.34 and the presence of birds in the household (AOR 2.99, 95% CI 1.16-7.73; specifically C. hominis (p = 0.03 and C. meleagridis (p<0.001 were associated with the presence of birds. The use of soap was protective against Giardia infection (OR 0.74, 95% CI 0.58-0.95.This is the first report to describe the different Cryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both anthroponotic and zoonotic transmission in this population. Interventions to improve

  14. Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

    Science.gov (United States)

    Moore, Catrin E.; Elwin, Kristin; Phot, Nget; Seng, Chanthou; Mao, Saroeun; Suy, Kuong; Kumar, Varun; Nader, Johanna; Bousfield, Rachel; Perera, Sanuki; Bailey, J. Wendi; Beeching, Nicholas J.; Day, Nicholas P. J.; Parry, Christopher M.; Chalmers, Rachel M.

    2016-01-01

    Background In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays. Methodology/Principal Findings Cryptosporidium oocysts were detected in 2.2% (11/498) of samples using microscopy and in 7.7% (38/498) with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498) by microscopy and 27.7% (138/498) by molecular tests; 82% of the positive samples (by either method) were from children aged 1–10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2%) samples, followed by Cryptosporidium meleagridis in 9 (23.7%), Cryptosporidium parvum in 8 (21.1%), Cryptosporidium canis in 5 (13.2%), and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples) was most common followed by A (12.3%) and mixed infections (5.1%). Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67–55.46), chronic medical diagnoses (AOR 4.51, 95% CI 1.79–11.34) and the presence of birds in the household (AOR 2.99, 95% CI 1.16–7.73); specifically C. hominis (p = 0.03) and C. meleagridis (p<0.001) were associated with the presence of birds. The use of soap was protective against Giardia infection (OR 0.74, 95% CI 0.58–0.95). Conclusions/Significance This is the first report to describe the different Cryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both

  15. Prevalence of Giardia Duodenalis Infection in Household Cats of Ahvaz District, South-West of Iran

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    AR Alborzi

    2010-09-01

    Full Text Available Background: The occurrence of Giardia duodenalis in cats is of potential significance from both clinical and public health perspectives. The object of this study was antigenic detection of G. duo­denalis in house­hold cats of Ahvaz district, South-West of Iran.Methods: The prevalence of G. duodenalis was determined in fecal samples by two techniques: centrifuga­tion-flotation and a commercial Giardia Antigen Test Kit (immunochromatography assay in 150 household cats of different ages among referred cases to Veterinary Hospital of Ahvaz University from January 2008 to February 2010.Results: Five out of 150 fecal samples (3.33% were positive for antigen of G. duodenalis by immunochromatography assay. The prevalence was significantly higher in young cats less than 6 months (15.79% compared with adult cats 6 months - 3 years (1.37% (P=0.027 and above 3 years (1.72% (P=0.044. The infection had more prevalence in diarrheic cats (17.39% com­pared with non-diarrheic cats (0.79% and the difference was significant (P=0.02 as well. The prevalence was higher in male cats (3.41% than females (3.23% and in the season of autumn (6.06%, but the difference was not significant between the prevalence of infection relative to host gender and season (P>0.05. Microscopy examination on fecal samples showed that 2% of the studied cats were positive.Conclusion: The parasite antigen was present as a zoonotic infection in Ahvaz district, South-west of Iran. More sensitive techniques, such as immunochromatography assay, might yield more reli­able results, in the detection of low levels of Giardia in fecal samples of cats.

  16. Evaluation of soil homogenization techniques for detection of Giardia duodenalis cysts

    OpenAIRE

    Oliveira, Clarisse M. Boni de; Franco, Regina M Bueno

    2012-01-01

    O solo é uma das vias de transmissão do protozoário Giardia duodenalis, que pode causar a doença global giardiose, gastroenterite de severidade variável. O objetivo deste trabalho foi avaliar diferentes formas de homogeneização de amostras de solo visando saber qual delas é mais eficiente para detecção e recuperação de cistos. Foram realizados quatro tratamentos de homogeneização com quatro réplicas cada. Cada réplica continha 10 g de solo contaminado com uma concentração 500 cistos, umedecid...

  17. Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children.

    Science.gov (United States)

    Moore, Catrin E; Elwin, Kristin; Phot, Nget; Seng, Chanthou; Mao, Saroeun; Suy, Kuong; Kumar, Varun; Nader, Johanna; Bousfield, Rachel; Perera, Sanuki; Bailey, J Wendi; Beeching, Nicholas J; Day, Nicholas P J; Parry, Christopher M; Chalmers, Rachel M

    2016-07-01

    In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays. Cryptosporidium oocysts were detected in 2.2% (11/498) of samples using microscopy and in 7.7% (38/498) with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498) by microscopy and 27.7% (138/498) by molecular tests; 82% of the positive samples (by either method) were from children aged 1-10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2%) samples, followed by Cryptosporidium meleagridis in 9 (23.7%), Cryptosporidium parvum in 8 (21.1%), Cryptosporidium canis in 5 (13.2%), and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples) was most common followed by A (12.3%) and mixed infections (5.1%). Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67-55.46), chronic medical diagnoses (AOR 4.51, 95% CI 1.79-11.34) and the presence of birds in the household (AOR 2.99, 95% CI 1.16-7.73); specifically C. hominis (p = 0.03) and C. meleagridis (pCryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both anthroponotic and zoonotic transmission in this population. Interventions to improve sanitation, increase hand washing after defecation and before preparing food and promote drinking boiled water may reduce the burden of these two parasites.

  18. Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

    Science.gov (United States)

    Moreira, Maria João; Soares, Sónia; de Lurdes Delgado, Maria; Figueiredo, João; Silva, Elisabete; Castro, António; Cosa, José Manuel Correida Da

    2010-01-01

    Cryptosporidium and Giardia are 2 protozoan parasites responsible for waterborne diseases outbreaks worldwide. In order to assess the prevalence of these protozoans in drinking water samples in the northern part of Portugal and the risk of human infection, we have established a long term program aiming at pinpointing the sources of surface water, drinking water, and environmental contamination, working with the water-supply industry. Total 43 sources of drinking water samples were selected, and a total of 167 samples were analyzed using the Method 1623. Sensitivity assays regarding the genetic characterization by PCR and sequencing of the genes, 18S SSU rRNA, for Cryptosporidium spp. and β,-giardin for G. duodenalis were set in the laboratory. According to the defined criteria, molecular analysis was performed over 4 samples. Environmental stages of the protozoa were detected in 25.7% (43 out of 167) of the water samples, 8.4% (14 out of 167) with cysts of Giardia, 10.2% (17 out of 167) with oocysts of Cryptosporidium and 7.2% (12 out of 167) for both species. The mean concentrations were 0.1-12.7 oocysts of Cryptosporidium spp. per 10 L and 0.1-108.3 cysts of Giardia duodenalis per 10 L. Our results suggest that the efficiency in drinking water plants must be ameliorated in their efficiency in reducing the levels of contamination. We suggest the implementation of systematic monitoring programs for both protozoa. To authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in drinking water samples in the northern part of Portugal. PMID:20333284

  19. Presence of Cryptosporidium spp. and Giardia duodenalis in drinking water samples in the north of Portugal.

    Science.gov (United States)

    Almeida, André; Moreira, Maria João; Soares, Sónia; Delgado, Maria de Lurdes; Figueiredo, João; Silva, Elisabete; Castro, António; Cosa, José Manuel Correida Da

    2010-03-01

    Cryptosporidium and Giardia are 2 protozoan parasites responsible for waterborne diseases outbreaks worldwide. In order to assess the prevalence of these protozoans in drinking water samples in the northern part of Portugal and the risk of human infection, we have established a long term program aiming at pinpointing the sources of surface water, drinking water, and environmental contamination, working with the water-supply industry. Total 43 sources of drinking water samples were selected, and a total of 167 samples were analyzed using the Method 1623. Sensitivity assays regarding the genetic characterization by PCR and sequencing of the genes, 18S SSU rRNA, for Cryptosporidium spp. and beta,-giardin for G. duodenalis were set in the laboratory. According to the defined criteria, molecular analysis was performed over 4 samples. Environmental stages of the protozoa were detected in 25.7% (43 out of 167) of the water samples, 8.4% (14 out of 167) with cysts of Giardia, 10.2% (17 out of 167) with oocysts of Cryptosporidium and 7.2% (12 out of 167) for both species. The mean concentrations were 0.1-12.7 oocysts of Cryptosporidium spp. per 10 L and 0.1-108.3 cysts of Giardia duodenalis per 10 L. Our results suggest that the efficiency in drinking water plants must be ameliorated in their efficiency in reducing the levels of contamination. We suggest the implementation of systematic monitoring programs for both protozoa. To authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in drinking water samples in the northern part of Portugal.

  20. Molecular detection and prevalence of Giardia duodenalis and Cryptosporidium spp. among long-tailed macaques (Macaca fascicularis) in Thailand.

    Science.gov (United States)

    Sricharern, Wanat; Inpankaew, Tawin; Keawmongkol, Sarawan; Supanam, Juthamas; Stich, Roger W; Jittapalapong, Sathaporn

    2016-06-01

    Giardia duodenalis and Cryptosporidium spp. are divergent protozoal intestinal parasites that infect human beings and other animals, including non-human primates. Although long-tailed macaques (Macaca fascicularis) reside in human communities in Thailand, the prevalence of Giardia spp. and Cryptosporidium spp. in these primates has not been previously investigated. The objective of this study was to evaluate long-tailed macaques living near human communities as possible hosts of these intestinal parasites. In 2014, 200 fecal samples were randomly collected from long-tailed macaques living in different areas of Lopburi province, Thailand, and tested with a panel of PCR assays for Giardia spp. and Cryptosporidium spp. G. duodenalis assemblage B was most frequently detected (6%), while assemblage A and an inconclusive assemblage were detected in single samples, for a total G. duodenalis infection rate of 7%. Two samples (1%) tested positive for Cryptosporidium spp., which were both classified as monkey genotypes. No significant associations were found between G. duodenalis infection and sex or location of macaques. This study indicates that long-tailed macaques can carry G. duodenalis and, to a lesser extent, Cryptosporidium spp. monkey genotype. These results warrant education of residents and tourists to limit contact with long-tailed macaques and to take hygienic precautions to mitigate risk of zoonotic and anthroponotic transmission of these parasites between people and macaques. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Introducing a simple and economical method to purify Giardia ...

    African Journals Online (AJOL)

    To prepare anti-Giardia lamblia antibodies needed to design diagnostic kits as well as parasite culture and other molecular studies, we require purification of the parasite cysts. In this study, we designed a rapid, simple and inexpensive method to purify parasite cysts from fecal samples of the patients suffering from ...

  2. Coinfection with Tritrichomonas foetus and Giardia duodenalis in Two Cats with Chronic Diarrhea

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    Sergio A. Zanzani

    2016-01-01

    Full Text Available A Tritrichomonas foetus and Giardia duodenalis mixed infection was diagnosed in two Maine Coon cats aged six months. One of them presented a history of chronic liquid diarrhea and of several unsuccessful treatments. In both cats, G. duodenalis and trichomonads were detected in fecal smears from freshly voided feces; the presence of T. foetus was confirmed by a real-time PCR assay. The cats completely recovered after treatment with ronidazole. In a refrigerated fecal sample collected from the cat with chronic diarrhea, drop-shaped trichomonad pseudocysts smaller than G. duodenalis cysts were detected. They appeared brownish or light-bluish when stained with Lugol’s solution or with Giemsa stain, respectively, and their morphological features were similar to those expressed by bovine T. foetus pseudocysts in vitro. Existence of pseudocysts even in feline trichomonads is noteworthy as they could represent a form of protozoan resistance due to unfavorable conditions whose detection in refrigerated feces can be a useful clue for clinicians.

  3. Prevalence and molecular typing of Giardia duodenalis in wildlife from eastern Poland.

    Science.gov (United States)

    Stojecki, Krzysztof; Sroka, Jacek; Caccio, Simone M; Cencek, Tomasz; Dutkiewicz, Jacek; Kusyk, Pawel

    2015-07-30

    Faecal samples from 162 wild animals were collected from 32 distinct sites of Łęczyńsko-Włodawskie Lakeland (eastern Poland). The presence of Giardia duodenalis (Stiles, 1902) was assessed by a Direct Fluorescence Assay (DFA) and by Polymerase Chain Reaction (PCR) and sequencing of a fragment of the beta-giardin gene. DFA showed the presence of cysts of G. duodenalis in 12 of 162 faecal samples (7%), namely in four wild boars (15%), four foxes (19%), two roe deer (4%), and two wolves (29%). PCR identified 34 of the 162 (21%) samples as positive, including 11 wild boars (41%), five red deer (18%), 11 roe deer (23%), four moose (17%), two wolves (29%) and a single sample from the European badger. Thus, PCR detected a significantly higher number of infection than DFA (P = 0.0005). However, 14 of 34 PCR products could not be sequenced because of their insufficient amount; the low number of cysts, poor conservation of the faeces or presence of PCR inhibitors may have contributed to weak DNA amplification. Sequence analysis of the remaining 20 products showed the presence of assemblage B in wild boars, red deer and roe deer, whereas samples from wolves were identified as assemblage D. This is the first detection of assemblage B in wild boars and deer. As assemblage B has zoonotic potential, wild animals from eastern Poland may act as reservoirs of cysts of G. duodenalis infectious for humans.

  4. First genotyping of Giardia duodenalis and prevalence of enteroparasites in children from Tetouan (Morocco

    Directory of Open Access Journals (Sweden)

    El Fatni Chadia

    2014-01-01

    Full Text Available Intestinal parasites are common in the Moroccan population. Enteroparasites in children from four schools in urban and rural areas of Tetouan (Morocco were studied to treat these children and to design prevention and control programs. A total of 673 children were examined. The prevalence of parasitized children was 51%. The average number of enteroparasites was half in urban areas than in rural areas. Multiple parasitism appeared in 30% of the samples presenting two, three, or four parasites. The most prevalent parasite was Blastocystis hominis (64%. Giardia duodenalis was the most frequent pathogen, with an overall prevalence of 20% (24% in rural areas and 16% in urban areas. Other pathogenic enteroparasites were Cyclospora cayetanensis (5% in rural and urban areas, Iodamoeba butschlii, Hymenolepis spp., Trichuris trichiura and Enterobius vermicularis, with prevalence lower than 2%. In this work, G. duodenalis genotypes were molecularly characterized by a study of the glutamate dehydrogenase (gdh and 18S rRNA genes. This is the first study of molecular characterization of G. duodenalis in Moroccan children, and the sequence analysis revealed both Assemblage A (AII and Assemblage B (BIII, BIV, with the predominance of Assemblage BIV (73%.

  5. First genotyping of Giardia duodenalis and prevalence of enteroparasites in children from Tetouan (Morocco).

    Science.gov (United States)

    El Fatni, Chadia; Olmo, Francisco; El Fatni, Hoummad; Romero, Desiré; Rosales, Maria José

    2014-01-01

    Intestinal parasites are common in the Moroccan population. Enteroparasites in children from four schools in urban and rural areas of Tetouan (Morocco) were studied to treat these children and to design prevention and control programs. A total of 673 children were examined. The prevalence of parasitized children was 51%. The average number of enteroparasites was half in urban areas than in rural areas. Multiple parasitism appeared in 30% of the samples presenting two, three, or four parasites. The most prevalent parasite was Blastocystis hominis (64%). Giardia duodenalis was the most frequent pathogen, with an overall prevalence of 20% (24% in rural areas and 16% in urban areas). Other pathogenic enteroparasites were Cyclospora cayetanensis (5% in rural and urban areas), Iodamoeba butschlii, Hymenolepis spp., Trichuris trichiura and Enterobius vermicularis, with prevalence lower than 2%. In this work, G. duodenalis genotypes were molecularly characterized by a study of the glutamate dehydrogenase (gdh) and 18S rRNA genes. This is the first study of molecular characterization of G. duodenalis in Moroccan children, and the sequence analysis revealed both Assemblage A (AII) and Assemblage B (BIII, BIV), with the predominance of Assemblage BIV (73%). C. El Fatni et al., published by EDP Sciences, 2014

  6. Extra-intestinal and long term consequences of Giardia duodenalis infections.

    Science.gov (United States)

    Halliez, Marie C M; Buret, André G

    2013-12-21

    Giardiasis is the most common waterborne parasitic infection of the human intestine worldwide. The etiological agent, Giardia duodenalis (syn. G. intestinalis, G. lamblia), is a flagellated, binucleated protozoan parasite which infects a wide array of mammalian hosts. Human giardiasis is a true cosmopolitan pathogen, with highest prevalence in developing countries. Giardiasis can present with a broad range of clinical manifestations from asymptomatic, to acute or chronic diarrheal disease associated with abdominal pain and nausea. Most infections are self-limiting, although re-infection and chronic infection can occur. Recent evidence indicating that Giardia may cause chronic post-infectious gastrointestinal complications have made it a topic of intense research. The causes of the post-infectious clinical manifestations due to Giardia, even after complete elimination of the parasite, remain obscure. This review offers a state-of-the-art discussion on the long-term consequences of Giardia infections, from extra-intestinal manifestations, growth and cognitive deficiencies, to post-infectious irritable bowel syndrome. The discussion also sheds light on some of the novel mechanisms recently implicated in the production of these post-infectious manifestations.

  7. Giardia duodenalis cathepsin B proteases degrade intestinal epithelial interleukin-8 and attenuate interleukin-8-induced neutrophil chemotaxis.

    Science.gov (United States)

    Cotton, James A; Bhargava, Amol; Ferraz, Jose G; Yates, Robin M; Beck, Paul L; Buret, Andre G

    2014-07-01

    Giardia duodenalis (syn. G. intestinalis, G. lamblia) infections are a leading cause of waterborne diarrheal disease that can also result in the development of postinfectious functional gastrointestinal disorders via mechanisms that remain unclear. Parasite numbers exceed 10(6) trophozoites per centimeter of gut at the height of an infection. Yet the intestinal mucosa of G. duodenalis-infected individuals is devoid of signs of overt inflammation. G. duodenalis infections can also occur concurrently with infections with other proinflammatory gastrointestinal pathogens. Little is known of whether and how this parasite can attenuate host inflammatory responses induced by other proinflammatory stimuli, such as a gastrointestinal pathogen. Identifying hitherto-unrecognized parasitic immunomodulatory pathways, the present studies demonstrated that G. duodenalis trophozoites attenuate secretion of the potent neutrophil chemoattractant interleukin-8 (CXCL8); these effects were observed in human small intestinal mucosal tissues and from intestinal epithelial monolayers, activated through administration of proinflammatory interleukin-1β or Salmonella enterica serovar Typhimurium. This attenuation is caused by the secretion of G. duodenalis cathepsin B cysteine proteases that degrade CXCL8 posttranscriptionally. Furthermore, the degradation of CXCL8 via G. duodenalis cathepsin B cysteine proteases attenuates CXCL8-induced chemotaxis of human neutrophils. Taken together, these data demonstrate for the first time that G. duodenalis trophozoite cathepsins are capable of attenuating a component of their host's proinflammatory response induced by a separate proinflammatory stimulus. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  8. Multilocus genotyping of Giardia duodenalis in captive non-human primates in Sichuan and Guizhou provinces, Southwestern China.

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    Zhijun Zhong

    Full Text Available Giardia duodenalis is a common human and animal pathogen. It has been increasingly reported in wild and captive non-human primates (NHPs in recent years. However, multilocus genotyping information for G. duodenalis infecting NHPs in southwestern China is limited. In the present study, the prevalence and multilocus genotypes (MLGs of G. duodenalis in captive NHPs in southwestern China were determined. We examined 207 fecal samples from NHPs in Sichuan and Guizhou provinces, and 16 specimens were positive for G. duodenalis. The overall infection rate was 7.7%, and only assemblage B was identified. G. duodenalis was detect positive in northern white-cheeked gibbon (14/36, 38.9%, crab-eating macaque (1/60, 1.7% and rhesus macaques (1/101, 0.9%. Multilocus sequence typing based on beta-giardin (bg, triose phosphate isomerase (tpi and glutamate dehydrogenase (gdh revealed nine different assemblage B MLGs (five known genotypes and four novel genotypes. Based on a phylogenetic analysis, one potentially zoonotic genotype of MLG SW7 was identified in a northern white-cheeked gibbon. A high degree of genetic diversity within assemblage B was observed in captive northern white-cheeked gibbons in Southwestern China, including a potentially zoonotic genotype, MLG SW7. To the best of our knowledge, this is the first report using a MLGs approach to identify G. duodenalis in captive NHPs in Southwestern China.

  9. Microarray analysis of the intestinal host response in Giardia duodenalis assemblage E infected calves.

    Science.gov (United States)

    Dreesen, Leentje; Rinaldi, Manuela; Chiers, Koen; Li, Robert; Geurden, Thomas; Van den Broeck, Wim; Goddeeris, Bruno; Vercruysse, Jozef; Claerebout, Edwin; Geldhof, Peter

    2012-01-01

    Despite Giardia duodenalis being one of the most commonly found intestinal pathogens in humans and animals, little is known about the host-parasite interactions in its natural hosts. Therefore, the objective of this study was to investigate the intestinal response in calves following a G. duodenalis infection, using a bovine high-density oligo microarray to analyze global gene expression in the small intestine. The resulting microarray data suggested a decrease in inflammation, immune response, and immune cell migration in infected animals. These findings were examined in more detail by histological analyses combined with quantitative real-time PCR on a panel of cytokines. The transcription levels of IL-6, IL-8, IL-13, IL-17, and IFN-γ showed a trend of being downregulated in the jejunum of infected animals compared to the negative controls. No immune cell recruitment could be seen after infection, and no intestinal pathologies, such as villus shortening or increased levels of apoptosis. Possible regulators of this intestinal response are the nuclear peroxisome proliferator-activated receptors alpha (PPARα), and gamma (PPARγ) and the enzyme adenosine deaminase (ADA), all for which an upregulated expression was found in the microarray and qRT-PCR analyses.

  10. Microarray analysis of the intestinal host response in Giardia duodenalis assemblage E infected calves.

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    Leentje Dreesen

    Full Text Available Despite Giardia duodenalis being one of the most commonly found intestinal pathogens in humans and animals, little is known about the host-parasite interactions in its natural hosts. Therefore, the objective of this study was to investigate the intestinal response in calves following a G. duodenalis infection, using a bovine high-density oligo microarray to analyze global gene expression in the small intestine. The resulting microarray data suggested a decrease in inflammation, immune response, and immune cell migration in infected animals. These findings were examined in more detail by histological analyses combined with quantitative real-time PCR on a panel of cytokines. The transcription levels of IL-6, IL-8, IL-13, IL-17, and IFN-γ showed a trend of being downregulated in the jejunum of infected animals compared to the negative controls. No immune cell recruitment could be seen after infection, and no intestinal pathologies, such as villus shortening or increased levels of apoptosis. Possible regulators of this intestinal response are the nuclear peroxisome proliferator-activated receptors alpha (PPARα, and gamma (PPARγ and the enzyme adenosine deaminase (ADA, all for which an upregulated expression was found in the microarray and qRT-PCR analyses.

  11. Co-infection with Ascaris lumbricoides modulates protective immune responses against Giardia duodenalis in school Venezuelan rural children.

    Science.gov (United States)

    Hagel, I; Cabrera, M; Puccio, F; Santaella, C; Buvat, E; Infante, B; Zabala, M; Cordero, R; Di Prisco, M C

    2011-03-01

    We evaluated the effect of Ascaris lumbricoides on Giardia duodenalis infection and TH1/TH2 type immune mechanisms toward this parasite in 251 rural parasitized and 70 urban non-parasitized school children. The children were classified according to light (0-5000 eggs/g faeces) or moderate (>5001-50,000 eggs/g faeces) A. lumbricoides infection. Anti G. duodenalis skin hyper-reactivity, IgE, IgG, IL-13, IFN γ, IL6 and IL-10 levels were compared among G. duodenalis infected and non-infected children according to light or moderate A. lumbricoides infection. It was found that 62% of the A. lumbricoides moderately infected children were co-infected by G. duodenalis compared to 45% of the lightly infected group. After treatment, 42% of the A. lumbricoides moderately group were infected with G. duodenalis compared to 11% of their lightly counterparts, being A. lumbricoides IL-10 levels higher (plumbricoides lightly parasitized children, G. duodenalis infection was associated to a significant increase (plumbricoides moderately parasitized group, being those levels similarly lower as those observed in the control group. Inverse correlations were found between the levels of anti G duodenalis antibodies, skin test hyper-reactivity and cytokines with the intensity of A. lumbricoides infection (p>0.0001) and A. lumbricoides IL-10 levels (p>0.0001), suggesting that co-infection with A. lumbricoides may affect both TH1 and TH2 type immunity against G. duodenalis that may play an important role in the susceptibility to the infection after chemotherapy in children from endemic areas. Copyright © 2010 Elsevier B.V. All rights reserved.

  12. Genetic characterizations of Giardia duodenalis in sheep and goats in Heilongjiang Province, China and possibility of zoonotic transmission.

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    Weizhe Zhang

    Full Text Available BACKGROUND: Giardia duodenalis is a widespread intestinal protozoan of both humans and mammals. To date, few epidemiological studies have assessed the potential and importance of zoonotic transmission; and the human giardiasis burden attributable to G. duodenalis of animal origin is unclear. No information about occurrence and genotyping data of sheep and goat giardiasis is available in China. The aim of the present study was to determine prevalence and distribution of G. duodenalis in sheep and goats in Heilongjiang Province, China, and to characterize G. duodenalis isolates and assess the possibility of zoonotic transmission. METHODOLOGY/PRINCIPAL FINDINGS: A total of 678 fecal specimens were collected from sheep and goats on six farms ranging in age from one month to four years in Heilongjiang Province, China. The average prevalence of G. duodenalis infection was 5.0% (34/678 by microscopy after Lugol's iodine staining, with 5.6% (30/539 for the sheep versus 2.9% (4/139 for the goats. Molecular analysis was conducted on 34 G. duodenalis isolates based on the triosephosphate isomerase (tpi gene. 29 tpi gene sequences were successfully obtained and identified as assemblages A (n = 4, B (n = 2 and E (n = 23. High heterogeneity was observed within assemblage E at the tpi locus, with five novel subtypes found out of seven subtypes. Two subtypes of assemblage A were detected, including subtype AI (n = 3 and a novel subtype (designated as subtype AIV (n = 1. Two assemblage B isolates were identical to each other in the tpi gene sequences. CONCLUSIONS/SIGNIFICANCE: This is the first report of G. duodenalis infections in sheep and goats in China. The present data revealed the unique endemicity on prevalence, distribution and genetic characterization of G. duodenalis in sheep and goats in Heilongjiang Province. The findings of assemblages A and B in sheep and goats implied the potential of zoonotic transmission.

  13. Giardia duodenalis and Cryptosporidium occurrence in Australian sea lions (Neophoca cinerea exposed to varied levels of human interaction

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    Tiffany C. Delport

    2014-12-01

    Full Text Available Giardia and Cryptosporidium are amongst the most common protozoan parasites identified as causing enteric disease in pinnipeds. A number of Giardia assemblages and Cryptosporidium species and genotypes are common in humans and terrestrial mammals and have also been identified in marine mammals. To investigate the occurrence of these parasites in an endangered marine mammal, the Australian sea lion (Neophoca cinerea, genomic DNA was extracted from faecal samples collected from wild populations (n = 271 in Southern and Western Australia and three Australian captive populations (n = 19. These were screened using PCR targeting the 18S rRNA of Giardia and Cryptosporidium. Giardia duodenalis was detected in 28 wild sea lions and in seven captive individuals. Successful sequencing of the 18S rRNA gene assigned 27 Giardia isolates to assemblage B and one to assemblage A, both assemblages commonly found in humans. Subsequent screening at the gdh and β-giardin loci resulted in amplification of only one of the 35 18S rRNA positive samples at the β-giardin locus. Sequencing at the β-giardin locus assigned the assemblage B 18S rRNA confirmed isolate to assemblage AI. The geographic distribution of sea lion populations sampled in relation to human settlements indicated that Giardia presence in sea lions was highest in populations less than 25 km from humans. Cryptosporidium was not detected by PCR screening in either wild colonies or captive sea lion populations. These data suggest that the presence of G. duodenalis in the endangered Australian sea lion is likely the result of dispersal from human sources. Multilocus molecular analyses are essential for the determination of G. duodenalis assemblages and subsequent inferences on transmission routes to endangered marine mammal populations.

  14. Molecular Genotyping of Giardia duodenalis Isolates from Symptomatic Individuals Attending Two Major Public Hospitals in Madrid, Spain.

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    Aida de Lucio

    Full Text Available The flagellate protozoan Giardia duodenalis is an enteric parasite causing human giardiasis, a major gastrointestinal disease of global distribution affecting both developing and industrialised countries. In Spain, sporadic cases of giardiasis have been regularly identified, particularly in pediatric and immigrant populations. However, there is limited information on the genetic variability of circulating G. duodenalis isolates in the country.In this longitudinal molecular epidemiological study we report the diversity and frequency of the G. duodenalis assemblages and sub-assemblages identified in 199 stool samples collected from 184 individual with symptoms compatible with giardiasis presenting to two major public hospitals in Madrid for the period December 2013-January 2015. G. duodenalis cysts were initially detected by conventional microscopy and/or immunochomatography on stool samples. Confirmation of the infection was performed by direct immunofluorescence and real-time PCR methods. G. duodenalis assemblages and sub-assemblages were determined by multi-locus genotyping of the glutamate dehydrogenase (GDH and β-giardin (BG genes of the parasite. Sociodemographic and clinical features of patients infected with G. duodenalis were also analysed.Of 188 confirmed positive samples from 178 giardiasis cases a total of 124 G. duodenalis isolates were successfully typed at the GDH and/or the BG loci, revealing the presence of sub-assemblages BIV (62.1%, AII (15.3%, BIII (4.0%, AI (0.8%, and AIII (0.8%. Additionally, 6.5% of the isolates were only characterised at the assemblage level, being all of them assigned to assemblage B. Discordant genotype results AII/AIII or BIII/BIV were also observed in 10.5% of DNA isolates. A large number of multi-locus genotypes were identified in G. duodenalis assemblage B, but not assemblage A, isolates at both the GDH and BG loci, confirming the high degree of genetic variability observed in other molecular surveys

  15. Axenic cultivation and comparative phospholipase A2 activity of Giardia duodenalis in a serum-free medium.

    Science.gov (United States)

    Mata-Cárdenas, Benito David; Hernández-García, Magda Elizabeth; González-Salazar, Francisco; Garza-González, Jesús Norberto; Palacios-Corona, Rebeca; Cortés-Gutiérrez, Elva I; Vargas-Villarreal, Javier

    2012-09-01

    Mammalian serum is essential for the growth of Giardia duodenalis cultivated under axenic conditions. Unfortunately, some factors present in bovine serum used as supplement in the culture medium may inhibit protozoal growth and activity. TYI-33-PACSR is a TYI medium supplemented with a serum replacement (PACSR) made up of Earle's amino acid solution, Diamond's vitamin-tween 80 mixtures and LCR (a lipid-cholesterol - rich mixture). PACSR was previously used in the culture media for axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. The main objective of this work was to demonstrate that TYI-33-PACSR is useful for axenic cultivation of G. duodenalis. Additionally, the activity of phospholipase A(2) (PLA A(2)) in the sub-cellular vesicular fraction (P30) of G. duodenalis grown in TYI-S-33 and TYI-33-PACSR was compared. All strains of Giardia grown in TYI-33-PACSR reached relative cellular densities of 91 to 95% compared to controls growing in serum-supplemented TYI-S-33 medium. Additionally, PLA A(2) activity was similar in the P30 sub-cellular fraction obtained from trophozoites growing in TYI-S-33 and TYI-33-PACSR. Thus, TYI-33-PACSR could be useful in analyzing the biological properties of G. duodenalis in the absence of serum.

  16. Molecular identification of Giardia duodenalis in Ecuador by polymerase chain reaction-restriction fragment length polymorphism

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    Richard Atherton

    2013-06-01

    Full Text Available The aim of this study was to determine the genetic diversity of Giardia duodenalis present in a human population living in a northern Ecuadorian rain forest. All Giardia positive samples (based on an ELISA assay were analysed using a semi-nested polymerase chain reaction-restriction fragment length polymorphism assay that targets the glutamate dehydrogenase (gdh gene; those amplified were subsequently genotyped using NlaIV and RsaI enzymes. The gdh gene was successfully amplified in 74 of 154 ELISA positive samples; 69 of the 74 samples were subsequently genotyped. Of these 69 samples, 42 (61% were classified as assemblage B (26 as BIII and 16 as BIV, 22 (32% as assemblage A (3 as AI and 19 as AII and five (7% as mixed AII and BIII types. In this study site we observe similar diversity in genotypes to other regions in Latin America, though in contrast to some previous studies, we found similar levels of diarrheal symptoms in those individuals infected with assemblage B compared with those infected with assemblage A.

  17. Induction of virulence factors in Giardia duodenalis independent of host attachment

    Science.gov (United States)

    Emery, Samantha J.; Mirzaei, Mehdi; Vuong, Daniel; Pascovici, Dana; Chick, Joel M.; Lacey, Ernest; Haynes, Paul A.

    2016-01-01

    Giardia duodenalis is responsible for the majority of parasitic gastroenteritis in humans worldwide. Host-parasite interaction models in vitro provide insights into disease and virulence and help us to understand pathogenesis. Using HT-29 intestinal epithelial cells (IEC) as a model we have demonstrated that initial sensitisation by host secretions reduces proclivity for trophozoite attachment, while inducing virulence factors. Host soluble factors triggered up-regulation of membrane and secreted proteins, including Tenascins, Cathepsin-B precursor, cystatin, and numerous Variant-specific Surface Proteins (VSPs). By comparison, host-cell attached trophozoites up-regulated intracellular pathways for ubiquitination, reactive oxygen species (ROS) detoxification and production of pyridoxal phosphate (PLP). We reason that these results demonstrate early pathogenesis in Giardia involves two independent host-parasite interactions. Motile trophozoites respond to soluble secreted signals, which deter attachment and induce expression of virulence factors. Trophozoites attached to host cells, in contrast, respond by up-regulating intracellular pathways involved in clearance of ROS, thus anticipating the host defence response. PMID:26867958

  18. Single dose of secnidazole treatment against naturally occuring Giardia duodenalis infection in Sakiz lambs

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    Kerem Ural

    2014-03-01

    Full Text Available Objective. The purposes of this study were to determine whether secnidazole administered at a single dose of 10 mg/kg, orally, lessens or eliminates Giardia cyst shedding, and to validate the benefit of secnidazole treatment on clinical signs and faecal consistency in lambs naturally infected with Giardia duodenalis. Materials and methods. To this extent weaned 12 weeks of age lambs were selected and randomly assigned into two groups based on placebo (group C, n=7 untreated control group or treatment (group S, n=10 lambs treated with a single dose of secnidazole at 10 mg/kg. Results. On days 0 and 10, before and after treatment, respectively, reduction in cyst excretion was determined. The faecal consistency and general health were recorded at each sampling day. Conclusions. During the study there was a high (99.98% reduction in cyst excretion in the secnidazol treatment group compared to the positive control group on day 10, resulting in a significant (p<0.001 reduction, making secnidazole highly effective treatment option.

  19. Predominance of Giardia duodenalis Assemblage AII in Fresh Leafy Vegetables from a Market in Southern Brazil.

    Science.gov (United States)

    Tiyo, Rogerio; de Souza, Carla Zangari; Arruda Piovesani, Ana Flávia; Tiyo, Bruna Tiaki; Colli, Cristiane Maria; Marchioro, Ariella Andrade; Gomes, Monica Lucia; Falavigna-Guilherme, Ana Lucia

    2016-06-01

    We investigated the presence of Giardia duodenalis cysts and its genotypes in raw leafy vegetables sold in a Brazilian market. These products are different from those sold in most street markets because the producers themselves display and sell their products and rely on specialized technical and sanitary assistance. Vegetable and water samples were collected from 14 (80%) producers who cultivated vegetables that are typically consumed raw for sale at the market, obtained at the market and farms, respectively. A total of 128 samples of leafy greens (chives, parsley, cabbage, arugula, watercress, and chicory) and 14 water samples were analyzed by direct immunofluorescence and PCR techniques. The positive samples were genotyped (GHD gene) using PCR and restriction fragment length polymorphism. The analyses indicated that 16 (12.5%) of 128 samples were positive by PCR, while 1 (0.8%) of 128 samples were positive by immunofluorescence. Giardia cysts were not detected in the water samples obtained at the farms. The molecular technique revealed a genotype with zoonotic potential, which underscores the challenge in the control of giardiasis dissemination via the consumption of raw vegetables.

  20. Prevalence and molecular characterization of Cryptosporidium spp. and Giardia duodenalis in diarrhoeic patients in the Qikiqtani Region, Nunavut, Canada

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    Asma Iqbal

    2015-06-01

    Full Text Available Background: Although the prevalences of infection with the protozoan parasites Cryptosporidium spp. and Giardia duodenalis in humans appear to be relatively high in the Canadian North, their transmission patterns are poorly understood. Objective: To determine the detection rate and the molecular characteristics of Cryptosporidium spp. and Giardia duodenalis in diarrhoeic patients in the Qikiqtani (Baffin Island Region of Nunavut, Canada, in order to better understand the burden of illness and the potential mechanisms of transmission. Study design/methods: Diarrhoeal stool specimens (n=108 submitted to the Qikiqtani General Hospital for clinical testing were also tested for the presence of Cryptosporidium spp. and Giardia duodenalis using epifluorescence microscopy and polymerase chain reaction (PCR. DNA sequencing and restriction fragment length polymorphism (RFLP analyses were performed on PCR-positive specimens to determine the species, genotypes and sub-genotypes of the parasites. Results: Cryptosporidium was detected in 15.7% of the diarrhoeic patients, while Giardia was detected in 4.6%. DNA sequencing of a fragment of the small subunit rRNA gene indicated that all of the Cryptosporidium amplicons had a 100% homology to C. parvum, and a gp60 assay showed that all aligned with C. parvum sub-genotype IIa. Microsatellite analysis revealed 3 cases of sub-genotype IIaA15G2R1, 2 of IIaA15G1R and 1 case each of sub-genotypes IIaA16G1R1 and IIaA15R1. For Giardia, results based on the amplification of both the 16S rRNA gene and the gdh gene were generally in agreement, and both DNA sequencing and RFLP demonstrated the presence of the G. duodenalis Assemblage B genotype. Conclusions: Both C. parvum and G. duodenalis Assemblage B were present in human diarrhoeal stool specimens from Nunavut, which was suggestive of zoonotic transmission, although human-to-human transmission cannot be ruled out. To fully understand the public health significance of the

  1. Disruptions of Host Immunity and Inflammation by Giardia Duodenalis: Potential Consequences for Co-Infections in the Gastro-Intestinal Tract

    OpenAIRE

    James A. Cotton; Christina B. Amat; Andre G. Buret

    2015-01-01

    Giardia duodenalis (syn. G. intestinalis, or G. lamblia) is a leading cause of waterborne diarrheal disease that infects hundreds of millions of people annually. Research on Giardia has greatly expanded within the last few years, and our understanding of the pathophysiology and immunology on this parasite is ever increasing. At peak infection, Giardia trophozoites induce pathophysiological responses that culminate in the development of diarrheal disease. However, human data has suggested tha...

  2. Giardia duodenalis Infection Reduces Granulocyte Infiltration in an In Vivo Model of Bacterial Toxin-Induced Colitis and Attenuates Inflammation in Human Intestinal Tissue

    OpenAIRE

    Cotton, James A.; Motta, Jean-Paul; Schenck, L. Patrick; Hirota, Simon A.; Beck, Paul L.; Buret, Andre G.

    2014-01-01

    Giardia duodenalis (syn. G. intestinalis, G. lamblia) is a predominant cause of waterborne diarrheal disease that may lead to post-infectious functional gastrointestinal disorders. Although Giardia-infected individuals could carry as much as 106 trophozoites per centimetre of gut, their intestinal mucosa is devoid of overt signs of inflammation. Recent studies have shown that in endemic countries where bacterial infectious diseases are common, Giardia infections can protect against the develo...

  3. Biological and Genetic Characterization of Cryptosporidium spp. and Giardia duodenalis Isolates from Five Hydrographical Basins in Northern Portugal

    Science.gov (United States)

    Moreira, Maria João; Soares, Sónia; de Lurdes Delgado, Maria; Figueiredo, João; Magalhães, Elisabete Silva; Castro, António; Viana Da Costa, Alexandra; Correia da Costa, José Manuel

    2010-01-01

    To understand the situation of water contamination with Cryptosporidium spp. and Giardia spp. in the northern region of Portugal, we have established a long-term program aimed at pinpointing the sources of surface water and environmental contamination, working with the water-supply industry. Here, we describe the results obtained with raw water samples collected in rivers of the 5 hydrographical basins. A total of 283 samples were analyzed using the Method 1623 EPA, USA. Genetic characterization was performed by PCR and sequencing of genes 18S rRNA of Cryptosporidium spp. and β-giardin of Giardia spp. Infectious stages of the protozoa were detected in 72.8% (206 of 283) of the water samples, with 15.2% (43 of 283) positive for Giardia duodenalis cysts, 9.5% (27 of 283) positive for Cryptosporidium spp. oocysts, and 48.1% (136 of 283) samples positive for both parasites. The most common zoonotic species found were G. duodenalis assemblages A-I, A-II, B, and E genotypes, and Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis, and Cryptosporidium muris. These results suggest that cryptosporidiosis and giardiasis are important public health issues in northern Portugal. To the authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in raw water samples in the northern region of Portugal. PMID:20585525

  4. Genotypes of Cryptosporidium spp., Enterocytozoon bieneusi and Giardia duodenalis in dogs and cats in Shanghai, China.

    Science.gov (United States)

    Xu, Hailing; Jin, Yue; Wu, Wenxian; Li, Pei; Wang, Lin; Li, Na; Feng, Yaoyu; Xiao, Lihua

    2016-03-01

    Controversies exist on the potential role of companion animals in the transmission of enteric pathogens in humans. This study was conducted to examine the genotype distribution of Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis in companion animals in Shanghai, China, and to assess their zoonotic potential. Fecal specimens from 485 dogs and 160 cats were examined for the occurrence and genotype distribution of the three pathogens by PCR. PCR products were sequenced to determine the species and genotypes. The χ(2) test was used to compare differences in infection rates between living conditions or age groups. Cryptosporidium spp., E. bieneusi and G. duodenalis were found in 39 (8.0 %), 29 (6.0 %) and 127 (26.2 %) of dogs, and 6 (3.8 %), 9 (5.6 %) and 21 (13.1 %) of cats, respectively. Infection rates of the pathogens in dogs from pet shops and a clinic were higher than those in household dogs, and higher in cats from one animal shelter than from pet shops. No significant differences in infection rates were detected among age groups. Cryptosporidium canis and C. felis were the only Cryptosporidium species found in dogs and cats, respectively. Enterocytozoon bieneusi genotype PtEb IX was the dominant genotype in dogs, whereas Type IV and D were the most common ones in cats. Multi-locus sequence typing at the glutamate dehydrogenase, β-giardin, and triosephosphate isomerase loci revealed the presence of G. duodenalis assemblages A (n = 23), B (n = 1), C (n = 26), and D (n = 58) in dogs (only A in household dogs) and assemblages A (n = 2), B (n = 6), C (n = 2), D (n = 1), and F (n = 7) in cats. Co-infection was detected in 24 dogs and 5 cats, especially those living in crowded conditions. Living condition is a major risk factor affecting the occurrence of enteric protists in companion animals in China, and although dogs and cats can be potential sources of human infections, the different distribution of

  5. Giardia duodenalis and Cryptosporidium spp. in the intestinal contents of ringed seals (Phoca hispida) and bearded seals (Erignathus barbatus) in Nunavik, Quebec, Canada.

    Science.gov (United States)

    Dixon, Brent R; Parrington, Lorna J; Parenteau, Monique; Leclair, Daniel; Santín, Mónica; Fayer, Ronald

    2008-10-01

    The prevalence of Giardia duodenalis and Cryptosporidium spp. was determined for ringed and bearded seals harvested for food in the Nunavik region in northern Quebec, Canada. Flow cytometric results demonstrated that G. duodenalis was present in the intestinal contents of 80% of the ringed seals and 75% of the bearded seals tested, while Cryptosporidium spp. were present in 9% of the ringed seals and none of the bearded seals. Prevalence of both parasites was highest in animals less than 1 yr of age. Giardia sp. isolates from ringed seals were identified as G. duodenalis Assemblage B, which is commonly identified in human infections. The high prevalence of G. duodenalis in ringed seals, and the presence of Assemblage B in these animals, highlights the potential for zoonotic transmission to the Inuit people, who consume dried seal intestines and uncooked seal meat.

  6. Giardia duodenalis assemblages and Entamoeba species infecting non-human primates in an Italian zoological garden: zoonotic potential and management traits

    OpenAIRE

    Berrilli, F; Prisco, C; Friedrich, K; Di Cerbo, P; Di Cave, D; De Liberato, C

    2011-01-01

    Abstract Background Giardia duodenalis and Entamoeba spp. are among the most common intestinal human protozoan parasites worldwide and they are frequently reported in captive non-human primates (NHP). From a public health point of view, infected animals in zoos constitute a risk for animal caretakers and visitors. In this study we carried out the molecular identification of G. duodenalis and Entamoeba spp. from nine species of primates housed in the zoological garden of Rome, to better ascert...

  7. Development of a chromogenic in situ hybridization for Giardia duodenalis and its application in canine, feline, and porcine intestinal tissues samples.

    Science.gov (United States)

    Weissenböck, Herbert; Ondrovics, Martina; Gurtner, Susanne; Schiessl, Peter; Mostegl, Meike M; Richter, Barbara

    2011-05-01

    In the present study, a chromogenic in situ hybridization for the identification of Giardia duodenalis in paraffin-embedded tissue samples was developed. The sensitivity and specificity of the probe was validated by testing it on cultured reference samples of different assemblages of G. duodenalis as well as culture and tissue samples containing other protozoa and infectious agents. The probe gave a positive reaction with the Giardia samples and a negative reaction with all other samples. Further, the probe was used for screening of histological slides of intestine from different animal species (99 canine samples, 85 feline samples, and 202 porcine samples) for the presence of G. duodenalis trophozoites. With this assay, the parasites were detected in samples from 8 dogs (8.08%), 6 cats (7.06%), and zero pigs. The results clearly indicate that the described method is useful for detection of Giardia trophozoites in routinely processed intestinal tissue of different animal species. © 2011 The Author(s)

  8. A new quantitative in vitro microculture method for Giardia duodenalis trophozoites.

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    Bénéré, Ely; da Luz, Raquel Andreia Inocêncio; Vermeersch, Marieke; Cos, Paul; Maes, Louis

    2007-11-01

    A reliable, rapid and low-cost method for drug sensitivity determination of Giardia duodenalis trophozoites (WB-strain) was developed in a 96-well plate. Using a standard inoculum of 5 x 10(4) trophozoites per well (300 microl), good growth was obtained after sealing the plate with an air-tight adhesive tape and incubation at 37 degrees C for 72 h in modified TYI-S-33 medium. Viable burdens were quantified using the formazan dyes MTT (100 microg/well) and XTT (20 microg/well) and the fluorescent substrate resazurin (2.5 microg/well). Prior removal of the culture medium is required since it causes spontaneous reduction of the substrate. Resazurin proved to be far superior to MTT and XTT with a level of sensitivity of about 3 x 10(4) trophozoites. Inhibitory concentrations (IC(50)) of several anti-giardial reference drugs were within the range of published values: metronidazole 2.25 microM, tinidazole 1.75 microM, albendazole 0.10 microM, furazolidone 2.00 microM and quinacrine 0.32 microM. The broad-spectrum antibiotics chloramphenicol, rifampicin, penicillin G+streptomycin and gentamycin were devoid of any inhibitory activity and are considered suitable for decontamination during excystation experiments.

  9. The efficacy of chloroquine treatment against naturally occuring Giardia duodenalis infection in lambs

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    Umit Karademir

    2016-05-01

    Full Text Available Objective. The aim of the present study was to determine the efficacy of Chloroquine (Cq, an antimalarial medicine, administered at a dose of 2.5 mg/kg, orally, during 5 consecutive days, in Sakiz and Merino lambs naturally infected with Giardia duodenalis. Materials and methods. To this extent weaned 10 weeks of aged lambs were enrolled and randomly assigned into two groups based on treatment (group C, n=18 lambs treated with Cq and placebo (group P, n=8 untreated control animals. Diagnosis was based on detection of trophozoit and/or cysts on fecal flotation. Cyst count per gram feces (days 0, 3, 7 and 10 was analyzed among groups. Results. During the trial, regarding the efficacy of Cq on days 3., 7., and 10. There was significant (p<0.01 reduction in cyst excretion; whereas evaluation of mean geometric cyst excretion revealed 100% reduction. Conclusions. There was a very high (100% reduction in cyst excretion in the Cq treatment group compared to the positive untreated control group on days 3, 7 and 10, resulting in a significant (p<0.001 reduction, making Cq, safety, and thus highly effective treatment option of lambs with naturally occuring giardiasis.

  10. Occurrence and molecular genotyping of Giardia duodenalis and Cryptosporidium spp. in wild mesocarnivores in Spain.

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    Mateo, Marta; de Mingo, Marta Hernández; de Lucio, Aida; Morales, Lucía; Balseiro, Ana; Espí, Alberto; Barral, Marta; Lima Barbero, José Francisco; Habela, Miguel Ángel; Fernández-García, José L; Bernal, Rafael Calero; Köster, Pamela C; Cardona, Guillermo A; Carmena, David

    2017-02-15

    There is a surprisingly scarce amount of epidemiological and molecular data on the prevalence, frequency, and diversity of the intestinal protozoan parasites Giardia duodenalis and Cryptosporidium spp. in wildlife in general and mesocarnivore species in particular. Consequently, the extent of the cyst/oocyst environmental contamination attributable to these wild host species and their potential implications for public veterinary health remain largely unknown. In this molecular epidemiological survey a total of 193 individual faecal samples from badgers (Meles meles, n=70), ferrets (Mustela putorius furo, n=2), genets (Genetta genetta, n=6), Iberian lynxes (Lynx pardinus, n=6), beech martens (Martes foina, n=8), mongooses (Herpestes ichneumon, n=2), otters (Lutra lutra, n=2), polecats (Mustela putorius, n=2), red foxes (Vulpes vulpes, n=87), wildcats (Felis silvestris, n=2), and wolves (Canis lupus, n=6) were obtained from road-killed, hunted, and accidentally found carcasses, and from camera-trap surveys or animals entering rescue shelters, during the period December 2003-April 2016. Investigated specimens were collected in five Spanish autonomous regions including Andalusia (n=1), Asturias (n=69), Basque Country (n=49), Castile-La Mancha (n=38), and Extremadura (n=36). The presence of cysts/oocysts was confirmed by PCR-based methods targeting the small subunit (ssu) ribosomal RNA gene of these parasite species. Genotyping of the obtained isolates were attempted at appropriate markers including the glutamate dehydrogenase (G. duodenalis) and the 60-kDa glycoprotein (C. parvum and C. ubiquitum) loci. Overall, G. duodenalis was detected in 8% (7/87) of red foxes, a single beech marten, and a single wolf, respectively. Cryptosporidium was identified in 3% (2/70) of badgers, 8% (7/87) of red foxes, a single genet, and a single mongoose, respectively. None of the nine G. duodenalis isolates generated could be genotyped at the assemblage/sub-assemblage level. Out of the

  11. Detection of Giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies

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    Sofía Duque-Beltrán

    2002-12-01

    Full Text Available The present study developed and standardized an enzime-linked immunosorbent assay (ELISA to detect Giardia antigen in feces using rabbit polyclonal antibodies. Giardia cysts were purified from human fecal samples by sucrose and percoll gradients. Gerbils (Meriones unguiculatus were infected to obtain trophozoites. Rabbits were inoculated with either cyst or trophozoite antigens of 14 Colombian Giardia isolates to develop antibodies against the respective stages. The IgG anti-Giardia were purified by sequential caprylic acid and ammonium sulfate precipitation. A portion of these polyclonal antibodies was linked to alkaline phosphatase (conjugate. One hundred and ninety six samples of human feces, from different patients, were tested by parasitologic diagnosis: 69 were positive for Giardia cysts, 56 had no Giardia parasites, and 71 revealed parasites other than Giardia. The optimal concentration of polyclonal antibodies for antigen capture was 40 µg/ml and the optimal conjugate dilution was 1:100. The absorbance cut-off value was 0.24. The parameters of the ELISA test for Giardia antigen detection were: sensitivity, 100% (95% CI: 93.4-100%; specificity, 95% (95% CI: 88.6-97.6%; positive predictive value, 91% (95% CI: 81.4-95.9%; and negative predictive value, 100% (95% CI: 96.1-100%. This ELISA will improve the diagnosis of Giardia infections in Colombia and will be useful in following patients after treatment.

  12. Molecular surveillance of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi by genotyping and subtyping parasites in wastewater.

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    Na Li

    Full Text Available BACKGROUND: Despite their wide occurrence, cryptosporidiosis and giardiasis are considered neglected diseases by the World Health Organization. The epidemiology of these diseases and microsporidiosis in humans in developing countries is poorly understood. The high concentration of pathogens in raw sewage makes the characterization of the transmission of these pathogens simple through the genotype and subtype analysis of a small number of samples. METHODOLOGY/PRINCIPAL FINDINGS: The distribution of genotypes and subtypes of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in 386 samples of combined sewer systems from Shanghai, Nanjing and Wuhan and the sewer system in Qingdao in China was determined using PCR-sequencing tools. Eimeria spp. were also genotyped to assess the contribution of domestic animals to Cryptosporidium spp., G. duodenalis, and E. bieneusi in wastewater. The high occurrence of Cryptosporidium spp. (56.2%, G. duodenalis (82.6%, E. bieneusi (87.6%, and Eimeria/Cyclospora (80.3% made the source attribution possible. As expected, several human-pathogenic species/genotypes, including Cryptosporidium hominis, Cryptosporidium meleagridis, G. duodenalis sub-assemblage A-II, and E. bieneusi genotype D, were the dominant parasites in wastewater. In addition to humans, the common presence of Cryptosporidium spp. and Eimeria spp. from rodents indicated that rodents might have contributed to the occurrence of E. bieneusi genotype D in samples. Likewise, the finding of Eimeria spp. and Cryptosporidium baileyi from birds indicated that C. meleagridis might be of both human and bird origins. CONCLUSIONS/SIGNIFICANCE: The distribution of Cryptosporidium species, G. duodenalis genotypes and subtypes, and E. bieneusi genotypes in urban wastewater indicates that anthroponotic transmission appeared to be important in epidemiology of cryptosporidiosis, giardiasis, and microsporidiosis in the study areas. The finding of

  13. Cryptosporidium spp. and Giardia duodenalis as pathogenic contaminants of water in Galicia, Spain: the need for safe drinking water.

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    Castro-Hermida, José Antonio; González-Warleta, Marta; Mezo, Mercedes

    2015-01-01

    The objectives of this cross-sectional study were to detect the presence of Cryptosporidium spp. and Giardia duodenalis in drinking water treatments plants (DWTPs) in Galicia (NW Spain) and to identify which species and genotype of these pathogenic protozoans are present in the water. Samples of untreated water (surface or ground water sources) and of treated drinking water (in total, 254 samples) were collected from 127 DWTPs and analysed by an immunofluorescence antibody test (IFAT) and by PCR. Considering the untreated water samples, Cryptosporidium spp. were detected in 69 samples (54.3%) by IFAT, and DNA of this parasite was detected in 57 samples (44.8%) by PCR, whereas G. duodenalis was detected in 76 samples (59.8%) by IFAT and in 56 samples (44.0%) by PCR. Considering the treated drinking water samples, Cryptosporidium spp. was detected in 52 samples (40.9%) by IFAT, and the parasite DNA was detected in 51 samples (40.1%) by PCR, whereas G. duodenalis was detected in 58 samples (45.6%) by IFAT and in 43 samples (33.8%) by PCR. The percentage viability of the (oo)cysts ranged between 90.0% and 95.0% in all samples analysed. Cryptosporidium andersoni, C. hominis, C. parvum and assemblages A-I, A-II, E of G. duodenalis were identified. The results indicate that Cryptosporidium spp. and G. duodenalis are widespread in the environment and that DWTPs are largely ineffective in reducing/inactivating these pathogens in drinking water destined for human and animal consumption in Galicia. In conclusion, the findings suggest the need for better monitoring of water quality and identification of sources of contamination. Copyright © 2014 Elsevier GmbH. All rights reserved.

  14. Strong genetic structure revealed by multilocus patterns of variation in Giardia duodenalis isolates of patients from Galicia (NW-Iberian Peninsula).

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    Gabín-García, Luis B; Bartolomé, Carolina; Abal-Fabeiro, José L; Méndez, Santiago; Llovo, José; Maside, Xulio

    2017-03-01

    We report a survey of genetic variation at three coding loci in Giardia duodenalis of assemblages A and B obtained from stool samples of patients from Santiago de Compostela (Galicia, NW-Iberian Peninsula). The mean pooled synonymous diversity for assemblage A was nearly five times lower than for assemblage B (0.77%±0.30% and 4.14%±1.65%, respectively). Synonymous variation in both assemblages was in mutation-drift equilibrium and an excess of low-frequency nonsynonymous variants suggested the action of purifying selection at the three loci. Differences between isolates contributed to 40% and 60% of total genetic variance in assemblages A and B, respectively, which revealed a significant genetic structure. These results, together with the lack of evidence for recombination, support that (i) Giardia assemblages A and B are in demographic equilibrium and behave as two genetically isolated populations, (ii) infections are initiated by a reduced number of individuals, which may be genetically diverse and even belong to different assemblages, and (iii) parasites reproduce clonally within the host. However, the observation of invariant loci in some isolates means that mechanisms for the homogenization of the genetic content of the two diploid nuclei in each individual must exist. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Genetic analysis of the Gdh and Bg genes of animal-derived Giardia duodenalis isolates in Northeastern China and evaluation of zoonotic transmission potential.

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    Aiqin Liu

    Full Text Available BACKGROUND: Giardia duodenalis is a common intestinal parasite that infects humans and many other mammals, mainly distributing in some areas with poor sanitation. The proportion of the human giardiasis burden attributable to G. duodenalis of animal origin differs in different geographical areas. In Mainland China, genetic data of the gdh and bg genes of G. duodenalis from animals are only limited in dogs and cats. The aim of the study was to provide information on the genetic characterizations of animal-derived G. duodenalis isolates (from rabbits, sheep and cattle at both loci in Heilongjiang Province, Northeastern China, and to assess the potential for zoonotic transmission. METHODOLOGY/PRINCIPAL FINDINGS: 61 G. duodenalis isolates from animal feces (dairy and beef cattle, sheep and rabbits in Heilongjiang Province were characterized at the gdh and bg loci in the present study. The gdh and bg gene sequences of sheep-derived G. duodenalis assemblage AI, and the gdh sequences of rabbit-derived G. duodenalis assemblage B had 100% similarity with those from humans, respectively. Novel subtypes of G. duodenalis were identified, with one and seven subtypes for assemblages A and E at the gdh locus, and two and three subtypes for assemblages B and E at the bg locus, respectively. Three pairs of the same bg sequences of assemblage E were observed in sheep and cattle. CONCLUSIONS/SIGNIFICANCE: This is the first description of genetic characterizations of the gdh and bg genes of G. duodenalis from rabbits, sheep and cattle in Mainland China. Homology analysis of assemblages AI and B implied the possibility of zoonotic transmission. The novel subtypes of assemblages of G. duodenalis may represent the endemic genetic characteristics of G. duodenalis in Heilongjiang Province, China.

  16. Detection and molecular diversity of Giardia duodenalis and Cryptosporidium spp. in sheltered dogs and cats in Northern Spain.

    Science.gov (United States)

    Gil, Horacio; Cano, Lourdes; de Lucio, Aida; Bailo, Begoña; de Mingo, Marta Hernández; Cardona, Guillermo A; Fernández-Basterra, José A; Aramburu-Aguirre, Juan; López-Molina, Nuria; Carmena, David

    2017-06-01

    Domestic dogs and cats may act as natural reservoirs of a large number of zoonotic pathogens, including the enteric parasites Giardia duodenalis and Cryptosporidium spp., the most relevant protozoan species causing gastrointestinal disease worldwide. A cross-sectional epidemiological study aiming to assess the prevalence and molecular diversity of G. duodenalis and Cryptosporidium spp. was conducted in an animal rescue centre in the province of Álava (Northern Spain). A total of 194 and 65 faecal dropping samples from individual dogs and cats, respectively, were collected between November 2013 and June 2016. G. duodenalis cysts and Cryptosporidium spp. oocysts were detected by direct fluorescence microscopy and PCR-based methods targeting the small subunit ribosomal RNA gene of these parasites. Overall, G. duodenalis and Cryptosporidium spp. were detected in 33% (63/194) and 4.1% (8/194) of dogs, and 9.2% (6/65) and 4.6% (3/65) of cats, respectively. G. duodenalis and Cryptosporidium co-infections were observed in 1.5% (3/194) of dogs, but not in cats. No significant differences in infection rates could be demonstrated among dogs or cats according to their sex, age group, status, or geographical origin. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 19 canine isolates that were unambiguously assigned to sub-assemblages AII (n=7), BIII (n=1), and BIV (n=7), and assemblages C (n=3) and D (n=1). Two feline isolates were genotyped as assemblages A and F, respectively. No mixed assemblage or sub-assemblage infections were identified. C. canis (n=5) and C. hominis (n=1) were the Cryptosporidium species found in dogs, whereas C. felis (n=1) was identified in cats. The finding of G. duodenalis sub-assemblages AII, BIII, and BIV circulating in dogs (but not cats) may have zoonotic potential, although most of the AII and BIV isolates sub-genotyped corresponded to genetic variants not

  17. Identification of a novel assemblage G subgenotype and a zoonotic assemblage B in rodent isolates of Giardia duodenalis in the Canary Islands, Spain.

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    Fernández-Álvarez, Ángela; Martín-Alonso, Aarón; Abreu-Acosta, Néstor; Feliu, Carlos; Hugot, Jean-Pierre; Valladares, Basilio; Foronda, Pilar

    2014-02-01

    The 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.

  18. Clonal diversity of the glutamate dehydrogenase gene in Giardia duodenalis from Thai Isolates: evidence of genetic exchange or Mixed Infections?

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    Saksirisampant Wilai

    2011-09-01

    Full Text Available Abstract Background The glutamate dehydrogenase gene (gdh is one of the most popular and useful genetic markers for the genotypic analysis of Giardia duodenalis (syn. G. lamblia, G. intestinalis, the protozoan that widely causes enteric disease in humans. To determine the distribution of genotypes of G. duodenalis in Thai populations and to investigate the extent of sequence variation at this locus, 42 fecal samples were collected from 3 regions of Thailand i.e., Central, Northern, and Eastern regions. All specimens were analyzed using PCR-based genotyping and recombinant subcloning methods. Results The results showed that the prevalence of assemblages A and B among these populations was approximately equal, 20 (47.6% and 22 (52.4%, respectively. Sequence analysis revealed that the nucleotide diversity of assemblage B was significantly greater than that in assemblage A. Among all assemblage B positive specimens, the allelic sequence divergence within isolates was detected. Nine isolates showed mixed alleles, ranged from three to nine distinct alleles per isolate. Statistical analysis demonstrated the occurrence of genetic recombination within subassemblages BIII and BIV was likely. Conclusion This study supports increasing evidence that G. duodenalis has the potential for genetic exchange.

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

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    Oliveira-Arbex, A P; David, E B; Oliveira-Sequeira, T C G; Bittencourt, G N; Guimarães, S

    2016-05-01

    Giardia 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.

  20. Potential role of beavers (Castor fiber in contamination of water in the Masurian Lake District (north-eastern Poland with protozoan parasites Cryptosporidium spp. and Giardia duodenalis

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    Sroka Jacek

    2015-06-01

    Full Text Available The purpose of this study was to assess the possible influence of beavers on the contamination of lake water with zoonotic parasites Giardia duodenalis and Cryptosporidium spp., with respect to the risk to human health. A total of 79 water samples were taken around the habitats of beavers from 14 localities situated in the recreational Masurian Lake District (north-eastern Poland. Water was sampled in the spring and autumn seasons, at different distances from beavers’ lodges (0-2, 10, 30, and 50 m. The samples were examined for the presence of (oocysts of zoonotic protozoa Giardia duodenalis and Cryptosporidium spp. by direct fluorescence assay (DFA and by nested and real time PCR. By DFA, the presence of Giardia cysts was found in 36 samples (45.6% and the presence of Cryptosporidium oocysts in 26 samples (32.9%. Numbers of Giardia cysts, Cryptosporidium oocysts, and summarised (oocysts of both parasites showed a significant variation depending on locality. The numbers of Giardia cysts significantly decreased with the distance from beavers’ lodges while the numbers of Cryptosporidium oocysts did not show such dependence. The amount of Giardia cysts in samples collected in spring was approximately 3 times higher than in autumn. Conversely, a larger number of Cryptosporidium oocysts were detected in samples collected in autumn than in spring. By PCR, Giardia DNA was found in 38 samples (48.1% whereas DNA of Cryptosporidium was found in only 7 samples (8.9%. Eleven Giardia isolates were subjected to phylogenetic analysis by restriction fragment length polymorphism PCR or sequencing which evidenced their belonging to zoonotic assemblages: A (3 isolates and B (8 isolates. In conclusion, water in the vicinity of beavers’ lodges in the tested region was markedly contaminated with (oocysts of Giardia duodenalis and Cryptosporidium spp., which confirms the potential role of beavers as a reservoir of these parasites and indicates a need for

  1. The first report of Cryptosporidium bovis, C. ryanae and Giardia duodenalis sub-assemblage A-II in roe deer (Capreolus capreolus) in Spain.

    Science.gov (United States)

    García-Presedo, Ignacio; Pedraza-Díaz, Susana; González-Warleta, Marta; Mezo, Mercedes; Gómez-Bautista, Mercedes; Ortega-Mora, Luis Miguel; Castro-Hermida, José Antonio

    2013-11-08

    The aim of the present study was to identify the species and/or genotypes of Cryptosporidium and Giardia duodenalis infecting roe deer (Capreolus capreolus) in Galicia (NW Spain). The presence of both enteropathogens was investigated in 212 faecal samples from roe deer shot in diverse game preserves in three different areas of Galicia. The samples were analyzed by immunofluorescence microscopy and PCR amplification, and fragments of the 18S SSU rRNA gene of Cryptosporidium and the β-giardin gene of G. duodenalis were sequenced. In total, 9 samples (4.2%) were positive for Cryptosporidium and 19 samples (8.9%) for G. duodenalis. These samples tested positive with both techniques. However, gene sequencing was only possible for Cryptosporidium in 6 of the samples and for G. duodenalis in 7 of the samples. Cryptosporidium bovis was identified in 3 samples and C. ryanae oocysts were detected in another 3 samples. Sequencing of the amplicons identified G. duodenalis sub-assemblage A-II in 7 samples. Both Cryptosporidium and G. duodenalis infections were more prevalent in juvenile than in adult animals, although the differences were not significant. G. duodenalis was more prevalent than Cryptosporidium in both age groups, although again the differences were not statistically significant. The mean intensity of infection by Cryptosporidium and G. duodenalis was similar in both age groups and ranged between 5 and 225 oocysts/g and 5 and 320 cysts/g of faeces, respectively. This study represents the first molecular characterization of these parasites in Spanish roe deer. Identification of C. bovis and G. duodenalis sub-assemblage A-II indicates that zoonotic transmission of these enteropathogens between roe deer and humans is possible and that cross transmission of some Cryptosporidium species and G. duodenalis (sub-assemblage A-II) may occur between related animal species sharing the same habitats. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Giardia duodenalis genotypes among schoolchildren and their families and pets in urban and rural areas of Sinaloa, Mexico.

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    García-Cervantes, Patricia Catalina; Báez-Flores, María Elena; Delgado-Vargas, Francisco; Ponce-Macotela, Martha; Nawa, Yukifumi; De-la-Cruz-Otero, María Del-Carmen; Martínez-Gordillo, Mario N; Díaz-Camacho, Sylvia Páz

    2017-02-28

    Giardiasis is a human health concern worldwide, especially among schoolchildren. Giardia duodenalis genotypes A and B are infective to humans, but their zoonotic potential remains controversial. In Mexico, the most prevalent genotype is A, but B was also detected in southeastern Mexico. In Sinaloa state, northwestern Mexico, giardiasis is highly prevalent, but Giardia genotypes have been poorly studied. This study aimed to investigate the distribution and clinical-epidemiological correlation of G. duodenalis genotypes in schoolchildren and their families and pets in urban and rural areas of Sinaloa state, Mexico. Among 395 schoolchildren (274 urban, 121 rural), 76 (49 urban, 27 rural) were infected with G. duodenalis. In total, 22 families (15 urban, 7 rural) of infected schoolchildren, consisting of 60 family members (41 urban, 19 rural) and 21 pet dogs (15 urban, 6 rural) were examined; 10 family members (5 urban, 5 rural) and 5 pet dogs (3 urban, 2 rural) of 10 families (6 urban, 4 rural) were infected. After PCR-RFLP analyses of vsp417 and gdh genes, genotype prevalence among infected urban schoolchildren was 79.5% AI, 12.8% AII, and 7.7% mixed AI+B. However, only AI genotype was found in family members and pets. In the rural area, only the AI genotype was detected. Genotypes were not correlated with clinical manifestations. This paper shows the presence of B genotype in northwestern Mexico for the first time. Detection of AI genotype in dogs suggested the possible role of dogs as the reservoir for human giardiasis in Sinaloa, Mexico.

  3. Zoonotic potential of Giardia duodenalis and Cryptosporidium spp. and prevalence of intestinal parasites in young dogs from different populations on Prince Edward Island, Canada.

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    Uehlinger, Fabienne D; Greenwood, Spencer J; McClure, J Trenton; Conboy, Gary; O'Handley, Ryan; Barkema, Herman W

    2013-09-23

    The prevalence of Giardia duodenalis, Cryptosporidium spp. and other intestinal parasites was determined in dogs Intestinal parasites isolated included G. duodenalis, Cryptosporidium spp., Toxocara canis, Isospora spp. and Uncinaria stenocephala. To estimate the zoonotic risk associated with these infections, genotypes of G. duodenalis and Cryptosporidium spp. were determined using 16S rRNA and Hsp70 gene sequencing, respectively. Dogs from the pet store had the highest prevalence of intestinal parasites (78%, 95% CI: 68-88%), followed by the private veterinary clinics (49%, 95% CI: 37-60%), and the local animal shelter (34%, 95% CI: 22-46%). The majority G. duodenalis belonged to host-adapted assemblages D (47%, 95% CI: 31-64%) and C (26%, 95% CI: 13-43%), respectively. Zoonotic assemblages A and B were isolated alone or in mixed infections from 16% (95% CI: 6-31%) of G. duodenalis-positive dogs. All Cryptosporidium spp. were the host-adapted C. canis. While host-adapted, non-zoonotic G. duodenalis genotypes were more common, the presence of G. duodenalis assemblages A and B, T. canis, and U. stenocephala suggests that these dogs may present a zoonotic risk. The zoonotic risk from Cryptosporidium-infected dogs was minimal. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Reducing occurrence of Giardia duodenalis in children living in semiarid regions: impact of a large scale rainwater harvesting initiative.

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    Fonseca, Jacqueline Evangelista; Carneiro, Mariângela; Pena, João Luiz; Colosimo, Enrico A; da Silva, Nívea Bispo; da Costa, André Gabriel F C; Moreira, Luciano E; Cairncross, Sandy; Heller, Léo

    2014-06-01

    In Brazil, about two million people living in rural semiarid regions were benefited with the construction of rainwater cement cisterns, as an initiative from the program "One Million Cisterns" (P1MC). Nevertheless, few epidemiological studies have been conducted to assess health risks or protection effects associated with consumption of this water source. The aim of this study was to evaluate whether access to rainwater harvesting cisterns is associated with the decrease in the occurrence of Giardia duodenalis infections in children, compared to other children living in households supplied by other water sources. A quasi-experimental study with two concurrent cohorts was developed in two rural municipalities of the semiarid region of Brazil. A sample of 664 children, aged between 4 months and 5 years old, was followed up, of which 332 had access to rainwater cisterns (cistern group) and 332 did not, having water supplied from alternative sources (comparison group). In a period of approximately one year (2010) intestinal parasites were investigated in feces three times. The prevalence of G. duodenalis in children from the cistern group ranged from 4.8 to 10.5%, while the prevalence in the comparison group ranged from 7.6 to 16.7%. Multivariate analysis (GEE) showed a higher risk of G. duodenalis infection in children who did not have access to rainwater cisterns, when compared to children who did (OR 1.72; 95% CI 1.14-2.59). The other variables associated with G. duodenalis infection were: number of rooms per house (OR 0.89; 95% CI 0.80-0.99); family income (OR0.48; 95% CI 0.26-0.88); birth order (OR 1.72; 95% CI 1.17-2.51); preterm children (OR 1.70; 95% CI 1.19-2.43); and improper hand hygiene prior to food preparation (OR 4.78; 95% CI 1.95-11.76). Ownership of a rainwater cistern is associated with a lower prevalence of G. duodenalis infection in children after adjustment for environmental and family-related factors. Nevertheless, the study suggests the necessity

  5. Reducing occurrence of Giardia duodenalis in children living in semiarid regions: impact of a large scale rainwater harvesting initiative.

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    Jacqueline Evangelista Fonseca

    2014-06-01

    Full Text Available BACKGROUND: In Brazil, about two million people living in rural semiarid regions were benefited with the construction of rainwater cement cisterns, as an initiative from the program "One Million Cisterns" (P1MC. Nevertheless, few epidemiological studies have been conducted to assess health risks or protection effects associated with consumption of this water source. The aim of this study was to evaluate whether access to rainwater harvesting cisterns is associated with the decrease in the occurrence of Giardia duodenalis infections in children, compared to other children living in households supplied by other water sources. METHODOLOGY/PRINCIPAL FINDINGS: A quasi-experimental study with two concurrent cohorts was developed in two rural municipalities of the semiarid region of Brazil. A sample of 664 children, aged between 4 months and 5 years old, was followed up, of which 332 had access to rainwater cisterns (cistern group and 332 did not, having water supplied from alternative sources (comparison group. In a period of approximately one year (2010 intestinal parasites were investigated in feces three times. The prevalence of G. duodenalis in children from the cistern group ranged from 4.8 to 10.5%, while the prevalence in the comparison group ranged from 7.6 to 16.7%. Multivariate analysis (GEE showed a higher risk of G. duodenalis infection in children who did not have access to rainwater cisterns, when compared to children who did (OR 1.72; 95% CI 1.14-2.59. The other variables associated with G. duodenalis infection were: number of rooms per house (OR 0.89; 95% CI 0.80-0.99; family income (OR0.48; 95% CI 0.26-0.88; birth order (OR 1.72; 95% CI 1.17-2.51; preterm children (OR 1.70; 95% CI 1.19-2.43; and improper hand hygiene prior to food preparation (OR 4.78; 95% CI 1.95-11.76. CONCLUSIONS/SIGNIFICANCE: Ownership of a rainwater cistern is associated with a lower prevalence of G. duodenalis infection in children after adjustment for

  6. Infection rate of Giardia duodenalis, Cryptosporidium spp. and Enterocytozoon bieneusi in cashmere, dairy and meat goats in China.

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    Peng, Xian-Qi; Tian, Ge-Ru; Ren, Guan-Jing; Yu, Zheng-Qing; Lok, James Barron; Zhang, Long-Xian; Wang, Xue-Ting; Song, Jun-Ke; Zhao, Guang-Hui

    2016-07-01

    Cryptosporidiosis, microsporidiosis, and giardiasis contribute significantly to the high burden of zoonotic diarrhea worldwide. Goats constitute an important species in animal agriculture by providing cashmere wool, meat, and dairy products for human consumption. However, zoonotic pathogens with the potential to cause morbidity and to degrade production have been reported frequently in goats recently. The present study examined 629 fecal specimens from goats, including 315 cashmere goats, 170 dairy goats and 144 meat goats, in multiple cities of Shaanxi and Henan provinces, northwestern and central China, to investigate the infection rate and species/assemblages/genotypes of Giardia duodenalis, Cryptosporidium spp. and Enterocytozoon bieneusi. Of these samples, 274 (43.6%) were positive for three zoonotic pathogens, including 80 (12.7%), 104 (16.5%) and 179 (28.5%) for G. duodenalis, Cryptosporidium spp. and E. bieneusi, respectively. Infections with G. duodenalis, Cryptosporidium spp. and E. bieneusi existed in meat, dairy and cashmere goats, with the highest infection rate of each pathogen being observed in meat goats. DNA sequencing of the SSU rRNA gene from 104 Cryptosporidium-positive specimens revealed existence of Cryptosporidium xiaoi, and the zoonotic parasites Cryptosporidium parvum and Cryptosporidium ubiquitum. Genotyping of G. duodenalis based on the triosephosphate isomerase (TPI) gene identified parasites from zoonotic assemblage A in four cashmere goats and the animal-adapted assemblage E in a group of 76 goats that included cashmere, dairy and meat animals. Polymorphisms in the ribosomal internal transcribed spacer characterized E. bieneusi genotype CHG1 and a novel genotype named as SX1 in both dairy and cashmere goats, genotypes CHS7 and COSI in meat goats, the genotype CHG2 in dairy goats, and the human-pathogenic genotype BEB6 in dairy and meat goats. This is the first detailed study to compare infection rate of the zoonotic protozoan pathogens

  7. Giardia duodenalis-induced alterations of commensal bacteria kill Caenorhabditis elegans: a new model to study microbial-microbial interactions in the gut.

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    Gerbaba, Teklu K; Gupta, Pratyush; Rioux, Kevin; Hansen, Dave; Buret, Andre G

    2015-03-15

    Giardia duodenalis is the most common cause of parasitic diarrhea worldwide and a well-established risk factor for postinfectious irritable bowel syndrome. We hypothesized that Giardia-induced disruptions in host-microbiota interactions may play a role in the pathogenesis of giardiasis and in postgiardiasis disease. Functional changes induced by Giardia in commensal bacteria and the resulting effects on Caenorhabditis elegans were determined. Although Giardia or bacteria alone did not affect worm viability, combining commensal Escherichia coli bacteria with Giardia became lethal to C. elegans. Giardia also induced killing of C. elegans with attenuated Citrobacter rodentium espF and map mutant strains, human microbiota from a healthy donor, and microbiota from inflamed colonic sites of ulcerative colitis patient. In contrast, combinations of Giardia with microbiota from noninflamed sites of the same patient allowed for worm survival. The synergistic lethal effects of Giardia and E. coli required the presence of live bacteria and were associated with the facilitation of bacterial colonization in the C. elegans intestine. Exposure to C. elegans and/or Giardia altered the expression of 172 genes in E. coli. The genes affected by Giardia included hydrogen sulfide biosynthesis (HSB) genes, and deletion of a positive regulator of HSB genes, cysB, was sufficient to kill C. elegans even in the absence of Giardia. Our findings indicate that Giardia induces functional changes in commensal bacteria, possibly making them opportunistic pathogens, and alters host-microbe homeostatic interactions. This report describes the use of a novel in vivo model to assess the toxicity of human microbiota. Copyright © 2015 the American Physiological Society.

  8. Disruptions of Host Immunity and Inflammation by Giardia Duodenalis: Potential Consequences for Co-Infections in the Gastro-Intestinal Tract.

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    Cotton, James A; Amat, Christina B; Buret, Andre G

    2015-11-10

    Giardia duodenalis (syn. G. intestinalis, or G. lamblia) is a leading cause of waterborne diarrheal disease that infects hundreds of millions of people annually. Research on Giardia has greatly expanded within the last few years, and our understanding of the pathophysiology and immunology on this parasite is ever increasing. At peak infection, Giardia trophozoites induce pathophysiological responses that culminate in the development of diarrheal disease. However, human data has suggested that the intestinal mucosa of Giardia-infected individuals is devoid of signs of overt intestinal inflammation, an observation that is reproduced in animal models. Thus, our understanding of host inflammatory responses to the parasite remain incompletely understood and human studies and experimental data have produced conflicting results. It is now also apparent that certain Giardia infections contain mechanisms capable of modulating their host's immune responses. As the oral route of Giardia infection is shared with many other gastrointestinal (GI) pathogens, co-infections may often occur, especially in places with poor sanitation and/or improper treatment of drinking water. Moreover, Giardia infections may modulate host immune responses and have been found to protect against the development of diarrheal disease in developing countries. The following review summarizes our current understanding of the immunomodulatory mechanisms of Giardia infections and their consequences for the host, and highlights areas for future research. Potential implications of these immunomodulatory effects during GI co-infection are also discussed.

  9. Giardia duodenalis in Captive Tigers (Panthera tigris), Palawan Bearcats (Arctictis binturong whitei) and Asian Palm Civet (Paradoxurus hermaphroditus) at a Wildlife Facility in Manila, Philippines.

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    Velante, Nick Angelo P; Oronan, Rey B; Reyes, Marco F; Divina, Billy P

    2017-01-01

    This study was conducted to determine the prevalence of Giardia duodenalis in captive animals in a wildlife facility. This is the first study conducted in these animals from the facility. Eight captive tigers ( Panthera tigris ), two Palawan bearcats ( Arctictis binturong whitei ) and one Asian Palm Civet ( Paradoxurus hermaphroditus ) currently housed at a wildlife facility in Manila, Philippines were considered in 2012. These animals were apparently healthy with no signs of disease during the study. Sample collection was done twice at two months interval where freshly voided fecal samples were grossly examined, characterized and preserved in Sodium Acetate Formalin (SAF). The samples were used to determine the presence of G. duodenalis using modified flotation-sedimentation and commercially available immuno-chromatographic assay test kit. All fecal samples tested were negative for the presence of G. duodenalis trophozoites, and cysts using the former. Furthermore, none of the samples tested positive for and G. duodenalis antigen using immune-chromatographic assay. There is no existing infection of G. duodenalis among captive tigers, Palawan Bearcats and Asian palm civet housed at the wildlife facility.

  10. Giardia duodenalis in Captive Tigers (Panthera tigris, Palawan Bearcats (Arctictis binturong whitei and Asian Palm Civet (Paradoxurus hermaphroditus at a Wildlife Facility in Manila, Philippines

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    Nick Angelo P. VELANTE

    2017-09-01

    Full Text Available Background: This study was conducted to determine the prevalence of Giardia duodenalis in captive animals in a wildlife facility. This is the first study conducted in these animals from the facility.Methods: Eight captive tigers (Panthera tigris, two Palawan bearcats (Arctictis binturong whitei and one Asian Palm Civet (Paradoxurus hermaphroditus currently housed at a wildlife facility in Manila, Philippines were considered in 2012. These animals were apparently healthy with no signs of disease during the study. Sample collection was done twice at two months interval where freshly voided fecal samples were grossly examined, characterized and preserved in Sodium Acetate Formalin (SAF. The samples were used to determine the presence of G. duodenalis using modified flotation-sedimentation and commercially available immuno-chromatographic assay test kit.Results: All fecal samples tested were negative for the presence of G. duodenalis trophozoites, and cysts using the former. Furthermore, none of the samples tested positive for and G. duodenalis antigen using immune-chromatographic assay.Conclusion: There is no existing infection of G. duodenalis among captive tigers, Palawan Bearcats and Asian palm civet housed at the wildlife facility. 

  11. Food handlers as a link in the chain of transmission of Giardia duodenalis and other protozoa in public schools in southern Brazil.

    Science.gov (United States)

    Colli, Cristiane Maria; Bezagio, Renata Coltro; Nishi, Letícia; Ferreira, Érika Cristina; Falavigna-Guilherme, Ana Lúcia; Gomes, Mônica Lúcia

    2015-09-01

    Food handlers (FHs) may facilitate transmission and dissemination of pathogens. The importance of FHs as a link in the epidemiological chain of transmission of Giardia duodenalis and other intestinal protozoa was assessed. Fecal and subungual material from 27 FHs were analyzed using parasitological methods. G. duodenalis was identified by direct immunofluorescence and genotyped by PCR-RFLP for the bg and gdh genes, and gdh was sequenced. At least one protozoan was detected in 30% (8/27) of the FHs and G. duodenalis (19%; 5/27) was the most common species. The AII and BIV genotypes were found in 20% (1/5) and 60% (3/5) of FHs infected with G. duodenalis, respectively. FHs can be involved in the chain of transmission of G. duodenalis and other protozoa. KJ741310 - KJ741313. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Comportamiento de la infección experimental por aislamientos colombianos de Giardia duodenalis en el modelo animal del gerbo (Meriones unguiculatus.

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    Adriana Arévalo

    2005-09-01

    Full Text Available Introducción. Se han informado infecciones naturales y experimentales con Giardia sp. en bovinos, equinos, caprinos, caninos, felinos y roedores como ratones, ratas y gerbos; estos últimos son el modelo más adecuado para estudios de la infección por Giardia duodenalis y Giardia muris. Objetivo. Establecer el comportamiento de la infección con aislamientos colombianos de Giardia duodenalis en el modelo animal del gerbo. Materiales y métodos. Se purificaron mediante gradientes de sacarosa y percoll quistes del parásito obtenidos a partir de heces de pacientes sintomáticos infectados. La inoculación a los animales se realizó mediante sonda gástrica con 5x103 quistes. El curso de la infección se estableció mediante recuento diario de quistes y semanal de trofozoítos durante treinta días. Resultados. La eliminación de quistes presentó un patrón intermitente de excreción, con ausencia en la primera y cuarta semanas de infección, y presencia constante durante la segunda y tercera semanas, en número variable con promedio mínimo de 79 y máximo de 17.943 quistes liberados en heces recolectadas en un período de dos horas. Se observó colonización de los trofozoítos en el intestino delgado, en número que osciló entre 15.000 y 6'577.778 trofozoítos por ml. Conclusiones. En gerbos infectados con aislamientos de Giardia duodenalis circulantes en otras regiones geográficas, la resolución natural de la infección oscila entre 86 y 114 días mientras que los gerbos infectados con aislamientos colombianos del parásito la resuelven al día 30. El gerbo constituye un modelo animal adecuado para la infección con aislamientos colombianos de G. duodenalis. La infección experimental por Giardia en gerbo permite obtener quistes y trofozoítos del parásito en cantidades suficientes con la finalidad de ser utilizados como antígenos para la inmunización de animales y para la obtención de anticuerpos que puedan utilizarse para la detecci

  13. Giardia duodenalis infection reduces granulocyte infiltration in an in vivo model of bacterial toxin-induced colitis and attenuates inflammation in human intestinal tissue.

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    Cotton, James A; Motta, Jean-Paul; Schenck, L Patrick; Hirota, Simon A; Beck, Paul L; Buret, Andre G

    2014-01-01

    Giardia duodenalis (syn. G. intestinalis, G. lamblia) is a predominant cause of waterborne diarrheal disease that may lead to post-infectious functional gastrointestinal disorders. Although Giardia-infected individuals could carry as much as 106 trophozoites per centimetre of gut, their intestinal mucosa is devoid of overt signs of inflammation. Recent studies have shown that in endemic countries where bacterial infectious diseases are common, Giardia infections can protect against the development of diarrheal disease and fever. Conversely, separate observations have indicated Giardia infections may enhance the severity of diarrheal disease from a co-infecting pathogen. Polymorphonuclear leukocytes or neutrophils (PMNs) are granulocytic, innate immune cells characteristic of acute intestinal inflammatory responses against bacterial pathogens that contribute to the development of diarrheal disease following recruitment into intestinal tissues. Giardia cathepsin B cysteine proteases have been shown to attenuate PMN chemotaxis towards IL-8/CXCL8, suggesting Giardia targets PMN accumulation. However, the ability of Giardia infections to attenuate PMN accumulation in vivo and how in turn this effect may alter the host inflammatory response in the intestine has yet to be demonstrated. Herein, we report that Giardia infection attenuates granulocyte tissue infiltration induced by intra-rectal instillation of Clostridium difficile toxin A and B in an isolate-dependent manner. This attenuation of granulocyte infiltration into colonic tissues paralled decreased expression of several cytokines associated with the recruitment of PMNs. Giardia trophozoite isolates that attenuated granulocyte infiltration in vivo also decreased protein expression of cytokines released from inflamed mucosal biopsy tissues collected from patients with active Crohn's disease, including several cytokines associated with PMN recruitment. These results demonstrate for the first time that certain

  14. Giardia duodenalis infection reduces granulocyte infiltration in an in vivo model of bacterial toxin-induced colitis and attenuates inflammation in human intestinal tissue.

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    James A Cotton

    Full Text Available Giardia duodenalis (syn. G. intestinalis, G. lamblia is a predominant cause of waterborne diarrheal disease that may lead to post-infectious functional gastrointestinal disorders. Although Giardia-infected individuals could carry as much as 106 trophozoites per centimetre of gut, their intestinal mucosa is devoid of overt signs of inflammation. Recent studies have shown that in endemic countries where bacterial infectious diseases are common, Giardia infections can protect against the development of diarrheal disease and fever. Conversely, separate observations have indicated Giardia infections may enhance the severity of diarrheal disease from a co-infecting pathogen. Polymorphonuclear leukocytes or neutrophils (PMNs are granulocytic, innate immune cells characteristic of acute intestinal inflammatory responses against bacterial pathogens that contribute to the development of diarrheal disease following recruitment into intestinal tissues. Giardia cathepsin B cysteine proteases have been shown to attenuate PMN chemotaxis towards IL-8/CXCL8, suggesting Giardia targets PMN accumulation. However, the ability of Giardia infections to attenuate PMN accumulation in vivo and how in turn this effect may alter the host inflammatory response in the intestine has yet to be demonstrated. Herein, we report that Giardia infection attenuates granulocyte tissue infiltration induced by intra-rectal instillation of Clostridium difficile toxin A and B in an isolate-dependent manner. This attenuation of granulocyte infiltration into colonic tissues paralled decreased expression of several cytokines associated with the recruitment of PMNs. Giardia trophozoite isolates that attenuated granulocyte infiltration in vivo also decreased protein expression of cytokines released from inflamed mucosal biopsy tissues collected from patients with active Crohn's disease, including several cytokines associated with PMN recruitment. These results demonstrate for the first time

  15. Identical assemblage of Giardia duodenalis in humans, animals and vegetables in an urban area in southern Brazil indicates a relationship among them.

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    Cristiane Maria Colli

    Full Text Available Giardia duodenalis infects humans and other mammals by ingestion of cysts in contaminated water or food, or directly in environments with poor hygiene. Eight assemblages, designated A-H, are described for this species.We investigated by microscopy or by direct immunofluorescence technique the occurrence of G. duodenalis in 380 humans, 34 animals, 44 samples of water and 11 of vegetables. G. duodenalis cysts present in samples were genotyped through PCR-RFLP of β giardin and glutamate dehydrogenase (gdh genes and sequencing of gdh. The gdh gene was amplified in 76.5% (26/34 of the human faeces samples with positive microscopy and in 2.9% (1/34 of negative samples. In 70.4% (19/27 of the positive samples were found BIV assemblage. In two samples from dogs with positive microscopy and one negative sample, assemblages BIV, C, and D were found. Cysts of Giardia were not detected in water samples, but three samples used for vegetable irrigation showed total coliforms above the allowed limit, and Escherichia coli was observed in one sample. G. duodenalis BIV was detected in two samples of Lactuca sativa irrigated with this sample of water. BIV was a common genotype, with 100% similarity, between different sources or hosts (humans, animals and vegetables, and the one most often found in humans.This is the first study in Brazil that reports the connection among humans, dogs and vegetables in the transmission dynamics of G. duodenalis in the same geographic area finding identical assemblage. BIV assemblage was the most frequently observed among these different links in the epidemiological chain.

  16. Assessment of the diagnostic performance of four methods for the detection of Giardia duodenalis in fecal samples from human, canine and feline carriers.

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    Uchôa, Flávia Fernandes de Mendonça; Sudré, Adriana Pittella; Campos, Sabrina Destri Emmerick; Almosny, Nádia Regina Pereira

    2018-02-01

    Enteric parasitic diseases including giardiasis are of public health concern. Different methods are available for the diagnosis of this parasitic infection in fecal samples such as the identification of protozoan cysts and trophozoites by light microscopy, detection of specific antigens by ELISA, and amplification of DNA fragments by PCR. The present study aimed at assessing the performance of four laboratory tests for the detection of Giardia duodenalis in fecal specimens from three different host species with a previous diagnosis of giardiasis; canine, feline and human patients provided new stool samples to be retested for Giardia before initiating treatment with antiprotozoal drugs. For this purpose, triplicate fecal specimens from 54 humans, 24 dogs and 18 cats living in the city of Niterói, RJ, southeast Brazil, were analysed by light microscopy, ELISA, immunochromatography, and nested PCR. The centrifugal-flotation method detected Giardia cysts in 89.6% (86/96) of the fecal samples. The protozoan parasite was detected via immunochromatography in 87.5% (84/96) of these samples. Giardia was detected by ELISA in 69.8% (67/96) of the stool specimens from carriers with a previous diagnosis of Giardia infection. Giardia was detected by PCR in only 39.6% (38/96) of the fecal specimens. Based on these findings, we suggest that, among the four assays that were used in this study, the zinc sulphate flotation technique (Faust et al., 1939) is the best diagnostic assay in terms of sensitivity and specificity to detect G. duodenalis on serially collected samples from dogs, cats and humans. Copyright © 2018 Elsevier B.V. All rights reserved.

  17. The Microbiota Contributes to CD8+ T Cell Activation and Nutrient Malabsorption following Intestinal Infection with Giardia duodenalis.

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    Keselman, Aleksander; Li, Erqiu; Maloney, Jenny; Singer, Steven M

    2016-10-01

    Giardia duodenalis is a noninvasive luminal pathogen that impairs digestive function in its host in part by reducing intestinal disaccharidase activity. This enzyme deficiency has been shown in mice to require CD8(+) T cells. We recently showed that both host immune responses and parasite strain affected disaccharidase levels during murine giardiasis. However, high doses of antibiotics were used to facilitate infections in that study, and we therefore decided to systematically examine the effects of antibiotic use on pathogenesis and immune responses in the mouse model of giardiasis. We found that antibiotic treatment did not overtly increase the parasite burden but significantly limited the disaccharidase deficiency observed in infected mice. Moreover, while infected mice had more activated CD8(+) αβ T cells in the small intestinal lamina propria, this increase was absent in antibiotic-treated mice. Infection also led to increased numbers of CD4(+) αβ T cells in the lamina propria and activation of T cell receptor γδ-expressing intraepithelial lymphocytes (IEL), but these changes were not affected by antibiotics. Finally, we show that activated CD8(+) T cells express gamma interferon (IFN-γ) and granzymes but that granzymes are not required for sucrase deficiency. We conclude that CD8(+) T cells become activated in giardiasis through an antibiotic-sensitive process and contribute to reduced sucrase activity. These are the first data directly demonstrating activation of CD8(+) T cells and γδ T cells during Giardia infections. These data also demonstrate that disruption of the intestinal microbiota by antibiotic treatment prevents pathological CD8(+) T cell activation in giardiasis. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  18. Giardia duodenalis assemblage-specific induction of apoptosis and tight junction disruption in human intestinal epithelial cells: effects of mixed infections.

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    Koh, Wan Hon; Geurden, Thomas; Paget, Tim; O'Handley, Ryan; Steuart, Robert F; Thompson, R C Andrew; Buret, Andre G

    2013-04-01

    In view of the interest in genotype-specific pathogenesis in Giardia duodenalis , the aim of the present study was to examine the effects of infection with different, or mixed, G. duodenalis assemblages on the integrity of human intestinal epithelia. To that end, human epithelial cells (HCT-8) were cultured and exposed to different G. duodenalis assemblages (A, B, and E) or a combination of these assemblages. Epithelial disruption and apoptosis were evaluated by fluorescent microscopy and apoptotic oligonucleosome quantification. The results indicate that infection with trophozoites disrupts epithelial tight junctions and induces varying degrees of enterocyte apoptosis, depending on the infecting assemblage. All disruptions were caspase-3 dependent and were more pronounced when caused by a non-host specific assemblage. Furthermore, infections by isolates in combination with isolates from another assemblage enhanced the epithelial disruption and apoptosis. Further studies in vitro and in vivo are required to confirm the mechanisms of enhanced pathogenicity of mixed or non-host specific (or both) G. duodenalis infections. Findings in the present study point to the potential pathogenic importance of intra-species polyparasitism in giardiasis.

  19. Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods

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    Rousseau Angélique

    2018-01-01

    Full Text Available Giardia duodenalis, Cryptosporidium spp. and Toxoplasma gondii are protozoan parasites that have been highlighted as emerging foodborne pathogens by the Food and Agriculture Organization of the United Nations and the World Health Organization. According to the European Food Safety Authority, 4786 foodborne and waterborne outbreaks were reported in Europe in 2016, of which 0.4% were attributed to parasites including Cryptosporidium, Giardia and Trichinella. Until 2016, no standardized methods were available to detect Giardia, Cryptosporidium and Toxoplasma (oocysts in food. Therefore, no regulation exists regarding these biohazards. Nevertheless, considering their low infective dose, ingestion of foodstuffs contaminated by low quantities of these three parasites can lead to human infection. To evaluate the risk of protozoan parasites in food, efforts must be made towards exposure assessment to estimate the contamination along the food chain, from raw products to consumers. This requires determining: (i the occurrence of infective protozoan (oocysts in foods, and (ii the efficacy of control measures to eliminate this contamination. In order to conduct such assessments, methods for identification of viable (i.e. live and infective parasites are required. This review describes the methods currently available to evaluate infectivity and viability of G. duodenalis cysts, Cryptosporidium spp. and T. gondii oocysts, and their potential for application in exposure assessment to determine the presence of the infective protozoa and/or to characterize the efficacy of control measures. Advantages and limits of each method are highlighted and an analytical strategy is proposed to assess exposure to these protozoa.

  20. Giardia duodenalis assemblages and Entamoeba species infecting non-human primates in an Italian zoological garden: zoonotic potential and management traits

    Science.gov (United States)

    2011-01-01

    Background Giardia duodenalis and Entamoeba spp. are among the most common intestinal human protozoan parasites worldwide and they are frequently reported in captive non-human primates (NHP). From a public health point of view, infected animals in zoos constitute a risk for animal caretakers and visitors. In this study we carried out the molecular identification of G. duodenalis and Entamoeba spp. from nine species of primates housed in the zoological garden of Rome, to better ascertain their occurrence and zoonotic potential. Results G. duodenalis was found only in Lemur catta (47.0%). Entamoeba spp. were detected in all species studied, with the exception of Eulemur macaco and Varecia rubra. The number of positive pools ranged from 5.9% in L. catta to 81.2% in Mandrillus sphinx; in Pan troglodytes the observed prevalence was 53.6%. A mixed Entamoeba-Giardia infection was recorded only in one sample of L. catta. All G. duodenalis isolates belonged to the zoonotic assemblage B, sub assemblage BIV. Three Entamoeba species were identified: E. hartmanni, E. coli and E. dispar. Conclusions Our results highlight the importance of regularly testing animals kept in zoos for the diagnosis of zoonotic parasites, in order to evaluate their pathogenic role in the housed animals and the zoonotic risk linked to their presence. A quick detection of the arrival of pathogens into the enclosures could also be a prerequisite to limit their spread into the structure via the introduction of specific control strategies. The need for molecular identification of some parasite species/genotype in order to better define the zoonotic risk is also highlighted. PMID:21988762

  1. Giardia duodenalis assemblages and Entamoeba species infecting non-human primates in an Italian zoological garden: zoonotic potential and management traits

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    Di Cave David

    2011-10-01

    Full Text Available Abstract Background Giardia duodenalis and Entamoeba spp. are among the most common intestinal human protozoan parasites worldwide and they are frequently reported in captive non-human primates (NHP. From a public health point of view, infected animals in zoos constitute a risk for animal caretakers and visitors. In this study we carried out the molecular identification of G. duodenalis and Entamoeba spp. from nine species of primates housed in the zoological garden of Rome, to better ascertain their occurrence and zoonotic potential. Results G. duodenalis was found only in Lemur catta (47.0%. Entamoeba spp. were detected in all species studied, with the exception of Eulemur macaco and Varecia rubra. The number of positive pools ranged from 5.9% in L. catta to 81.2% in Mandrillus sphinx; in Pan troglodytes the observed prevalence was 53.6%. A mixed Entamoeba-Giardia infection was recorded only in one sample of L. catta. All G. duodenalis isolates belonged to the zoonotic assemblage B, sub assemblage BIV. Three Entamoeba species were identified: E. hartmanni, E. coli and E. dispar. Conclusions Our results highlight the importance of regularly testing animals kept in zoos for the diagnosis of zoonotic parasites, in order to evaluate their pathogenic role in the housed animals and the zoonotic risk linked to their presence. A quick detection of the arrival of pathogens into the enclosures could also be a prerequisite to limit their spread into the structure via the introduction of specific control strategies. The need for molecular identification of some parasite species/genotype in order to better define the zoonotic risk is also highlighted.

  2. Giardia duodenalis assemblages and Entamoeba species infecting non-human primates in an Italian zoological garden: zoonotic potential and management traits.

    Science.gov (United States)

    Berrilli, Federica; Prisco, Cristina; Friedrich, Klaus G; Di Cerbo, Pilar; Di Cave, David; De Liberato, Claudio

    2011-10-12

    Giardia duodenalis and Entamoeba spp. are among the most common intestinal human protozoan parasites worldwide and they are frequently reported in captive non-human primates (NHP). From a public health point of view, infected animals in zoos constitute a risk for animal caretakers and visitors. In this study we carried out the molecular identification of G. duodenalis and Entamoeba spp. from nine species of primates housed in the zoological garden of Rome, to better ascertain their occurrence and zoonotic potential. G. duodenalis was found only in Lemur catta (47.0%). Entamoeba spp. were detected in all species studied, with the exception of Eulemur macaco and Varecia rubra. The number of positive pools ranged from 5.9% in L. catta to 81.2% in Mandrillus sphinx; in Pan troglodytes the observed prevalence was 53.6%. A mixed Entamoeba-Giardia infection was recorded only in one sample of L. catta. All G. duodenalis isolates belonged to the zoonotic assemblage B, sub assemblage BIV. Three Entamoeba species were identified: E. hartmanni, E. coli and E. dispar. Our results highlight the importance of regularly testing animals kept in zoos for the diagnosis of zoonotic parasites, in order to evaluate their pathogenic role in the housed animals and the zoonotic risk linked to their presence. A quick detection of the arrival of pathogens into the enclosures could also be a prerequisite to limit their spread into the structure via the introduction of specific control strategies. The need for molecular identification of some parasite species/genotype in order to better define the zoonotic risk is also highlighted.

  3. Molecular identification of Giardia duodenalis isolates from domestic dogs and cats in Wroclaw, Poland

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    Jolanta Piekarska

    2016-07-01

    The prevalence of G. duodenalis was 21.1% (27/128 in dogs and 15.1% (5/33 in cats. In dogs, C assemblage was present in 18 (81% positive stool samples, D assemblage in 2 (9% samples, B assemblage present in one (4.5%, and mixed assemblages (C and D occurred in one (4.5% sample. F assemblage was found in 4 (80% cats’ positive stool samples and A assemblage occurred in one case (20%. Confirmation of the presence of A and B zoonotic assemblages suggests that infected pets can be a threat to human health. This study describes for the first time the presence of mixed infections within host-specific C and D assemblages in dogs in Poland.

  4. Evaluation of the BD MAX Enteric Parasite Panel for the detection of Cryptosporidium parvum/hominis, Giardia duodenalis and Entamoeba histolytica.

    Science.gov (United States)

    Perry, Michael D; Corden, Sally A; Lewis White, P

    2017-08-01

    Conventional laboratory detection methods for gastrointestinal parasites are time consuming, require considerable technical expertise and may suffer from poor analytical sensitivity. This study sought to evaluate the automated BD MAX Enteric Parasite Panel (EPP) for the detection of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia duodenalis.Methodolgy. A total of 104 known positive samples (43 Cryptosporidium parvum/hominis and 61 G. duodenalis), 15 simulated samples (E. histolytica and other Entamoeba species) and 745 patient stool samples, submitted for enteric pathogen culture and microscopy, were inoculated into BD MAX EPP sample buffer tubes (SBTs). All specimens were blinded and tested within 7 days of SBT inoculation using the BD MAX EPP assay with results compared to those generated by microscopy.Results/Key findings. Combining the results from the known positive samples and anonymously tested patient samples, the sensitivity of the BD MAX EPP assay was 100 % for both Cryptosporidium spp. and G. duodenalis. Specificities of 99.7 and 98.9 % were calculated for the detection of Cryptosporidium spp. and G. duodenalis respectively. Insufficient clinical specimen data was available to determine the performance of the assay for E. histolytica detection. The findings of this study indicate that the BD MAX EPP is suitable for the detection of Cryptosporidium parvum/hominis and G. duodenalis from clinical specimens with reduced hands-on time and complexity compared to microscopy. Results for the detection of E. histolytica were promising although further work is required to evaluate the assay for the detection of this pathogen.

  5. Molecular characterization of Giardia duodenalis cysts in the Oreto River (Sicily, Southern Italy

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    Maria Antonella Di Benedetto

    2010-03-01

    Full Text Available

    Background: The presence of Giardia was investigated along the Oreto river between January 2008 and May 2009 with the aim of understanding the source of contamination by molecular typing of cysts.

    Methods: A total of 38 water samples (10 collected from the river mouth, 24 from the whole Oreto basin and 4 sewage samples from the Monreale treatment plant were processed. In addition, 22 faecal samples of livestock living close to the Oreto area, were analyzed. The presence of Giardia was determined by immunofluorescence assay and their genetic characterization was achieved by a nested PCR assay targeting the triosephosphate isomerase gene.

    Results: All water samples from the river mouth were positive for Giardia, even if the concentration of cysts fluctuated considerably among sampling occasions. Our investigation showed that the Vadduneddu and Altofonte torrents, two influents of the river, were the principal sources of contamination. Moreover, the genotypes of Vadduneddu torrent were the same as those detected in human wastewater taken from the activated sludge plant of Monreale city. Assemblages A and B were found in water samples with a predominance of Assemblage A, subtype AII. Assemblage E was only found in a single calf isolate.

    Conclusions: The data show that the high cyst counts regularly detected in the Oreto river are due to contamination with wastewater of human origin. This finding is relevant for public health, particularly because river water is used for agricultural purposes.

  6. Molecular typing of Giardia duodenalis isolates from nonhuman primates housed IN a Brazilian zoo.

    Science.gov (United States)

    David, Érica Boarato; Patti, Mariella; Coradi, Silvana Torossian; Oliveira-Sequeira, Teresa Cristina Goulart; Ribolla, Paulo Eduardo Martins; Guimarães, Semíramis

    2014-01-01

    Giardia infections in captive nonhuman primates (NHP) housed at a Brazilian zoo were investigated in order to address their zoonotic potential. Fresh fecal samples were collected from the floors of 22 enclosures where 47 primates of 18 different species were housed. The diagnosis of intestinal parasites after concentration by sedimentation and flotation methods revealed the following parasites and their frequencies: Giardia (18%); Entamoeba spp. (18%); Endolimax nana (4.5%); Iodamoeba spp. (4.5%); Oxyurid (4.5%) and Strongylid (4.5%). Genomic DNA extracted from all samples was processed by PCR methods in order to amplify fragments of gdh and tpi genes of Giardia. Amplicons were obtained from samples of Ateles belzebuth, Alouatta caraya, Alouatta fusca and Alouatta seniculus. Clear sequences were only obtained for the isolates from Ateles belzebuth (BA1), Alouatta fusca (BA2) and Alouatta caraya (BA3). According to the phenetic analyses of these sequences, all were classified as assemblage A. For the tpi gene, all three isolates were grouped into sub-assemblage AII (BA1, BA2 and BA3) whereas for the gdh gene, only BA3 was sub-assemblage AII, and the BA1 and BA2 were sub-assemblage AI. Considering the zoonotic potential of the assemblage A, and that the animals of the present study show no clinical signs of infection, the data obtained here stresses that regular coproparasitological surveys are necessary to implement preventive measures and safeguard the health of the captive animals, of their caretakers and of people visiting the zoological gardens.

  7. Prevalence and Genetic Diversity of Giardia duodenalis and Cryptosporidium spp. among School Children in a Rural Area of the Amhara Region, North-West Ethiopia.

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    Aida de Lucio

    Full Text Available Giardia duodenalis and Cryptosporidium spp. are enteric protozoan causing gastrointestinal illness in humans and animals. Giardiasis and cryptosporidiosis are not formally considered as neglected tropical diseases, but belong to the group of poverty-related infectious diseases that impair the development and socio-economic potential of infected individuals in developing countries.We report here the prevalence and genetic diversity of G. duodenalis and Cryptosporidium spp. in children attending rural primary schools in the Bahir Dar district of the Amhara Region, Ethiopia. Stool samples were collected from 393 children and analysed by molecular methods. G. duodenalis was detected by real-time PCR, and the assemblages and sub-assemblages were determined by multilocus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Detection and identification of Cryptosporidium species was carried out by sequencing of a partial fragment of the small-subunit ribosomal RNA gene.The PCR-based prevalences of G. duodenalis and Cryptosporidium spp. were 55.0% (216/393 and 4.6% (18/393, respectively. A total of 78 G. duodenalis isolates were successfully characterized, revealing the presence of sub-assemblages AII (10.3%, BIII (28.2%, and BIV (32.0%. Discordant typing results AII/AIII and BIII/BIV were identified in 7.7% and 15.4% of the isolates, respectively. An additional five (6.4% isolates were assigned to assemblage B. No mixed infections of assemblages A+B were found. Extensive genetic variation at the nucleotide level was observed within assemblage B (but no within assemblage A, resulting in the identification of a large number of sub-types. Cryptosporidium diversity was demonstrated by the occurrence of C. hominis, C. parvum, and C. viatorum in the population under study.Our data suggest an epidemiological scenario with an elevated transmission intensity of a wide range of G. duodenalis genetic variants. Importantly

  8. Molecular diversity and frequency of the diarrheagenic enteric protozoan Giardia duodenalis and Cryptosporidium spp. in a hospital setting in Northern Spain.

    Science.gov (United States)

    Azcona-Gutiérrez, José Manuel; de Lucio, Aida; Hernández-de-Mingo, Marta; García-García, Concepción; Soria-Blanco, Luis Miguel; Morales, Lucía; Aguilera, María; Fuentes, Isabel; Carmena, David

    2017-01-01

    Human giardiosis and cryptosporidiosis are caused by the enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. Both pathogens are major contributors to the global burden of diarrhoeal disease, affecting primarily children and immunodebilitated individuals in resource-poor settings. Giardiosis and cryptosporidiosis also represent an important, often underestimate, public health threat in developed countries. In Spain only limited information is currently available on the epidemiology of these infections. Molecular data on the diversity, frequency, geographical distribution, and seasonality of G. duodenalis assemblages/sub-assemblages and Cryptosporidium species/sub-genotypes are particularly scarce. A longitudinal molecular epidemiological survey was conducted between July 2015 to September 2016 in patients referred to or attended at the Hospital San Pedro (La Rioja, Northern Spain) that tested positive for G. duodenalis (N = 106) or Cryptosporidium spp. (N = 103) by direct microscopy and/or a rapid lateral flow immunochromatographic assay. G. duodenalis infections were subsequently confirmed by real-time PCR and positive isolates assessed by multi-locus sequence genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Cryptosporidium species and sub-genotypes were investigated at the 60 kDa glycoprotein or the small subunit ribosomal RNA genes of the parasite. Sociodemographic and clinical parameters of infected patients were also gathered and analysed. Out of 90 G. duodenalis-positive isolates by real-time PCR a total of 16 isolates were successfully typed. AII (44%, 7/16) was the most prevalent sub-assemblage found, followed by BIV (31%, 5/16) and BIII (19%, 3/16). A discordant genotype result AII/AIII was identified in an additional (6%, 1/16) isolate. No mixed infections A+B were detected. Similarly, a total of 81 Cryptosporidium spp. isolates were successfully typed, revealing the presence of C. hominis (81%, 66/81) and

  9. Molecular diversity and frequency of the diarrheagenic enteric protozoan Giardia duodenalis and Cryptosporidium spp. in a hospital setting in Northern Spain.

    Directory of Open Access Journals (Sweden)

    José Manuel Azcona-Gutiérrez

    Full Text Available Human giardiosis and cryptosporidiosis are caused by the enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. Both pathogens are major contributors to the global burden of diarrhoeal disease, affecting primarily children and immunodebilitated individuals in resource-poor settings. Giardiosis and cryptosporidiosis also represent an important, often underestimate, public health threat in developed countries. In Spain only limited information is currently available on the epidemiology of these infections. Molecular data on the diversity, frequency, geographical distribution, and seasonality of G. duodenalis assemblages/sub-assemblages and Cryptosporidium species/sub-genotypes are particularly scarce.A longitudinal molecular epidemiological survey was conducted between July 2015 to September 2016 in patients referred to or attended at the Hospital San Pedro (La Rioja, Northern Spain that tested positive for G. duodenalis (N = 106 or Cryptosporidium spp. (N = 103 by direct microscopy and/or a rapid lateral flow immunochromatographic assay. G. duodenalis infections were subsequently confirmed by real-time PCR and positive isolates assessed by multi-locus sequence genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Cryptosporidium species and sub-genotypes were investigated at the 60 kDa glycoprotein or the small subunit ribosomal RNA genes of the parasite. Sociodemographic and clinical parameters of infected patients were also gathered and analysed.Out of 90 G. duodenalis-positive isolates by real-time PCR a total of 16 isolates were successfully typed. AII (44%, 7/16 was the most prevalent sub-assemblage found, followed by BIV (31%, 5/16 and BIII (19%, 3/16. A discordant genotype result AII/AIII was identified in an additional (6%, 1/16 isolate. No mixed infections A+B were detected. Similarly, a total of 81 Cryptosporidium spp. isolates were successfully typed, revealing the presence of C. hominis (81

  10. A meta-analysis of the effectiveness of albendazole compared with metronidazole as treatments for infections with Giardia duodenalis.

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    Shahram Solaymani-Mohammadi

    2010-05-01

    Full Text Available Metronidazole is the most commonly used drug for the treatment of giardiasis in humans. In spite of its therapeutic efficacy for giardiasis, low patient compliance, especially in children, side effects, and the emergence of metronidazole-resistant strains may restrict its use. Albendazole has been used to treat Giardia duodenalis infections in recent years. However, efficacy studies in vivo and in vitro have produced diverse results as to its effectiveness. A moderately benign side effect profile, combined with established efficacy against many helminths, renders it promising for treatment of giardiasis in humans.We performed a search in the PubMed, Scopus, EMBASE, the ISI Web of Science, LILIACS, and Cochrane Controlled Trials Register for trials published before February 2010 as well as in references of relevant research and review articles. Eight randomized clinical trials (including 900 patients comparing the effectiveness of albendazole with that of metronidazole were included in meta-analysis. After extracting and validating the data, the pooled risk ratio (RR was calculated using an inverse-variance random-effects model. Albendazole was found to be equally as effective as metronidazole in the treatment of giardiasis in humans (RR 0.97; 95% CI, 0.93, 1.01. In addition, safety analysis suggested that patients treated with albendazole had a lower risk of adverse effects compared with those who received metronidazole (RR 0.36; 95% CI, 0.10, 1.34, but limitations of the sample size precluded a definite conclusion.The effectiveness of albendazole, when given as a single dose of 400 mg/day for 5 days, was comparable to that of metronidazole. Patients treated with albendazole tended to have fewer side effects compared with those who took metronidazole. Given the safety, effectiveness, and low costs of albendazole, this drug could be potentially used as an alternative and/or a replacement for the existing metronidazole therapy protocols in the

  11. Seasonal prevalence and incidence of Cryptosporidium spp. and Giardia duodenalis and associated diarrhoea in children attending pre-schools in Kafue, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I.G.K.; Enemark, Heidi L.

    2011-01-01

    Prevalence, incidence and seasonal variation of Cryptosporidium and Giardia duodenalis were studied over a 12-month period in 100 children from four pre-schools in Kafue, Zambia. Questionnaire data and a single stool sample were collected monthly from each child. Samples were processed using...... a commercial kit (Meridian Diagnostics Inc., USA) and oo(cysts) visualised by immunofluorescence microscopy. Cryptosporidium was detected in 30.7% (241/786; 95% CI = 27.5-33.9) while G. duodenalis was detected in 29.0% (228/786; 95% CI = 25.8-32.2). A total of 86% experienced one or more episodes...... of cryptosporidiosis while 75% had giardiasis. Cumulative incidence per 100 children was 75.4 for Cryptosporidium and 49.0 for G. duodenalis. Both infections were significantly more common in the wet compared to the dry season (34.8%, 162/466 vs. 24.7%, 79/320, P = 0.003 and 35.2%, 164/466 vs. 20.0%, 64/320, P

  12. Evaluation of the Roche LightMix®Gastro Parasites multiplex PCR assay detecting Giardia duodenalis, Entamoeba histolytica, cryptosporidia, Dientamoeba fragilis, and Blastocystis hominis.

    Science.gov (United States)

    Friesen, Johannes; Fuhrmann, Jörg; Kietzmann, Heike; Tannich, Egbert; Müller, Michael; Ignatius, Ralf

    2018-03-23

    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. 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 analyzed by in-house PCR. D. fragilis or B. hominis were detected by LMAGP in 131 (14.4%) and 179 (19.9%; 16 samples positive by microscopy; P 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 analyzed 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). The data call for routine use of multiplex PCR assays for the detection of enteric protozoan parasites in laboratory diagnostics. Copyright © 2018. Published by Elsevier Ltd.

  13. Prevalence of Dientamoeba fragilis, Giardia duodenalis, Entamoeba histolytica/dispar, and Cryptosporidium spp in Da Nang, Vietnam, detected by a multiplex real-time PCR.

    Science.gov (United States)

    Ögren, Jessica; Van Nguyen, Song; Nguyen, Minh Khac; Dimberg, Jan; Matussek, Andreas

    2016-06-01

    We surveyed the prevalence of Dientamoeba fragilis, Giardia duodenalis, Entamoeba histolytica, Entamoeba dispar, and Cryptosporidium spp in individuals with and without gastrointestinal (GI) symptoms residing in and around Da Nang city, Vietnam. Fecal samples were collected from children (n = 100) and adults (n = 80) with GI symptoms and from healthy individuals (n = 88) reporting no GI symptoms. Parasite detection was performed by multiplex real-time PCR. Overall, except for G. duodenalis, we found a low prevalence (<5%) of D. fragilis and E. dispar and no detection of E. histolytica and C. spp in all participants with GI symptoms. Specifically for D. fragilis this contrasts with findings in European populations of children with GI symptoms showing prevalence up to 73%. Moreover, our results indicate that the prevalence of G. duodenalis is higher in patients with GI symptoms compared to asymptomatic individuals and this difference is most obvious in young patients. © 2016 The Authors. APMIS published by John Wiley & Sons Ltd on behalf of Scandinavian Societies for Medical Microbiology and Pathology.

  14. Gut microbiota related to Giardia duodenalis, Entamoeba spp. and Blastocystis hominis infections in humans from Côte d'Ivoire.

    Science.gov (United States)

    Iebba, Valerio; Santangelo, Floriana; Totino, Valentina; Pantanella, Fabrizio; Monsia, Anatole; Di Cristanziano, Veronica; Di Cave, David; Schippa, Serena; Berrilli, Federica; D'Alfonso, Rossella

    2016-09-30

    Literature data provide little information about protozoa infections and gut microbiota compositional shifts in humans. This preliminary study aimed to describe the fecal bacterial community composition of people from Côte d'Ivoire harboring Giardia duodenalis, Entamoeba spp., and Blastocystis hominis, in trying to discover possible alterations in their fecal microbiota structure related to the presence of such parasites. Twenty fecal samples were collected from people inhabiting three different localities of Côte d'Ivoire for copromicroscopic analysis and molecular identification of G. duodenalis, Entamoeba spp., and B. hominis. Temporal temperature gradient gel electrophoresis (TTGE) was used to obtain a fingerprint of the overall bacterial community; quantitative polymerase chain reaction (qPCR) was used to define the relative abundances of selected bacterial species/group, and multivariate statistical analyses were employed to correlate all data. Cluster analysis revealed a significant separation of TTGE profiles into four clusters (p Entamoeba spp./B. hominis-positive subjects were linked to a eubiotic condition, as shown by a significantly higher Faecalibacterium prausnitzii-Escherichia coli ratio. This preliminary investigation demonstrates a differential fecal microbiota structure in subjects infected with G. duodenalis or Entamoeba spp./B. hominis, paving the way for using further next-generation DNA technologies to better understand host-parasite-bacteria interactions, aimed at identifying potential indicators of microbiota changes.

  15. [Giardia duodenalis genotypes found in the Instituto Colombiano de Bienestar Familiar day care centers and dogs in Ibagué, Colombia].

    Science.gov (United States)

    Rodríguez, Victoria; Espinosa, Oneida; Carranza, Julio César; Duque, Sofía; Arévalo, Adriana; Clavijo, Jairo Alfonso; Urrea, Daniel Alfonso; Vallejo, Gustavo Adolfo

    2014-01-01

    Eight Giardia duodenalis genotypes (A-H) have been described to date. Genotypes A and B have been isolated from humans and a wide range of mammals; however, genotypes C-H have shown greater host specificity. Identifying G. duodenalis genotypes from cysts in faeces obtained from children attending the Instituto Colombiano de Bienestar Familiar (ICBF) day care centres and from dogs in Ibagué by PCR-RFLP targeting both the b -giardin and glutamate dehydrogenase genes. Cysts from G. duodenalis positive samples were concentrated, DNA was extracted and the b -giardin and glutamate dehydrogenase genes were analysed by PCR-RFLP. The MHOM/CO/04/G40 strain was used as positive control (this was obtained from the Grupo de Parasitología at the Instituto Nacional de Salud ). Of the total human samples, 11/23 (48%) were genotyped as A and 12/23 (52%) as B; PCR-RFLP revealed that four canine samples were genotypes C and D, these being host-specific. Only genotypes associated with human infection (AII, BIII and BIV) were found in the children and host-specific genotypes were observed in canines (C and D). No interaction could be established between animal and human transmission cycles due to the small canine sample size and as the former did not come into contact with children attending ICBF day-care centres.

  16. Comparison of sensitivity and specificity of 4 methods for detection of Giardia duodenalis in feces: immunofluorescence and PCR are superior to microscopy of concentrated iodine-stained samples.

    Science.gov (United States)

    Gotfred-Rasmussen, Helle; Lund, Marianne; Enemark, Heidi L; Erlandsen, Mogens; Petersen, Eskild

    2016-03-01

    For decades, microscopy of feces after formol-ethylacetate (FEA) concentration and iodine staining has been the routine test for intestinal protozoa. Lately, polymerase chain reaction or fluorescence-labeled parasite-specific antibodies have been introduced, but their place in everyday routine diagnostics has not yet been established. We compared FEA and salt-sugar flotation (SSF) concentration followed by microscopy of iodine-stained concentrate and immunofluorescence assay (IFA) and real-time polymerase chain reaction (qPCR) for detection of Giardia duodenalis in human feces. The median number of Giardia cysts found by FEA in 19 Giardia-positive samples was 50 cysts per gram (CPG), by SSF 350 CPG, by IFA 76,700 CPG, and by qPCR 316,000 CPG. We next tested 455 consecutive samples for presence of Giardia cysts. Using IFA as reference, qPCR had a sensitivity of 91%, specificity of 95.1%, a false-positive rate of 50%, a false-negative rate of 0.48%, a positive predictive value of 50%, and a negative predictive value of 99.5%. In conclusion, qPCR and IFA were significantly more sensitive than microscopy of iodine-stained concentrates using either FEA or SSF. We suggest, when using qPCR, that positive samples are verified by IFA to prevent false-positive results. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  17. Performance of Glutamate Dehydrogenase and Triose Phosphate Isomerase Genes in the Analysis of Genotypic Variability of Isolates of Giardia duodenalis from Livestocks

    Science.gov (United States)

    Fava, Natália M. N.; Soares, Rodrigo M.; Scalia, Luana A. M.; Kalapothakis, Evanguedes; Pena, Isabella F.; Vieira, Carlos U.; Faria, Elaine S. M.; Cunha, Maria J.; Couto, Talles R.; Cury, Márcia Cristina

    2013-01-01

    Giardia duodenalis is a small intestinal protozoan parasite of several terrestrial vertebrates. This work aims to assess the genotypic variability of Giardia duodenalis isolates from cattle, sheep and pigs in the Southeast of Brazil, by comparing the standard characterization between glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) primers. Fecal samples from the three groups of animals were analyzed using the zinc sulphate centrifugal flotation technique. Out of 59 positive samples, 30 were from cattle, 26 from sheep and 3 from pigs. Cyst pellets were stored and submitted to PCR and nested-PCR reactions with gdh and tpi primers. Fragment amplification of gdh and tpi genes was observed in 25 (42.4%) and 36 (61.0%) samples, respectively. Regarding the sequencing, 24 sequences were obtained with gdh and 20 with tpi. For both genes, there was a prevalence of E specific species assemblage, although some isolates have been identified as A and B, by the tpi sequencing. This has also shown a larger number of heterogeneous sequences, which have been attribute to mixed infections between assemblages B and E. The largest variability of inter-assemblage associated to the frequency of heterogeneity provided by tpi sequencing reinforces the polymorphic nature of this gene and makes it an excellent target for studies on molecular epidemiology. PMID:24308010

  18. Identificación de antígenos de aislamientos colombianos de Giardia duodenalis reconocidos por IgG total y subclases.

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    Jenny Fabiola Hernández

    2003-09-01

    Full Text Available Se conocen investigaciones realizadas sobre la respuesta inmune humoral en giardiosis. Sin embargo, estudios sobre perfiles de antígenos de quistes y de trofozoítos de Giardia duodenalis reconocidos por IgG y subclases (IgG1, IgG2, IgG3, IgG4 anti-G. duodenalis no se han realizado en la misma magnitud. Con el fin de determinar los antígenos de los aislamientos colombianos de G. duodenalis reconocidos por IgG y sus subclases anti-G. duodenalis, se utilizó la metodología de Western blot. Quistes y trofozoítos del parásito, independientemente, se sometieron a separación de proteínas mediante SDS-PAGE. Las proteínas separadas fueron transferidas a membrana de nitrocelulosa mediante inmunoelectrotransferencia y su antigenicidad determinada confrontando éstas con IgG y sus subclases anti-G. duodenalis presentes en el suero de pacientes con giardiosis comprobada parasitológicamente. La unión antígeno-anticuerpo se detectó con conjugados anti-inmunoglobulina específicos unidos a fosfatasa alcalina, la cual permitió evidenciar los polipéptidos antigénicos cuando se adicionó el sustrato 5-bromo-4-cloro-3-indolil-fosfato/azul de nitrotetrazolio. Se leyeron y analizaron las bandas mediante análisis de regresión lineal, utilizando el programa Quantity One®. Se reconocieron 32 antígenos, simultáneamente en quistes y trofozoítos de aislamientos colombianos del parásito por IgG total anti-G. duodenalis, que oscilaron entre 22-185 kDa y 19 en un rango de 42 a 180 kDa, reconocidos tanto por IgG1 como por IgG3 anti-G. duodenalis. Las IgG2 e IgG4 anti-parásito no reconocieron antígenos en ninguno de los dos estadios. Los antígenos de 27, 30, 31, 33, 45, 49, 57, 78, 89 y 170 kDa son compartidos con aislamientos de G. duodenalis circulantes en otras regiones geográficas. El reconocimiento de antígenos de quistes y trofozoítos de los aislamientos colombianos de G. duodenalis por IgG, IgG1 e IgG3 anti-G. duodenalis de pacientes

  19. Deconjugated bile salts produced by extracellular bile-salt hydrolase-like activities from the probiotic Lactobacillus johnsonii La1 inhibit Giardia duodenalis in vitro growth

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    Marie-Agnès Travers

    2016-09-01

    Full Text Available Giardiasis, currently considered a neglected disease, is caused by the intestinal protozoan parasite Giardia duodenalis and is widely spread in human as well as domestic and wild animals. The lack of appropriate medications and the spread of resistant parasite strains urgently call for the development of novel therapeutic strategies. Host microbiota or certain probiotic strains have the capacity to provide some protection against giardiasis. By combining biological and biochemical approaches, we have been able to decipher a molecular mechanism used by the probiotic strain Lactobacillus johnsonii La1 to prevent Giardia growth in vitro. We provide evidence that the supernatant of this strain contains active principle(s not directly toxic to Giardia but able to convert non-toxic components of bile into components highly toxic to Giardia. By using bile acid profiling, these components were identified as deconjugated bile-salts. A bacterial bile-salt-hydrolase of commercial origin was able to mimic the properties of the supernatant. Mass spectrometric analysis of the bacterial supernatant identified two of the three bile-salt-hydrolases encoded in the genome of this probiotic strain. These observations document a possible mechanism by which L. johnsonii La1, by secreting or releasing BSH-like activity(ies in the vicinity of replicating Giardia in an environment where bile is present and abundant, can fight this parasite. This discovery has both fundamental and applied outcomes to fight giardiasis, based on local delivery of deconjugated bile salts, enzyme deconjugation of bile components, or natural or recombinant probiotic strains that secrete or release such deconjugating activities in a compartment where both bile salts and Giardia are present.

  20. Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi and other intestinal parasites in young children in Lobata province, Democratic Republic of São Tomé and Principe.

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    Maria Luísa Lobo

    Full Text Available Rare systemic studies concerning prevalence of intestinal parasites in children have been conducted in the second smallest country in Africa, the Democratic Republic of São Tomé and Príncipe. Fecal specimens from 348 children (214 in-hospital attending the Aires de Menezes Hospital and 134 from Agostinho Neto village in São Tome Island were studied by parasitological and molecular methods. Of the 134 children from Agostinho Neto, 52.2% presented intestinal parasites. 32.1% and 20.2% of these children had monoparasitism and polyparasitism, respectively. Ascaris lumbricoides (27.6%, G. duodenalis (7.5%, T. trichiura (4.5% and Entamoeba coli (10.5% were the more frequent species identified in the children of this village. Giardia duodenalis (7.5% and E. bieneusi (5.2% were identified by PCR. Nested-PCR targeting G. duodenalis TPI identified Assemblage A (60% and Assemblage B (40%. The E. bieneusi ITS-based sequence identified genotypes K (57.1%, KIN1 (28.6% and KIN3 (14.3%. Among the 214 in-hospital children, 29.4% presented intestinal parasites. In 22.4% and 7.0% of the parasitized children, respectively, one or more species were concurrently detected. By microscopy, A. lumbricoides (10.3% and Trichiuris trichiura (6.5% were the most prevalent species among these children, and Cryptosporidium was detected by PCR in 8.9% of children. GP60 locus analysis identified 6.5% of C. hominis (subtypes IaA27R3 [35.7%], IaA23R3 [14.3%], IeA11G3T3 [28.6%] and IeA11G3T3R1 [21.4%] and 2.3% of C. parvum (subtypes IIaA16G2R1 [20.0%], IIaA15G2R1 [20.0%], IIdA26G1 [40.0%] and IIdA21G1a [20.0%]. G. duodenalis and E. bieneusi were identified in 0.5% and 8.9% of the in-hospital children, respectively. G. duodenalis Assemblage B was characterized. The E. bieneusi genotypes K (52.6%, D (26.4%, A (10.5% and KIN1 (10.5% were identified. Although further studies are required to clarify the epidemiology of these infectious diseases in this endemic region the significance

  1. Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi and other intestinal parasites in young children in Lobata province, Democratic Republic of São Tomé and Principe.

    Science.gov (United States)

    Lobo, Maria Luísa; Augusto, João; Antunes, Francisco; Ceita, José; Xiao, Lihua; Codices, Vera; Matos, Olga

    2014-01-01

    Rare systemic studies concerning prevalence of intestinal parasites in children have been conducted in the second smallest country in Africa, the Democratic Republic of São Tomé and Príncipe. Fecal specimens from 348 children (214 in-hospital attending the Aires de Menezes Hospital and 134 from Agostinho Neto village) in São Tome Island were studied by parasitological and molecular methods. Of the 134 children from Agostinho Neto, 52.2% presented intestinal parasites. 32.1% and 20.2% of these children had monoparasitism and polyparasitism, respectively. Ascaris lumbricoides (27.6%), G. duodenalis (7.5%), T. trichiura (4.5%) and Entamoeba coli (10.5%) were the more frequent species identified in the children of this village. Giardia duodenalis (7.5%) and E. bieneusi (5.2%) were identified by PCR. Nested-PCR targeting G. duodenalis TPI identified Assemblage A (60%) and Assemblage B (40%). The E. bieneusi ITS-based sequence identified genotypes K (57.1%), KIN1 (28.6%) and KIN3 (14.3%). Among the 214 in-hospital children, 29.4% presented intestinal parasites. In 22.4% and 7.0% of the parasitized children, respectively, one or more species were concurrently detected. By microscopy, A. lumbricoides (10.3%) and Trichiuris trichiura (6.5%) were the most prevalent species among these children, and Cryptosporidium was detected by PCR in 8.9% of children. GP60 locus analysis identified 6.5% of C. hominis (subtypes IaA27R3 [35.7%], IaA23R3 [14.3%], IeA11G3T3 [28.6%] and IeA11G3T3R1 [21.4%]) and 2.3% of C. parvum (subtypes IIaA16G2R1 [20.0%], IIaA15G2R1 [20.0%], IIdA26G1 [40.0%] and IIdA21G1a [20.0%]). G. duodenalis and E. bieneusi were identified in 0.5% and 8.9% of the in-hospital children, respectively. G. duodenalis Assemblage B was characterized. The E. bieneusi genotypes K (52.6%), D (26.4%), A (10.5%) and KIN1 (10.5%) were identified. Although further studies are required to clarify the epidemiology of these infectious diseases in this endemic region the significance

  2. Concurrent infections of Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile in children during a cryptosporidiosis outbreak in a pediatric hospital in China.

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    Lin Wang

    Full Text Available BACKGROUND: Over 200 cryptosporidiosis outbreaks have been reported, but little is known if other enteric pathogens were also involved in some of these outbreaks. Recently, an outbreak of cryptosporidiosis linked to poor hygiene by two Cryptosporidium hominis subtypes occurred in a pediatric hospital ward (Ward A in China, lasting for more than 14 months. In this study, the concurrence during the outbreak of three other enteric pathogens with a similar transmission route, Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile, was assessed. METHODS/PRINCIPAL FINDINGS: The occurrence of G. duodenalis, E. bieneusi, and C. difficile in 78 inpatients from Ward A and 283 and 216 inpatients from two control wards (Wards C and D in the same hospital was examined using molecular diagnostic tools. Significantly higher infection rates were found in children in Ward A for all study pathogens than in Wards C and D (P<0.01: 9.5% versus 1.4% and 0% for G. duodenalis, 10.8% versus 2.8% and 3.7% for E. bieneusi, and 60.8% versus 37.8% and 27.8% for C. difficile, respectively. These differences were mostly seen in children ≤ 12 months. Enteric pathogen-positive children in Ward A (31/58 or 53.4% were more likely to have mixed infections than those in Ward C (4/119 or 3.4% or D (5/68, 7.4%; P<0.01. Having cryptosporidiosis was a risk factor for G. duodenalis (OR = 4.3; P = 0.08, E. bieneusi (OR = 3.1; P = 0.04, and C. difficile (OR = 4.7; P<0.01 infection. In addition, a lower diversity of G. duodenalis, E. bieneusi, and C. difficile genotypes/subtypes was observed in Ward A. CONCLUSIONS/SIGNIFICANCE: Data from this study suggest that multiple pathogens were concurrently present during the previous cryptosporidiosis outbreak. Examination of multiple enteric pathogens should be conducted when poor hygiene is the likely cause of outbreaks of diarrhea.

  3. Multicenter Evaluation of BD Max Enteric Parasite Real-Time PCR Assay for Detection of Giardia duodenalis, Cryptosporidium hominis, Cryptosporidium parvum, and Entamoeba histolytica.

    Science.gov (United States)

    Madison-Antenucci, S; Relich, R F; Doyle, L; Espina, N; Fuller, D; Karchmer, T; Lainesse, A; Mortensen, J E; Pancholi, P; Veros, W; Harrington, S M

    2016-11-01

    Common causes of chronic diarrhea among travelers worldwide include protozoan parasites. The majority of parasitic infections are caused by Giardia duodenalis, Entamoeba histolytica, Cryptosporidium parvum, and Cryptosporidium hominis Similarly, these species cause the majority of parasitic diarrhea acquired in the United States. Detection of parasites by gold standard microscopic methods is time-consuming and requires considerable expertise; enzyme immunoassays and direct fluorescent-antibody (DFA) stains have lowered hands-on time for testing, but improvements in sensitivity and technical time may be possible with a PCR assay. We performed a clinical evaluation of a multiplex PCR panel, the enteric parasite panel (EPP), for the detection of these common parasites using the BD Max instrument, which performs automated extraction and amplification. A total of 2,495 compliant specimens were enrolled, including 2,104 (84%) specimens collected prospectively and 391 (16%) specimens collected retrospectively. Approximately equal numbers were received in 10% formalin (1,273 specimens) and unpreserved (1,222 specimens). The results from the EPP were compared to those from alternate PCR and bidirectional sequencing (APCR), as well as DFA (G. duodenalis and C. parvum or C. hominis) or trichrome stain (E. histolytica). The sensitivity and specificity for prospective and retrospective specimens combined were 98.2% and 99.5% for G. duodenalis, 95.5% and 99.6 for C. parvum or C. hominis, and 100% and 100% for E. histolytica, respectively. The performance of the FDA-approved BD Max EPP compared well to the reference methods and may be an appropriate substitute for microscopic examination or immunoassays. Copyright © 2016 Madison-Antenucci et al.

  4. Genotyping and subtyping Cryptosporidium parvum and Giardia duodenalis carried by flies on dairy farms in Henan, China

    OpenAIRE

    Zhao, Zifang; Dong, Haiju; Wang, Rongjun; Zhao, Wei; Chen, Gongyi; Li, Shouyi; Qi, Meng; Zhang, Sumei; Jian, Fuchun; Zhao, Jinfeng; Zhang, Longxian; Wang, Haiyan; Liu, Aiqin

    2014-01-01

    Background Cryptosporidium and Giardia are important causes of diarrhea diseases in humans and animals worldwide, and both of them are transmitted by the fecal–oral route, either by direct contact or by the ingestion of contaminated food or water. The role of flies in the mechanical transmission of Cryptosporidium and Giardia has been receiving increasing attention. To date, no information is available in China about the occurrence of Cryptosporidium and Giardia in flies. We here investigated...

  5. Serodiagnóstico de giardiosis: identificación de inmunoglobulina G anti-Giardia duodenalis en suero mediante ELISA

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    Sofía Duque

    2001-09-01

    Full Text Available El diagnóstico de la infección por Giardia duodenalis se basa en la identificación de quistes o trofozoítos en materia fecal, alcanzando una sensibilidad máxima de 85% en muestras seriadas, debido a la excreción intermitente de quistes. Aunque la determinación de anticuerpos no permite diferenciar entre una infección pasada y una reciente, si podría ayudar en el diagnóstico diferencial. El presente estudio tuvo como objetivos estandarizar y evaluar una prueba inmunoenzimática (ELISA para la detección de anticuerpos tipo IgG anti-Giardia duodenalis en sueros de pacientes con la infección. La preparación del antigeno se hizo a partir de trofozoitos cultivados que, inicialmente, fueron obtenidos del intestino de gerbils (Meriones unguiculatus infectados experimentalmente con quistes purificados a partir de muestras positivas de materia fecal humana. Para la estandarización y evaluación de la prueba, se emplearon 60 sueros de pacientes con giardiosis comprobada parasitológicamente (muestras positivas y 47 sueros de cordón umbilical recién cortado (muestras negativas. Como conjugado, se utilizó anti-lgG humana ligada a fosfatasa alcalina. La concentración óptima de antígeno de trofozoíto de G. duodenalis fue de 15 ug/ml y las diluciones óptimas de suero y conjugado fueron 1:25 y 1:400, respectivamente. El valor del punto de corte (absorbancia fue 0,300. Los parámetros de la prueba fueron: sensibilidad, 98,3% (intervalo de confianza de 95% (lC 95%: 89,9%-99,974; especificidad, 957% (IC 95%:84,3%-99,3%; valor predictivo positivo, 96,7% (IC 95%: 87,6%-99,4%, y valor predictivo negativo, 97,8% (IC 95%: 87,0%-99,9%. La prueba ELlSA descrita contribuirá a mejorar el diagnóstico de giardiosis y podrá ser empleada para estudios epidemiológicos de seroprevalencia.

  6. Giardia duodenalis and soil-transmitted helminths infections in children in São Tomé and Príncipe: do we think Giardia when addressing parasite control?

    Science.gov (United States)

    Ferreira, Filipa Santana; Baptista-Fernandes, Teresa; Oliveira, Dinamene; Rodrigues, Rúben; Neves, Edgar; Lima, António; Garrido, Eduardo; Afonso, Guilherme; Zaky, Ahmed; Telles de Freitas, Paulo; Atouguia, Jorge; Centeno-Lima, Sónia

    2015-04-01

    Giardia duodenalis prevalence is commonly as high as soil-transmitted helminths (STH), nevertheless is not considered for large-scale chemotherapy through mass drug administration (MDA) due to its short incubation period and frequent reinfections, its control being associated to improving access to water and sanitation. A study enrolling 444 children attending preschools was conducted in May 2011 during a deworming campaign. Faecal samples were obtained and analysed through microscopy of wet mounting and after Kato-Katz and formol-ether concentration techniques. The majority of children were infected with at least one pathogenic parasite (86.7%, 385 of 444). Ascaris lumbricoides and Trichuris trichiura (56.3%, 250 of 444 and 52.5%, 233 of 444, respectively) were the most frequent parasites followed by G. duodenalis infecting 41.7% (185 of 444) of the children. The present work aimed at obtaining updated information concerning intestinal parasite infections in children attending preschools in São Tomé and Príncipe and to contribute for the adequate management of the enteric infections. © The Author [2015]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. No molecular epidemiological evidence supporting household transmission of zoonotic Giardia duodenalis and Cryptosporidium spp. from pet dogs and cats in the province of Álava, Northern Spain.

    Science.gov (United States)

    de Lucio, Aida; Bailo, Begoña; Aguilera, María; Cardona, Guillermo A; Fernández-Crespo, Juan C; Carmena, David

    2017-06-01

    The role of pet dogs and cats as suitable source of human infections by the diarrheagenic protozoan parasites Giardia duodenalis and Cryptosporidium spp. has been a topic of intense debate for long time and still remains a largely unsolved problem. In this cross-sectional molecular epidemiological survey we attempted to investigate whether zoonotic (or zooanthroponotic) disease transmission was occurring among humans and domestic dogs and cats sharing the same spatial and temporal setting in both rural and urban areas of the province of Álava, Northern Spain. A total of 268 (including 179 human, 55 canine, and 34 feline) individual faecal specimens were obtained from 63 family households during February-March and November-December 2014. Detection of G. duodenalis cysts and Cryptosporidium spp. oocysts was achieved by direct fluorescence microscopy (DFAT) and PCR-based methods targeting the small subunit (SSU) ribosomal RNA gene of the parasites. Giardia-positive isolates were subsequently sub-genotyped at the glutamate dehydrogenase (GDH) and β-giardin (BG) genes. Overall, G. duodenalis infections were identified in 3.4% (6/179) of humans, 29% (16/55) of dogs, and 5.9% (2/34) of cats, respectively. Cryptosporidium spp. infections were detected in 1.1% (2/179) of humans, 5.5% (3/55) of dogs, and 8.8% (3/34) of cats, respectively. Simultaneous infections in human and canine/feline hosts by G. duodenalis or Cryptosporidium spp. were only demonstrated in a single household in which a cat and its owner tested positive for Cryptosporidium by DFAT, but this result could not be confirmed by SSU-PCR. Infections were homogeneously distributed among the studied human or animal populations irrespectively of their sex, age group, or geographical region of origin. Inadequate washing of raw vegetables and fruits was the only risk factor significantly associated to a higher likelihood of having human giardiosis/cryptosporidiosis. Molecular characterization of G. duodenalis

  8. Prevalence of Giardia duodenalis and Cryptosporidium ssp. in females of the Holstein breed during the post weaning phase in the herd in the southern state of Minas Gerais during the fall-winter season of 2008

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    Fabiana Alves Demeu

    2011-01-01

    Full Text Available This study aimed to evaluate the frequency of the species of Giardia duodenalis species and the genus Cryptosporidium spp. in dairy calves at different ages during the post-weaning phase in a herd located in Boa Esperança, South of Minas Gerais State, Brazil, during the fallwinter season. It were analyzed 254 samples from 37 cows of Holstein breed, aged between three and 10 months in the period from May to September 2008, collected directly from the rectum every 30 days. To search cysts or trophozoites of Giardia duodenalis was used the flotation in zinc sulfate solution to 33% technique, and for detection of Cryptosporidium was used the Ziehl-Neelsen modified technique. Of the 254 samples, 37.79% were positive for cysts or trophozoites of Giardia and 8.93% for Cryptosporidium spp. The comparison of age and positivity to G. duodenalis and Cryptosporidium, it was observed that for both parasites, the age between three and four months is the most affected in the post-weaning phase in cattle. Whereas the post-weaning is a critical point in the growing cattle, this would indicate the pathogenic potential for these parasites in dairy cattle in southern Minas Gerais.

  9. Prebiotic inulin supplementation modulates the immune response and restores gut morphology in Giardia duodenalis-infected malnourished mice.

    Science.gov (United States)

    Shukla, Geeta; Bhatia, Ruchika; Sharma, Anuj

    2016-11-01

    Malnutrition induces a state of growth retardation and immunologic depression, enhancing the host susceptibility to various infections. In the present study, it was observed that prebiotic supplementation either prior or simultaneously with Giardia infection in malnourished mice significantly reduced the severity of giardiasis and increased the body and small intestine mass, along with increased lactobacilli counts in faeces compared with malnourished-Giardia-infected mice. More specifically, prebiotic supplementation significantly increased the levels of anti-giardial IgG and IgA antibodies and anti-inflammatory cytokines IL-6 and IL-10 and reduced the pro-inflammatory cytokine TNF-α, along with increased levels of nitric oxide in both the serum and intestinal fluid of malnourished-prebiotic-Giardia-infected mice compared with malnourished-Giardia-infected mice. Histopathology and scanning electron microscopy of the small intestine also revealed less cellular and mucosal damage in the microvilli of prebiotic-supplemented malnourished-Giardia-infected mice compared with severely damaged mummified and blunted villi of malnourished-Giardia-infected mice. This is the first study to report that prebiotic supplementation modulated the gut morphology and improved the immune status even in malnourished-Giardia-infected mice.

  10. Occurrence of enteroparasites in day care centers in Botucatu (São Paulo State, Brazil with emphasis on Cryptosporidium sp., Giardia duodenalis and Enterobius vermicularis Ocorrência de enteroparasitas em creches de Botucatu (Estado de São Paulo, Brasil com ênfase em Cryptosporidium sp., Giardia duodenalis e Enterobius vermicularis

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    Thaís Batista de Carvalho

    2006-10-01

    Full Text Available The occurrence of the enteroparasites was verified in 279 children (0 to 6 years of four municipal day cares of Botucatu/SP. Three samples of each child's feces were collected and processed by the methods of Hoffman-Pons-Janner, Faust and Ritchie and subsequent coloration of the fecal smear by the methods of Auramina-O and Ziehl-Neelsen modified for diagnosis of Cryptosporidium sp. and Graham method for diagnosis of Enterobius vermicularis. Of the analyzed children we verified a prevalence of intestinal parasitism in 53.40%, and the most frequent parasite was Giardia duodenalis (26.88%. Significant association was verified among enteroparasitosis, family income, maternal education and age; the lowest enteroparasite frequency occurred in children of families with larger income and higher education. It was observed that G. duodenalis is more prevalent in children from 0 to 4 years and E. vermicularis is more frequent in children between three and four years old. The high enteroparasite prevalence in day cares suggests complex structure in its epidemiology, where factors beyond sanitation should be considered.Verificou-se a prevalência dos enteroparasitas em 279 crianças (0 a 6 anos de quatro creches municipais de Botucatu/SP. Foram coletadas três amostras de fezes de cada criança e processadas pelos métodos Hoffman, Faust e Ritchie e posterior coloração do esfregaço fecal pelos métodos de Auramina-O e Ziehl-Neelsen modificado para diagnóstico de Cryptosporidium sp. e método da fita gomada para diagnóstico de Enterobius vermicularis. Das crianças analisadas apresentaram-se parasitadas 53.40%, sendo que o parasita mais freqüente foi Giardia duodenalis (26.88%. Verificou-se associação significativa entre enteroparasitose, renda familiar, escolaridade materna e idade; quanto maior a renda e o grau escolar, menor a freqüência de enteroparasitas. Observou-se que G. duodenalis é mais prevalente em crianças de 0 a 4 anos e E

  11. The crystal structure of Giardia duodenalis 14-3-3 in the apo form: when protein post-translational modifications make the difference.

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    Annarita Fiorillo

    Full Text Available The 14-3-3s are a family of dimeric evolutionary conserved pSer/pThr binding proteins that play a key role in multiple biological processes by interacting with a plethora of client proteins. Giardia duodenalis is a flagellated protozoan that affects millions of people worldwide causing an acute and chronic diarrheal disease. The single giardial 14-3-3 isoform (g14-3-3, unique in the 14-3-3 family, needs the constitutive phosphorylation of Thr214 and the polyglycylation of its C-terminus to be fully functional in vivo. Alteration of the phosphorylation and polyglycylation status affects the parasite differentiation into the cyst stage. To further investigate the role of these post-translational modifications, the crystal structure of the g14-3-3 was solved in the unmodified apo form. Oligomers of g14-3-3 were observed due to domain swapping events at the protein C-terminus. The formation of filaments was supported by TEM. Mutational analysis, in combination with native PAGE and chemical cross-linking, proved that polyglycylation prevents oligomerization. In silico phosphorylation and molecular dynamics simulations supported a structural role for the phosphorylation of Thr214 in promoting target binding. Our findings highlight unique structural features of g14-3-3 opening novel perspectives on the evolutionary history of this protein family and envisaging the possibility to develop anti-giardial drugs targeting g14-3-3.

  12. The crystal structure of Giardia duodenalis 14-3-3 in the apo form: when protein post-translational modifications make the difference.

    KAUST Repository

    Fiorillo, Annarita

    2014-03-21

    The 14-3-3s are a family of dimeric evolutionary conserved pSer/pThr binding proteins that play a key role in multiple biological processes by interacting with a plethora of client proteins. Giardia duodenalis is a flagellated protozoan that affects millions of people worldwide causing an acute and chronic diarrheal disease. The single giardial 14-3-3 isoform (g14-3-3), unique in the 14-3-3 family, needs the constitutive phosphorylation of Thr214 and the polyglycylation of its C-terminus to be fully functional in vivo. Alteration of the phosphorylation and polyglycylation status affects the parasite differentiation into the cyst stage. To further investigate the role of these post-translational modifications, the crystal structure of the g14-3-3 was solved in the unmodified apo form. Oligomers of g14-3-3 were observed due to domain swapping events at the protein C-terminus. The formation of filaments was supported by TEM. Mutational analysis, in combination with native PAGE and chemical cross-linking, proved that polyglycylation prevents oligomerization. In silico phosphorylation and molecular dynamics simulations supported a structural role for the phosphorylation of Thr214 in promoting target binding. Our findings highlight unique structural features of g14-3-3 opening novel perspectives on the evolutionary history of this protein family and envisaging the possibility to develop anti-giardial drugs targeting g14-3-3.

  13. Obtención, purificación y caracterización de anticuerpos policlonales IgY desarrollados en gallina, dirigidos contra aislamientos colombianos de Giardia duodenalis.

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    Dabeiba Adriana García

    2005-12-01

    Full Text Available Introducción. El desarrollo de anticuerpos policlonales requiere de animales de laboratorio, generalmente, el conejo, que deben sangrarse para su obtención. Como alternativa, se han utilizado las gallinas. Objetivo. Desarrollar anticuerpos policlonales IgY anti-Giardia duodenalis y evaluar diferentes métodos para su purificación a partir de yema de huevo. Materiales y métodos. Se inmunizaron tres gallinas intramuscularmente con trofozoítos del parásito: las inmunizaciones se realizaron a los 0, 15, 30, 45, 60, 90 y 120 días. Se recolectaron huevos en cada etapa de la inmunización y se purificó la IgY por deslipidación (D y precipitación (P mediante cinco protocolos diferentes: M1: (P: sulfato de amonio/D: dextrán sulfato-cloruro de calcio, M2: (D: dextrán sulfato-cloruro de calcio/P: sulfato de amonio, M3: (D: cloroformo/ P: sulfato de amonio 50%, M4: (D: solución A/P:solución B y M5: (D: cloroformo/P: sulfato de amonio 30%. Se realizó evaluación inmunoquímica de la IgY anti-Giardia duodenalis mediante inmunodifusión, contrainmunoelectroforesis e inmunoelectrotransferencia (Western blot; la pureza de IgY por SDS-PAGE en presencia y ausencia de reductor y la concentración de la inmunoglobulina (mg/mL por espectrofotometría y densitometría. Resultados. La IgY anti-G. duodenalis mediante evaluación inmunoquímica presentó títulos hasta de 1:32. La inmunoglobulina en ausencia de reductor mostró una banda de 180 kd y en su presencia bandas de 30 y 68 kd, características de sus cadenas liviana y pesada, respectivamente. Las mayores concentraciones de inmunoglobulina se recuperaron con el método dos (M2, 4,6 mg de IgY por mL de yema de partida. Conclusiones. Por su facilidad y economía de producción en gallinas, los anticuerpos policlonales IgY anti-Giardia así obtenidos podrán utilizarse para desarrollar inmunoensayos que detecten el parásito en eluídos de heces.

  14. Genotyping of Giardia in Dutch patients and animals: a phylogenetic analysis of human and animal isolates.

    NARCIS (Netherlands)

    Giessen, J W B van der; Vries, A de; Roos, M; Wielinga, Peter; Kortbeek, L M; Mank, T G

    2006-01-01

    Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis) is a protozoan organism that can infect the intestinal tract of many animal species including mammals. Genetic heterogeneity of G. duodenalis is well described but the zoonotic potential is still not clear. In this study, we analysed

  15. Detection and molecular characterisation of Giardia duodenalis, Cryptosporidium spp. and Entamoeba spp. among patients with gastrointestinal symptoms in Gambo Hospital, Oromia Region, southern Ethiopia.

    Science.gov (United States)

    Flecha, María J; Benavides, Cynthia M; Tissiano, Gabriel; Tesfamariam, Abraham; Cuadros, Juan; de Lucio, Aida; Bailo, Begoña; Cano, Lourdes; Fuentes, Isabel; Carmena, David

    2015-09-01

    To assess the prevalence and genetic diversity of the enteric protozoa species G. duodenalis, Cryptosporidium spp. and Entamoeba histolytica in individuals with gastrointestinal symptoms compatible with infections by these pathogens seeking medical attention in a rural area in southern Ethiopia. A total of 92 stool samples were initially screened by direct microscopy and immunochromatography and further confirmed by molecular methods. G. duodenalis-positive samples were molecularly characterised by multilocus genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. PCR and DNA sequence analysis of the gene encoding the 60-kDa glycoprotein was used for the subtyping of Cryptosporidium isolates. Detection and differential diagnosis of E. histolytica/dispar were conducted by real-time PCR. PCR-based prevalences were 10.9% for G. duodenalis, 1.1% for Cryptosporidium spp. and 3.3% for Entamoeba spp. Seven (four novel and three known) subtypes of G. duodenalis assemblage B were identified at the GDH locus and 5 (one novel and four known) at the BG locus. A novel variant of C. hominis subtype IbA9G3 was also identified. Two Entamoeba isolates were assigned to E. dispar and an additional one to E. histolytica. Although preliminary, our results strongly suggest that giardiasis, cryptosporidiosis and amoebiasis represent a significant burden in Ethiopian rural population. © 2015 John Wiley & Sons Ltd.

  16. Genotipos de Giardia duodenalis en muestras de niños de las guarderías del Instituto Colombiano de Bienestar Familiar y de perros en Ibagué, Colombia

    Directory of Open Access Journals (Sweden)

    Victoria Rodríguez

    2014-06-01

    Full Text Available Introducción. Se han descrito ocho genotipos de Giardia duodenalis, del A al H. Los genotipos A y B se han aislado de humanos y de una gran variedad de mamíferos; sin embargo, los genotipos del C al H han mostrado mayor especificidad de huésped. Objetivo. Identificar los genotipos de G. duodenalis a partir de quistes obtenidos en heces de niños de las guarderías del Instituto Colombiano de Bienestar Familiar (ICBF y de perros en Ibagué, mediante PCR-RFLP de los genes de la beta giardina y la glutamato deshidrogenasa. Materiales y métodos. Los quistes de las muestras positivas para G. duodenalis fueron sometidos a concentración; se extrajo su ADN y se efectuó el análisis de PCR-RFLP de los genes de la beta giardina y de la glutamato deshidrogenasa. Como control positivo se utilizó la cepa MHOM/CO/04/G40 procedente del Grupo de Parasitología del Instituto Nacional de Salud. Resultados. De las muestras tomadas de niños, 11/23 (48 % correspondieron al genotipo A y, 12/23 (52 %, al genotipo B. Cuatro muestras de perros presentaron los genotipos C y D, específicos de este huésped. Conclusiones. En los niños solamente se encontraron los genotipos asociados a infecciones humanas (AII, BIII y BIV y en los perros, los genotipos específicos para este huésped (C y D. Debido al reducido tamaño de las muestras analizadas provenientes de perros, y dado que estos no estuvieron en contacto con los niños de las guarderías del ICBF, no fue posible determinar una interacción entre el ciclo de transmisión de los humanos y el de los animales.

  17. Genotipagem de isolados de Giardia duodenalis de primatas não humanos mantidos em zoológico do Brasil

    OpenAIRE

    David, Erica Boarato; Patti, Mariella; Coradi, Silvana Torossian; Oliveira-Sequeira, Teresa Cristina Goulart; Ribolla, Paulo Eduardo Martins; Guimaraes, Semiramis

    2014-01-01

    A pesquisa de infecções por Giardia e a caracterização genotípica deste protozoário foi realizada em primatas não humanos (PNH) mantidos em Zoológico a fim de avaliar o seu potencial zoonótico. As amostras dos animais consistiram de fezes colhidas do piso de 22 baias onde eram mantidos 47 primatas de 18 diferentes espécies. Exames coproparasitológicos foram realizados pelos métodos de concentração por sedimentação e centrífugo-flutuação e revelaram a presença dos seguintes parasitas e suas re...

  18. Intestinal growth and pathology of Giardia duodenalis assemblage subtype A(I), A(II), B and E in the gerbil model.

    Science.gov (United States)

    Bénéré, E; Van Assche, T; Van Ginneken, C; Peulen, O; Cos, P; Maes, L

    2012-04-01

    This study investigated the significance of the genetic differences between assemblages A, B and E on intestinal growth and virulence. Intestinal growth and virulence were studied in 2 laboratory (A(I): WB and B: GS/M-83-H7) and 6 field isolates of assemblage subtype A(I), A(II), B and E(III). Intestinal trophozoite burdens, body weight and faecal consistency were monitored until day 29 post-infection (p.i.), morphological (mucosal architecture and inflammation) and functional (disaccharidase and alkaline phosphatase enzyme activity) damage to the small intestine were evaluated on days 7 and 18 p.i. The assemblage subtypes A(I) and B were more infectious and produced higher trophozoite loads for a longer period compared to the subtypes A(II) and E(III). The body weight of infected gerbils was significantly reduced compared to uninfected controls, but did not differ between the assemblage subtypes. Consistent softening of the faeces was only observed with assemblage B. Assemblage B next to assemblage subtype A(I) elicited relatively higher pathogenicity, characterized by more extensive damage to mucosal architecture, decreased brush-border enzyme function and infiltration of inflammatory cells. Assemblage E(III) and A(II) isolates showed relatively low virulence. The Giardia assemblage subtypes exhibit different levels of growth and virulence in the gerbil model.

  19. Zoonotic potential of Giardia.

    Science.gov (United States)

    Ryan, Una; Cacciò, Simone M

    2013-11-01

    Giardia duodenalis (syn. Giardia lamblia and Giardia intestinalis) is a common intestinal parasite of humans and mammals worldwide. Assessing the zoonotic transmission of the infection requires molecular characterization as there is considerable genetic variation within G. duodenalis. To date eight major genetic groups (assemblages) have been identified, two of which (A and B) are found in both humans and animals, whereas the remaining six (C to H) are host-specific and do not infect humans. Sequence-based surveys of single loci have identified a number of genetic variants (genotypes) within assemblages A and B in animal species, some of which may have zoonotic potential. Multi-locus typing data, however, has shown that in most cases, animals do not share identical multi-locus types with humans. Furthermore, interpretation of genotyping data is complicated by the presence of multiple alleles that generate "double peaks" in sequencing files from PCR products, and by the potential exchange of genetic material among isolates, which may account for the non-concordance in the assignment of isolates to specific assemblages. Therefore, a better understanding of the genetics of this parasite is required to allow the design of more sensitive and variable subtyping tools, that in turn may help unravel the complex epidemiology of this infection. Copyright © 2013. Published by Elsevier Ltd.

  20. First record of Giardia assemblage D infection in farmed raccoon dogs (Nyctereutes procyonoides

    Directory of Open Access Journals (Sweden)

    Piotr Solarczyk

    2016-09-01

    Full Text Available The presence of Giardia genotypes was investigated in 18 raccoon dogs ( Nyctereutes procyonoides and 80 red foxes ( Vulpes vulpes on one farm. To demonstrate Giardia cysts, fresh and trichrome stained smears were microscopically screened. Two molecular markers were used for Giardia genotyping: a fragment of the beta-giardin gene and a fragment of the glutamate dehydrogenase gene. All faecal samples obtained from red foxes were negative. Giardia cysts were identified only in 2 of the 18 raccoon dogs. The result of genotyping and phylogenetic analysis showed that the G. duodenalis from both raccoon dogs belonged to the D assemblage. This finding of a new animal reservoir of G. duodenalis canids-specific genotypes is important in order to eliminate the risk of infecting other animals bred for fur. Further molecular analyses of Giardia isolates in raccoon dogs are required. The present study represents the first contribution to knowledge of G. duodenalis genotypes in raccoon dogs.

  1. Occurrence of Giardia in Swedish Red Foxes ( Vulpes vulpes ).

    Science.gov (United States)

    Debenham, John J; Landuyt, Hanne; Troell, Karin; Tysnes, Kristoffer; Robertson, Lucy J

    2017-07-01

    Giardia duodenalis is an intestinal protozoan capable of causing gastrointestinal disease in a range of vertebrate hosts. It is transmitted via the fecal-oral route. Understanding the epidemiology of G. duodenalis in animals is important, both for public health and for the health of the animals it infects. We investigated the occurrence of G. duodenalis in wild Swedish red foxes ( Vulpes vulpes ), with the aim of providing preliminary information on how this abundant predator might be involved in the transmission and epidemiology of G. duodenalis . Fecal samples (n=104) were analysed for G. duodenalis using a commercially available direct immunofluorescent antibody test. Giardia duodenalis cysts were found in 44% (46/104) of samples, with foxes excreting 100 to 140,500 cysts per gram of feces (mean, 4,930; median, 600). Molecular analysis, using PCR with sequencing of PCR amplicons, was performed on 14 samples, all containing over 2,000 cysts per gram feces. Amplification only occurred in four samples at the tpi gene, sequencing of which revealed assemblage B in all four samples. This study provides baseline information on the role of red foxes in the transmission dynamics of G. duodenalis in Sweden.

  2. Giardia muris infection in mice is associated with a protective interleukin 17A response and induction of peroxisome proliferator-activated receptor alpha

    NARCIS (Netherlands)

    L. Dreesen (Leentje); L. Bosscher (Lisette); K.J. Grit (Kor); B. Staels (Bart); E.W. Lubberts (Erik); E. Bauge (Eric); P. Geldhof (Peter)

    2014-01-01

    textabstractThe protozoan parasite Giardia duodenalis (Giardia lamblia) is one of the most commonly found intestinal pathogens in mammals, including humans. In the current study, a Giardia muris-mouse model was used to analyze cytokine transcription patterns and histological changes in intestinal

  3. Giardia assemblage A: human genotype in muskoxen in the Canadian Arctic

    Directory of Open Access Journals (Sweden)

    Nagy John

    2008-09-01

    Full Text Available Abstract As part of an ongoing program assessing the biodiversity and impacts of parasites in Arctic ungulates we examined 72 fecal samples from muskoxen on Banks Island, Northwest Territories, Canada for Giardia and Cryptosporidium. Cryptosporidium spp. were not detected, but 21% of the samples were positive for Giardia. Sequencing of four isolates of Giardia demonstrated G. duodenalis, Assemblage A, a zoonotic genotype.

  4. Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand.

    Science.gov (United States)

    Tangtrongsup, Sahatchai; Scorza, A Valeria; Reif, John S; Ballweber, Lora R; Lappin, Michael R; Salman, Mo D

    2017-05-10

    The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA), PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70), and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA). Giardia duodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and generic and dog-specific assays of triosephosphate isomerase (tpi) genes. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium -positive samples and 21 Giardia -positive samples revealed the presence of C. canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis . Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance.

  5. Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand

    Directory of Open Access Journals (Sweden)

    Sahatchai Tangtrongsup

    2017-05-01

    Full Text Available The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA, PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70, and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA. Giardia duodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh, beta-giardin (bg, and generic and dog-specific assays of triosephosphate isomerase (tpi genes. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium-positive samples and 21 Giardia-positive samples revealed the presence of C. canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis. Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance.

  6. Giardia infection

    Science.gov (United States)

    Giardia, or giardiasis, is an infection of the small intestine. A tiny parasite called Giardia lamblia causes it. ... from some of the medicines used to treat giardia are: Metallic ... used to treat the infection can be harmful to the unborn baby.

  7. Genotyping of Giardia isolates in Scotland: a descriptive epidemiological study.

    Science.gov (United States)

    Alexander, C; Jones, B; Inverarity, D; Pollock, K G J

    2014-08-01

    Giardiasis, caused by the intestinal protozoan parasite Giardia intestinalis (synonyms: G. lamblia, G. duodenalis), is one of the most frequent parasites to infect the Scottish population. Transmission of the infective cysts in faecal matter is commonly via food and/or water. Giardia is subdivided into assemblages, where clinical and epidemiological differences have been described between assemblages A and B. This snapshot descriptive epidemiological study examines 30 positive cases of Giardia of which 72% (n = 21) were shown to be assemblage A, 14% (n = 4) assemblage B and 10% (n = 3) mixed assemblages (A and B). There was a 2:3 female:male ratio of affected individuals with foreign travel recorded in 22 of these cases. The commonest symptom was diarrhoea which was reported in 80% of cases followed by tiredness. Five cases required hospitalization emphasizing the importance of gaining a greater understanding of how Giardia assemblages influence clinical outcomes to assist in formulating guidelines to manage potential Giardia outbreaks.

  8. Giardia & Pets

    Science.gov (United States)

    ... Giardia spread? Anything that comes into contact with feces (poop) from infected humans or animals can become contaminated ... get infected by: Being in contact with infected feces (poop) from another dog or cat Rolling and ...

  9. Recent advances in the genomic and molecular biology of Giardia.

    Science.gov (United States)

    Ortega-Pierres, M Guadalupe; Jex, Aaron R; Ansell, Brendan R E; Svärd, Staffan G

    2017-09-06

    Giardia duodenalis is the most common gastrointestinal protozoan parasite of humans and a significant contributor to the global burden of both diarrheal disease and post-infectious chronic disorders. Robust tools for analyzing gene function in this parasite have been developed and a range of genetic tools are now available. These together with public databases have provided insights on the function of different genes in Giardia. In this review we provide a current perspective on different molecular aspects of Giardia related to genomics, regulation of encystation, trophozoite transcriptional responses to physiological and xenobiotic (drug-induced) stress, and mechanisms of drug resistance. We also examine recent insights that have contributed to gain knowledge in the study of VSPs, antigenic variation, epigenetics, DNA repair and in the direct manipulation of gene function in Giardia, with a particular focus on the inducible Cre/loxP system. Copyright © 2017. Published by Elsevier B.V.

  10. Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp.

    Science.gov (United States)

    Paulos, Silvia; Mateo, Marta; de Lucio, Aida; Hernández-de Mingo, Marta; Bailo, Begoña; Saugar, José M; Cardona, Guillermo A; Fuentes, Isabel; Mateo, María; Carmena, David

    2016-08-01

    High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n=29), giardiasis (n=47) and amoebiasis by Entamoeba histolytica (n=3) or E. dispar (n=10) and apparently healthy subjects (n=24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56°C were proven more efficient for the release of DNA from Cryptosporidium oocysts. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Giardia lamblia.

    OpenAIRE

    Rodríguez Padilla, Eva María

    2015-01-01

    Las infecciones parasitarias son un verdadero problema para la salud pública especialmente en países con niveles socioeconómicos bajos. La infección por Giardia es la responsable de producir un 2-3% de las diarreas del viajero, muy relacionada también con la diarrea infecciosa no viral en niños. El reservorio de Giardia es el hombre, ya sea enfermo o en estado de portador asintomático, la infección puede ser frecuente entre animales domésticos, mamíferos salvajes y aves....

  12. Occurrence of Giardia and Cryptosporidium in wild birds in Galicia (Northwest Spain).

    Science.gov (United States)

    Reboredo-Fernández, Aurora; Ares-Mazás, Elvira; Cacciò, Simone M; Gómez-Couso, Hipólito

    2015-06-01

    Faecal samples were obtained from 433 wild birds being treated in wildlife recovery centres in Galicia (Northwest Spain), between February 2007 and September 2009. The birds belonged to 64 species representing 17 different orders. Giardia cysts and Cryptosporidium oocysts were detected by an immunofluorescence antibody test and identified at the molecular level by established PCR-sequencing methods. The overall prevalence of Giardia was 2·1% and that of Cryptosporidium, 8·3%. To our knowledge, this is the first description of Giardia sp. in Tyto alba and Caprimulgus europaeus; and of Cryptosporidium sp. in Apus apus, Athene noctua, C. europaeus, Falco tinnunculus, Morus bassanus, Parabuteo unicinctus and Strix aluco. Furthermore, the first PCR-sequence confirmed detection of Giardia duodenalis assemblage B in, Buteo buteo, Coturnix coturnix and Pica pica; G. duodenalis assemblage D in Garrulus glandarius; and G. duodenalis assemblage F in Anas platyrhynchos; Cryptosporidium parvum in Accipiter nisus, B. buteo, Milvus migrans, Pernis apivorus and P. pica; and Cryptosporidium meleagridis in Streptopelia turtur. The study findings demonstrate the wide spread of Giardia and Cryptosporidium between wild birds.

  13. Giardia Infection (Giardiasis)

    Science.gov (United States)

    Giardia infection (giardiasis) Overview Giardia infection is an intestinal infection marked by abdominal cramps, bloating, nausea and bouts of watery diarrhea. Giardia infection is caused by a microscopic parasite ...

  14. Ethanol and isopropanol in concentrations present in hand sanitizers sharply reduce excystation of Giardia and Entamoeba and eliminate oral infectivity of Giardia cysts in gerbils.

    Science.gov (United States)

    Chatterjee, Aparajita; Bandini, Giulia; Motari, Edwin; Samuelson, John

    2015-11-01

    Enteric protozoan parasites, which are spread by the fecal-oral route, are important causes of diarrhea (Giardia duodenalis) and amebic dysentery (Entamoeba histolytica). Cyst walls of Giardia and Entamoeba have a single layer composed of fibrils of β-1,3-linked GalNAc and β-1,4-linked GlcNAc (chitin), respectively. The goal here was to determine whether hand sanitizers that contain ethanol or isopropanol as the active microbicide might reduce transmission of these parasites. We found that treatment with these alcohols with or without drying in a rotary evaporator (to model rapid evaporation of sanitizers on hands) kills 85 to 100% of cysts of G. duodenalis and 90 to 100% of cysts of Entamoeba invadens (a nonpathogenic model for E. histolytica), as shown by nuclear labeling with propidium iodide and failure to excyst in vitro. Alcohols with or without drying collapsed the cyst walls of Giardia but did not collapse the cyst walls of Entamoeba. To validate the in vitro results, we showed that treatment with alcohols eliminated oral infection of gerbils by 1,000 G. duodenalis cysts, while a commercial hand sanitizer (Purell) killed E. invadens cysts that were directly applied to the hands. These results suggest that expanded use of alcohol-based hand sanitizers might reduce the transmission of Giardia and Entamoeba. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  15. Saliva and sera IgA and IgG in Egyptian Giardia-infected children.

    Science.gov (United States)

    El-Gebaly, Naglaa Saad M; Halawa, Eman Fawzy; Moussa, Hanaa M Ezzat; Rabia, Ibrahim; Abu-Zekry, Maha

    2012-08-01

    Giardiasis is a gastrointestinal infection of wide distribution that is more prevalent in childhood. Easy and rapid diagnosis of giardiasis is essential for reduction of this infection. This cross-sectional study included 62 children in which collection of saliva, stool and serum samples was performed. An enzyme-linked immunosorbent assay (ELISA) technique was evaluated to detect IgA and IgG responses in both saliva and serum samples. Twenty-two children were positive for Giardia duodenalis infection by direct examination of faecal specimens, 20 non-infected and 20 infected with other parasites. Salivary and serum IgA and IgG responses against G. duodenalis infection were significantly higher in Giardia parasitized than non-Giardia parasitized children (p < 0.001). This concludes that specific salivary IgA may serve as a diagnostic tool and specific salivary IgG as a screening tool in monitoring the exposure of various populations to Giardia duodenalis. The advantage of salivary assays over serum immunoglobulin assay is being easy and non-invasive in sampling technique which is important especially for young children.

  16. Giardia and Cryptosporidium in harp and hooded seals from the Gulf of St. Lawrence, Canada.

    Science.gov (United States)

    Appelbee, A J; Thompson, R C A; Measures, L M; Olson, M E

    2010-10-11

    Giardia and Cryptosporidium are protozoan parasites known to cause enteric disease in terrestrial wildlife species (mammals, reptiles and birds). Few surveys for Giardia and Cryptosporidium in marine wildlife species, such as pinnipeds, have been reported. The objective of this study was to determine the prevalence and genotype of Giardia and Cryptosporidium in two species of pinnipeds, harp seal (Phoca groenlandica) and hooded seal (Cystophora cristata), from the Gulf of St. Lawrence, Canada. Faecal samples were collected from pup and adult seals and examined for the presence of cysts of Giardia and oocysts of Cryptosporidium using microscopy and immunofluorescent staining. Tissues from the small intestine of adult seals were also collected and examined for infections using the polymerase chain reaction (PCR) technique. Giardia cysts were found in the faeces of 42% (16/38) of adult harp seals, but in none of the harp seal pups (0/20). Although Giardia cysts were not detected in faeces of adult hooded seals (0/10) using microscopy, 80% tested positive for Giardia using PCR of intestinal tissue indicative of a true replicating infection. Both harp and hooded seals harboured infections with the zoonotic strain, Giardia duodenalis Assemblage A, as determined using a nested-PCR technique to amplify a small subunit ribosomal (SSU-rRNA) gene of Giardia. Cryptosporidium was not detected by microscopy, nor using the PCR technique on intestinal tissues from any of the 68 seals examined. Copyright 2010 Elsevier B.V. All rights reserved.

  17. Modulation of human dendritic cell activity by Giardia and helminth antigens

    DEFF Research Database (Denmark)

    Summan, Anneka; Nejsum, Peter; Williams, Andrew R

    2018-01-01

    Giardia duodenalis is a common intestinal protozoan parasite known to modulate host immune responses, including dendritic cell (DC) function. Co-infections of intestinal pathogens are common, and thus DCs may be concurrently exposed to antigens from multiple parasites. Here, we investigated...... and tumour necrosis factor (TNF)-α secretion. G. duodenalis and T. suis products also consistently up-regulated IL-10 production. Despite a similar modulation of cytokine secretion, additive effects between Giardia and helminth products were not observed, indicating a dominant effect of a single parasite...... by modulating cytokine secretion and/or inducing apoptosis, which may be a parasite driven mechanism to dampen host immunity and establish chronic infections. The differential mechanisms of DC modulation by intestinal parasites warrant further attention. This article is protected by copyright. All rights...

  18. Giardia co-infection promotes the secretion of antimicrobial peptides beta-defensin 2 and trefoil factor 3 and attenuates attaching and effacing bacteria-induced intestinal disease.

    Science.gov (United States)

    Manko, Anna; Motta, Jean-Paul; Cotton, James A; Feener, Troy; Oyeyemi, Ayodele; Vallance, Bruce A; Wallace, John L; Buret, Andre G

    2017-01-01

    Our understanding of polymicrobial gastrointestinal infections and their effects on host biology remains incompletely understood. Giardia duodenalis is an ubiquitous intestinal protozoan parasite infecting animals and humans. Concomitant infections with Giardia and other gastrointestinal pathogens commonly occur. In countries with poor sanitation, Giardia infection has been associated with decreased incidence of diarrheal disease and fever, and reduced serum inflammatory markers release, via mechanisms that remain obscure. This study analyzed Giardia spp. co-infections with attaching and effacing (A/E) pathogens, and assessed whether and how the presence of Giardia modulates host responses to A/E enteropathogens, and alters intestinal disease outcome. In mice infected with the A/E pathogen Citrobacter rodentium, co-infection with Giardia muris significantly attenuated weight loss, macro- and microscopic signs of colitis, bacterial colonization and translocation, while concurrently enhancing the production and secretion of antimicrobial peptides (AMPs) mouse β-defensin 3 and trefoil factor 3 (TFF3). Co-infection of human intestinal epithelial cells (Caco-2) monolayers with G. duodenalis trophozoites and enteropathogenic Escherichia coli (EPEC) enhanced the production of the AMPs human β-defensin 2 (HBD-2) and TFF3; this effect was inhibited with treatment of G. duodenalis with cysteine protease inhibitors. Collectively, these results suggest that Giardia infections are capable of reducing enteropathogen-induced colitis while increasing production of host AMPs. Additional studies also demonstrated that Giardia was able to directly inhibit the growth of pathogenic bacteria. These results reveal novel mechanisms whereby Giardia may protect against gastrointestinal disease induced by a co-infecting A/E enteropathogen. Our findings shed new light on how microbial-microbial interactions in the gut may protect a host during concomitant infections.

  19. Assessment of zoonotic transmission of Giardia and Cryptosporidium between cattle and humans in rural villages in Bangladesh.

    Directory of Open Access Journals (Sweden)

    Amimul M Ehsan

    Full Text Available Giardia and Cryptosporidium are important causes of diarrhoea in Bangladesh. The high prevalence of both parasites in humans and cattle in rural Bangladesh and the common use of water ponds by village inhabitants and their animals suggest a potential for zoonotic transmission. Direct transmission of Giardia and Cryptosporidium between cattle and their handlers and indirect transmission through water ponds was investigated. Faecal/stool samples were collected from 623 calves and 125 calf handlers in a cross-sectional survey. In two villages, water samples were collected monthly from water ponds and faecal/stool samples were collected monthly from inhabitants and their cattle. Giardia cysts and Cryptosporidium oocysts were detected in water samples and in faecal/stool samples and positive samples were genotyped, to determine their human or animal origin. The prevalence of Giardia and Cryptosporidium in calves was 22% and 5% respectively. In calf handlers, the prevalence of Giardia and Cryptosporidium was 11.2% and 3.2% respectively. Both in the cross-sectional survey and in the longitudinal study in the villages, G. duodenalis assemblage E was most prevalent in calves, while in humans assemblage AII, BIII and BIV were found. In cattle, Cryptosporidium parvum, C. bovis and C. andersoni were identified, but no Cryptosporidium sequences were obtained from humans. Giardia and Cryptosporidium were detected in 14/24 and 12/24 water samples respectively. G. duodenalis assemblage E and BIV (-like, as well as C. andersoni and C. hominis were identified. Although the presence of Giardia and Cryptosporidium in both water ponds suggests that water-borne transmission of Giardia and Cryptosporidium is possible, the genotyping results indicate that there is no significant direct or indirect (water-borne transmission of Giardia between cattle and people in this area of rural Bangladesh. No conclusions could be drawn for Cryptosporidium, because of the low number

  20. Giardia y giardiasis Giardia and giardiasis

    OpenAIRE

    Hugo D. Luján

    2006-01-01

    Giardia lamblia es un protozoario parásito que habita el intestino delgado de los seres humanos y de muchos otros vertebrados y es una de las más comunes causas de diarrea en todo el mundo. Durante su ciclo de vida Giardia sufre significativos cambios bioquímicos y morfológicos que le permiten sobrevivir en ambientes y condiciones que de otro modo lo destruirían. Para sobrevivir fuera del intestino del hospedador, los trofozoítos de Giardia se diferencian a quistes, los que se caracterizan po...

  1. Giardia Assemblages A and B in Diarrheic Patients: A Comparative Study in Egyptian Children and Adults.

    Science.gov (United States)

    El Basha, Noussa R; Zaki, Mayssa M; Hassanin, Omayma M; Rehan, Mohamed K; Omran, Dalia

    2016-02-01

    Giardia duodenalis is considered the most common intestinal parasite in humans worldwide. Children are especially affected, with more severe consequences than adults. The present study was designed to determine the distribution of assemblages A and B Giardia infection in children and adults, with the use of light microscopy and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as diagnostic procedures, and to investigate its associations with clinical and epidemiological data collected from children and adult groups. This cross-sectional study was conducted from October 2012 to October 2013 by collecting fecal samples from 200 children and 200 adults complaining of diarrhea. Samples were subjected to parasitological examination by direct wet smear and formol-ether methods. Genotyping of G. doudenalis samples was conducted by PCR-RFLP analysis. Giardia duodenalis infection caused by assemblages A and B was identified in 60 samples, 34 from children and 26 from adults. Assemblage B was detected in 38 patients (63.34%), and assemblage A was detected in 22 patients (36.66%). Assemblage A was significantly more frequent in children with age range 2-8 yr, and assemblage B was higher in children with age range 6-16 yr old. Diarrhea frequency/day and recurrences per month affected patients infected with assemblage A (P value Giardia assemblages A and B were identified in children and adults, assemblage A infected younger children more frequently and was more closely related to severe clinical manifestations than assemblage B.

  2. Giardia in a selected population of dogs and cats in Germany - diagnostics, coinfections and assemblages.

    Science.gov (United States)

    Sommer, M F; Rupp, P; Pietsch, M; Kaspar, A; Beelitz, P

    2018-01-15

    Infections with the zoonotic endoparasite Giardia duodenalis are widely spread among dogs and cats worldwide. Since the question whether the infection might be transmitted from domestic animals to their owners is still an important topic, a reliable detection of patent Giardia infections and the determination of the associated Giardia assemblages is of major concern. The objectives of the present study were to determine the prevalence of Giardia infections in dogs and cats living in Germany using different diagnostic tests and to identify the Giardia assemblages of infected animals. Furthermore, a possible correlation of coinfections with other endoparasites was analysed. All samples were investigated by enzyme-linked immunosorbent assay (ELISA), merthiolate-iodine-formalin concentration technique (MIFC) and zinc chloride flotation. ELISA-positive samples were additionally screened with a direct immunofluorescence assay (IFA). Faecal DNA was extracted from all Giardia cyst-positive samples and used for multilocus sequence typing with nested PCRs targeting the following gene loci: SSU rRNA (SSU), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). Samples from dogs and cats tested positive for Giardia coproantigen (ELISA) in 30.6% and 17.9%, respectively. The MIFC technique revealed Giardia cysts in 33.9% of canine and in 34.6% of feline ELISA-positive samples, while using IFA, cysts were present in 90.4% of canine and in 76.9% of feline ELISA-positive samples. Coinfections with other endoparasites besides Giardia were found in both dogs and cats, yet a statistically significant correlation could solely be drawn for the canine samples. The success rate of the different PCR protocols varied between 23.1% (tpi) and 91.3% (SSU) for dogs and between 25.0% (gdh) and 90.0% (SSU) for cats. Dog-specific Giardia assemblages C and D were detected in 42 and 55 canine isolates, respectively. The cat-specific Giardia assemblage F was detected in 14 feline isolates

  3. Occurrence of Cryptosporidium and Giardia in domestic animals in peri-urban communities of Kafue district, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I. G. K.; Enemark, Heidi L.

    2013-01-01

    Cryptosporidium spp. and Giardia duodenalis are important parasites infecting a wide range of domestic animals worldwide. The aim of the present study was to determine the occurrence of Cryptosporidium spp. and Giardia parasites in different domestic animals living in close contact with humans...... within rural/semiurban communities in Kafue district in Zambia. A single faecal sample per animal was collected from pigs, goats, dogs, ducks, chickens and pigeons and analysed by Merifluor Cryptosporidium/Giardia immunofluorescence antibody assay for the simultaneous detection of these parasites....... The faecal consistency was noted and scored as non-diarrhoeic or diarrhoeic. A total of 236 samples were collected. Cryptosporidium spp. oocysts were detected in pigs (11.5%, 17/148), goats (5.9%; 1/17), ducks (10.0%; 3/30) and chickens (14.3%; 2/14) while Giardia cysts were detected in pigs (8.1%; 12...

  4. Purifying Nanomaterials

    Science.gov (United States)

    Hung, Ching-Cheh (Inventor); Hurst, Janet (Inventor)

    2014-01-01

    A method of purifying a nanomaterial and the resultant purified nanomaterial in which a salt, such as ferric chloride, at or near its liquid phase temperature, is used to penetrate and wet the internal surfaces of a nanomaterial to dissolve impurities that may be present, for example, from processes used in the manufacture of the nanomaterial.

  5. Immunological aspects of Giardia infections

    OpenAIRE

    Heyworth, Martin F.

    2014-01-01

    Immunodeficiency, particularly antibody deficiency, predisposes to increased intensity and persistence of Giardia infections. Giardia-infected immunocompetent hosts produce serum and intestinal antibodies against Giardia trophozoites. The number of Giardia muris trophozoites, in mice with G. muris infection, is reduced by intra-duodenal administration of anti-G. muris antibody. Giardia intestinalis antigens that are recognised by human anti-trophozoite antibodies include variable (variant-spe...

  6. Giardia y giardiasis Giardia and giardiasis

    Directory of Open Access Journals (Sweden)

    Hugo D. Luján

    2006-02-01

    Full Text Available Giardia lamblia es un protozoario parásito que habita el intestino delgado de los seres humanos y de muchos otros vertebrados y es una de las más comunes causas de diarrea en todo el mundo. Durante su ciclo de vida Giardia sufre significativos cambios bioquímicos y morfológicos que le permiten sobrevivir en ambientes y condiciones que de otro modo lo destruirían. Para sobrevivir fuera del intestino del hospedador, los trofozoítos de Giardia se diferencian a quistes, los que se caracterizan por poseer una rígida pared glicoproteica externa que les permiten sobrevivir inclusive frente a la acción de los desinfectantes más comunes. Otro de los mecanismos de adaptación de este parásito es la variación de los antígenos de superficie que le permite a los trofozoítos evadir la respuesta inmune del huésped y generar infecciones tanto agudas como crónicas o recurrentes en individuos infectados. Durante los últimos años se han producido importantes avances en el conocimiento de las bases moleculares de los procesos de enquistamiento y variación antigénica en Giardia que pronostican el pronto hallazgo de nuevos agentes quimioterapéuticos y/o inmunoprofilácticos contra este importante parásito intestinal.Giardia lamblia is a protozoan parasite that inhabits the upper small intestine of humans and several other vertebrates and one of the most frequent cause of intestinal disease worldwide. During its life cycle, G. lamblia undergoes significant biochemical and morphological changes that allow the parasite to survive under hostile environmental conditions. To survive outside the host’s intestine, Giardia differentiates into resistant cysts, which are characterized by a rigid extracellular cyst wall that protect the parasite even to the action of the most common chemical disinfectants. Another adaptive mechanism of Giardia is the switching of its surface antigens, a mechanism that allows the trophozoites to evade the host’s immune

  7. Giardia: Illness & Symptoms

    Science.gov (United States)

    ... Illness & Symptoms Recommend on Facebook Tweet Share Compartir Giardia trophozoites stained with trichrome. Credit: Waterborne Disease Prevention Branch, CDC Giardiasis is the most frequently diagnosed intestinal parasitic disease in the United States and among ...

  8. Giardia: General Information

    Science.gov (United States)

    ... lives in the intestines and is passed in feces (poop). Once outside the body, Giardia can sometimes survive ... changing tables, diaper pails, or toys) that contain feces (poop) from an infected person or animal Drinking ...

  9. Genotyping and subtyping of Giardia and Cryptosporidium isolates from commensal rodents in China.

    Science.gov (United States)

    Zhao, Z; Wang, R; Zhao, W; Qi, M; Zhao, J; Zhang, L; Li, J; Liu, A

    2015-05-01

    Cryptosporidium and Giardia are two important zoonotic intestinal parasites responsible for diarrhoea in humans and other animals worldwide. Rodents, as reservoirs or carriers of Cryptosporidium and Giardia, are abundant and globally widespread. In the present study, we collected 232 fecal specimens from commensal rodents captured in animal farms and farm neighbourhoods in China. We collected 33 Asian house rats, 168 brown rats and 31 house mice. 6.0% (14/232) and 8.2% (19/232) of these rodents were microscopy-positive for Giardia cysts and Cryptosporidium oocysts, respectively. All 14 Giardia isolates were identified as Giardia duodenalis assemblage G at a minimum of one or maximum of three gene loci (tpi, gdh and bg). By small subunit rRNA (SSU rRNA) gene sequencing, Cryptosporidium parvum (n = 12) and Cryptosporidium muris (n = 7) were identified. The gp60 gene encoding the 60-kDa glycoprotein was successfully amplified and sequenced in nine C. parvum isolates, all of which belonged to the IIdA15G1 subtype. Observation of the same IIdA15G1 subtype in humans (previously) and in rodents (here) suggests that rodents infected with Cryptosporidium have the potential to transmit cryptosporidiosis to humans.

  10. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

    OpenAIRE

    Rogerio TIYO; Carla Zangari de SOUZA; Letícia NISHI; Camila Fernanda BRUSTOLIN; Bianca Altrão RATTI; Ana Lucia FALAVIGNA GUILHERME

    2015-01-01

    SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analy...

  11. Immunological aspects of Giardia infections.

    Science.gov (United States)

    Heyworth, Martin F

    2014-01-01

    Immunodeficiency, particularly antibody deficiency, predisposes to increased intensity and persistence of Giardia infections. Giardia-infected immunocompetent hosts produce serum and intestinal antibodies against Giardia trophozoites. The number of Giardia muris trophozoites, in mice with G. muris infection, is reduced by intra-duodenal administration of anti-G. muris antibody. Giardia intestinalis antigens that are recognised by human anti-trophozoite antibodies include variable (variant-specific) and invariant proteins. Nitric oxide (NO) appears to contribute to host clearance of Giardia trophozoites. Arginine is a precursor of NO and is metabolised by Giardia trophozoites, possibly reducing its availability for generation of NO by the host. Work with mice suggests that T lymphocytes and interleukin-6 (IL-6) contribute to clearance of Giardia infection via mechanisms independent of antibodies. © M.F. Heyworth, published by EDP Sciences, 2014.

  12. Interactions of Giardia sp. with the intestinal barrier: Epithelium, mucus, and microbiota.

    Science.gov (United States)

    Allain, Thibault; Amat, Christina B; Motta, Jean-Paul; Manko, Anna; Buret, André G

    2017-01-02

    Understanding how intestinal enteropathogens cause acute and chronic alterations has direct animal and human health perspectives. Significant advances have been made on this field by studies focusing on the dynamic crosstalk between the intestinal protozoan parasite model Giardia duodenalis and the host intestinal mucosa. The concept of intestinal barrier function is of the highest importance in the context of many gastrointestinal diseases such as infectious enteritis, inflammatory bowel disease, and post-infectious gastrointestinal disorders. This crucial function relies on 3 biotic and abiotic components, first the commensal microbiota organized as a biofilm, then an overlaying mucus layer, and finally the tightly structured intestinal epithelium. Herein we review multiple strategies used by Giardia parasite to circumvent these 3 components. We will summarize what is known and discuss preliminary observations suggesting how such enteropathogen directly and/ or indirectly impairs commensal microbiota biofilm architecture, disrupts mucus layer and damages host epithelium physiology and survival.

  13. Giardia disrupts the arrangement of tight, adherens and desmosomal junction proteins of intestinal cells.

    Science.gov (United States)

    Maia-Brigagão, C; Morgado-Díaz, J A; De Souza, W

    2012-06-01

    Giardia duodenalis is a parasitic protozoan that causes diarrhea and other symptoms which together constitute a disease known as giardiasis. Although the disease has been well defined, the mechanisms involving the establishment of the infection have not yet been fully elucidated. In this study, we show that after 24h of interaction between parasites and intestinal Caco-2 cells, there was an alteration of the paracellular permeability, as observed by an approximate 42% of reduction in the transepithelial electrical resistance and permeation to ruthenium red, which was concomitant with ultrastructural changes. Nevertheless, epithelium viability was not affected. We also demonstrate that there was no change in expression of junctional proteins (tight and adherens) but that the distribution of these proteins in Caco-2 cells after parasite adhesion was significantly altered, as observed via laser scanning confocal microscopy 3D reconstruction. The present work shows that adhesion of Giardia duodenalis trophozoites to intestinal cells in vitro induces disturbances of the tight, adherens and desmosomal junctions. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  14. Giardia survey in live-trapped small domestic and wild mammals in four regions in the southwest region of the State of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Maria Inês L. Sogayar

    1995-12-01

    Full Text Available For the first time, a survey on Giardia in the live-trapped small domestic and wild mammals was perfomed in four regions of state of the São Paulo, Brazil, with special attention to the parasitism of Rattus rattus rattus by Giardia. This species was found infected in all studied sites: Botucatu (15.4%, Conchas (28.5%, Itaporanga (38.7% and São Roque (100 %. Two new hosts and their frequency of infection were described for Giardia in Nectomys squamipes, an aquatic rodent and in Bolomys lasiurus, a forest rodent (100 % and 14.3 %, respectively. Both G. muris and G. duodenalis groups were found in scrapings of intestinal mucosa of those rodents. Mixed infection was observed in some animals. It is important to emphasize the infection by G. duodenalis in the black rat as this species lives as a comensal with man and in N. squamipes as it may contaminate small streams used for domestic consumption. Therefore, further investigation will be necessary to elucidate the potential of these rodents to act as reservoirs of Giardia for man.

  15. Introducing a simple and economical method to purify Giardia ...

    African Journals Online (AJOL)

    Jane

    2011-08-08

    Aug 8, 2011 ... Then they solved in distilled water. The second step of purification was done by adding them on 0.85 M sucrose and centrifugation at. 1600 g and 4°C for 10 min. In this step, fecal particles remained from the first step of purification were trapped in the middle phase and the cysts settled at bottom of the tubes ...

  16. Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil

    Directory of Open Access Journals (Sweden)

    Jonatas Campos Almeida

    Full Text Available The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR and sequencing. In the immunofluorescence, 2/24 (8.33% samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33% samples of raw water were positive for Giardia spp., and 2/24 (8.33% samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.

  17. Occurrence of Giardia and Cryptosporidium in captive chimpanzees (Pan troglodytes), mandrills (Mandrillus sphinx) and wild Zanzibar red colobus monkeys (Procolobus kirkii).

    Science.gov (United States)

    Debenham, John J; Atencia, Rebeca; Midtgaard, Fred; Robertson, Lucy J

    2015-04-01

    The aim of this study was to investigate the occurrence of Giardia duodenalis and Cryptosporidium spp. in primates and determine their zoonotic or anthropozoonotic potential. Direct immunofluorescence was used to identify Giardia and Cryptosporidium from faecal samples. PCR and DNA sequencing was performed on positive results. Giardia cysts were identified from 5.5% (5/90) of captive chimpanzees and 0% (0/11) of captive mandrills in the Republic of Congo; 0% (0/10) of captive chimpanzees in Norway; and 0% of faecal samples (n = 49) from wild Zanzibar red colobus monkeys. Two Giardia positive samples were also positive on PCR, and sequencing revealed identical isolates of Assemblage B. Cryptosporidium oocysts were not detected in any of the samples. In these primate groups, in which interactions with humans and human environments are quite substantial, Giardia and Cryptosporidium are rare pathogens. In chimpanzees, Giardia may have a zoonotic or anthropozoonotic potential. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Host defences against Giardia lamblia.

    Science.gov (United States)

    Lopez-Romero, G; Quintero, J; Astiazarán-García, H; Velazquez, C

    2015-08-01

    Giardia spp. is a protozoan parasite that inhabits the upper small intestine of mammals and other species and is the aetiological agent of giardiasis. It has been demonstrated that nitric oxide, mast cells and dendritic cells are the first line of defence against Giardia. IL-6 and IL-17 play an important role during infection. Several cytokines possess overlapping functions in regulating innate and adaptive immune responses. IgA and CD4(+) T cells are fundamental to the process of Giardia clearance. It has been suggested that CD4(+) T cells play a double role during the anti-Giardia immune response. First, they activate and stimulate the differentiation of B cells to generate Giardia-specific antibodies. Second, they act through a B-cell-independent mechanism that is probably mediated by Th17 cells. Several Giardia proteins that stimulate humoral and cellular immune responses have been described. Variant surface proteins, α-1 giardin, and cyst wall protein 2 can induce host protective responses to future Giardia challenges. The characterization and evaluation of the protective potential of the immunogenic proteins that are associated with Giardia will offer new insights into host-parasite interactions and may aid in the development of an effective vaccine against the parasite. © 2015 John Wiley & Sons Ltd.

  19. An audit of Cryptosporidium and Giardia detection in Scottish National Health Service Diagnostic Microbiology Laboratories.

    Science.gov (United States)

    Alexander, C L; Currie, S; Pollock, K; Smith-Palmer, A; Jones, B L

    2017-06-01

    Giardia duodenalis and Cryptosporidium species are protozoan parasites capable of causing gastrointestinal disease in humans and animals through the ingestion of infective faeces. Whereas Cryptosporidium species can be acquired locally or through foreign travel, there is the mis-conception that giardiasis is considered to be largely travel-associated, which results in differences in laboratory testing algorithms. In order to determine the level of variation in testing criteria and detection methods between diagnostic laboratories for both pathogens across Scotland, an audit was performed. Twenty Scottish diagnostic microbiology laboratories were invited to participate with questions on sample acceptance criteria, testing methods, testing rates and future plans for pathogen detection. Reponses were received from 19 of the 20 laboratories representing each of the 14 territorial Health Boards. Detection methods varied between laboratories with the majority performing microscopy, one using a lateral flow immunochromatographic antigen assay, another using a manually washed plate-based enzyme immunoassay (EIA) and one laboratory trialling a plate-based EIA automated with an EIA plate washer. Whereas all laboratories except one screened every stool for Cryptosporidium species, an important finding was that significant variation in the testing algorithm for detecting Giardia was noted with only four laboratories testing all diagnostic stools. The most common criteria were 'travel history' (11 laboratories) and/or 'when requested' (14 laboratories). Despite only a small proportion of stools being examined in 15 laboratories for Giardia (2%-18% of the total number of stools submitted), of interest is the finding that a higher positivity rate was observed for Giardia than Cryptosporidium in 10 of these 15 laboratories. These findings highlight that the underreporting of Giardia in Scotland is likely based on current selection and testing algorithms.

  20. Multilocus sequence analysis of Giardia spp. isolated from patients with diarrhea in Austria.

    Science.gov (United States)

    Lee, Mellesia F; Auer, Herbert; Lindo, John F; Walochnik, Julia

    2017-02-01

    Giardia duodenalis is a protozoan parasite causing intestinal infections in a wide range of mammals. Two distinct assemblages, A and B, infect humans predominantly; however, both are believed to be generally zoonotic. Giardia strains associated with infections in Austria have not been investigated at the molecular level. In this study, 65 human stool samples microscopically positive for Giardia spp. were subjected to DNA isolation and nested PCR targeting fragments of the glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), and beta-gardin (bg) genes. A total of 52 samples were successfully analyzed using PCR and DNA sequencing. Assemblage B was detected most frequently and accounted for 65.4% (34/52) of infections, while Assemblage A accounted for 34.6% (18/52). There was a high level of genetic diversity among the isolates with 46.2% designated as sub-assemblage BIV (24/52), 25% sub-assemblage AII (13/52), 19.2% sub-assemblage BIII (10/52), and 9.6% sub-assemblage AI (5/52). No mixed infections were detected. The results suggest that the majority of infections were imported and that endemic anthroponotic transmission plays a minor role in Austria.

  1. First survey of Cryptosporidium, Giardia and Enterocytozoon in diarrhoeic children from Wuhan, China.

    Science.gov (United States)

    Wang, Tao; Fan, Yingying; Koehler, Anson V; Ma, Guangxu; Li, Tao; Hu, Min; Gasser, Robin B

    2017-07-01

    Intestinal protozoan pathogens cause significant diarrhoeal diseases in children. However, to date, there has been limited genetic study of the intestinal pathogens Cryptosporidium, Giardia and Enterocytozoon in humans in China, with the exception of research in a small number of cities/provinces. In the present study, PCR-based tools were used to detect and characterise these protistan parasites from 500 children with a history of diarrhoea in Wuhan and environs, Hubei province, China. Genomic DNAs from faecal samples were screened for the particular protists by PCR utilising regions in the small subunit (SSU) of the nuclear ribosomal RNA, the 60kDa glycoprotein (gp60), the internal transcribed spacer of nuclear ribosomal DNA (ITS) and/or the triose phosphate isomerase (tpi) genes as markers. Cryptosporidium meleagridis subtype IIIb (10/500, 2.0%), Giardia duodenalis assemblage A (7/500, 1.4%) and Enterocytozoon bieneusi genotype D (1/500, 0.2%) were identified in small percentages of the 500 samples. No significant gender- or age-associated differences in the prevalence of Cryptosporidium and Giardia infections were found. Future studies might focus on the occurrence of these protists in children as well as animals, with an emphasis on Cryptosporidium meleagridis in pets and agriculturally important birds, in different parts of Hubei province. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. The Superoxide Reductase from the Early Diverging Eukaryote Giardia Intestinalis

    International Nuclear Information System (INIS)

    Cabelli, D.E.; Testa, F.; Mastronicola, D.; Bordi, E.; Pucillo, L.P.; Sarti, P.; Saraiva, L.M.; Giuffre, A.; Teixeira, M.

    2011-01-01

    Unlike superoxide dismutases (SODs), superoxidereductases (SORs) eliminate superoxide anion (O 2 # sm b ullet# - ) not through its dismutation, but via reduction to hydrogen peroxide (H 2 O 2 ) in the presence of an electron donor. The microaerobic protist Giardia intestinalis, responsible for a common intestinal disease in humans, though lacking SOD and other canonical reactive oxygen species-detoxifying systems, is among the very few eukaryotes encoding a SOR yet identified. In this study, the recombinant SOR from Giardia (SOR Gi ) was purified and characterized by pulse radiolysis and stopped-flow spectrophotometry. The protein, isolated in the reduced state, after oxidation by superoxide or hexachloroiridate(IV), yields a resting species (T final ) with Fe 3+ ligated to glutamate or hydroxide depending on pH (apparent pK a = 8.7). Although showing negligible SOD activity, reduced SOR Gi reacts with O 2 # sm b ullet# - with a pH-independent second-order rate constant k 1 = 1.0 x 10 9 M -1 s -1 and yields the ferric-(hydro)peroxo intermediate T 1 ; this in turn rapidly decays to the T final state with pH-dependent rates, without populating other detectable intermediates. Immunoblotting assays show that SOR Gi is expressed in the disease-causing trophozoite of Giardia. We propose that the superoxide-scavenging activity of SOR in Giardia may promote the survival of this air-sensitive parasite in the fairly aerobic proximal human small intestine during infection.

  3. Molecular and phylogenetic characterization of Cryptosporidium and Giardia from pigs and cattle in Denmark

    DEFF Research Database (Denmark)

    Langkjær, Rikke Breinhold; Vigre, Håkan; Enemark, Heidi L.

    2007-01-01

    The genetic diversity of Cryptosporidium spp. and Giardia duodenalis from dairy cattle and pigs in Denmark was determined in the present study. Faecal samples from 1237 pigs and 1150 cattle originating from 50 sow herds and 50 dairy herds, respectively, were analysed for the presence of the two...... parasites by immunofluorescence microscopy. A large proportion of the (oo)cyst containing samples were selected for molecular characterization. Sequencing and phylogenetic analysis of the 18S rDNA locus and/or the HSP70 gene of 183 pig and 154 cattle isolates of Cryptosporidium revealed the presence of C....... suis, pig genotype II, C. parvum (cattle genotype), C. bovis, Cryptosporidium deer-like genotype and a novel C. suis-like genotype. For both cattle and pigs, a host age-related change in distribution of species/genotypes was observed. The zoonotic C. parvum (cattle genotype) was most prevalent in young...

  4. Cryptosporidium and Giardia in Danish organic pig farms: seasonal and age-related variation in prevalence, infection intensity and species/genotypes

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Jianmin, Wang; Katakam, Kiran K.

    2015-01-01

    Although pigs are commonly infected with Cryptosporidium spp. and Giardia duodenalis, including potentially zoonotic species or genotypes, little is known about age-related infection levels, seasonal differences and genetic variation in naturally infected pigs raised in organic management systems....... Therefore, the current study was conducted to assess seasonal and age-related variations in prevalence and infection intensity of Cryptosporidium and Giardia, evaluate zoonotic potential and uncover correlations between species/genotypes, infection intensity and faecal consistency. Shedding of oocysts...... and cysts ((oo-) cysts) was monitored at quarterly intervals (September 2011 to June 2012) in piglets (n=152), starter pigs (n=234), fatteners (n=230) and sows (n=240) from three organic farms in Denmark. (Oo-) cysts were quantified by immunofluorescence microscopy; and 56/75 subsamples from Cryptosporidium...

  5. Detection and molecular characterization of Giardia and Cryptosporidium in common dolphins (Delphinus delphis) stranded along the Galician coast (Northwest Spain).

    Science.gov (United States)

    Reboredo-Fernández, A; Gómez-Couso, H; Martínez-Cedeira, J A; Cacciò, S M; Ares-Mazás, E

    2014-05-28

    The ubiquitous protozoan parasites Giardia and Cryptosporidium have been detected from many species of captive and free-living wildlife, representing most mammalian orders. Twenty species of marine mammals have been reported to inhabit Galician waters and the region has one of the highest rates of stranding in Europe. Evidence from stranding, reported by-catches and sightings, suggests that the common dolphin (Delphinus delphis) is the most abundant cetacean on the Galician coast (Northwest Spain). The objective of this study was to detect and molecularly characterize isolates of Giardia and Cryptosporidium obtained from common dolphins stranded in this area. Between 2005 and 2012, sections of large intestine from 133 common dolphins stranded along the Galician coast were collected by the personnel of the Galician Stranding Network (Coordinadora para o Estudo dos Mamíferos Mariños, CEMMA). Using direct immunofluorescence antibody test (IFAT) and PCR amplification and sequencing of the SSU-rDNA, β-giardin genes and the ITS1-5.8S-ITS2 region, Giardia and Cryptosporidium were detected in 8 (6.0%) and 12 samples (9.0%), respectively. In two samples, co-infection by both parasites was observed. The molecular characterization revealed the presence of Giardia duodenalis assemblages A (genotypes A1 and A2) and B and Cryptosporidium parvum in these samples. This constitutes the first study in which the presence of Giardia and Cryptosporidium has been investigated in common dolphins on the European Atlantic coast, and it is also the first report of C. parvum in this host. Our findings indicate that these animals could act as reservoir of these waterborne parasites or could be victims of the contamination originated by anthropogenic activities. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Cryptosporidium,Giardia, Cryptococcus, Pneumocystis genetic variability: cryptic biological species or clonal near-clades?

    Directory of Open Access Journals (Sweden)

    Michel Tibayrenc

    2014-04-01

    Full Text Available An abundant literature dealing with the population genetics and taxonomy of Giardia duodenalis, Cryptosporidium spp., Pneumocystis spp., and Cryptococcus spp., pathogens of high medical and veterinary relevance, has been produced in recent years. We have analyzed these data in the light of new population genetic concepts dealing with predominant clonal evolution (PCE recently proposed by us. In spite of the considerable phylogenetic diversity that exists among these pathogens, we have found striking similarities among them. The two main PCE features described by us, namely highly significant linkage disequilibrium and near-clading (stable phylogenetic clustering clouded by occasional recombination, are clearly observed in Cryptococcus and Giardia, and more limited indication of them is also present in Cryptosporidium and Pneumocystis. Moreover, in several cases, these features still obtain when the near-clades that subdivide the species are analyzed separately ("Russian doll pattern". Lastly, several sets of data undermine the notion that certain microbes form clonal lineages simply owing to a lack of opportunity to outcross due to low transmission rates leading to lack of multiclonal infections ("starving sex hypothesis". We propose that the divergent taxonomic and population genetic inferences advanced by various authors about these pathogens may not correspond to true evolutionary differences and could be, rather, the reflection of idiosyncratic practices among compartmentalized scientific communities. The PCE model provides an opportunity to revise the taxonomy and applied research dealing with these pathogens and others, such as viruses, bacteria, parasitic protozoa, and fungi.

  7. Giardia and Vilem Dusan Lambl

    Czech Academy of Sciences Publication Activity Database

    Lipoldová, Marie

    2014-01-01

    Roč. 8, č. 5 (2014), e2686 ISSN 1935-2735 R&D Projects: GA MŠk LH12049 Institutional support: RVO:68378050 Keywords : History of Science * Giardia lamblia * Infectious disease Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.446, year: 2014

  8. New system for higher recovery rate of water borne Cryptosporidium oocysts and Giardia cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, Jens; Klinting, Mette

    2012-01-01

    Background: The two most common water borne pathogenic protozoa, Cryptosporidium and Giardia, cause diarrhea worldwide. Detecting these parasites in water samples depends on effective parasite recovery from the water matrix. The reported low recovery rates of the currently used filter methods...... motivate the development of systems with higher recovery rates. Materials and methods: Five replicates of IMS purified Cryptosporidium oocysts and Giardia cysts (N=2x103) were injected into a specially coated filter unit with a carefully chosen pore size. Following filtration, sonication was performed...... at predetermined time intervals to loosen all parasites and other particles from the filter and break up clusters. The concentrated parasite suspension was backwashed and counted after immuno-fluorescence staining. Results and discussion: Without sonication the recovery rates of both Cryptosporidium and Giardia...

  9. The Superoxide Reductase from the Early Diverging Eukaryote Giardia Intestinalis

    Energy Technology Data Exchange (ETDEWEB)

    Cabelli, D.E.; Testa, F.; Mastronicola, D.; Bordi, E.; Pucillo, L.P.; Sarti, P.; Saraiva, L.M.; Giuffre, A.; Teixeira, M.

    2011-10-15

    Unlike superoxide dismutases (SODs), superoxidereductases (SORs) eliminate superoxide anion (O{sub 2}{sup {sm_bullet}-}) not through its dismutation, but via reduction to hydrogen peroxide (H{sub 2}O{sub 2}) in the presence of an electron donor. The microaerobic protist Giardia intestinalis, responsible for a common intestinal disease in humans, though lacking SOD and other canonical reactive oxygen species-detoxifying systems, is among the very few eukaryotes encoding a SOR yet identified. In this study, the recombinant SOR from Giardia (SOR{sub Gi}) was purified and characterized by pulse radiolysis and stopped-flow spectrophotometry. The protein, isolated in the reduced state, after oxidation by superoxide or hexachloroiridate(IV), yields a resting species (T{sub final}) with Fe{sup 3+} ligated to glutamate or hydroxide depending on pH (apparent pK{sub a} = 8.7). Although showing negligible SOD activity, reduced SOR{sub Gi} reacts with O{sub 2}{sup {sm_bullet}-} with a pH-independent second-order rate constant k{sub 1} = 1.0 x 10{sup 9} M{sup -1} s{sup -1} and yields the ferric-(hydro)peroxo intermediate T{sub 1}; this in turn rapidly decays to the T{sub final} state with pH-dependent rates, without populating other detectable intermediates. Immunoblotting assays show that SOR{sub Gi} is expressed in the disease-causing trophozoite of Giardia. We propose that the superoxide-scavenging activity of SOR in Giardia may promote the survival of this air-sensitive parasite in the fairly aerobic proximal human small intestine during infection.

  10. Giardia cyst wall protein 1 is a lectin that binds to curled fibrils of the GalNAc homopolymer.

    Directory of Open Access Journals (Sweden)

    Aparajita Chatterjee

    2010-08-01

    Full Text Available The infectious and diagnostic stage of Giardia lamblia (also known as G. intestinalis or G. duodenalis is the cyst. The Giardia cyst wall contains fibrils of a unique beta-1,3-linked N-acetylgalactosamine (GalNAc homopolymer and at least three cyst wall proteins (CWPs composed of Leu-rich repeats (CWP(LRR and a C-terminal conserved Cys-rich region (CWP(CRR. Our goals were to dissect the structure of the cyst wall and determine how it is disrupted during excystation. The intact Giardia cyst wall is thin (approximately 400 nm, easily fractured by sonication, and impermeable to small molecules. Curled fibrils of the GalNAc homopolymer are restricted to a narrow plane and are coated with linear arrays of oval-shaped protein complex. In contrast, cyst walls of Giardia treated with hot alkali to deproteinate fibrils of the GalNAc homopolymer are thick (approximately 1.2 microm, resistant to sonication, and permeable. The deproteinated GalNAc homopolymer, which forms a loose lattice of curled fibrils, is bound by native CWP1 and CWP2, as well as by maltose-binding protein (MBP-fusions containing the full-length CWP1 or CWP1(LRR. In contrast, neither MBP alone nor MBP fused to CWP1(CRR bind to the GalNAc homopolymer. Recombinant CWP1 binds to the GalNAc homopolymer within secretory vesicles of Giardia encysting in vitro. Fibrils of the GalNAc homopolymer are exposed during excystation or by treatment of heat-killed cysts with chymotrypsin, while deproteinated fibrils of the GalNAc homopolymer are degraded by extracts of Giardia cysts but not trophozoites. These results show the Leu-rich repeat domain of CWP1 is a lectin that binds to curled fibrils of the GalNAc homopolymer. During excystation, host and Giardia proteases appear to degrade bound CWPs, exposing fibrils of the GalNAc homopolymer that are digested by a stage-specific glycohydrolase.

  11. Occurrence and potential health risk of Cryptosporidium and Giardia in different water catchments in Belgium.

    Science.gov (United States)

    Ehsan, Amimul; Geurden, Thomas; Casaert, Stijn; Paulussen, Jef; De Coster, Lut; Schoemaker, Toon; Chalmers, Rachel; Grit, Grietje; Vercruysse, Jozef; Claerebout, Edwin

    2015-02-01

    Human wastewater and livestock can contribute to contamination of surface water with Cryptosporidium and Giardia. In countries where a substantial proportion of drinking water is produced from surface water, e.g., Belgium, this poses a constant threat on drinking water safety. Our objective was to monitor the presence of Cryptosporidium and Giardia in different water catchment sites in Belgium and to discriminate between (oo)cysts from human or animal origin using genotyping. Monthly samples were collected from raw water and purified drinking water at four catchment sites. Cryptosporidium and Giardia were detected using USEPA method 1623 and positive samples were genotyped. No contamination was found in purified water at any site. In three catchments, only low numbers of (oo)cysts were recovered from raw water samples (agriculture may be a more important source of surface water contamination than human waste in this catchment. In catchment sites with contaminated surface water, such as the Blankaart, continuous monitoring of treated water for the presence of Cryptosporidium and Giardia would be justified and (point) sources of surface water contamination should be identified.

  12. Plasmid vectors for proteomic analyses in Giardia: purification of virulence factors and analysis of the proteasome.

    Science.gov (United States)

    Jerlström-Hultqvist, Jon; Stadelmann, Britta; Birkestedt, Sandra; Hellman, Ulf; Svärd, Staffan G

    2012-07-01

    In recent years, proteomics has come of age with the development of efficient tools for purification, identification, and characterization of gene products predicted by genome projects. The intestinal protozoan Giardia intestinalis can be transfected, but there is only a limited set of vectors available, and most of them are not user friendly. This work delineates the construction of a suite of cassette-based expression vectors for use in Giardia. Expression is provided by the strong constitutive ornithine carbamoyltransferase (OCT) promoter, and tagging is possible in both N- and C-terminal configurations. Taken together, the vectors are capable of providing protein localization and production of recombinant proteins, followed by efficient purification by a novel affinity tag combination, streptavidin binding peptide-glutathione S-transferase (SBP-GST). The option of removing the tags from purified proteins was provided by the inclusion of a PreScission protease site. The efficiency and feasibility of producing and purifying endogenous recombinant Giardia proteins with the developed vectors was demonstrated by the purification of active recombinant arginine deiminase (ADI) and OCT from stably transfected trophozoites. Moreover, we describe the tagging, purification by StrepTactin affinity chromatography, and compositional analysis by mass spectrometry of the G. intestinalis 26S proteasome by employing the Strep II-FLAG-tandem affinity purification (SF-TAP) tag. This is the first report of efficient production and purification of recombinant proteins in and from Giardia, which will allow the study of specific parasite proteins and protein complexes.

  13. Efficacy of some metronidazole derivatives against Giardia lamblia, in vivo study

    Directory of Open Access Journals (Sweden)

    Mohammad Hossein Motazedian

    2015-09-01

    Full Text Available Giardiasis is a protozoal infection of small intestine caused by Giardia lamblia. This disease is usually asymptomatic, though it can present as acute or chronic diarrhea. Giardiasis, which is a major cause of intestinal infection, is endemic in Iran. Despite reports about drug resistance, long course treatment and various side effects of metronidazole, it is the drug of choice for giardiasis. In this study, we investigated the in vivo effects of five new derivatives (a-e of metronidazole (MTZ on the Giardia lamblia trophozoite in infected mice. Giardia intestinalis cysts were isolated from a patient and purified by sucrose gradient method. Fifty purified cysts were fed to the mice. After development of infection, the new metronidazole derivatives were given to the mice and the results were compared with metronidazole as the positive control group. Compounds a and b showed desirable antigiardiasis activity and could destroy the cyst and trophozoite of Giardia lamblia in mice after both two and four days, but the activity of the other compounds appeared only after 4 days.

  14. [In vitro effect against Giardia of 14 plant extracts].

    Science.gov (United States)

    Ponce-Macotela, M; Navarro-Alegría, I; Martínez-Gordillo, M N; Alvarez-Chacón, R

    1994-01-01

    To investigate antigiardiasic activity in plants used in Mexico as antidiarrheics and/or antiparasitics. Fourteen species were evaluated. The antigiardiasic activity was measured in vitro in a blinded fashion using trophozoites of Giardia duodenalis incubated with plant extracts. The viability of trophozoites was ascertained using MTT (3-[4,5-dimethylthiazol-2-il]-2, 5-diphenyl tetrazolium bromide) which is reduced to MTT-formazan by the activity of live trophozoites. The reduced MTT was extracted with an acidified alcohol (2-propanol with HCI 0.04 M) and measured in a spectrophotometer at 570 nm. Negative (trophozoites without extract) and positive controls (incubated with tinadazol) were included. The scientific and trivial names of the plants are given (trivials in Spanish marked by an asterisk). They had the following trophozoite mortality (mean +/- SD in percent): Justicia spicigera (muicle*) = 91 +/- 0.5; Lipia beriandieri (oregano) = 90 +/- 0.6; Psidium guajava (guava) = 87 +/- 1.0; positive control of tinidazol = 79 +/- 1.9; Punica granutus (granado*) = 78 +/- 1.3; Magnifera indica (mango) = 77 +/- 1.0; Plantago major (lante*) = 76 +/- 1.2; Cupressus semperbirens (cipres) = 73 +/- 1.2; Castella tormentosa (chaparro amargoso*) = 70 +/- 0.7; Hematoxilon campechanum (palo de Campeche*) = 67 +/- 1.2. Without or with a low mean activity were Prosopis juliflora (mesquite*) and Rizophora mangle (mangle*) with 0%, Oriza sativa (rice) with 5%, Capsicum annum (pimiento*) with 21% and Persea americana (avocado) with 23%. There were no associations of the antigiardiasic effect with concentration or osmolality of the extracts. A clear in vitro antigiardiasic effects was seen in nine species. Three of them were superior to tinidazol which is a drug of common use in the treatment of giardiasis.

  15. Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms

    Science.gov (United States)

    Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; dos Santos, Hannah Lia Ettiene Peruch Lemos; dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

    2017-01-01

    The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were

  16. Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

    Directory of Open Access Journals (Sweden)

    Roberta Dos Santos Toledo

    Full Text Available The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oocysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%, C. ryanae in eight (12.5%, C. bovis in four (6.3%, C. andersoni in five (7.8%, and a mixed infection in 20 samples (31.3%. These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%. Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2 in the fecal samples and one (IIaA17G2R1 in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G

  17. Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

    Science.gov (United States)

    Toledo, Roberta Dos Santos; Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; Dos Santos, Hannah Lia Ettiene Peruch Lemos; Dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

    2017-01-01

    The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were

  18. Handbook of purified gases

    CERN Document Server

    Schoen, Helmut

    2015-01-01

    Technical gases are used in almost every field of industry, science and medicine and also as a means of control by government authorities and institutions and are regarded as indispensable means of assistance. In this complete handbook of purified gases the physical foundations of purified gases and mixtures as well as their manufacturing, purification, analysis, storage, handling and transport are presented in a comprehensive way. This important reference work is accompanied with a large number of Data Sheets dedicated to the most important purified gases.  

  19. Giardia and Drinking Water from Private Wells

    Science.gov (United States)

    ... Once an animal or person is infected with Giardia , the parasite lives in the intestine and is passed in feces. Because the parasite is protected by an outer shell, it can survive outside the body and in the ... past 30 years, Giardia infection has become recognized as a common cause ...

  20. Modification of the Alere GIARDIA Ag TEST immunochromatography KIT methodology for its use in frozen fecal sediment of dogs and cats

    Directory of Open Access Journals (Sweden)

    VIVIANE A.N. COSTA

    Full Text Available ABSTRACT Giardia duodenalis is a worldwide intestinal parasite and is one of the most frequent protozoa species infecting dogs and cats. This study aimed to modify the methodology of Alere GIARDIA Ag TEST KIT for its use in frozen fecal sediments with different storage times in a freezer (-20°C, thus expanding the range of use of this methodology. One hundred fecal sediments from dogs (n=50 and cats (n=50 previously examined by optical microscopy for Giardia cysts were selected for this study. The agreement between the modified immunochromatography and microscopy results was calculated by Kappa coefficient. To evaluate the performance of the modified immunochromatography assay on samples with different storage time, the fecal sediments were divided into three groups according to the time of storage in a freezer: (a ≤ 1 year (n=37; (b > 1 year and ≤ 3 years (n=39; (c > 10 years (max. 13 years (n=24. The results obtained by the modified immunochromatography assay demonstrates a higher sensitivity of this technique when compared with microscopy, regardless of the frozen storage time. These results allow for the use of this methodology in a greater scope of analysis, especially in frozen fecal sediment triage in sample collections, enabling epidemiological and comparative analysis along different decades.

  1. Prevalence of Entamoeba histolytica , Giardia lamblia , and ...

    African Journals Online (AJOL)

    Introduction: The intestinal protozoa Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. are the causative agents of giardiasis, amebiasis, and cryptosporidiosis, respectively. Adequate knowledge of the geographical distribution of parasites and the demographic variables that influence their prevalence is ...

  2. Quantification of viable Giardia cysts and Cryptosporidium oocysts in wastewater using propidium monoazide quantitative real-time PCR.

    Science.gov (United States)

    Alonso, José L; Amorós, Inmaculada; Guy, Rebecca A

    2014-07-01

    Real-time PCR (qPCR) is a rapid tool to quantify pathogens in the aquatic environment; however, it quantifies all pathogens, including both viable and nonviable. Propidium monoazide (PMA) is a membrane-impairment dye that penetrates only membrane-damaged cells. Once inside the cell, PMA is covalently cross-linked to DNA through light photoactivation, and PCR amplification is strongly inhibited. The goal of this study was to evaluate PMA-qPCR assays for rapid quantification of viable and heat-treated Giardia cysts and Cryptosporidium oocysts in wastewater. We observed a reduction in detection of heat-treated Giardia duodenalis cysts of 83.2, 89.9, 98.2, or 97% with PMA-qPCR assays amplifying a 75 base-pair (bp) β-giardin target, 77-bp triosephosphate isomerase (tpi), 133-bp glutamate dehydrogenase (GDH), and 143-bp β-giardin gene target, respectively. Thus, the exclusion of dead cysts was more effective when qPCR assays that produced larger amplicons were used. The PMA treatment of Cryptosporidium oocysts plus/minus heat treatment abolished the fluorescent signal for dead oocysts with a PMA-qPCR assay amplifying a Cryptosporidium parvum (150-bp) oocyst wall protein (COWP) gene. The PMA-qPCR 143-bp β-giardin assay for Giardia and the PMA-qPCR 150-bp COWP assay for Cryptosporidium accurately quantified live oo(cysts), and failed to detect dead oo(cysts), when phosphate-buffered saline and tertiary effluent wastewater were spiked with concentrations of 10(3) or 10(2) dead oo(cysts), respectively. Therefore, these assays are suitable for the detection of viable parasites that are typically present in tertiary wastewater effluents at concentrations of <10(3) oo(cysts)/l and can provide rapid risk assessments of environmental water.

  3. Prevalence of intestinal parasites in companion dogs with diarrhea in Beijing, China, and genetic characteristics of Giardia and Cryptosporidium species.

    Science.gov (United States)

    Yu, Zhongjia; Ruan, Yang; Zhou, Mengjie; Chen, Siyuan; Zhang, Yinxin; Wang, Liya; Zhu, Guan; Yu, Yonglan

    2018-01-01

    Companion animals including dogs are one of the important components in One Health. Parasites may cause not only diseases in pet animals but also many zoonotic diseases infecting humans. In this study, we performed a survey of intestinal parasites in fecal specimens (n = 485) collected from outpatient pet dogs with diarrhea in Beijing, China, for the entire year of 2015 by microscopic examination (all parasites) and SSU rRNA-based nested PCR detection (Giardia and Cryptosporidium). We observed a total of 124 (25.6%) parasite-positive specimens that contained one or more parasites, including Giardia duodenalis (12.8%), Cryptosporidium spp. (4.9%), Cystoisospora spp. (4.3%), trichomonads (4.3%), Toxocara canis (3.5%), Trichuris vulpis (0.6%), and Dipylidium caninum (0.2%). Among the 55 dog breeds, infection rates were significantly higher in border collies and bulldogs, but lower in poodles (p Giardia-positive specimens, 21 were successfully assigned into assemblages using glutamate dehydrogenase (gdh) and/or beta-giardin (bg) genes, including assemblage D (n = 15), C (n = 5), and F (n = 1). Among the 24 Cryptosporidium-positive specimens by SSU rRNA PCR, 20 PCR amplicons could be sequenced and identified as Cryptosporidium canis (n = 20). Collectively, this study indicates that parasites are a significant group of pathogens in companion dogs in Beijing, and companion dogs may potentially transmit certain zoonotic parasites to humans, particularly those with weak or weakened immunity.

  4. Purified water quality study

    International Nuclear Information System (INIS)

    Spinka, H.; Jackowski, P.

    2000-01-01

    Argonne National Laboratory (HEP) is examining the use of purified water for the detection medium in cosmic ray sensors. These sensors are to be deployed in a remote location in Argentina. The purpose of this study is to provide information and preliminary analysis of available water treatment options and associated costs. This information, along with the technical requirements of the sensors, will allow the project team to determine the required water quality to meet the overall project goals

  5. Individual Water Purifier Study

    Science.gov (United States)

    2007-11-01

    study participants (Appendix A) who contributed their time and expertise to the analysis described in this report. 5 Blank 6 CONTENTS 1. INTRODUCTION ...and Weaknesses of Individual Devices ....................... 44 8 INDIVIDUAL WATER PURIFIER STUDY 1. INTRODUCTION This study was performed to evaluate...Data were gathered on each device and documented in a database. A Multi-Criteria Decision Model ( MCDM ) was developed, and a panel of experts assessed

  6. Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents

    Science.gov (United States)

    Cryptosporidiosis, giardiasis, and microsporidiosis are important waterborne diseases. In the standard for wastewater treatment plant (WWTP) effluents in China and other countries, fecal coliform is the only microbial indicator, raising concerns about the potential for pathogen t...

  7. Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents

    Data.gov (United States)

    U.S. Environmental Protection Agency — Cryptosporidiosis, giardiasis, and microsporidiosis are important waterborne diseases. In the standard for wastewater treatment plant (WWTP) effluents in China and...

  8. Giardia muris infection in mice is associated with a protective interleukin 17A response and induction of peroxisome proliferator-activated receptor alpha.

    Science.gov (United States)

    Dreesen, Leentje; De Bosscher, Karolien; Grit, Grietje; Staels, Bart; Lubberts, Erik; Bauge, Eric; Geldhof, Peter

    2014-08-01

    The protozoan parasite Giardia duodenalis (Giardia lamblia) is one of the most commonly found intestinal pathogens in mammals, including humans. In the current study, a Giardia muris-mouse model was used to analyze cytokine transcription patterns and histological changes in intestinal tissue at different time points during infection in C57BL/6 mice. Since earlier work revealed the upregulation of peroxisome proliferator-activated receptors (PPARs) in Giardia-infected calves, a second aim was to investigate the potential activation of PPARs in the intestines of infected mice. The most important observation in all mice was a strong upregulation of il17a starting around 1 week postinfection. The significance of interleukin 17A (IL-17A) in orchestrating a protective immune response was further demonstrated in an infection trial or experiment using IL-17 receptor A (IL-17RA) knockout (KO) mice: whereas in wild-type (WT) mice, cyst secretion dropped significantly after 3 weeks of infection, the IL-17RA KO mice were unable to clear the infection. Analysis of the intestinal response further indicated peroxisome proliferator-activated receptor alpha (PPARα) induction soon after the initial contact with the parasite, as characterized by the transcriptional upregulation of ppara itself and several downstream target genes such as pltp and cpt1. Overall, PPARα did not seem to have any influence on the immune response against G. muris, since PPARα KO animals expressed il-17a and could clear the infection similar to WT controls. In conclusion, this study shows for the first time the importance of IL-17 production in the clearance of a G. muris infection together with an early induction of PPARα. The effect of the latter, however, is still unclear. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  9. Process for purifying gases

    Energy Technology Data Exchange (ETDEWEB)

    Nogami, Y.; Ooka, I.; Hamamoto, T.; Shibano, H.; Miyao, K.

    1977-12-13

    An improved process is described for purifying a gas containing at least one of hydrogen cyanide and hydrogen sulfide as a harmful component by fixing the harmful component in water as an aqueous solution of nitrogen-containing sulfur compound so as to render the gas free of the harmful component. The improvement consists of: (1) burning the aqueous solution of nitrogen-containing sulfur compound to obtain an exhaust gas containing sulfur dioxide; (2) separating the sulfur dioxide from the exhaust gas for recovery; and (3) exhausting the gas after the separation of the sulfur dioxide.

  10. Process for purifying molybdenum

    International Nuclear Information System (INIS)

    Cheresnowsky, J.

    1989-01-01

    This patent describes a process for purifying molybdenum containing arsenic and phosphorus. The process comprising: adding to an acidic slurry of molybdenum trioxide, a source of magnesium ions in a solid form, with the amount of magnesium and the magnesium ion concentration in the subsequently formed ammonium molybdate solution being sufficient to subsequently form insoluble compounds containing greater than about 80% by weight of the arsenic and greater than about 80% by weight of the phosphorus, and ammonia in an amount sufficient to subsequently dissolve the molybdenum and subsequently form the insoluble compounds, with the source of magnesium ions being added prior to the addition of the ammonia; digesting the resulting ammoniated slurry at a temperature sufficient to dissolve the molybdenum and form an ammonium molybdate solution while the pH is maintained at from bout 9 to about 10 to form a solid containing the insoluble compounds; and separating the solid from the ammonium molybdate solution

  11. Tonic shock induces detachment of Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Wendy R Hansen

    Full Text Available BACKGROUND: The parasite Giardia lamblia must remain attached to the host small intestine in order to proliferate and subsequently cause disease. However, little is known about the factors that may cause detachment in vivo, such as changes in the aqueous environment. Osmolality within the proximal small intestine can vary by nearly an order of magnitude between host fed and fasted states, while pH can vary by several orders of magnitude. Giardia cells are known to regulate their volume when exposed to changes in osmolality, but the short-timescale effects of osmolality and pH on parasite attachment are not known. METHODOLOGY AND PRINCIPAL FINDINGS: We used a closed flow chamber assay to test the effects of rapid changes in media osmolality, tonicity, and pH on Giardia attachment to both glass and C2(Bbe-1 intestinal cell monolayer surfaces. We found that Giardia detach from both surfaces in a tonicity-dependent manner, where tonicity is the effective osmolality experienced by the cell. Detachment occurs with a characteristic time constant of 25 seconds (SD = 10 sec, n = 17 in both hypo- and hypertonic media but is otherwise insensitive to physiologically relevant changes in media composition and pH. Interestingly, cells that remain attached are able to adapt to moderate changes in tonicity. By exposing cells to a timed pattern of tonicity variations and adjustment periods, we found that it is possible to maximize the tonicity change experienced by the cells, overcoming the adaptive response and resulting in extensive detachment. CONCLUSIONS AND SIGNIFICANCE: These results, conducted with human-infecting Giardia on human intestinal epithelial monolayers, highlight the ability of Giardia to adapt to the changing intestinal environment and suggest new possibilities for treatment of giardiasis by manipulation of tonicity in the intestinal lumen.

  12. Pathogenic Mechanisms of Cryptosporidium and Giardia.

    Science.gov (United States)

    Certad, Gabriela; Viscogliosi, Eric; Chabé, Magali; Cacciò, Simone M

    2017-07-01

    Intestinal protozoa are important etiological agents of diarrhea, particularly in children, yet the public health risk they pose is often neglected. Results from the Global Enteric Multicenter Study (GEMS) showed that Cryptosporidium is among the leading causes of moderate to severe diarrhea in children under 2 years. Likewise, Giardia infects approximately 200 million individuals worldwide, and causes acute diarrhea in children under 5 years. Despite this recognized role as pathogens, the question is why and how these parasites cause disease in some individuals but not in others. This review focuses on known pathogenic mechanisms of Cryptosporidium and Giardia, and infection progress towards disease. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Checking the detail in retail: Occurrence of Cryptosporidium and Giardia on vegetables sold across different counters in Chandigarh, India.

    Science.gov (United States)

    Utaaker, Kjersti Selstad; Kumar, Anil; Joshi, Himanshu; Chaudhary, Suman; Robertson, Lucy J

    2017-12-18

    Fresh produce has been recognized as a vehicle of infection for protozoan parasites, particularly Cryptosporidium, and, to a lesser extent, Giardia. For both parasites, outbreaks associated with fresh produce have been documented. Although documented outbreaks tend to be from industrialized countries, contamination of fresh produce with these parasites is a global issue. In developing countries, infections with these parasites are often endemic in the community, and basic infrastructure and hygiene measures may be inadequate, thus the likelihood of contamination of fresh produce with these parasites may be higher. Realization of the importance of this transmission route comes against a backdrop of raw salads and more Western culinary habits gaining a foothold, and fresh produce being encouraged as part of the diet due to their associated health benefits. However, if consumption of uncooked fresh produce is going to increase its market sector in India, it is important that it is safe. In this study, various types of fresh produce obtained from three types of vendors in Chandigarh, a major city in Northern India, were analyzed for contamination with Cryptosporidium oocysts and Giardia cysts using a method that has been previously validated in inter-laboratory spiking experiments. A total of 284 samples of different fresh produce items were analyzed, obtained from the different retailers situated in different societal layers of the city. The overall prevalence of contamination of fresh produce with these parasites was just under 11%, with 6% of the vegetables contaminated with Cryptosporidium oocysts, and 5% with Giardia cysts. Contaminated vegetables included turnip, cabbage, carrot, chili, coriander, cucumber, radishes, and tomatoes. Molecular analyses identified contamination with Cryptosporidium parvum and Giardia duodenalis of Assemblage A and Assemblage D, indicating that contamination from animals may be of relevance. Although the prevalence of contamination is

  14. Prevalence of Cryptosporidium species and Giardia intestinalis ...

    African Journals Online (AJOL)

    Cryptosporidium species and Giardia intestinalis cause diarrheal infections in humans and other vertebrate animals globally and are considered to be of great public health importance. The study was conducted to determine the prevalence Cryptosporidium species and G. intestinalis infections among patients attending ...

  15. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

    Schrijven JF; Bruin HAM de; Engels GB; Leenen EJTM; MGB

    1999-01-01

    In this study, the relative contributions of the pathogenic protozoa Cryptosporidium and Giardia by manure of farm animals in The Netherlands to the total yearly environmental load was studied. Manure of veal calves forms a very large source of Cryptosporidium (1.5 m 10 square 16 oocysts per year)

  16. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

    Schrijven JF; Bruin HAM de; Engels GB; Leenen EJTM; MGB

    1999-01-01

    Het hier gepresenteerde deelonderzoek richt zich op de relatieve bijdrage van verschillende populaties landbouwhuisdieren via mest en afvalwater aan de totale emissie van Cryptosporidium en Giardia in Nederland. Vleeskalveren vormen per jaar in Nederland via hun mest een grote emissiebron van

  17. Prevalence of Entamoeba histolytica, Giardia lamblia, and ...

    African Journals Online (AJOL)

    2016-06-29

    Jun 29, 2016 ... Introduction: The intestinal protozoa Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. are ... prevalence rate of the three organisms was found according to gender, but most of infections were observed in children aged 10 ..... In developing countries, poor hygiene and the use of untreated ...

  18. Functional characterization of peroxiredoxins from the human protozoan parasite Giardia intestinalis.

    Directory of Open Access Journals (Sweden)

    Daniela Mastronicola

    Full Text Available The microaerophilic protozoan parasite Giardia intestinalis, causative of one of the most common human intestinal diseases worldwide, infects the mucosa of the proximal small intestine, where it has to cope with O2 and nitric oxide (NO. Elucidating the antioxidant defense system of this pathogen lacking catalase and other conventional antioxidant enzymes is thus important to unveil novel potential drug targets. Enzymes metabolizing O2, NO and superoxide anion (O2 (-• have been recently reported for Giardia, but it is yet unknown how the parasite copes with H2O2 and peroxynitrite (ONOO(-. Giardia encodes two yet uncharacterized 2-cys peroxiredoxins (Prxs, GiPrx1a and GiPrx1b. Peroxiredoxins are peroxidases implicated in virulence and drug resistance in several parasitic protozoa, able to protect from nitroxidative stress and repair oxidatively damaged molecules. GiPrx1a and a truncated form of GiPrx1b (deltaGiPrx1b were expressed in Escherichia coli, purified and functionally characterized. Both Prxs effectively metabolize H2O2 and alkyl-hydroperoxides (cumyl- and tert-butyl-hydroperoxide in the presence of NADPH and E. coli thioredoxin reductase/thioredoxin as the reducing system. Stopped-flow experiments show that both proteins in the reduced state react with ONOO(- rapidly (k = 4×10(5 M(-1 s(-1 and 2×10(5 M(-1 s(-1 at 4°C, for GiPrx1a and deltaGiPrx1b, respectively. Consistent with a protective role against oxidative stress, expression of GiPrx1a (but not deltaGiPrx1b is induced in parasitic cells exposed to air O2 for 24 h. Based on these results, GiPrx1a and deltaGiPrx1b are suggested to play an important role in the antioxidant defense of Giardia, possibly contributing to pathogenesis.

  19. Cytokines, antibodies and histopathological profiles during Giardia infection and VSP-based vaccination.

    Science.gov (United States)

    Serradell, Marianela C; Gargantini, Pablo R; Saura, Alicia; Oms, Sergio R; Rupil, Lucía L; Berod, Luciana; Sparwasser, Tim; Luján, Hugo D

    2018-03-19

    Giardiasis is one of the most common human intestinal diseases worldwide. Several experimental animals have been used to evaluate Giardia infections, with gerbils ( Meriones unguiculatus ) being the most valuable model due to their high susceptibility to Giardia infection, abundant shedding of cysts, and pathophysiological alterations and signs of disease similar to those observed in humans. Here we report cytokine and antibody profiles both during the course of Giardia infection in gerbils and after immunization with a novel oral vaccine comprising a mixture of purified Variant-specific Surface Proteins (VSPs). Transcript levels of representative cytokines of different immune profiles as well as macro- and micro-tissue alterations were assessed in Peyer's patches, mesenteric lymph nodes and spleens. During infection, cytokine responses showed a biphasic profile: an early induction of Th1 (IFN-γ, IL-1β, IL-6 and TNF), Th17 (IL-17) and Th2 (IL-4) cytokines, together with intestinal alterations typical of inflammation, followed by a shift toward a predominant Th2 (IL-5) response, likely associated with a counter-regulatory mechanism. Conversely, immunization with an oral vaccine comprising the entire repertoire of VSPs specifically showed high levels of IL-17, IL-6, IL-4 and IL-5, without obvious signs of inflammation. Both immunized and infected animals developed local (intestinal S-IgA) and systemic (serum IgG) humoral immune responses against VSPs; however, only infected animals showed evident signs of giardiasis. This is the first comprehensive report of cytokine expression and anti- Giardia antibody production during infection and VSP vaccination in gerbils, a reliable model of the human disease. Copyright © 2018 American Society for Microbiology.

  20. Ocorrência de Giardia spp. em bezerros leiteiros no Brasil Occurrence of Giardia spp. in dairy calves in Brazil

    Directory of Open Access Journals (Sweden)

    A.M. Guimarães

    2001-12-01

    Full Text Available Fecal samples were collected from 50 female Holstein calves (1-90 days old from a commercial dairy cattle farm located in Lavras, Minas Gerais, Brazil and examined for presence of Giardia cysts using the zinc sulfate flotation method. A total of 120 samples were collected from March 1999 to April 2000, and fecal consistency (normal or diarrheic was noted prior to the examination for Giardia cysts. Giardia spp. cysts were found in 11 (9% of the 120 faecal samples of calves co-infected with Eimeria spp., in calves from 19 to 58 days of age (38±19. In 82% (9/11 calves assessed for mixed infections with Cryptosporidium spp. and Giardia, oocysts of Cryptosporidium were found in three out of four diarrheic calves (75% positive for Giardia. Giardia cysts (n= 30 sized 13-16 x 10-12µm (mean of 15 x 11µm. They appeared to belong to the morphological group of G. intestinallis according to the size and morphological characteristic of the cysts. This is the first detection of Giardia spp. in calves in Brazil. Considering that Giardia has the potential to cause clinical disease in calves and that organisms infecting humans and domestic ruminants are morphologically and antigenically similar, and calves can shed Giardia cysts potentially infective for humans, the parasite in calves may be of major epidemiological significance, and suggests that naturally infected calves may be reservoirs of Giardia infections for man.

  1. Occurrence of Giardia, Cryptosporidium, and Entamoeba in wild rhesus macaques (Macaca mulatta living in urban and semi-rural North-West India

    Directory of Open Access Journals (Sweden)

    John J. Debenham

    2017-04-01

    Full Text Available Giardia duodenalis, Cryptosporidium spp., and Entamoeba spp. are intestinal protozoa capable of infecting a range of host species, and are important causes of human morbidity and mortality. Understanding their epidemiology is important, both for public health and for the health of the animals they infect. This study investigated the occurrence of these protozoans in rhesus macaques (Macaca mulatta in India, with the aim of providing preliminary information on the potential for transmission of these pathogens between macaques and humans. Faecal samples (n = 170 were collected from rhesus macaques from four districts of North-West India. Samples were analysed for Giardia/Cryptosporidium using a commercially available direct immunofluorescent antibody test after purification via immunomagnetic separation. Positive samples were characterised by sequencing of PCR products. Occurrence of Entamoeba was investigated first by using a genus-specific PCR, and positive samples further investigated via species-specific PCRs for Entamoeba coli, Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii. Giardia cysts were found in 31% of macaque samples, with all isolates belonging to Assemblage B. Cryptosporidium oocysts were found in 1 sample, however this sample did not result in amplification by PCR. Entamoeba spp. were found in 79% of samples, 49% of which were positive for E. coli. Multiplex PCR for E. histolytica, E. dispar and E. moshkovskii, did not result in amplification in any of the samples. Thus in 51% of the samples positive at the genus specific PCR, the Entamoeba species was not identified. This study provides baseline information on the potential for transmission of these zoonotic parasites at the wildlife-human interface.

  2. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

    Science.gov (United States)

    Tiyo, Rogerio; de Souza, Carla Zangari; Nishi, Letícia; Brustolin, Camila Fernanda; Ratti, Bianca Altrão; Falavigna Guilherme, Ana Lucia

    2015-01-01

    The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

  3. Molecular identification of Giardia and Cryptosporidium from dogs and cats

    Science.gov (United States)

    Sotiriadou, Isaia; Pantchev, Nikola; Gassmann, Doreen; Karanis, Panagiotis

    2013-01-01

    The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR) and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU) rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH) locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans. PMID:23477297

  4. Molecular identification of Giardia and Cryptosporidium from dogs and cats

    Directory of Open Access Journals (Sweden)

    Sotiriadou Isaia

    2013-01-01

    Full Text Available The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans.

  5. Advances in understanding Giardia: determinants and mechanisms of chronic sequelae

    Science.gov (United States)

    Sartor, R. Balfour

    2015-01-01

    Giardia lamblia is a flagellated protozoan that is the most common cause of intestinal parasitic infection in children living in resource-limited settings. The pathogenicity of Giardia has been debated since the parasite was first identified, and clinical outcomes vary across studies. Among recent perplexing findings are diametrically opposed associations between Giardia and acute versus persistent diarrhea and a poorly understood potential for long-term sequelae, including impaired child growth and cognitive development. The mechanisms driving these protean clinical outcomes remain elusive, but recent advances suggest that variability in Giardia strains, host nutritional status, the composition of microbiota, co-infecting enteropathogens, host genetically determined mucosal immune responses, and immune modulation by Giardia are all relevant factors influencing disease manifestations after Giardia infection. PMID:26097735

  6. Attachment of Giardia lamblia to rat intestinal epithelial cells.

    OpenAIRE

    Inge, P M; Edson, C M; Farthing, M J

    1988-01-01

    The human enteric protozoan, Giardia lamblia, has surface membrane lectin activity which mediates parasite adherence to erythrocytes. To determine whether an intestinal binding site exists for this lectin we have studied the interaction in vitro between axenically cultured Giardia trophozoites and isolated rat intestinal epithelial cells. Scanning electron microscopy showed that Giardia attached to the apical microvillus membrane and basolateral membrane of rat enterocytes. Any location on th...

  7. Giardia fatty acyl-CoA synthetases as potential drug targets

    Directory of Open Access Journals (Sweden)

    Fengguang eGuo

    2015-07-01

    Full Text Available Giardiasis caused by Giardia intestinalis (syn. G. lamblia, G. duodenalis is one of the leading causes of diarrheal parasitic diseases worldwide. Although limited drugs to treat giardiasis are available, there are concerns regarding toxicity in some patients and the emerging drug resistance. By data-mining genome sequences, we observed that G. intestinalis is incapable of synthesizing fatty acids de novo. However, this parasite has five long-chain fatty acyl-CoA synthetases (GiACS1 to GiACS5 to activate fatty acids scavenged from the host. ACS is an essential enzyme because fatty acids need to be activated to form acyl-CoA thioesters before they can enter subsequent metabolism. In the present study, we performed experiments to explore whether some GiACS enzymes could serve as drug targets in Giardia. Based on the high-throughput datasets and protein modeling analyses, we initially studied the GiACS1 and GiACS2, because genes encoding these two enzymes were found to be more consistently expressed in varied parasite life cycle stages and when interacting with host cells based on previously reported transcriptome data. These two proteins were cloned and expressed as recombinant proteins. Biochemical analysis revealed that both had apparent substrate preference towards palmitic acid (C16:0 and myristic acid (C14:0, and allosteric or Michaelis-Menten kinetics on palmitic acid or ATP. The ACS inhibitor triacsin C inhibited the activity of both enzymes (IC50 = 1.56 µM, Ki = 0.18 µM for GiACS1 and IC50 = 2.28 µM, Ki = 0.23 µM for GiACS2, respectively and the growth of G. intestinalis in vitro (IC50 = 0.8 µM. As expected from giardial evolutionary characteristics, both GiACSs displayed differences in overall folding structure as compared with their human counterparts. These observations support the notion that some of the GiACS enzymes may be explored as drug targets in this parasite.

  8. Análisis de los patrones de variación genética de las poblaciones de Giardia que parasitan humanos en el Área Sanitaria de Santiago de Compostela (Galicia)

    OpenAIRE

    Gabín García, Luis Basilio

    2017-01-01

    Giardia duodenalis es un protozoo parásito de humanos y otros animales de interés económico como el ganado. Su complejidad genética (dos núcleos diploides) junto a las limitaciones de las técnicas moleculares utilizadas en su estudio han impedido el avance en la comprensión de su epidemiología (potencial zoonótico, recombinación,...). La utilización de un protocolo de amplificación, clonación y secuenciación permitió realizar una descripción detallada de la diversidad genética de los ensambla...

  9. Ubiquity of the water-borne pathogens, Cryptosporidium and Giardia ...

    African Journals Online (AJOL)

    The prevalence of the diarrhoea disease caused by the water-borne pathogens Cryptosporidium and Giardia in KwaZulu-Natal, was determined from pathology laboratory data. Cryptosporidium and Giardia were found to be endemic in KwaZulu-Natal with laboratory-confirmed incidences ranging from 2.9 to 3.7% and 2.9 to ...

  10. What Entamoeba histolytica and Giardia lamblia tell us about the ...

    Indian Academy of Sciences (India)

    Unknown

    Entamoeba histolytica and Giardia lamblia are microaerophilic protists, which have long been considered models of ancient pre-mitochondriate eukaryotes. As transitional eukaryotes, amoebae and giardia appeared to lack organelles of higher eukaryotes and to depend upon energy metabolism appropriate for anaerobic ...

  11. Mechanisms of adaptation in the intestinal parasite Giardia lamblia.

    Science.gov (United States)

    Lujan, Hugo D

    2011-01-01

    Giardia lamblia, a parasite of humans, is a major source of waterborne diarrhoeal disease. Giardia is also an excellent system to study basic biochemical processes because it is a single-celled eukaryote with a small genome and its entire life cycle can be replicated in vitro. Giardia trophozoites undergo fundamental changes to survive outside the intestine of their host by differentiating into infective cysts. Encystation entails the synthesis, processing, transport, secretion and extracellular assembly of cyst wall components. To survive within the intestine, Giardia undergoes antigenic variation, a process by which the parasite continuously switches its major surface molecules, allowing the parasite to evade the host's immune response and produce chronic and recurrent infections. The objective of the present chapter is to provide a better understanding of the molecular mechanisms involved in adaptation and differentiation in Giardia, with a particular focus on the process of encystation and antigenic variation of this interesting micro-organism.

  12. Genetic Variation of Giardia lamblia Isolates from Food-handlers in Kashan, Central Iran

    Directory of Open Access Journals (Sweden)

    Hossein HOOSHYAR

    2017-02-01

    Full Text Available Background: Based on genotyping study of human isolates of Giardia lamblia; humans are mainly infected by two assemblages A and B. The present study was carried out to determine the sub-assemblages of G. lamblia isolated from food handlers referred to Kashan health centers, central Iran, 2015.Methods: In this cross-sectional study, 3653 stool samples collected from food-handlers that annually refer to health center for getting a health certification and examined microscopically for G. lamblia cyst. Totally, 44 isolates were selected from 47 Giardia positive samples. Cysts were partially purified by the sucrose density gradient method. After freeze-thaw cycles, genomic DNA was extracted using QIAamp Stool Mini kit. A single step PCR-RFLP method was used to amplify a 458bp fragment at the glutamate dehydrogenase (gdh locus, restriction enzymes BspLI and RsaI were used for distinguish between genotypes A and B and their subgroups.Results: Of 44 isolates, 24(54.5% were sub-assemblage AII, 9(20.5% group B including 7(15.9% BIII and 2(4.6% BIV sub-assemblage and 11(25% isolates showed a mixed pattern of AII and B. Sub-assemblage AI was not detected in this study.Conclusion: The higher rate of sub-assemblage AII demonstrated an anthroponotic origin of the infection so infected food-handlers could directly transmit this protozoan to consumers via contaminated food and water. For finding of pattern of transmission and distribution of Giardia assemblages and sub-assemblage, more studies in human and animal population in different regions are necessary.

  13. Immunological interaction between Giardia cyst extract and experimental toxoplasmosis.

    Science.gov (United States)

    Ashour, D S; Saad, A E; Dawood, L M; Zamzam, Y

    2018-01-01

    Toxoplasmosis is mostly associated with other intestinal parasitic infections especially Giardia due to shared mode of peroral infection. Toxoplasma and Giardia induce a strong T-helper 1- immune response. Our aim was to induce a protective immune response that results in significant impact on intestinal and extra-intestinal phases of Toxoplasma infection. This study was conducted in experimental animals and assessment of Giardia cyst extract effect on Toxoplasma infection was investigated by histopathological examination of small intestine and brain, Toxoplasma cyst count and iNOS staining of the brain, measurement of IFN-γ and TGF-β in intestinal tissues. Results showed that the brain Toxoplasma cyst number was decreased in mice infected with Toxoplasma then received Giardia cyst extract as compared to mice infected with Toxoplasma only. This effect was produced because Giardia cyst extract augmented the immune response to Toxoplasma infection as evidenced by severe inflammatory reaction in the intestinal and brain tissues, increased levels of IFN-γ and TGF-β in intestinal tissues and strong iNOS staining of the brain. In conclusion, Giardia cyst extract generated a protective response against T. gondii infection. Therefore, Giardia antigen will be a suitable candidate for further researches as an immunomodulatory agent against Toxoplasma infection. © 2017 John Wiley & Sons Ltd.

  14. Mine water purify from radium

    International Nuclear Information System (INIS)

    Lebecka, J.

    1996-01-01

    The article describes purification of radium containing water in coal mines. Author concludes that water purification is relatively simple and effective way to decrease environmental pollution caused by coal mining. The amount of radium disposed with type A radium water has been significantly decreased. The results of investigations show that it will be soon possible to purify also type B radium water. Article compares the amounts of radium disposed by coal mines in 1990, 1995 and forecast for 2000

  15. Binding properties and immunolocalization of a fatty acid-binding protein in Giardia lamblia.

    Science.gov (United States)

    Hassan, S M T; Maache, M; de la Guardia, R Díaz; Córdova, O M; García, J R Gil; Galiana, M; Acuña Castroviejo, D; Martins, M; Osuna, Antonio

    2005-04-01

    We describe here a fatty acid-binding protein (FABP) isolated and purified from the parasitic protozoon Giardia lamblia. The protein has a molecular mass of 8 kDa and an isoelectric point of 4.96. A Scatchard analysis of the data at equilibrium revealed a dissociation constant of 3.12 x 10(-8) M when the labeled oleic acid was displaced by a 10-fold greater concentration of unlabeled oleic acid. Testosterone, sodium desoxycholate, taurocholate, metronidazol, and alpha-tocopherol, together with butyric, arachidonic, palmitic, retinoic, and glycocholic acids, were also bound to the protein. Assays with polyclonal antibodies revealed that the protein is located in the ventral disk and also appears in the dorsal membrane, the cytoplasm, and in the vicinity of the lipid vacuoles.

  16. Accurate diagnosis of Giardia spp and proper fecal examination procedures.

    Science.gov (United States)

    Dryden, M W; Payne, P A; Smith, V

    2006-01-01

    A series of investigations evaluated the ability of different testing methods - a swing-head centrifugation technique using two flotation solutions (1.18-specific gravity zinc sulfate and 1.27-specific gravity Sheather's sugar solution), a passive commercial flotation technique, and the SNAP Giardia Test Kit from IDEXX Laboratories - to identify Giardia-positive dogs and recover the eggs of other intestinal parasites. It was determined that the SNAP Giardia test can improve a practice's ability to identify Giardia-infected dogs. Because of its higher specific gravity, the sugar solution was better for recovering heavy parasite eggs, such as Taenia spp, and thus is the flotation solution of choice when conducting routine centrifugation fecal examinations.

  17. New insights regarding the biology of Giardia lamblia.

    Science.gov (United States)

    Carranza, Pedro G; Lujan, Hugo D

    2010-01-01

    Giardia lamblia is one of the most common causes of intestinal disease in humans. To adapt to environments both inside and outside of the host's small intestine, this protozoan parasite undergoes significant developmental changes during its life cycle. In this review, we analyze and discuss the most recent findings regarding the process of Giardia trophozoites differentiation into infective cysts as well as the mechanism of antigenic variation, which allows the parasite to cause chronic and recurrent infections in susceptible individuals.

  18. Transient transfection and expression of firefly luciferase in Giardia lamblia.

    OpenAIRE

    Yee, J; Nash, T E

    1995-01-01

    We have developed a gene transfer system for the protozoan parasite Giardia lamblia. This organism is responsible for many cases of diarrhea worldwide and is considered to be one of the most primitive eukaryotes. Expression of a heterologous gene was detected in this parasite after electroporation with appropriate DNA constructs. We constructed a series of transfection plasmids using flanking sequences of the Giardia glutamate dehydrogenase (GDH) gene to drive expression of the firefly lucife...

  19. Giardia Alters Commensal Microbial Diversity throughout the Murine Gut.

    Science.gov (United States)

    Barash, N R; Maloney, J G; Singer, S M; Dawson, S C

    2017-06-01

    Giardia lamblia is the most frequently identified protozoan cause of intestinal infection. Over 200 million people are estimated to have acute or chronic giardiasis, with infection rates approaching 90% in areas where Giardia is endemic. Despite its significance in global health, the mechanisms of pathogenesis associated with giardiasis remain unclear, as the parasite neither produces a known toxin nor induces a robust inflammatory response. Giardia colonization and proliferation in the small intestine of the host may, however, disrupt the ecological homeostasis of gastrointestinal commensal microbes and contribute to diarrheal disease associated with giardiasis. To evaluate the impact of Giardia infection on the host microbiota, we used culture-independent methods to quantify shifts in the diversity of commensal microbes throughout the gastrointestinal tract in mice infected with Giardia We discovered that Giardia 's colonization of the small intestine causes a systemic dysbiosis of aerobic and anaerobic commensal bacteria. Specifically, Giardia colonization is typified by both expansions in aerobic Proteobacteria and decreases in anaerobic Firmicutes and Melainabacteria in the murine foregut and hindgut. Based on these shifts, we created a quantitative index of murine Giardia -induced microbial dysbiosis. This index increased at all gut regions during the duration of infection, including both the proximal small intestine and the colon. Giardiasis could be an ecological disease, and the observed dysbiosis may be mediated directly via the parasite's unique anaerobic fermentative metabolism or indirectly via parasite induction of gut inflammation. This systemic alteration of murine gut commensal diversity may be the cause or the consequence of inflammatory and metabolic changes throughout the gut. Shifts in the commensal microbiota may explain observed variations in giardiasis between hosts with respect to host pathology, degree of parasite colonization, infection

  20. Purification and characterization of a protein capable of binding to fatty acids and bile salts in Giardia lamblia.

    Science.gov (United States)

    de la Guardia, R Diaz; Lopez, M B; Burgos, M; Osuna, A

    2011-08-01

    A specific fatty acid binding protein was isolated from Giardia lamblia, using an affinity column with butyric acid acting as a ligand in place of stearic acid. This method has proved to be more efficient than the one previously described using stearic acid as ligand. The purified fraction showed 8 electrophoretic bands of proteins, with molecular weights ranging between 8 and 80 kDa. This pattern is a consequence of the aggregation of a protein with a molecular weight of 8,215 Da, corresponding to the lower molecular weight band, the only one capable of binding to fatty acids. The labeled oleic acid bound to these purified proteins was replaced by a 100-fold greater concentration of taurocholate, glycocholate, deoxycholate, palmitic acid, and arachidonic acid, having a greater displacement of the bile salts than the free fatty acids.

  1. Development of functional gastrointestinal disorders after Giardia lamblia infection

    Directory of Open Access Journals (Sweden)

    Hanevik Kurt

    2009-04-01

    Full Text Available Abstract Background Functional gastrointestinal disorders (FGID may occur following acute gastroenteritis. This long-term complication has previously not been described after infection with the non-invasive protozoan Giardia lamblia. This study aims to characterize persistent abdominal symptoms elicited by Giardia infection according to Rome II criteria and symptoms scores. Methods Structured interview and questionnaires 12–30 months after the onset of Giardia infection, and at least 6 months after Giardia eradication, among 82 patients with persisting abdominal symptoms elicited by the Giardia infection. All had been evaluated to exclude other causes. Results We found that 66 (80.5% of the 82 patients had symptoms consistent with irritable bowel syndrome (IBS and 17 (24.3% patients had functional dyspepsia (FD according to Rome II criteria. IBS was sub classified into D-IBS (47.0%, A-IBS (45.5% and C-IBS (7.6%. Bloating, diarrhoea and abdominal pain were reported to be most severe. Symptoms exacerbation related to specific foods were reported by 45 (57.7% patients and to physical or mental stress by 34 (44.7% patients. Conclusion In the presence of an IBS-subtype pattern consistent with post-infectious IBS (PI-IBS, and in the absence of any other plausible causes, we conclude that acute Giardia infection may elicit functional gastrointestinal diseases with food and stress related symptoms similar to FGID patients in general.

  2. Reconstruction of Sugar Metabolic Pathways of Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Jian Han

    2012-01-01

    Full Text Available Giardia lamblia is an “important” pathogen of humans, but as a diplomonad excavate it is evolutionarily distant from other eukaryotes and relatively little is known about its core metabolic pathways. KEGG, the widely referenced site for providing information of metabolism, does not yet include many enzymes from Giardia species. Here we identify Giardia’s core sugar metabolism using standard bioinformatic approaches. By comparing Giardia proteomes with known enzymes from other species, we have identified enzymes in the glycolysis pathway, as well as some enzymes involved in the TCA cycle and oxidative phosphorylation. However, the majority of enzymes from the latter two pathways were not identifiable, indicating the likely absence of these functionalities. We have also found enzymes from the Giardia glycolysis pathway that appear more similar to those from bacteria. Because these enzymes are different from those found in mammals, the host organisms for Giardia, we raise the possibility that these bacteria-like enzymes could be novel drug targets for treating Giardia infections.

  3. Home drinking-water purifiers

    International Nuclear Information System (INIS)

    Pizzichini, Massimo; Pozio, Alfonso; Russo, Claudio

    2005-01-01

    To salve the widespread problem of contaminated drinking water, home purifiers are now sold in Italy as well as other countries. This article describes how these devices work, how safe they are to use and how safe the water they produce, in the broad context of regulations on drinking water and mineral water. A new device being developed by ENEA to treat municipal water and ground water could provide greater chemical and bacteriological safety. However, the appearance of these new systems makes it necessary to update existing regulations [it

  4. Process for purifying zirconium sponge

    International Nuclear Information System (INIS)

    Abodishish, H.A.M.; Kimball, L.S.

    1992-01-01

    This patent describes a Kroll reduction process wherein a zirconium sponge contaminated with unreacted magnesium and by-product magnesium chloride is produced as a regulus, a process for purifying the zirconium sponge. It comprises: distilling magnesium and magnesium chloride from: a regulus containing a zirconium sponge and magnesium and magnesium chloride at a temperature above about 800 degrees C and at an absolute pressure less than about 10 mmHg in a distillation vessel to purify the zirconium sponge; condensing the magnesium and the magnesium chloride distilled from the zirconium sponge in a condenser; and then backfilling the vessel containing the zirconium sponge and the condenser containing the magnesium and the magnesium chloride with a gas; recirculating the gas between the vessel and the condenser to cool the zirconium sponge from above about 800 degrees C to below about 300 degrees C; and cooling the recirculating gas in the condenser containing the condensed magnesium and the condensed magnesium chloride as the gas cools the zirconium sponge to below about 300 degrees C

  5. Methods for Purifying Enzymes for Mycoremediation

    Science.gov (United States)

    Cullings, Kenneth W. (Inventor); DeSimone, Julia C. (Inventor); Paavola, Chad D. (Inventor)

    2014-01-01

    A process for purifying laccase from an ectomycorrhizal fruiting body is disclosed. The process includes steps of homogenization, sonication, centrifugation, filtration, affinity chromatography, ion exchange chromatography, and gel filtration. Purified laccase can also be separated into isomers.

  6. Endurance Pump Test with MIL-PRF-83282 Hydraulic Fluid, Purified with Malabar Purifier

    National Research Council Canada - National Science Library

    Sharma, Shashi

    2004-01-01

    .... Endurance aircraft hydraulic pump tests under carefully controlled conditions were previously conducted using hydraulic fluid purified with a rotating-disk and vacuum type purifier, the portable...

  7. Tendencia de la prevalencia y factores asociados a la infección por Giardia duodenalis en escolares y preescolares de una zona rural de Cundinamarca

    Directory of Open Access Journals (Sweden)

    María del Pilar Chaves

    2007-09-01

    Conclusión. La giardiasis continúa siendo un problema de salud pública en esta población; mientras persistan los factores ambientales y socioeconómicos asociados con la infección, esta protozoosis intestinal no podrá ser controlada.

  8. Evaluation of a Screening Test for Detection of Giardia and Cryptosporidium Parasites ▿

    OpenAIRE

    Youn, Sojin; Kabir, Mamun; Haque, Rashidul; Petri, William A.

    2008-01-01

    The Giardia/Cryptosporidium Chek test (TechLab, Inc.), a screening test for Giardia and Cryptosporidium, was evaluated with 136 fecal samples. Using the results of the Giardia II test and Cryptosporidium II test as gold standards, it was 98.4% sensitive and 100% specific and had positive and negative predictive values of 98.7% and 99.3%.

  9. Crosstalk between Zinc Status and Giardia Infection: A New Approach

    Directory of Open Access Journals (Sweden)

    Humberto Astiazarán-García

    2015-06-01

    Full Text Available Zinc supplementation has been shown to reduce the incidence and prevalence of diarrhea; however, its anti-diarrheal effect remains only partially understood. There is now growing evidence that zinc can have pathogen-specific protective effects. Giardiasis is a common yet neglected cause of acute-chronic diarrheal illness worldwide which causes disturbances in zinc metabolism of infected children, representing a risk factor for zinc deficiency. How zinc metabolism is compromised by Giardia is not well understood; zinc status could be altered by intestinal malabsorption, organ redistribution or host-pathogen competition. The potential metal-binding properties of Giardia suggest unusual ways that the parasite may interact with its host. Zinc supplementation was recently found to reduce the rate of diarrhea caused by Giardia in children and to upregulate humoral immune response in Giardia-infected mice; in vitro and in vivo, zinc-salts enhanced the activity of bacitracin in a zinc-dose-dependent way, and this was not due to zinc toxicity. These findings reflect biological effect of zinc that may impact significantly public health in endemic areas of infection. In this paper, we shall explore one direction of this complex interaction, discussing recent information regarding zinc status and its possible contribution to the outcome of the encounter between the host and Giardia.

  10. Crosstalk between Zinc Status and Giardia Infection: A New Approach.

    Science.gov (United States)

    Astiazarán-García, Humberto; Iñigo-Figueroa, Gemma; Quihui-Cota, Luis; Anduro-Corona, Iván

    2015-06-03

    Zinc supplementation has been shown to reduce the incidence and prevalence of diarrhea; however, its anti-diarrheal effect remains only partially understood. There is now growing evidence that zinc can have pathogen-specific protective effects. Giardiasis is a common yet neglected cause of acute-chronic diarrheal illness worldwide which causes disturbances in zinc metabolism of infected children, representing a risk factor for zinc deficiency. How zinc metabolism is compromised by Giardia is not well understood; zinc status could be altered by intestinal malabsorption, organ redistribution or host-pathogen competition. The potential metal-binding properties of Giardia suggest unusual ways that the parasite may interact with its host. Zinc supplementation was recently found to reduce the rate of diarrhea caused by Giardia in children and to upregulate humoral immune response in Giardia-infected mice; in vitro and in vivo, zinc-salts enhanced the activity of bacitracin in a zinc-dose-dependent way, and this was not due to zinc toxicity. These findings reflect biological effect of zinc that may impact significantly public health in endemic areas of infection. In this paper, we shall explore one direction of this complex interaction, discussing recent information regarding zinc status and its possible contribution to the outcome of the encounter between the host and Giardia.

  11. Role of nitric oxide and superoxide in Giardia lamblia killing

    Directory of Open Access Journals (Sweden)

    P.D. Fernandes

    1997-01-01

    Full Text Available Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine. Activated macrophages were cytotoxic to Giardia trophozoites (~60% dead trophozoites. This effect was inhibited (>90% by an NO synthase inhibitor (200 µM and unaffected by superoxide dismutase (SOD, 300 U/ml. Giardia trophozoites were killed by the NO donors, S-nitroso-acetyl-penicillamine (SNAP and sodium nitroprusside (SNP in a dose-dependent manner (LD50 300 and 50 µM, respectively. A dual NO-superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1, did not have a killing effect in concentrations up to 1 mM. However, when SOD (300 U/ml was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (~35% dead trophozoites at 1 mM. The mixture of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM. These results indicate that NO accounts for Giardia trophozoite killing and this effect is not mediated by peroxynitrite

  12. Prokaryotic Expression ofα-13 Giardin Gene and Its Intracellular Localization inGiardia lamblia.

    Science.gov (United States)

    Yu, Xingang; Abdullahi, Auwalu Yusuf; Wu, Sheng; Pan, Weida; Shi, Xianli; Hu, Wei; Tan, Liping; Li, Kangxin; Wang, Zhen; Li, Guoqing

    2017-01-01

    To study prokaryotic expression and subcellular localization of α -13 giardin in Giardia lamblia trophozoites, α -13 giardin gene was amplified and cloned into prokaryotic expression vector pET-28a(+). The positive recombinant plasmid was transformed into E. coli BL21(DE3) for expression by using IPTG and autoinduction expression system (ZYM-5052). The target protein was validated by SDS-PAGE and Western blotting and purified by Ni-NTA Resin. Rabbits were immunized with purified fusion proteins for preparation of polyclonal antibody; then the intracellular location of α -13 giardin was determined by fluorescence immunoassay. The results showed that the length of α -13 giardin gene was 1038 bp, encoding a polypeptide of 345 amino acids. The expressed product was a fusion protein with about 40 kDa largely present in soluble form. The target protein accounted for 21.0% of total proteins after being induced with IPTG, while it accounted for 28.8% with ZYM-5052. The anti- α 13-giardin polyclonal antibody possessed good antigenic specificity as well as excellent binding activity with recombinant α -13 giardin. Immunofluorescence assays revealed that α -13 giardin was localized in the cytoplasm of G. lamblia trophozoite, suggesting that it is a cytoplasm-associated protein. The present study may lay a foundation for further functional research on α -13 giardin of G. lamblia .

  13. Genomic minimalism in the early diverging intestinal parasite Giardia lamblia.

    Science.gov (United States)

    Morrison, Hilary G; McArthur, Andrew G; Gillin, Frances D; Aley, Stephen B; Adam, Rodney D; Olsen, Gary J; Best, Aaron A; Cande, W Zacheus; Chen, Feng; Cipriano, Michael J; Davids, Barbara J; Dawson, Scott C; Elmendorf, Heidi G; Hehl, Adrian B; Holder, Michael E; Huse, Susan M; Kim, Ulandt U; Lasek-Nesselquist, Erica; Manning, Gerard; Nigam, Anuranjini; Nixon, Julie E J; Palm, Daniel; Passamaneck, Nora E; Prabhu, Anjali; Reich, Claudia I; Reiner, David S; Samuelson, John; Svard, Staffan G; Sogin, Mitchell L

    2007-09-28

    The genome of the eukaryotic protist Giardia lamblia, an important human intestinal parasite, is compact in structure and content, contains few introns or mitochondrial relics, and has simplified machinery for DNA replication, transcription, RNA processing, and most metabolic pathways. Protein kinases comprise the single largest protein class and reflect Giardia's requirement for a complex signal transduction network for coordinating differentiation. Lateral gene transfer from bacterial and archaeal donors has shaped Giardia's genome, and previously unknown gene families, for example, cysteine-rich structural proteins, have been discovered. Unexpectedly, the genome shows little evidence of heterozygosity, supporting recent speculations that this organism is sexual. This genome sequence will not only be valuable for investigating the evolution of eukaryotes, but will also be applied to the search for new therapeutics for this parasite.

  14. Simultaneous detection of the protozoan parasites Toxoplasma, Cryptosporidium and Giardia in food matrices and their persistence on basil leaves.

    Science.gov (United States)

    Hohweyer, Jeanne; Cazeaux, Catherine; Travaillé, Emmanuelle; Languet, Emilie; Dumètre, Aurélien; Aubert, Dominique; Terryn, Christine; Dubey, Jitender P; Azas, Nadine; Houssin, Maryline; Loïc, Favennec; Villena, Isabelle; La Carbona, Stéphanie

    2016-08-01

    Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis are emerging pathogen parasites in the food domain. However, without standardized methods for their detection in food matrices, parasitic foodborne outbreaks remain neglected. In this study, a new immunomagnetic separation assay (IMS Toxo) targeting the oocyst's wall of T. gondii was developed using a specific purified monoclonal antibody. Performance of this IMS Toxo coupled to microscopic and qPCR analyses was evaluated in terms of limit of detection (LOD) and recovery rate (RR) on: i) simple matrix (LOD = 5 oocysts; RR between 5 and 56%); ii) raspberries and basil (LOD = 33 oocysts/g; RR between 0.2 and 35%). Finally, to simultaneously extract the three protozoa from these food matrices, T. gondii oocysts were directly concentrated (without IMS Toxo) from the supernatant of the IMS of Cryptosporidium and Giardia (oo)cysts. This strategy associated to qPCR detection led to LOD <1 to 3 (oo)cysts/g and RR between 2 and 35%. This procedure was coupled to RT-qPCR analyses and showed that the three protozoa persisted on the leaves of basil and remained viable following storage at 4 °C for 8 days. These data strengthen the need to consider these protozoa in food safety. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Hydrogen purifier module with membrane support

    Science.gov (United States)

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

    2012-07-24

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

  16. Effect of Piper betle on Giardia intestinalis infection in vivo

    Czech Academy of Sciences Publication Activity Database

    Pecková, R.; Doležal, Karel; Sak, Bohumil; Květoňová, Dana; Kváč, Martin; Nurcahyo, W.; Foitová, I.

    2018-01-01

    Roč. 184, JAN (2018), s. 39-45 ISSN 0014-4894 R&D Projects: GA ČR(CZ) GAP505/11/1163 Institutional support: RVO:61389030 ; RVO:60077344 Keywords : Antigiardial activity * Drug of choice * Giardia intestinalis * Natural antiparasitics * Parasites * Piper betle Subject RIV: EF - Botanics OBOR OECD: Plant sciences, botany Impact factor: 1.724, year: 2016

  17. Molecular identification of Giardia lamblia isolates from adult human ...

    African Journals Online (AJOL)

    ARL

    2013-02-27

    Feb 27, 2013 ... Giardia lamblia is a flagellated protozoa that infects the intestinal tract of a wide range of mammalian hosts, including both wild and domestic animals as well as humans. ... Person-to-person, zoonotic, foodborne and waterborne transmissions may occur through the fecal-oral route after contact with the ...

  18. Prevalence of Giardia intestinalis and Hymenolepis nana in Afghan ...

    African Journals Online (AJOL)

    Background: Present study aimed to investigate prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugees visiting Central Health Unit (CHU), Kot ... are common among Afghan refugees and serious preventive measures should be implemented to promote the safety and healthy lifestyle of these people.

  19. What Entamoeba histolytica and Giardia lamblia tell us about the ...

    Indian Academy of Sciences (India)

    However, our studies have shown that amoebae and giardia contain splicoeosomal introns, ras-family signal-transduction proteins, ATP-binding casettes (ABC)-family drug transporters, Golgi, and a mitochondrion-derived organelle (amoebae only). These results suggest that most of the organelles of higher eukaryotes ...

  20. Eliminatie van virussen, Cryptosporidium en Giardia door drinkwaterzuiveringsprocessen

    NARCIS (Netherlands)

    Medema GJ; Theunissen J; MGB

    1996-01-01

    A study on the removal efficiency of drinking water treatment processes for viruses and protozoa (Cryptosporidium/Giardia). The description is based on the best available Dutch and, if data on the Dutch situation are absent, international research data. The approach is valid for well-designed and

  1. (ELISA) kit for diagnosis copro-antigens of Giardia lamblia

    African Journals Online (AJOL)

    STORAGESEVER

    2010-08-02

    Aug 2, 2010 ... methods based on antigen scanning of parasites such as enzyme linked immunosorbent assay (ELISA), ... samples. To design this method, a pure antibody against parasite as well as an antibody conjugated to a ..... school children in Santiago, Chile by capture ELISA for the detection of fecal Giardia ...

  2. Molecular identification of Giardia lamblia isolates from adult human ...

    African Journals Online (AJOL)

    ... reflect environmental contamination of water resources. Therefore, it seems further studies are needed to clarify the route of infection in the study area. Keywords: Giardia lamblia, glutamate dehydrogenase (gdh), semi-nested polymerase chain reaction (PCR), PCR-RFLP, Ahvaz, Iran African Journal of Biotechnology Vol.

  3. What Entamoeba histolytica and Giardia lamblia tell us about the ...

    Indian Academy of Sciences (India)

    Unknown

    have a single diploid nucleus, while giardia have two similar nuclei. ... bacterial species rather than being reinvented by each bacterium as the need .... Structures of Jacob lectins and peritrophins. Examples of convergent evolution. Figure 4. Coincident evolution of phagocytosis. Phagocytosis of bacteria. Phagocytosis of.

  4. The search for Cryptosporidium oocysts and Giardia cysts in source ...

    African Journals Online (AJOL)

    driniev

    associated with feacal pollution does not necessarily indicate the absence of Cryptosporidium or Giardia. Being a common cause of waterborne disease, the requirement for detecting the presence of these organisms in environmental and treated water samples is growing. Although cysts and oocysts can be found in natural ...

  5. Occurrence of Cryptosporidium and Giardia in domestic animals in ...

    African Journals Online (AJOL)

    A single faecal sample per animal was collected from pigs, goats, dogs, ducks, chickens and pigeons and analysed by Merifluor Cryptosporidium/Giardia immunofluorescence antibody assay for the simultaneous detection of these parasites. The faecal consistency was noted and scored as non-diarrhoeic or diarrhoeic.

  6. Host parasite interactions and pathophysiology in Giardia infections.

    Science.gov (United States)

    Cotton, James A; Beatty, Jennifer K; Buret, Andre G

    2011-08-01

    Giardia is a protozoan parasite of the small intestine, and a leading cause of diarrhoeal disease worldwide in a variety of animals, including humans. The host-parasite interaction and pathophysiological processes of giardiasis remain incompletely understood. Current research suggests that Giardia-induced diarrhoeal disease is mediated by small intestinal malabsorption and maldigestion, chloride hypersecretion and increased rates of small intestinal transit. Small intestinal malabsorption and maldigestion results from the CD8+ lymphocyte-induced diffuse shortening of brush border microvilli. Activation of CD8+ lymphocytes occurs secondary to small intestinal barrier dysfunction, which results from heightened rates of enterocyte apoptosis and disruption of epithelial tight junctions. Both host and parasite factors contribute to the pathogenesis of giardiasis and ongoing research in this field may elucidate genotype/assemblage-specific pathogenic mechanisms. Giardia infections can result in chronic gastrointestinal disorders such as post-infectious Irritable Bowel Syndrome and symptoms may manifest at extra-intestinal sites, even though the parasite does not disseminate beyond the gastrointestinal tract. The infection can cause failure to thrive in children. Furthermore, there is now evidence suggesting that Giardia symptoms may vary between industrialised and developing areas of the world, for reasons that remain obscure. More research is needed to improve our understanding of this parasitic infection which was recently included in the World Health Organisation "Neglected Disease Initiative". Copyright © 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  7. Prevalence of Giardia intestinalis and Hymenolepis nana in Afghan ...

    African Journals Online (AJOL)

    Background: Present study aimed to investigate prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugees visiting Central Health Unit (CHU), Kot Chandana (Mianwali, Northern Punjab) during two years period (February 2007 to December 2009). Methods: A total of 687 stool samples were collected from ...

  8. Cryptosporidium and Giardia: new challenges to the water industry

    NARCIS (Netherlands)

    Medema, Gerriet Jan

    1999-01-01

    The protozoan parasites Cryptosporidium parvum and Giardia intestinalis have emerged as significant waterborne pathogens over the past decades. Many outbreaks of waterborne cryptosporidiosis and giardiasis have been recorded,primarily in the United States and the United Kingdom.Chapter 1 gives an

  9. First record of Giardia in cattle in Denmark

    DEFF Research Database (Denmark)

    Iburg, T.; Gasser, R.B.; Henriksen, S.A.

    1996-01-01

    Faecal samples from asymptomatic dairy cows and calves from a farm on the Island Falster, Denmark, were examined by a sucrose gradient flotation technique. Giardia cysts were found in 7.6% of the 92 samples, and estimated cyst excretion rates ranged from 50-200 cysts per gram faeces. Given that G...

  10. Giardia intestinalis incorporates heme into cytosolic cytochrome b₅.

    Science.gov (United States)

    Pyrih, Jan; Harant, Karel; Martincová, Eva; Sutak, Robert; Lesuisse, Emmanuel; Hrdý, Ivan; Tachezy, Jan

    2014-02-01

    The anaerobic intestinal pathogen Giardia intestinalis does not possess enzymes for heme synthesis, and it also lacks the typical set of hemoproteins that are involved in mitochondrial respiration and cellular oxygen stress management. Nevertheless, G. intestinalis may require heme for the function of particular hemoproteins, such as cytochrome b5 (cytb5). We have analyzed the sequences of eukaryotic cytb5 proteins and identified three distinct cytb5 groups: group I, which consists of C-tail membrane-anchored cytb5 proteins; group II, which includes soluble cytb5 proteins; and group III, which comprises the fungal cytb5 proteins. The majority of eukaryotes possess both group I and II cytb5 proteins, whereas three Giardia paralogs belong to group II. We have identified a fourth Giardia cytb5 paralog (gCYTb5-IV) that is rather divergent and possesses an unusual 134-residue N-terminal extension. Recombinant Giardia cytb5 proteins, including gCYTb5-IV, were expressed in Escherichia coli and exhibited characteristic UV-visible spectra that corresponded to heme-loaded cytb5 proteins. The expression of the recombinant gCYTb5-IV in G. intestinalis resulted in the increased import of extracellular heme and its incorporation into the protein, whereas this effect was not observed when gCYTb5-IV containing a mutated heme-binding site was expressed. The electrons for Giardia cytb5 proteins may be provided by the NADPH-dependent Tah18-like oxidoreductase GiOR-1. Therefore, GiOR-1 and cytb5 may constitute a novel redox system in G. intestinalis. To our knowledge, G. intestinalis is the first anaerobic eukaryote in which the presence of heme has been directly demonstrated.

  11. Risk factors for Giardia infection among hospitalized children in Cuba.

    Science.gov (United States)

    Bello, J; Núñez, F A; González, O M; Fernández, R; Almirall, P; Escobedo, A A

    2011-01-01

    The risk factors associated with Giardia infection, in children hospitalized in Havana, Cuba, were recently explored. Children aged ≥5 years were more likely to be positive for Giardia infection than the younger children, with an odds ratio (OR) of 3·41 [95% confidence interval (CI) = 1·36-9·69]. The risk factors found to be associated with Giardia infection in univariate analyses were rural residence (OR = 3·01; CI = 1·23-7·35), belonging to a household that did not receive water from an aqueduct (OR = 3·27; CI = 1·21-8·91), drinking unboiled water (OR = 3·64; CI = 2·14-6·26), nail biting (OR = 3·47; CI = 1·97-6·08), eating unwashed vegetables raw (OR = 4·84; CI = 2·33-10·14), and a personal (OR = 3·23; CI = 1·58-6·59) or family history (OR = 3·96; CI = 1·53-10·47) of previous parasitic infection. In multivariate analyses, however, only two (modifiable) risk factors were found to be independently and significantly associated with Giardia infection: nail biting and eating unwashed vegetables raw. It therefore seems that, at least at the individual level, giardiasis-prevention activities in Havana should be focussed on health education to improve personal hygiene and food-related practices. If appropriately managed, the surveillance of drinking water and foodstuffs, for Giardia and other parasites, might also help to reduce the hospitalization of Cuban children.

  12. Evaluation of a Combination Rapid Immunoassay for Detection of Giardia and Cryptosporidium Antigens

    OpenAIRE

    Chan, Raymond; Chen, Jing; York, Mary K.; Setijono, Norman; Kaplan, Raymond L.; Graham, Fitzroy; Tanowitz, Herbert B.

    2000-01-01

    A combination cassette format nonenzymatic rapid immunoassay for detection of Giardia and Cryptosporidium antigens was evaluated by using 556 patient stool specimens from three clinical laboratories. This assay (Genzyme Diagnostics Contrast Giardia/Cryptosporidium), which can be used with fresh or formalin-fixed specimens, had unadjusted sensitivities and specificities of 96.1 and 98.5% for Giardia and 100 and 98.7% for Cryptosporidium, respectively, in this study.

  13. A simple fibril and lectin model for cyst walls of Entamoeba and perhaps Giardia

    Science.gov (United States)

    Samuelson, John; Robbins, Phillips

    2010-01-01

    Cyst walls of Entamoeba and Giardia protect them from environmental insults, stomach acids, and intestinal proteases. Each cyst wall contains a sugar homopolymer: chitin in Entamoeba and a unique N-acetylgalactosamine (GalNAc) homopolymer in Giardia. Entamoeba cyst wall proteins include Jacob lectins (carbohydrate-binding proteins) that cross-link chitin, chitinases that degrade chitin, and Jessie lectins that make walls impermeable. Giardia cyst wall proteins are also lectins that bind fibrils of the GalNAc homopolymer. While many of the details remain to be determined for the Giardia cyst wall, current data suggests a relatively simple fibril and lectin model for the Entamoeba cyst wall. PMID:20934911

  14. Evaluation of ColorPAC Giardia/Cryptosporidium Rapid Assay and ProSpecT Giardia/Cryptosporidium Microplate Assay for Detection of Giardia and Cryptosporidium in Fecal Specimens

    OpenAIRE

    Katanik, M. T.; Schneider, S. K.; Rosenblatt, J. E.; Hall, G. S.; Procop, G. W.

    2001-01-01

    Detection of Giardia and Cryptosporidium in clinical stool specimens using the ColorPAC and ProSpecT enzyme immunoassays revealed 98.7% agreement for Giardia detection and 98.1% agreement for Cryptosporidium detection. Sensitivities were uniformly 100%. The specificities of the ColorPAC immunoassay for Giardia and Cryptosporidium detection were 100 and 99.5%, respectively, and those for the ProSpecT assay were 98.4 and 98.6%, respectively. The false-positive reactions with the ProSpecT assay ...

  15. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

    Directory of Open Access Journals (Sweden)

    Rogerio TIYO

    2015-08-01

    Full Text Available SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis, bacteriological (total and thermotolerants coliforms and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r, Barueri, SP, Brazil. The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(rkit. From 20 rural properties investigated, 10 had artesian wells (40 samples, 10 had common wells (40 samples, and one had a mine (four samples, the latter contaminated by Cryptosporidiumspp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardiaspp. or Cryptosporidiumspp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidiumspp. and Giardiaspp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

  16. Antigenic variation in the intestinal parasite Giardia lamblia.

    Science.gov (United States)

    Gargantini, Pablo Rubén; Serradell, Marianela Del Carmen; Ríos, Diego Nicolás; Tenaglia, Albano Heraldo; Luján, Hugo Daniel

    2016-08-01

    Giardia lamblia trophozoites undergo antigenic variation, where one member of the Variant-specific Surface Protein (VSP) family is expressed on the surface of proliferating trophozoites and periodically replaced by another one. Two main questions have challenged researchers since antigenic switching was discovered in Giardia: What are the mechanisms involved? How are they influenced by other cellular processes or by the environment? Two molecular mechanisms have been proposed, both involving small non-coding RNAs. Here we postulate that (a) chromatin remodeling, triggered by environmental factors, also plays an important role in selecting the VSP that will be expressed and (b) the particular VSP structure may not only protect the parasite in the small intestine but also signal the need to exchange the existing VSP for another. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Gene duplication in the genome of parasitic Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Flores Roberto

    2010-02-01

    Full Text Available Abstract Background Giardia are a group of widespread intestinal protozoan parasites in a number of vertebrates. Much evidence from G. lamblia indicated they might be the most primitive extant eukaryotes. When and how such a group of the earliest branching unicellular eukaryotes developed the ability to successfully parasitize the latest branching higher eukaryotes (vertebrates is an intriguing question. Gene duplication has long been thought to be the most common mechanism in the production of primary resources for the origin of evolutionary novelties. In order to parse the evolutionary trajectory of Giardia parasitic lifestyle, here we carried out a genome-wide analysis about gene duplication patterns in G. lamblia. Results Although genomic comparison showed that in G. lamblia the contents of many fundamental biologic pathways are simplified and the whole genome is very compact, in our study 40% of its genes were identified as duplicated genes. Evolutionary distance analyses of these duplicated genes indicated two rounds of large scale duplication events had occurred in G. lamblia genome. Functional annotation of them further showed that the majority of recent duplicated genes are VSPs (Variant-specific Surface Proteins, which are essential for the successful parasitic life of Giardia in hosts. Based on evolutionary comparison with their hosts, it was found that the rapid expansion of VSPs in G. lamblia is consistent with the evolutionary radiation of placental mammals. Conclusions Based on the genome-wide analysis of duplicated genes in G. lamblia, we found that gene duplication was essential for the origin and evolution of Giardia parasitic lifestyle. The recent expansion of VSPs uniquely occurring in G. lamblia is consistent with the increment of its hosts. Therefore we proposed a hypothesis that the increment of Giradia hosts might be the driving force for the rapid expansion of VSPs.

  18. Giardia lamblia infection after pancreas-kidney transplantation.

    Science.gov (United States)

    Kristensen, Ann Abkjaer; Horneland, Rune; Birn, Henrik; Svensson, My

    2016-01-18

    Infection is a common complication of solid organ transplantation. It is associated with an increased risk of acute cellular rejection and loss of graft function. The most common infections are due to bacteria and viruses, including transmission of cytomegalovirus from donor to recipient. In the past years, an increasing number of parasitic infections have been documented in transplant recipients. We describe the first reported case of intestinal Giardia lamblia transmission following simultaneous pancreas and kidney transplantation. 2016 BMJ Publishing Group Ltd.

  19. Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water.

    Science.gov (United States)

    Osaki, Silvia Cristina; Costa, Adriana Oliveira; Troiano, Ludmilla Della Coletta; Kruger, Ernesto Renato; Pereira, Juliana Tracz; Fernandes, Nelson Luis Mello; Silva, Márcia Benedita de Oliveira; Soccol, Vanete Thomaz

    2011-10-01

    The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

  20. Purified azure B as a reticulocyte stain.

    Science.gov (United States)

    Marshall, P N; Bentley, S A; Lewis, S M

    1976-01-01

    A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two dyes, varying amounts of an extraneous, particulate dye deposit were present in these preparations, making accuracte counting both tedious and timeconsuming. Purified azure B, on the other hand, gave reproducibly stained, deposit-free preparations. Reticulocyte counts obtained from azure B preparations correlated almost exactly with those determined using new methylene blue. Purified azure B is therefore recommended as a convenient reticulocyte stain for routine use. Images PMID:64475

  1. Phase change liquid purifier and pump

    Science.gov (United States)

    Steinhour, Leif Alexi

    2017-05-23

    Systems, methods, and apparatus are provided for purifying and pumping liquids, and more particularly, for purifying and pumping water. The apparatus includes a chamber including a top portion and a bottom portion. A surface configured to be heated is proximate the bottom portion of the chamber. A baffle is disposed within the chamber and above the surface. The baffle is disposed at an angle relative to a vertical direction. The chamber further includes an inlet and a first outlet. The surface heats a liquid in the chamber, causing the liquid to boil. In operation, bubbles rise from the surface and are forced in a horizontal direction by the baffle disposed in the chamber.

  2. Method for purifying bidentate organophosphorous compounds

    International Nuclear Information System (INIS)

    Krupa, J.F.; Mcisaac, L.D.; Schroeder, N.C.

    1981-01-01

    Bidentate organophosphorous compounds are purified of undesirable impurities by contacting a solution of the compounds with a mercuric nitrate solution to form an insoluble mercuric bidentate compound which precipitates while the impurities remain in solution. The precipitate is washed and then contacted with a mixture of an aqueous solution of a strong mercuric ion complexing agent and an organic solvent to complex the mercuric ion away from the bidentate compound which then dissolves in the solvent. The purified bidentate compounds are useful for extracting the actinide elements from aqueous acidic nuclear waste solutions

  3. Studies on the prevalence of giardia cysts and cryptosporidium oocysts in South-African water

    CSIR Research Space (South Africa)

    Kfir, R

    1995-01-01

    Full Text Available per sample and the number of positive samples. Almost 50% of sewage samples studied contained Giardia cysts and 30% contained both Giardia cysts and Cryptosporium oocysts. Treatment of sewage resulted in a reduction in the percentage of samples...

  4. Cryptosporidium oocysts and giardia cysts on salad products irrigated with contaminated water.

    Science.gov (United States)

    Amorós, Inmaculada; Alonso, José L; Cuesta, Gonzalo

    2010-06-01

    A field study in Valencia, Spain, was done to determine the occurrence of Giardia and Cryptosporidium on salad products that are frequently eaten raw, such as lettuces and Chinese cabbage, and in irrigation waters. Four water samples were taken weekly 1 month before harvesting the vegetables. All water samples were analyzed using techniques included in the U.S. Environmental Protection Agency Method 1623. Standard methods for detecting protozoan parasites on salad vegetables are not available. Published techniques for the isolation of parasites from vegetables generally have low and variable recovery efficiencies. In this study, vegetables were analyzed using a recently reported method for detection of Cryptosporidium oocysts and Giardia cysts on salad products. The waters tested were positive for both Cryptosporidium and Giardia. Of 19 salad products studied, we observed Cryptosporidium in 12 samples and Giardia in 10 samples. Recoveries of the Texas Red-stained Cryptosporidium and Giardia, which were used as internal controls, were 24.5% +/- 3.5% for Cryptosporidium and 16.7% +/- 8.1% for Giardia (n = 8). This study provides data on the occurrence of Cryptosporidium and Giardia in salad products in Spain. The method was useful in the detection of Cryptosporidium oocysts and Giardia cysts on the vegetables tested, and it provides a useful analytical tool for occurrence monitoring.

  5. Drug-Free Approach To Study the Unusual Cell Cycle of Giardia intestinalis

    Science.gov (United States)

    Horlock-Roberts, Kathleen; Reaume, Chase; Dayer, Guillem; Ouellet, Christine; Cook, Nicholas

    2017-01-01

    ABSTRACT Giardia intestinalis is a protozoan parasite that causes giardiasis, a form of severe and infectious diarrhea. Despite the importance of the cell cycle in the control of proliferation and differentiation during a giardia infection, it has been difficult to study this process due to the absence of a synchronization procedure that would not induce cellular damage resulting in artifacts. We utilized counterflow centrifugal elutriation (CCE), a size-based separation technique, to successfully obtain fractions of giardia cultures enriched in G1, S, and G2. Unlike drug-induced synchronization of giardia cultures, CCE did not induce double-stranded DNA damage or endoreplication. We observed increases in the appearance and size of the median body in the cells from elutriation fractions corresponding to the progression of the cell cycle from early G1 to late G2. Consequently, CCE could be used to examine the dynamics of the median body and other structures and organelles in the giardia cell cycle. For the cell cycle gene expression studies, the actin-related gene was identified by the program geNorm as the most suitable normalizer for reverse transcription-quantitative PCR (RT-qPCR) analysis of the CCE samples. Ten of 11 suspected cell cycle-regulated genes in the CCE fractions have expression profiles in giardia that resemble those of higher eukaryotes. However, the RNA levels of these genes during the cell cycle differ less than 4-fold to 5-fold, which might indicate that large changes in gene expression are not required by giardia to regulate the cell cycle. IMPORTANCE Giardias are among the most commonly reported intestinal protozoa in the world, with infections seen in humans and over 40 species of animals. The life cycle of giardia alternates between the motile trophozoite and the infectious cyst. The regulation of the cell cycle controls the proliferation of giardia trophozoites during an active infection and contains the restriction point for the

  6. Occurrence of Cryptosporidium oocysts and Giardia cysts in water supplies of San Pedro Sula, Honduras

    Directory of Open Access Journals (Sweden)

    Solo-Gabriele Helena María

    1998-01-01

    Full Text Available During June 1996, water supplies of the city of San Pedro Sula, Honduras, were sampled to obtain an assessment of Cryptosporidium oocyst and Giardia cyst concentrations. Each sample was concentrated and stained with an indirect immunofluorescent antibody, and parasites were counted through microscopic analysis. In three surface water supplies, Cryptosporidium oocyst concentrations ranged from 58 to 260 oocysts per 100 L, and Giardia cysts were present in concentrations ranging from 380 to 2100 cysts per 100 L. Unlike the surface water samples, groundwater had a higher concentration of Cryptosporidium oocysts (26/100 L than Giardia cysts (6/100 L, suggesting that the groundwater aquifer protects the water supply more effectively from larger Giardia cysts. Cryptosporidium oocyst concentrations are within the typical range for surface water supplies in North America whereas Giardia cyst concentrations are elevated. Efforts should be made to protect raw water from sources of contamination.

  7. Home Air Purifiers Eradicate Harmful Pathogens

    Science.gov (United States)

    2014-01-01

    Marshall Space Flight Center funded the University of Madison-Wisconsin to develop ethylene scrubbers to keep produce fresh in space. Akida Holdings of Jacksonville, Florida, licensed the technology and developed Airocide, an air purifier that can kill airborne pathogens. Previously designed for industrial spaces, there is now a specially designed unit for home use.

  8. Antimicrobial action of purified raspberry flavonoid

    African Journals Online (AJOL)

    user

    2012-02-07

    Feb 7, 2012 ... In this paper, the bacteriostatic action of purified raspberry flavonoid was explored. A study was conducted on the effect of raspberry flavonoid on Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Saccharomyces cerevisiae, Rhizopus, Mucor and Penicillium. Raspberry flavonoid had the best.

  9. Prevalence of Cryptosporidium and Giardia in the water resources of the Kuang River catchment, Northern Thailand.

    Science.gov (United States)

    Chuah, C Joon; Mukhaidin, Nabila; Choy, Seow Huey; Smith, Gavin J D; Mendenhall, Ian H; Lim, Yvonne A L; Ziegler, Alan D

    2016-08-15

    A catchment-scale investigation of the prevalence of Cryptosporidium and Giardia in the Kuang River Basin was carried out during the dry and rainy seasons. Water samples were collected from the Kuang River and its tributaries as well as a major irrigation canal at the study site. We also investigated the prevalence of gastrointestinal parasitic infection among dairy and beef cattle hosts. Cryptosporidium and/or Giardia were detected in all the rivers considered for this study, reflecting their ubiquity within the Kuang River Basin. The high prevalence of Cryptosporidium/Giardia in the upper Kuang River and Lai River is of a particular concern as both drain into the Mae Kuang Reservoir, a vital source of drinking-water to many local towns and villages at the research area. We did not, however, detected neither Cryptosporidium nor Giardia were in the irrigation canal. The frequency of Cryptosporidium/Giardia detection nearly doubled during the rainy season compared to the dry season, highlighting the importance of water as an agent of transport. In addition to the overland transport of these protozoa from their land sources (e.g. cattle manure, cess pits), Cryptosporidium/Giardia may also be re-suspended from the streambeds (a potentially important repository) into the water column of rivers during storm events. Faecal samples from dairy and beef cattle showed high infection rates from various intestinal parasites - 97% and 94%, respectively. However, Cryptosporidium and Giardia were only detected in beef cattle. The difference in management style between beef (freeranging) and dairy cattle (confined) may account for this disparity. Finally, phylogenetic analyses revealed that the Cryptosporidium/Giardia-positive samples contained C. ryanae (non-zoonotic) as well as Giardia intestinalis assemblages B (zoonotic) and E (non-zoonotic). With only basic water treatment facilities afforded to them, the communities of the rural area relying on these water supplies are

  10. Therapeutic enhancement of newly derived bacteriocins against Giardia lamblia.

    Science.gov (United States)

    Amer, Eglal I; Mossallam, Shereen F; Mahrous, Hoda

    2014-11-01

    Trials for identifying efficient anti-giardial agents are still ongoing. Nowadays, bacteriocins have attracted the attention as potential antimicrobial compounds. For the first time, the current study evaluated the therapeutic efficacy of bacteriocins derived from newly isolated Egyptian strains of probiotics Lactobacilli; L. acidophilus (P106) and L. plantarum (P164) against Giardia lamblia. Bacteriocins' efficacy was evaluated both in vitro; by growth inhibition and adherence assays, and in vivo; through estimation of parasite density, intestinal histopathological examination and ultrastructural analysis of Giardia trophozoites. In vivo bacteriocins' clinical safety was assessed. In vitro results proved that 50 µg of L. acidophilus bacteriocin induced reduction of the mean Giardia lamblia trophozoites by 58.3 ± 4.04%, while at lower concentrations of 10 and 20 µg of both L. acidophilus and L. plantarum, non significant reduction of the mean parasite density was achieved. In vitro trophozoites adherence was susceptible to the tested bacteriocins at all studied concentrations with variable degrees, while the highest adherence reduction was demonstrated using 50 µg of L acidophilus bacteriocin. In vivo, oral inoculation of 50 µg/mouse L. acidophilus bacteriocin for 5 successive days resulted in a noteworthy decline of the intestinal parasite density, along with amelioration of intestinal pathology of infected mice. Ultrastructural examination proved thatfive doses of L. acidophilus bacteriocin showed marked changes in cellular architecture of the trophozoites with evident disorganization of the cell membrane, adhesive disc and cytoplasmic components. This is the first reported study of the safe anti-giardial efficacy of L. acidophilus (P106) derived bacteriocin, hence highlighting its great promise as a potential therapeutic safe alternative to existing commercial drugs. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Incidence of Giardia lamblia Subspecies by PCR-RFLP in Stool Specimens of Hospitalized Children at Urmia Mutahhari Hospital, West Azerbaijan Province, Iran.

    Directory of Open Access Journals (Sweden)

    Khosro Hazrati Tappeh

    2014-12-01

    Full Text Available Giardia lamblia is one of the most prevalent intestinal flagellate protozoa that infects a wide range of vertebrate hosts causing severe intestinal disorder in children.This study was performed to determine subspecies of G.lamblia by the PCR-RFLP method, targeting the glutamate dehydrogenase(gdhlocus, in hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province,Iran and determining the infection transformational storages in this area.Overall, 720 stool specimens were collected from the hospitalized children, 34 samples were positive and Giardia cysts were detected under the microscope. Cysts were partially purified by the sucrose density gradient method and then washed with sterile distilled water to remove effectively the PCR inhibitors. Genomic DNA of G. lamblia isolates was extracted by freeze-thaw cycles followed by phenol/ chloroform/isoamyl alcohol method. The single step PCR-RFLP assay was used to differentiate the assemblages between A and B, which were found in humans. In this method, 432 bp expected size was amplified, and then for detection of subspecies, specific restriction RsaI and BspLI enzymes were used.Totally 34 samples were positive in terms of Giardia cyst out of 720 examined samples microscopically, so the parasite spread rate is reported 4.72%. Analysis PCR-RFLP on these samples revealed that 28 samples (93.3% have the genotype BIII and 2 samples (6.7% belong to the subgroup BIV.PCR-RFLP is a proper analytical method for determining the genotype among parasite types, using the glutamate dehydrogenizes zone's genes. Based on the results, an animal origin of infection cycle is suggested.

  12. Echinococcus, Giardia and Cryptosporidium: observational studies challenging accepted dogma.

    Science.gov (United States)

    Thompson, R C A

    2009-10-01

    The development of in vitro culture systems that allow the maintenance, and support the development of Echinococcus, Giardia and Cryptosporidium in the laboratory have had a significant impact on their biology and taxonomy and the epidemiology of infections they cause. This short retrospective review demonstrates how radical shifts in our understanding have occurred as a result of being able to grow these organisms in culture, and how molecular tools have helped in the interpretation of such research that often reflects the observations of earlier workers.

  13. Unusual ribosomal RNA of the intestinal parasite Giardia lamblia.

    OpenAIRE

    Edlind, T D; Chakraborty, P R

    1987-01-01

    The anaerobic protozoan Giardia lamblia is a common intestinal parasite in humans, but is poorly defined at molecular and phylogenetic levels. We report here a structural characterization of the ribosomal RNA (rRNA) and rRNA genes of G. lamblia. Gel electrophoresis under native or non-denaturing conditions identified two high molecular weight rRNA species corresponding to the 16-18S and 23-28S rRNAs. Surprisingly, both species (1300 and 2300 nucleotides long, respectively) were considerably s...

  14. Flagellar generated flow mediates attachment of Giardia Lamblia

    Science.gov (United States)

    Picou, Theodore; Polackwich, Jamie; Burrola Gabilondo, Beatriz; McAllister, Ryan; Powers, Tom; Elmendorf, Heidi; Urbach, Jeff

    2011-11-01

    Giardia lamblia is a protozoan parasite responsible for widespread diarrheal disease in humans and animals worldwide. Attachment to the host intestinal mucosa and resistance to peristalsis is necessary for establishing infection, but the physical basis for this attachment is poorly understood. We report results from confocal fluorescence microscopy that demonstrate that the regular beating of the posterior flagella generate a flow through the ventral disk, a suction-cup shaped structure that is against the substrate during attachment. Finite element simulations show that the negative pressure generated by the flow is consistent with the measured force of attachement between the parasite and its substrate.

  15. Cryptosporidium and Giardia in Swimming Pools, Atlanta, Georgia

    Centers for Disease Control (CDC) Podcasts

    2008-05-29

    In this podcast, Dan Rutz speaks with Dr. Joan Shields, a guest researcher with the Healthy Swimming Program at CDC, about an article in June 2008 issue of Emerging Infectious Diseases reporting on the results of a test of swimming pools in the greater Atlanta, Georgia area. Dr. Shields tested 160 pools in metro Atlanta last year for Cryptosporidium and Giardia. These germs cause most recreational water associated outbreaks.  Created: 5/29/2008 by Emerging Infectious Diseases.   Date Released: 5/29/2008.

  16. Endurance Pump Test with MIL-PRF-83282 Hydraulic Fluid, Purified with Malabar Purifier

    National Research Council Canada - National Science Library

    Sharma, Shashi

    2004-01-01

    The possible deterioration of the fluid during use or the potential for the purifier to remove some of the performance improving additives from the fluid as well as the contaminants was investigated...

  17. Steroidogenesis in amlodipine treated purified Leydig cells

    Energy Technology Data Exchange (ETDEWEB)

    Latif, Rabia, E-mail: rabialatif08@hotmail.com [Department of Physiology, Army Medical College, National University of Sciences and Technology, Islamabad (Pakistan); Lodhi, Ghulam Mustafa, E-mail: drmustafa786@gmail.com [Department of Physiology, Wah Medical College, Wah (Pakistan); Hameed, Waqas, E-mail: waqham@hotmail.com [Department of Physiology, Rehman Medical College, Peshawar (Pakistan); Aslam, Muhammad, E-mail: professormaslam@yahoo.com [Department of Physiology, Shifa College of Medicine, Islamabad (Pakistan)

    2012-01-01

    Drugs have been shown to adversely affect male fertility and recently anti-hypertensive drugs were added to the list. The anti-fertility effects of amlodipine, a calcium channel blocker, are well-illustrated in in vivo experiments but lack an in vitro proof. The present study was designed to experimentally elucidate the effects of amlodipine on Leydig cell steroidogenesis and intracellular calcium in vitro. Leydig cells of Sprague–Dawley rats were isolated and purified by Percoll. Cells were incubated for 3 h with/without amlodipine in the presence/absence of LH, dbcAMP, Pregnenolone and 25-Hydroxycholesterol. Cytosolic calcium was measured in purified Leydig cells by fluorometric technique. The results showed significantly reduced (P < 0.05) steroidogenesis and intracellular calcium in amlodipine exposed rats. The site of amlodipine induced steroidogenic inhibition seems to be prior to the formation of Pregnenolone at the level of StAR protein. -- Highlights: ► Inhibition of steroidogenesis in isolated and purified Leydig cells by amlodipine. ► Site of inhibition was before Pregnenolone formation, at the level of StAR protein. ► Inhibition of LH stimulated rise in cytosolic calcium by amlodipine.

  18. Urban park-related risks for Giardia spp. infection in dogs.

    Science.gov (United States)

    Smith, A F; Rock, M; Neumann, N; Massolo, A

    2015-11-01

    Giardia spp. is a common gastrointestinal (GI) parasite of multiple host species, including dogs and humans, with the potential for zoonotic transmission. The risk of GI parasitism in dogs (including Giardia spp.) may increase with park use in urban areas. This study aimed to (1) determine whether park attendance is a risk factor for Giardia spp. infection in dogs and (2) characterize the behavioural and demographic risk factors for Giardia spp. infection in park-attending and non-park-attending dogs. From August to September 2012, a total of 1293 dog owners completed a survey and 860 corresponding dog faecal samples were collected. Dog faeces were screened for Giardia spp. using a direct immunofluorescence assay and associations assessed among behaviours, demographics, and Giardia spp. infection. Main results included off-leash and swimming frequencies within parks as significantly positively associated with Giardia spp. infection in dogs. Dog-owner age was negatively associated with off-leash and swimming frequencies in parks. The results suggest some recreational behaviours in parks and certain demographics are risk factors for parasitism in pet dogs.

  19. Giardia: a pathogen or commensal for children in high-prevalence settings?

    Science.gov (United States)

    Bartelt, Luther A; Platts-Mills, James A

    2016-10-01

    Giardia is a common intestinal parasite worldwide, and infection can be associated with clear and sometimes persistent symptomatology. However, in children in high-prevalence settings, it is not associated with or is perhaps even protective against acute diarrhea, and the association with long-term outcomes has been difficult to discern. Recent studies have made progress in helping us disentangle this apparent paradox. First, prospective, well-characterized cohort studies have added to the data on the association between Giardia and diarrhea in these settings and have further characterized associations between Giardia infection and nutrition, gut function, and growth. Second, animal models have further characterized the host response to Giardia and helped elucidate mechanisms by which Giardia could impair child development. Finally, new work has shed light on the heterogeneity of human Giardia strains, which may both explain discrepant findings in the literature and help guide higher-resolution analyses of this pathogen in the future. The true clinical impact of endemic pediatric giardiasis remains unclear, but recent prospective studies have confirmed a high prevalence of persistent, subclinical Giardia infections and associated growth shortfalls. Integrating how nutritional, microbial, metabolic, and pathogen-strain variables influence these outcomes could sharpen delineations between pathogenic and potentially beneficial attributes of this enigmatic parasite.

  20. Lactobacillus rhamnosus GG antagonizes Giardia intestinalis induced oxidative stress and intestinal disaccharidases: an experimental study.

    Science.gov (United States)

    Goyal, Nisha; Rishi, Praveen; Shukla, Geeta

    2013-06-01

    The present study describes the in vivo modulatory potential of Lactobacillus rhamnosus GG (LGG), an effective probiotic, in Giardia intestinalis-infected BALB/c mice. Experimentally, it was observed that oral administration of lactobacilli prior or simultaneous with Giardia trophozoites to mice, efficiently (p Giardia-infected mice, showed a significant increase in the levels of antioxidants [reduced glutathione (GSH) and superoxide dismutase (SOD)] and intestinal disaccharidases [sucrase and lactase] and decreased levels of oxidants in the small intestine, in comparison with Giardia-infected mice. Histopathological findings also revealed almost normal cellular morphology of the small intestine in probiotic-fed Giardia-infected mice compared with fused enterocytes, villous atrophy and increased infiltration of lymphocytes in Giardia-infected mice. The results of the present study has shed new light on the anti-oxidative properties of LGG in Giardia mediated tissue injury, thereby suggesting that the effects of probiotic LGG are biologically plausible and could be used as an alternative microbial interference therapy.

  1. Giardia: a pathogen or commensal for children in high prevalence settings?

    Science.gov (United States)

    Bartelt, Luther A.

    2016-01-01

    Purpose of review Giardia is a common intestinal parasite worldwide, and infection can be associated with clear and sometimes persistent symptomatology. However, in children in high prevalence settings, it is not associated with or is perhaps even protective against acute diarrhea, and the association with long-term outcomes has been difficult to discern. Recent findings Recent studies have made progress in helping us disentangle this apparent paradox. First, prospective, well-characterized cohort studies have added to the data on the association between Giardia and diarrhea in these settings and have further characterized associations between Giardia infection and nutrition, gut function, and growth. Second, animal models have further characterized the host response to Giardia and helped elucidate mechanisms by which Giardia could impair child development. Finally, new work has shed light on the heterogeneity of human Giardia strains, which may both explain discrepant findings in the literature and help guide higher-resolution analyses of this pathogen in the future. Summary The true clinical impact of endemic pediatric giardiasis remains unclear, but recent prospective studies have confirmed a high prevalence of persistent, subclinical Giardia infections and associated growth shortfalls. Integrating how nutritional, microbial, metabolic, and pathogen-strain variables influence these outcomes could sharpen delineations between pathogenic and potentially beneficial attributes of this enigmatic parasite. PMID:27479025

  2. Genome analysis and comparative genomics of a Giardia intestinalis assemblage E isolate

    Directory of Open Access Journals (Sweden)

    Andersson Jan O

    2010-10-01

    Full Text Available Abstract Background Giardia intestinalis is a protozoan parasite that causes diarrhea in a wide range of mammalian species. To further understand the genetic diversity between the Giardia intestinalis species, we have performed genome sequencing and analysis of a wild-type Giardia intestinalis sample from the assemblage E group, isolated from a pig. Results We identified 5012 protein coding genes, the majority of which are conserved compared to the previously sequenced genomes of the WB and GS strains in terms of microsynteny and sequence identity. Despite this, there is an unexpectedly large number of chromosomal rearrangements and several smaller structural changes that are present in all chromosomes. Novel members of the VSP, NEK Kinase and HCMP gene families were identified, which may reveal possible mechanisms for host specificity and new avenues for antigenic variation. We used comparative genomics of the three diverse Giardia intestinalis isolates P15, GS and WB to define a core proteome for this species complex and to identify lineage-specific genes. Extensive analyses of polymorphisms in the core proteome of Giardia revealed differential rates of divergence among cellular processes. Conclusions Our results indicate that despite a well conserved core of genes there is significant genome variation between Giardia isolates, both in terms of gene content, gene polymorphisms, structural chromosomal variations and surface molecule repertoires. This study improves the annotation of the Giardia genomes and enables the identification of functionally important variation.

  3. A study of two U.S. Army installation drinking water sources and treatment systems for the removal of Giardia and Cryptosporidium

    Energy Technology Data Exchange (ETDEWEB)

    Kelley, M.B. [United States Military Academy, West Point, NY (United States); Brokaw, J.K. [Rensselaer Polytechnic Institute, Troy, NY (United States); Brokaw, J.K. [Army Center for Health Promotion and Preventive Medicine, Aberdeen Proving Ground, MD (United States); Warrier, P.K. [Rensselaer Polytechnic Institute, Troy, NY (United States)] [and others

    1996-11-01

    This paper provides the results of a study of two U.S. Army installation drinking water sources and treatment systems for the removal of Giardia and Cryptosporidium. Sampling was conducted monthly for one year commencing in late March 1994 and concluding at the end of February 1995. Results of this detailed study include examination of turbidity, particle counts, and total and fecal coliforms as well as the enumeration of Giardia cysts and Cryptosporidium oocysts. Our goal, in addition to determining the levels of these pathogens in both raw and product waters, was to determine if typical operating parameters would be helpful in identifying either elevated raw water protozoa or breakthrough of either pathogen in the product water from the treatment facilities. A data summary for the results of the protozoa enumeration is a Table 1. Our results indicate frequent contamination of the raw waters at both sites by either or both pathogens. Further, we observed sporadic breakthrough of low levels of Cryptosporidium in the filtered waters of both sites. The method employed to concentrate, purify and enumerate the pathogenic protozoa is also discussed and comparisons are made to the proposed Information Collection Rule (ICR) method for detection of these microorganisms.

  4. Human Memory CD4+ T Cell Immune Responses against Giardia lamblia

    Science.gov (United States)

    Sørnes, Steinar; Peirasmaki, Dimitra; Svärd, Staffan; Langeland, Nina

    2015-01-01

    The intestinal protozoan parasite Giardia lamblia may cause severe prolonged diarrheal disease or pass unnoticed as an asymptomatic infection. T cells seem to play an important role in the immune response to Giardia infection, and memory responses may last years. Recently, TH17 responses have been found in three animal studies of Giardia infection. The aim of this study was to characterize the human CD4+ T cell responses to Giardia. Peripheral blood mononuclear cells (PBMCs) were obtained from 21 returning travelers with recent or ongoing giardiasis and 12 low-risk healthy controls and stimulated in vitro with Giardia lamblia proteins. Production of tumor necrosis factor alpha (TNF-α), gamma interferon, interleukin-17A (IL-17A), IL-10, and IL-4 was measured in CD4+ effector memory (EM) T cells after 24 h by flow cytometry. After 6 days of culture, activation and proliferation were measured by flow cytometry, while an array of inflammatory cytokine levels in supernatants were measured with multiplex assays. We found the number of IL-17A-producing CD4+ EM T cells, as well as that of cells simultaneously producing both IL-17A and TNF-α, to be significantly elevated in the Giardia-exposed individuals after 24 h of antigen stimulation. In supernatants of PBMCs stimulated with Giardia antigens for 6 days, we found inflammation-associated cytokines, including 1L-17A, as well as CD4+ T cell activation and proliferation, to be significantly elevated in the Giardia-exposed individuals. We conclude that symptomatic Giardia infection in humans induces a CD4+ EM T cell response of which IL-17A production seems to be an important component. PMID:26376930

  5. Genotyping of Giardia lamblia and Entamoeba spp from river waters in Iran.

    Science.gov (United States)

    Mahmoudi, Mohammad Reza; Nazemalhosseini-Mojarad, Ehsan; Karanis, Panagiotis

    2015-12-01

    In this study, DNA from 55 surface and river water samples, which were collected from some water sources of Tehran and the Guilan Province, Iran, were extracted and examined for Entamoeba spp. and Giardia lamblia by PCR and genotyping. Twenty-seven samples, which were concentrated using the immunomagnetic separation technology (IMS) method, were examined for Giardia alone. Twenty-eight samples, which were concentrated using the sucrose flotation (SF) method, were examined for both Giardia and Entamoeba species. The results showed that 27/55 (17/27 and 10/28) (49 %), 4 /28 (14.28 %) and 3/28 (10.7 %) of the samples were positive for Giardia lamblia, Entamoeba spp and mixed infections (Entamoeba spp. and Giardia spp.), respectively. Sixteen out of 55 samples were negative. Entamoeba genus-specific PCR primers in single-round PCR were used to differentiate between the Entamoeba spp. (E. histolytica, E. dispar and E. moshkovskii). With respect to the 7 samples that were positive for Entamoeba, (14.28 %) 4 out of 28 were positive for E. moshkovskii, (7.14 %), 2 out of 28 were positive for E. histolytica and (3.57 %) 1 out of 28 was positive for E. dispar. Genus-specific PCR primers in a semi-nested PCR assay was performed to genotype Giardia species. Of the 27 samples that were positive for Giardia, 10 samples were sequences. All 10 successfully sequenced samples contained assemblage B of Giardia lamblia.This is first study to investigate the G. lamblia genotypes in the water supply of the Tehran and Guilan provinces, and it is the first study to investigate Entamoeba species in the water supplies of Iran. The investigated river water supplies, which are used for agriculture, camping and animal farming, were heavily contaminated by the human pathogenic Entamoeba and Giardia parasites. There is a potential risk of waterborne outbreaks in humans and animals.

  6. Existence of Cryptosporidium spp. oocysts and Giardia spp. cysts in well water in Nineveh governorate

    OpenAIRE

    R. G. Mohammad

    2011-01-01

    The study included examination of 110 water samples from well distributed in Mosul city and few towns and villages around it from May 2009 to March 2010 for detection of Cryptosporidium spp oocysts and Giardia spp cysts in well water. The results revealed that the prevalence of Cryptosporidium oocysts was 16.36% while the prevalence of Giardia cysts was 12.72%. The percentages of prevalence with Cryptosporidium and Giardia were in high rate in Bartilla and some villages around it 20% for Cryp...

  7. Detection of Cryptosporidium and Giardia doudenalis in equines in Nineveh, Iraq

    Directory of Open Access Journals (Sweden)

    E. T. Butty

    2011-01-01

    Full Text Available A total of 107 fecal samples of equines from different regions in Nineveh, were colleted from January 2007 till December 2007 and examined for Cryptosporidium sp., and Giardia doudenalis by using different methods (wet mount, flotation, lugol's iodine, modified Ziehl Nelsecn (hot and Giemsa stain Just for Giardia doudenalis. The animal age examined ranged from 4 to 10 years. The total prevalence of cryptosporidium sp. was 27.10% (29 out of 107, while the prevalence of Giardia doudenalis was 19.63% (21 out of 107. This study represents the first trial to explore cryptosporidiosis, giardiasis in equines as in Nineveh there is no survey of these intestinal protozoa.

  8. Isolation and enumeration of Giardia cysts, cryptosporidium oocysts, and Ascaris eggs from fruits and vegetables.

    Science.gov (United States)

    Robertson, L J; Gjerde, B

    2000-06-01

    Published techniques for recovering parasites from fruit and vegetables are generally inadequate, with low and variable recovery efficiencies. Here we describe an improved methodology for analyzing fruit and vegetables for Giardia cysts, Cryptosporidium oocysts, and Ascaris eggs. The method includes washing procedures, sonication, and, for Giardia and Cryptosporidium, immunomagnetic separation. Identification is by immunofluorescence (Giardia and Cryptosporidium) or brightfield microscopy (Ascaris). Recovery efficiencies from lettuce, Chinese leaves, and strawberries were found to be approximately 67% for Giardia, 42% for Cryptosporidium, and 72% for Ascaris. Recovery efficiencies from bean sprouts tended to be more variable and lower. This could be due to material removed with the parasites during the washing procedures, which, in turn, appeared related to the age of the bean sprouts. It is therefore recommended that fruit and vegetables should be as fresh as possible when analyzed for parasites.

  9. [A medium available in Czechoslovakia for the culture of Entamoeba histolytica and Giardia intestinalis].

    Science.gov (United States)

    Cerva, L

    1989-07-01

    From nutritive media produced in the CSSR a modification of Diamond's medium TYI-S-33 was prepared which ensures the long-term axenic cultivation of Entamoeba histolytica and Giardia intestinalis. Details of the cultivation technique are also presented.

  10. Thermogelling properties of purified poloxamer 407

    Directory of Open Access Journals (Sweden)

    Amir Fakhari

    2017-08-01

    Full Text Available Poloxamers are triblock copolymers with a center block of hydrophobic polypropylene oxide (PPO flanked by two hydrophilic polyethyleneoxide (PEO blocks. Among this family of copolymers, poloxamer 407 is a non-ionic surfactant with reversible gelation properties above a particular polymer concentration and a particular temperature. Easy preparation of poloxamer 407 based sterile injectable formulations have made this copolymer a good candidate for drug delivery, specifically when controlled release of the drug is required. Previously, the applications of compendial poloxamer 407 preparations were demonstrated; however, low viscosity, poor elasticity, and sol-to-gel transition temperature (Tsol-gel over a wide temperature range were observed. A purification process was introduced to eliminate impurities and low molecular weight copolymer molecules from the compendial poloxamer 407 resulting in higher viscosity values with Tsol-gel in a narrow temperature range. Here, poloxamer 407 was purified based on the proposed process and the rheological and analytical evaluation of the purified poloxamer 407 was conducted and compared to unpurified, compendial poloxamer 407. Then, the impact of poloxamer 407 concentration on gel formation was evaluated. For drug delivery applications, the effect of relevant buffer salts and the effect of addition of ethanol to the poloxamer 407 solutions were rheologically evaluated.

  11. Giardia Colonizes and Encysts in High-Density Foci in the Murine Small Intestine

    Science.gov (United States)

    Barash, N. R.; Nosala, C.; Pham, J. K.; McInally, S. G.; Gourguechon, S.; McCarthy-Sinclair, B.

    2017-01-01

    ABSTRACT Giardia lamblia is a highly prevalent yet understudied protistan parasite causing significant diarrheal disease worldwide. Hosts ingest Giardia cysts from contaminated sources. In the gastrointestinal tract, cysts excyst to become motile trophozoites, colonizing and attaching to the gut epithelium. Trophozoites later differentiate into infectious cysts that are excreted and contaminate the environment. Due to the limited accessibility of the gut, the temporospatial dynamics of giardiasis in the host are largely inferred from laboratory culture and thus may not mirror Giardia physiology in the host. Here, we have developed bioluminescent imaging (BLI) to directly interrogate and quantify the in vivo temporospatial dynamics of Giardia infection, thereby providing an improved murine model to evaluate anti-Giardia drugs. Using BLI, we determined that parasites primarily colonize the proximal small intestine nonuniformly in high-density foci. By imaging encystation-specific bioreporters, we show that encystation initiates shortly after inoculation and continues throughout the duration of infection. Encystation also initiates in high-density foci in the proximal small intestine, and high density contributes to the initiation of encystation in laboratory culture. We suggest that these high-density in vivo foci of colonizing and encysting Giardia likely result in localized disruption to the epithelium. This more accurate visualization of giardiasis redefines the dynamics of the in vivo Giardia life cycle, paving the way for future mechanistic studies of density-dependent parasitic processes in the host. IMPORTANCE Giardia is a single-celled parasite causing significant diarrheal disease in several hundred million people worldwide. Due to limited access to the site of infection in the gastrointestinal tract, our understanding of the dynamics of Giardia infections in the host has remained limited and largely inferred from laboratory culture. To better understand

  12. Evaluation of Three Commercial Assays for Detection of Giardia and Cryptosporidium Organisms in Fecal Specimens

    OpenAIRE

    Johnston, Stephanie P.; Ballard, Melissa M.; Beach, Michael J.; Causer, Louise; Wilkins, Patricia P.

    2003-01-01

    There is an increasing demand for diagnostic testing for Giardia intestinalis (G. lamblia) and Cryptosporidium parvum, with a priority being placed on obtaining diagnostic results in an efficient and timely manner. Several commercial companies have developed rapid diagnostic tests that are simple to perform and can be completed in less time than traditional methods for detecting Giardia and Cryptosporidium. We compared one of these rapid tests, the ImmunoCard STAT! (Meridian Bioscience, Inc.)...

  13. Real-Time PCR for Quantification of Giardia and Cryptosporidium in Environmental Water Samples and Sewage

    OpenAIRE

    Guy, Rebecca A.; Payment, Pierre; Krull, Ulrich J.; Horgen, Paul A.

    2003-01-01

    The protozoan pathogens Giardia lamblia and Cryptosporidium parvum are major causes of waterborne enteric disease throughout the world. Improved detection methods that are very sensitive and rapid are urgently needed. This is especially the case for analysis of environmental water samples in which the densities of Giardia and Cryptosporidium are very low. Primers and TaqMan probes based on the β-giardin gene of G. lamblia and the COWP gene of C. parvum were developed and used to detect DNA co...

  14. Ozone Air Purifiers: Can They Improve Asthma Symptoms?

    Science.gov (United States)

    ... daughter has asthma. Would she benefit from an ozone air purifier in her room? Answers from James ... Li, M.D., Ph.D. Despite manufacturers' claims, ozone air purifiers don't remove asthma triggers from ...

  15. Temporal patterns of human and canine Giardia infection in the United States: 2003-2009.

    Science.gov (United States)

    Mohamed, Ahmed S; Levine, Michael; Camp, Joseph W; Lund, Elisabeth; Yoder, Jonathan S; Glickman, Larry T; Moore, George E

    2014-02-01

    Giardia protozoa have been suspected to be of zoonotic transmission, including transmission from companion animals such as pet dogs to humans. Patterns of infection have been previously described for dogs and humans, but such investigations have used different time periods and locations for these two species. Our objective was to describe and compare the overall trend and seasonality of Giardia species infection among dogs and humans in the United States from 2003 through 2009 in an ecological study using public health surveillance data and medical records of pet dogs visiting a large nationwide private veterinary hospital. Canine data were obtained from all dogs visiting Banfield hospitals in the United States with fecal test results for Giardia species, from January 2003 through December 2009. Incidence data of human cases from the same time period were obtained from the CDC. Descriptive time plots, a seasonal trend decomposition (STL) procedure, and seasonal autoregressive moving-average (SARIMA) models were used to assess the temporal characteristics of Giardia infection in the two species. Canine incidence showed a gradual decline from 2003 to 2009 with no significant/distinct regular seasonal component. By contrast, human incidence showed a stable annual rate with a significant regular seasonal cycle, peaking in August and September. Different temporal patterns in human and canine Giardia cases observed in this study suggest that the epidemiological disease processes underlying both series might be different, and Giardia transmission between humans and their companion dogs seems uncommon. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Development of Sensitive Detection of Cryptosporidium and Giardia from Surface Water in Iran

    Directory of Open Access Journals (Sweden)

    N Taghipour Lailabadi

    2011-09-01

    Full Text Available Background: The protozoan parasites Cryptosporidium spp. and Giardia are known to occur widely in both raw and drinking waters. They are two of the causative agents of waterborne out-breaks of gastroenteritis throughout the world. In the present study, a PCR assay and FA were developed for detection of Cryptosporidium oocysts and Giardia cyst in environmental samples. Methods: We have detected Cryptosporidium spp. oocysts and Giardia cysts in seeded and un-seeded environmental water samples by PCR method. Water samples were spiked with oocysts (50, 100,300,500 and filtrated with a 1.2-µm pore size cellulose nitrate and follow by DNA extrac¬tion and purification by QIAamp DNA mini kit. Nested-PCR assay amplified an 850 bp fragment of 18s rRNA gene specific for Cryptosporidium and 435 bp fragment of glutamate dehydrogenase (GDH target gene for Giardia. Also many river water from north of Iran, be checked by these methods. Results: Cryptosporidium and Giardia DNAs were detected in seeded water sample and Giardia was detected in all 5 water samples from river in north of Iran by nested- PCR and FA. Also in one river water sample, Cryptosporidium was detected.Conclusion: This protocol is effective for detection of these waterborne parasites in treated and untreated water samples. This study can also serve as a platform for further investigations and research water source in Iran.

  17. Existence of Cryptosporidium spp. oocysts and Giardia spp. cysts in well water in Nineveh governorate

    Directory of Open Access Journals (Sweden)

    R. G. Mohammad

    2011-01-01

    Full Text Available The study included examination of 110 water samples from well distributed in Mosul city and few towns and villages around it from May 2009 to March 2010 for detection of Cryptosporidium spp oocysts and Giardia spp cysts in well water. The results revealed that the prevalence of Cryptosporidium oocysts was 16.36% while the prevalence of Giardia cysts was 12.72%. The percentages of prevalence with Cryptosporidium and Giardia were in high rate in Bartilla and some villages around it 20% for Cryptosporidium and 17.14% for Giardia, the low rates were in Mosul city 10% for both protozoa. The highest prevalence rate of Cryptosporidium was in March 38.46% and the lowest was in November and July 0%. The highest prevalence rate of Giardia was in October 23.53% and the lowest rate in July 0%. This first study shows the presence of Cryptosporidium oocysts and Giardia cysts in well water (Ground water in Nineveh governorate.

  18. Diagnosis of Giardia infections by PCR-based methods in children of an endemic area

    Directory of Open Access Journals (Sweden)

    EB David

    2011-01-01

    Full Text Available The present study was designed to estimate the prevalence of Giardia infection in preschool- and school-aged children living in an endemic area. Fecal samples from 573 children were processed by zinc sulfate centrifugal flotation, centrifugal sedimentation (using a commercial device for fecal concentration - TF-Test kit® and polymerase chain reaction (PCR-based methods. Of the stool samples assessed, 277 (48.3% were positive for intestinal parasites and/or commensal protozoa. Centrifugal flotation presented the highest diagnostic sensitivity for Giardia infections. The kappa index revealed that both coproparasitological techniques closely agreed on the Giardia diagnosis (86% versus satisfactory (72% and poor (35% concordances for commensal protozoan and helminth infections, respectively. Concerning Giardia molecular diagnosis, from the 71 microscopy-positive samples, specific amplification of gdh and tpi fragments was noted in 68 (95.7% and 64 (90% samples, respectively. Amplification of gdh and tpi genes was observed, respectively, in 95.7% and 90% of microscopy-positive Giardia samples. For 144 microscopy-negative samples, gdh and tpi gene amplification products were obtained from 8.3% and 35.9% samples, respectively. The agreement between these genes was about 40%. The centrifuge-flotation based method was the most suitable means of Giardia diagnosis assessed in the present study by combining accuracy and low cost.

  19. Comparison of Cytokine Responses in Ecuadorian Children Infected with Giardia, Ascaris, or Both Parasites.

    Science.gov (United States)

    Weatherhead, Jill; Cortés, Andrea Arévalo; Sandoval, Carlos; Vaca, Maritza; Chico, Martha; Loor, Sophia; Cooper, Philip J; Mejia, Rojelio

    2017-06-01

    AbstractMore than 2 billion people are infected with parasites globally, and the majority have coinfections. Intestinal protozoa and helminths induce polarizing CD4 + T-helper cell 1 (Th1) mediated cytokine responses within the host. Such immune polarization may inhibit the ability of the host to mount an adequate immune response for pathogen clearance to concurrent pathogens. The current study evaluated the plasma cytokine profile in Ascaris and Giardia coinfected children compared with Giardia - and Ascaris -only infected children. Fecal samples and blood samples were collected from asymptomatic 3-year-old children living in the district of Quininde, Ecuador. Stool samples that tested positive for Giardia lamblia -only, Ascaris lumbricoides- only, or G. lamblia and A. lumbricoides coinfections were confirmed by quantitative real-time polymerase chain reaction. Plasma samples from the study subjects were used to quantitate cytokines. A total of 39 patients were evaluated. Children with coinfection had a significant decrease in Th1 cytokine production, interleukin 2 (IL-2) ( P Giardia -only infected children. Coinfected children had an increase in IL-10/interferon gamma (IFN-γ) ratio compared with uninfected ( P Giardia may impair the host's ability to eradicate Giardia infection leading to chronic giardiasis.

  20. The minimal kinome of Giardia lamblia illuminates early kinase evolution and unique parasite biology

    Science.gov (United States)

    2011-01-01

    Background The major human intestinal pathogen Giardia lamblia is a very early branching eukaryote with a minimal genome of broad evolutionary and biological interest. Results To explore early kinase evolution and regulation of Giardia biology, we cataloged the kinomes of three sequenced strains. Comparison with published kinomes and those of the excavates Trichomonas vaginalis and Leishmania major shows that Giardia's 80 core kinases constitute the smallest known core kinome of any eukaryote that can be grown in pure culture, reflecting both its early origin and secondary gene loss. Kinase losses in DNA repair, mitochondrial function, transcription, splicing, and stress response reflect this reduced genome, while the presence of other kinases helps define the kinome of the last common eukaryotic ancestor. Immunofluorescence analysis shows abundant phospho-staining in trophozoites, with phosphotyrosine abundant in the nuclei and phosphothreonine and phosphoserine in distinct cytoskeletal organelles. The Nek kinase family has been massively expanded, accounting for 198 of the 278 protein kinases in Giardia. Most Neks are catalytically inactive, have very divergent sequences and undergo extensive duplication and loss between strains. Many Neks are highly induced during development. We localized four catalytically active Neks to distinct parts of the cytoskeleton and one inactive Nek to the cytoplasm. Conclusions The reduced kinome of Giardia sheds new light on early kinase evolution, and its highly divergent sequences add to the definition of individual kinase families as well as offering specific drug targets. Giardia's massive Nek expansion may reflect its distinctive lifestyle, biphasic life cycle and complex cytoskeleton. PMID:21787419

  1. Prevalence of Giardia in dairy cattle in Lusaka and Chilanga districts, Zambia.

    Science.gov (United States)

    Kakandelwa, Cliff; Siwila, Joyce; Nalubamba, King S; Muma, John B; Phiri, Isaac G K

    2016-01-15

    Giardia is an intestinal protozoan parasite of mammals including humans. A cross-sectional study was conducted to estimate prevalence of Giardia infections in smallholder and commercial dairy herds in Chilanga and Lusaka districts of Zambia. A total of 377 calves aged from 1 to 365 days were sampled on 34 farms. All faecal samples were analyzed for Giardia antigen using a commercially available ELISA kit. Overall prevalence of Giardia was 34.5% (95% CI=29.7-39.3). Among smallholder farms, animal level prevalence ranged from 0 to 100% (mean=44.6±36.9 standard deviations) and 12.5 to 60.9% (mean=33.5±16.7 standard deviations) within commercial herds. Prevalence was highest in calves less than three months old (p=0.010), and there was no significant difference in the prevalence between smallholder and commercial farms (p=0.300). Giardia prevalence was not associated with occurrence of diarrhoea in the calves (p=0.205). The study demonstrates that Giardia infections are common in dairy herds in the study areas, especially in calves less than three months of age. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Measurement of Giardia lamblia adhesion force using an integrated microfluidic assay.

    Science.gov (United States)

    Lu, Ling; Zheng, Guo-Xia; Yang, Yu-Suo; Feng, Cheng-Yu; Liu, Fang-Fang; Wang, Yun-Hua

    2017-02-01

    The mechanisms how Giardias attach to the intestinal epithelium remain unclear. None of the methods currently being used to measure the attachment force could provide a continuous nutrition supply and a micro-aerobic atmosphere to the Giardia. Besides, they are all labor-intensive. In the present research, a microfluidic method based on electric circuit analogy was developed. The input fluid flowed through the inlet channel with different lengths and was distributed in four assay chambers. Shear force gradients were generated in chambers, too. This allowed an easy control of fluids and the shear forces. Most importantly, the shear stress large enough to detach Giardia could be generated in laminar flow regime. Moreover, analysis could be accomplished in one single test. By applying inlet flow rates of 30, 60, and 120 μL ml -1 , shear force gradients ranging from 19.47 to 60.50 Pa were generated. The adhesion forces of trophozoites were analyzed and the EC 50 of the force that caused 50% trophozoites detachment was calculated as 36.60 Pa. This paper presents a novel method for measurement of Giardia adhesion force. Graphical Abstract Measurement of Giardia adhesion force. Various of flow rates were applied to generate different shear forces and Giardia trophozoites remaining attached were counted (a-c). The percentages of attachment vs shear stress were plotted and the EC 50 of adhesion force was calculated (d).

  3. Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan.

    Science.gov (United States)

    Itoh, N; Itagaki, T; Kawabata, T; Konaka, T; Muraoka, N; Saeki, H; Kanai, K; Chikazawa, S; Hori, Y; Hoshi, F; Higuchi, S

    2011-02-28

    The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP®Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase (gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. The invasive potential of Giardia intestinalis in an in vivo model.

    Science.gov (United States)

    Reynoso-Robles, R; Ponce-Macotela, M; Rosas-López, L E; Ramos-Morales, A; Martínez-Gordillo, M N; González-Maciel, A

    2015-10-16

    Giardiasis is a neglected parasitic disease that affects primarily children, in whom it delays physical and mental development. The pathophysiology of giardiasis in not well understood, and most reports have identified Giardia intestinalis trophozoites only in the lumen and on the brush border of the small intestine. We identified Giardia trophozoites within the epithelium of the small intestine of a lactose intolerance patient. The Giardia trophozoites were obtained and cultured in vitro. In addition, we demonstrated Giardia trophozoite invasion in an animal model. Giardia trophozoites invaded the intestinal mucosa and submucosa of infected gerbils. The invasive trophozoites were observed at 21, 30 and 60 days age, and the average numbers of invaded sites were 17 ± 5, 15 ± 4, and 9 ± 3, respectively. We found trophozoites between epithelial cells, at the base of empty goblet cells, in lacteal vessels and within the submucosa. The morphological integrity of the invasive trophozoites was demonstrated via electron microscopy. The analysis of the gerbils infected with the trophozoites of the WB reference strain did not show intraepithelial trophozoites. These results demonstrate another Giardia pathogenic mechanism, opening the door to numerous future studies.

  5. Life with eight flagella: flagellar assembly and division in Giardia.

    Science.gov (United States)

    Dawson, Scott C; House, Susan A

    2010-08-01

    Flagellar movement in Giardia, a common intestinal parasitic protist, is crucial to its survival in the host. Each axoneme is unique in possessing a long, cytoplasmic portion as well as a membrane-bound portion. Intraflagellar transport (IFT) is required for the assembly of membrane-bound regions, yet the cytoplasmic regions may be assembled by IFT-independent mechanisms. Steady-state axoneme length is maintained by IFT and by intrinsic and active microtubule dynamics. Following mitosis and before their segregation, giardial flagella undergo a multigenerational division cycle in which the parental eight flagella migrate and reposition to different cellular locations; eight new flagella are assembled de novo. Each daughter cell thus inherits four mature and four newly synthesized flagella. Copyright 2010 Elsevier Ltd. All rights reserved.

  6. Proteomics of secretory and endocytic organelles in Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Petra B Wampfler

    Full Text Available Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs, mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs, for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694.

  7. Participation of actin on Giardia lamblia growth and encystation.

    Directory of Open Access Journals (Sweden)

    Araceli Castillo-Romero

    Full Text Available BACKGROUND: Microfilaments play a determinant role in different cell processes such as: motility, cell division, phagocytosis and intracellular transport; however, these structures are poorly understood in the parasite Giardia lamblia. METHODOLOGY AND PRINCIPAL FINDINGS: By confocal microscopy using TRITC-phalloidin, we found structured actin distributed in the entire trophozoite, the label stand out at the ventral disc, median body, flagella and around the nuclei. During Giardia encystation, a sequence of morphological changes concurrent to modifications on the distribution of structured actin and in the expression of actin mRNA were observed. To elucidate whether actin participates actively on growth and encystation, cells were treated with Cytochalasin D, Latrunculin A and Jasplakinolide and analyzed by confocal and scanning electron microscopy. All drugs caused a growth reduction (27 to 45% and changes on the distribution of actin. Besides, 60 to 80% of trophozoites treated with the drugs, exhibited damage at the caudal region, alterations in the flagella and wrinkles-like on the plasma membrane. The drugs also altered the cyst-yield and the morphology, scanning electron microscopy revealed diminished cytokinesis, cysts with damages in the wall and alterations in the size and on the intermembranal space. Furthermore, the drugs caused a significant reduction of the intensity of fluorescence-labeled CWP1 on ESV and on cyst wall, this was coincident with a reduction of CWP1 gene expression (34%. CONCLUSIONS AND SIGNIFICANCE: All our results, indicated an important role of actin in the morphology, growth and encystation and indirectly suggested an actin role in gene expression.

  8. Apparatus and methods for purifying lead

    Science.gov (United States)

    Tunison, Harmon M.

    2016-01-12

    Disclosed is an exemplary method of purifying lead which includes the steps of placing lead and a fluoride salt blend in a container; forming a first fluid of molten lead at a first temperature; forming a second fluid of the molten fluoride salt blend at a second temperature higher than the first temperature; mixing the first fluid and the second fluid together; separating the two fluids; solidifying the molten fluoride salt blend at a temperature above a melting point of the lead; and removing the molten lead from the container. In certain exemplary methods the molten lead is removed from the container by decanting. In still other exemplary methods the molten salt blend is a Lewis base fluoride eutectic salt blend, and in yet other exemplary methods the molten salt blend contains sodium fluoride, lithium fluoride, and potassium fluoride.

  9. Flow-through immunomagnetic separation system for waterborne pathogen isolation and detection: Application to Giardia and Cryptosporidium cell isolation

    Energy Technology Data Exchange (ETDEWEB)

    Ramadan, Qasem, E-mail: qasem.alramadan@epfl.ch [Bioelectronics Program, Institute of Microelectronics, 11 Science Park Road, Singapore 117685 (Singapore); Christophe, Lay; Teo, William; ShuJun, Li; Hua, Feng Han [Bioelectronics Program, Institute of Microelectronics, 11 Science Park Road, Singapore 117685 (Singapore)

    2010-07-12

    Simultaneous sample washing and concentration of two waterborne pathogen samples were demonstrated using a rotational magnetic system under continuous flow conditions. The rotation of periodically arranged small permanent magnets close to a fluidic channel carrying magnetic particle suspension allows the trapping and release of particles along the fluidic channel in a periodic manner. Each trapping and release event resembles one washing cycle. The performance of the magnetic separation system (MSS) was evaluated in order to test its functionality to isolate magnetic-labelled protozoan cells from filtered, concentrated tap water, secondary effluent water, and purified water. Experimental protocols described in US Environmental Protection Agency method 1623 which rely on the use of a magnetic particle concentrator, were applied to test and compare our continuous flow cell separation system to the standard magnetic bead-based isolation instruments. The recovery efficiencies for Giardia cysts using the magnetic tube holder and our magnetic separation system were 90.5% and 90.1%, respectively, from a tap water matrix and about 31% and 18.5%, respectively, from a spiked secondary effluent matrix. The recovery efficiencies for Cryptosporidium cells using the magnetic tube holder and our magnetic separation system were 90% and 83.3%, respectively, from a tap water matrix and about 38% and 36%, respectively, from a spiked secondary effluent matrix. Recoveries from all matrices with the continuous flow system were typically higher in glass tubing conduits than in molded plastic conduits.

  10. Cryptosporidium and Giardia in different age groups of Danish cattle and pigs - Occurrence and management associated risk factors

    DEFF Research Database (Denmark)

    Maddox-Hyttel, Charlotte; Langkjær, Rikke Breinhold; Enemark, Heidi L.

    2006-01-01

    and sow herds, respectively. Each herd was visited once for the collection of faecal samples and registration of basic management parameters. Faecal samples were collected from three different age groups of animals, i.e. 5 sows/cows. 10 nursing piglets/calves less than 1 month, and 10 weaner pigs 8-45 kg....../calves 1-12 months. The faecal samples were purified and the number of (oo)cysts quantified. The study revealed an age-specific herd prevalence of Cryptosporidium of 16, 31 and 100% for sows, piglets and weaners, respectively, and of 14, 96 and 84% for cows, young calves and older calves, respectively....... For Giardia the age-specific herd prevalence was 18, 22 and 84% for the sows, piglets and weaners, while for cattle herds the prevalence was 60, 82 and 100% for cows, young calves and older calves, correspondingly. The (oo)cyst excretion levels varied considerably both within and between herds for all age...

  11. Expression, purification, crystallization and preliminary X-ray diffraction analysis of α-11 giardin from Giardia lamblia

    International Nuclear Information System (INIS)

    Pathuri, Puja; Nguyen, Emily Tam; Luecke, Hartmut

    2006-01-01

    α-11 giardin from the intestinal protozoan parasite, G. lamblia has been cloned, expressed, purified and crystallized under two different conditions and in two different space groups. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group and crystals obtained in the second condition diffracted to 2.93 Å and belong to a primitive monoclinic space group. α-11 Giardin, a protein from the annexin superfamily, is a 35.0 kDa protein from the intestinal protozoan parasite Giardia lamblia which triggers a form of diarrhea called giardiasis. Here, the cloning, expression, purification and the crystallization of α-11 giardin under two different conditions and in two different space groups is reported. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group, while crystals from the second condition, which included calcium in the crystallization solution, diffracted to 2.93 Å and belong to a primitive monoclinic space group. Determination of the detailed atomic structure of α-11 giardin will provide a better insight into its biological function and might establish whether this class of proteins is a potential drug target against giardiasis

  12. Expression, purification, crystallization and preliminary X-ray diffraction analysis of α-11 giardin from Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Pathuri, Puja; Nguyen, Emily Tam [Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697 (United States); Luecke, Hartmut, E-mail: hudel@uci.edu [Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697 (United States); Department of Physiology and Biophysics, University of California, Irvine, CA 92697 (United States); Department of Information and Computer Sciences, University of California, Irvine, CA 92697 (United States)

    2006-11-01

    α-11 giardin from the intestinal protozoan parasite, G. lamblia has been cloned, expressed, purified and crystallized under two different conditions and in two different space groups. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group and crystals obtained in the second condition diffracted to 2.93 Å and belong to a primitive monoclinic space group. α-11 Giardin, a protein from the annexin superfamily, is a 35.0 kDa protein from the intestinal protozoan parasite Giardia lamblia which triggers a form of diarrhea called giardiasis. Here, the cloning, expression, purification and the crystallization of α-11 giardin under two different conditions and in two different space groups is reported. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group, while crystals from the second condition, which included calcium in the crystallization solution, diffracted to 2.93 Å and belong to a primitive monoclinic space group. Determination of the detailed atomic structure of α-11 giardin will provide a better insight into its biological function and might establish whether this class of proteins is a potential drug target against giardiasis.

  13. O2-Dependent Efficacy of Novel Piperidine- and Piperazine-Based Chalcones against the Human Parasite Giardia intestinalis

    Science.gov (United States)

    Bahadur, Vijay; Mastronicola, Daniela; Tiwari, Hemandra Kumar; Kumar, Yogesh; Falabella, Micol; Pucillo, Leopoldo Paolo; Sarti, Paolo

    2014-01-01

    Giardia intestinalis is the most frequent protozoan agent of intestinal diseases worldwide. Though commonly regarded as an anaerobic pathogen, it preferentially colonizes the fairly oxygen-rich mucosa of the proximal small intestine. Therefore, when testing new potential antigiardial drugs, O2 should be taken into account, since it also reduces the efficacy of metronidazole, the gold standard drug against giardiasis. In this study, 46 novel chalcones were synthesized by microwave-assisted Claisen-Schmidt condensation, purified, characterized by high-resolution mass spectrometry, 1H and 13C nuclear magnetic resonance, and infrared spectroscopy, and tested for their toxicity against G. intestinalis under standard anaerobic conditions. As a novel approach, compounds showing antigiardial activity under anaerobiosis were also assayed under microaerobic conditions, and their selectivity against parasitic cells was assessed in a counterscreen on human epithelial colorectal adenocarcinoma cells. Among the tested compounds, three [30(a), 31(e), and 33] were more effective in the presence of O2 than under anaerobic conditions and killed the parasite 2 to 4 times more efficiently than metronidazole under anaerobiosis. Two of them [30(a) and 31(e)] proved to be selective against parasitic cells, thus representing potential candidates for the design of novel antigiardial drugs. This study highlights the importance of testing new potential antigiardial agents not only under anaerobic conditions but also at low, more physiological O2 concentrations. PMID:24217695

  14. Detection of Giardia intestinalis infections in Polish soldiers deployed to Afghanistan.

    Science.gov (United States)

    Korzeniewski, Krzysztof; Konior, Monika; Augustynowicz, Alina; Lass, Anna; Kowalska, Ewa

    2016-01-01

    Members of the Polish Military Contingent (PMC) have been stationed in Afghanistan since 2002. They typically serve in areas characterised by low standards of sanitation which often leads to the development of food- and waterborne diseases. The aim of the study was to evaluate the prevalence of Giardia intestinalis infections among Polish soldiers deployed to Afghanistan. The research study was conducted as part of a programme for prevention of parasitic diseases of the gastrointestinal tract run by the Polish Armed Forces. The study was carried out in August 2011; it involved 630 asymptomatic Polish soldiers serving in the Forward Operational Base (FOB) Ghazni in eastern Afghanistan. Stool specimens obtained from members of the PMC were first tested in FOB Ghazni (detection of Giardia intestinalis by Rida Quick Giardia immunochromatographic tests and Ridascreen Giardia immunoenzymatic tests - single samples). Next, the same biological material and two other faecal specimens fixed in 10% formalin were transported to the Military Institute of Medicine in Poland, where they were tested for Giardia intestinalis under light microscopy (direct smear, decantation in distilled water). Parasitological tests performed under light microscopy showed that 2.7% (17/630) of the study group were infected with Giardia intestinalis. Some of these results were confirmed by immunochromatographic tests (6/630). In contrast, immunoenzymatic tests (ELISA) demonstrated a significantly higher detection rate reaching 18.1% (114/630). Immunoenzymatic tests confirmed all the positive results given by light microscopy and by immunochromatographic tests. The prevalence rate of Giardia intestinalis infections in Polish soldiers deployed to Afghanistan was found to be high. Microscopic methods exhibit low sensitivity and therefore may result in the underestimation of the true parasite prevalence. Immunoenzymatic tests (ELISA) showing a much higher sensitivity in comparison to light microscopy

  15. Study of hot corrosion of flakes of non purified graphite and of purified graphite

    International Nuclear Information System (INIS)

    Boule, Michel

    1967-01-01

    The author reports the study of hot corrosion of the Ticonderoga graphite. He reports the study of the defects of graphite flakes (structure defects due to impurities), the dosing of these impurities, and then their removal by purification. Flakes have then been oxidised by means of a specially designed apparatus. Based on photographs taken by optical and electronic microscopy, the author compares the oxidation features obtained in dry air and in humid air, between purified and non purified flakes. He also reports the study of the evolution of oxidation with respect to the initial rate of impurities, and the study of the evolution of oxidation features in humid air during oxidation. All these comparisons are made while taking the oxidation rate into account [fr

  16. Effect of Giardia infection on growth and psychomotor development of children aged 0-5 years.

    Science.gov (United States)

    Simsek, Z; Zeyrek, F Yildiz; Kurcer, M A

    2004-04-01

    Giardiasis, an intestinal protozoan infection caused by Giardia intestinalis, is common in southeastern Anatolia, Turkey. In this cross-sectional survey, to investigate the role of giardiasis on growth and psychomotor development, we studied 160 children aged 0-5 years. Data were collected using a structured questionnaire, anthropometry, Ankara Developmental Screening Inventory, and laboratory analysis of fecal samples. The results showed that 50 per cent of the subjects were infected with at least one pathogen of intestinal parasitic infections. Giardia intestinalis was the most frequent pathogenic parasite. Giardia-infected children had a risk for stunted (OR = 7.67, 95 per cent CI = 2.25-26.16; p = 0.001) and poor psychomotor development (OR = 2.68, 95 per cent CI = 1.09-6.58; p = 0.030). The data indicate that Giardia intestinalis infection has an adverse impact on child linear growth and psychomotor development. In the primary healthcare centers, during the programme of the monitoring growth and developmental status of children, following children in terms of Giardia, diagnosis and treatment will have a positive effect on child health.

  17. Drinking Water Quality and Occurrence of Giardia in Finnish Small Groundwater Supplies

    Directory of Open Access Journals (Sweden)

    Tarja Pitkänen

    2015-08-01

    Full Text Available The microbiological and chemical drinking water quality of 20 vulnerable Finnish small groundwater supplies was studied in relation to environmental risk factors associated with potential sources of contamination. The microbiological parameters analyzed included the following enteric pathogens: Giardia and Cryptosporidium, Campylobacter species, noroviruses, as well as indicator microbes (Escherichia coli, intestinal enterococci, coliform bacteria, Clostridium perfringens, Aeromonas spp. and heterotrophic bacteria. Chemical analyses included the determination of pH, conductivity, TOC, color, turbidity, and phosphorus, nitrate and nitrite nitrogen, iron, and manganese concentrations. Giardia intestinalis was detected from four of the water supplies, all of which had wastewater treatment activities in the neighborhood. Mesophilic Aeromonas salmonicida, coliform bacteria and E. coli were also detected. None of the samples were positive for both coliforms and Giardia. Low pH and high iron and manganese concentrations in some samples compromised the water quality. Giardia intestinalis was isolated for the first time in Finland in groundwater wells of public water works. In Europe, small water supplies are of great importance since they serve a significant sector of the population. In our study, the presence of fecal indicator bacteria, Aeromonas and Giardia revealed surface water access to the wells and health risks associated with small water supplies.

  18. Detection of Giardia lamblia and Cryptosporidium parvum by direct immunofluorescence assay in stool specimen.

    Science.gov (United States)

    Rahman, M M; Hossain, M A; Paul, S K; Ahmed, S; Islam, A; Ehsan, M A; Alam, M M; Kabir, M R; Sarkar, S R

    2014-07-01

    Giardia and Cryptosporidium are the pathogens which transmitted through contaminated soil and contaminated water are significant causes of diarrhea and nutritional disorders in institutional and community peoples. Children and immune compromise persons are more vulnerable for these infections. Both Giardiasis and Cryptosporidiosis were included in 2004 as WHO Neglected Disease. So this is a major public health problem in developing countries. The present study was carried out to detect the Giardia and Cryptosporidium from diarrheic or patient having loose stool by Direct Immunofluorescence assay. The study was conducted during July 20012 to February 2013 and the work was done in Mymensingh Medical College in the department of Microbiology and in Bangladesh Agricultural University in the department of Veterinary Medicine. A total of 100 loose stools were collected from school children of different area and hospital under sadar upazilla, Mymensingh. The detection of Giardia lamblia and Cryptosporidium parvum showed the individual prevalence 8% and 4% respectively. The highest cyst/oocyst count was 85,000 and 1,000/gm of stool and the lowest being 100 and 50/gm of stool for Giardiasis and Cryptosporidiosis respectively. The detection rate of Giardia and Cryptosporidium by Immunofluorescence assay was relatively higher than the previous study done in Bangladesh and this was the first report from Bangladesh over human stool specimen using Immunofluorescence assay. So, Immunofluorescence assay could be adapted for rapid and accurate detection of Giardia and Cryptosporidium.

  19. Isolation and partial characterization of an immunogenic antigen of Giardia lamblia.

    Science.gov (United States)

    Quintero, Jael; Valdez, Alejandra; Samaniego, Brenda; Lopez-Romero, Gloria; Astiazaran-Garcia, Humberto; Rascon, Lucila; Breci, Linda; Garibay-Escobar, Adriana; Robles-Zepeda, Ramón; Velazquez, Carlos

    2017-06-01

    Humoral and cellular immune responses play an important role during Giardia lamblia infection. Several Giardia proteins have been identified as immunogenic antigens based on their elicited humoral immune response. Poorly is known about Giardia antigens that stimulate a cellular immune response. The main purpose of this study was to isolate and partial characterize an immunogenic antigen (5G8) of G. lamblia. The 5G8 protein was isolated from G. lamblia trophozoite lysates by affinity chromatography using moAb 5G8-coupled CNBr-Sepharose. The isolated protein was analysed by electrospray tandem mass spectrometry (ESI-MS/MS), and by diverse bioinformatics tools (GiardiaDB, BLASTn, BLASTp and ExPASy). Additionally, several biochemical and immunological characteristics of the isolated protein were analysed. By ESI-MS/MS the amino acidic 5G8 sequence was deduced. The 5G8 antigen belongs to the VSP family proteins of G. lamblia. This protein is composed by one polypeptide chain (±71kDa). Using the algorithm SYFPHEITI, we identified candidate CD4 + T-cell epitopes from the 5G8 antigen, which can elicit cell-mediated immune responses. In this study, we have identified a G. lamblia protein that induces a strong immune response in infected mice. The biochemical and immunological characterization of the immunogenic 5G8 antigen may contribute to the rational design of a Giardia vaccine. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  20. [Investigation on contamination of Cryptosporidium and Giardia in drinking water and environmental water in Shanghai].

    Science.gov (United States)

    Zhang, Xiao-Ping; He, Yan-Yan; Zhu, Qian; Ma, Xiao-Jiang; Cai, Li

    2010-12-30

    To understand the contamination status of Cryptosporidium sp. and Giardia lamblia in drinking water, source water and environmental water in Shanghai. All water samples collected from drinking water, source water and environmental water were detected by a procedure of micromembrane filtration, immune magnetic separation (IMS), and immunofluorescent assay (IFA). Cryptosporidium oocysts and Giardia cysts were not found in 156 samples of the drinking water including finished water, tap water, or pipe water for directly drinking in communities. Among 70 samples either source water of water plants (15 samples), environmental water from Huangpu River(25), canal water around animal sheds(15), exit water from waste-water treatment plants(9), or waste water due to daily life(6), Cryptosporidium oocysts were detected in 1(6.7%), 2(8.0%), 7(46.7%), 1(11.1%), and 1(16.7%) samples, respectively; and Giardia cysts were detected in 1(6.7%), 3(12.0%), 6 (40.0%), 2(22.2%), and 2(33.3%), respectively. The positive rate of Cryptosporidium oocysts and Giardia cysts was 17.1% (12/70) and 20.0% (14/70), respectively. No Cryptosporidium oocysts and Giardia cysts have been detected in drinking water, but found in source water and environmental water samples in Shanghai.

  1. Interleukin-17 receptor A (IL-17RA) as a central regulator of the protective immune response against Giardia

    Science.gov (United States)

    The protozoan parasite Giardia is a highly prevalent intestinal pathogen with a wide host range. Data obtained in mice, cattle and humans revealed the importance of IL-17A in the development of a protective immune response against Giardia. The aim of this study was to further unravel the protective ...

  2. Interleukin-17 receptor A (IL-17RA) as a central regulator of the protective immune response against Giardia

    NARCIS (Netherlands)

    Paerewijck, O. (Oonagh); Maertens, B. (Brecht); L. Dreesen (Leentje); Van Meulder, F. (Frederik); Peelaers, I. (Iris); Ratman, D. (Dariusz); Li, R.W. (Robert W.); E.W. Lubberts (Erik); K. De Bosscher; P. Geldhof (Peter)

    2017-01-01

    textabstractThe protozoan parasite Giardia is a highly prevalent intestinal pathogen with a wide host range. Data obtained in mice, cattle and humans revealed the importance of IL-17A in the development of a protective immune response against Giardia. The aim of this study was to further unravel the

  3. Chitin synthetase in encysting Giardia lamblia and Entamoeba invadens

    International Nuclear Information System (INIS)

    Das, S.; Gillin, F.D.

    1987-01-01

    Giardia lamblia (Gl) and Entamoeba invadens (Ei) are protozoan parasites with two morphologic stages in their life cycles. Motile trophozoites colonize the intestine of humans and reptiles respectively. Water resistant cysts, which can survive outside the host, transmit infection. In vitro cyst formation of Ei from trophozoites has been reported, and the authors have recently induced in vitro encystation of Gl. Although the cyst walls of both parasites contain chitin, it synthesis by encysting trophozoites has not been reported. The authors now show that encystation conditions greatly increase chitin synthetase (CS) specific activity (incorporation of 3 H GlcNAc from UDP-GlcNAc into TCA-or alcohol-precipitable material). Extracts of encysting Gl incorporated 3.6 nmol/mg protein in 5 hr compared to < 0.005 in controls. Extracts of encysting Fi incorporated 4.8 n mol/mg protein, compared to 1.7 in the control. CS activity of both parasites requires preformed chitin. The Gl enzyme requires a reducing agent, is inhibited by digitonin and the CS inhibitors, polyoxin D and Nikkomycin, but not by tunicamycin. The product is digested by chitinase. Ei enzyme does not require a reducing agent and is stimulated by 1 mg/ml digitonin, but inhibited by higher concentrations. These studies demonstrate CS enzymes which may play important roles in encystation of Gl and Ei

  4. Chitin synthetase in encysting Giardia lamblia and Entamoeba invadens

    Energy Technology Data Exchange (ETDEWEB)

    Das, S.; Gillin, F.D.

    1987-05-01

    Giardia lamblia (Gl) and Entamoeba invadens (Ei) are protozoan parasites with two morphologic stages in their life cycles. Motile trophozoites colonize the intestine of humans and reptiles respectively. Water resistant cysts, which can survive outside the host, transmit infection. In vitro cyst formation of Ei from trophozoites has been reported, and the authors have recently induced in vitro encystation of Gl. Although the cyst walls of both parasites contain chitin, it synthesis by encysting trophozoites has not been reported. The authors now show that encystation conditions greatly increase chitin synthetase (CS) specific activity (incorporation of /sup 3/H GlcNAc from UDP-GlcNAc into TCA-or alcohol-precipitable material). Extracts of encysting Gl incorporated 3.6 nmol/mg protein in 5 hr compared to < 0.005 in controls. Extracts of encysting Fi incorporated 4.8 n mol/mg protein, compared to 1.7 in the control. CS activity of both parasites requires preformed chitin. The Gl enzyme requires a reducing agent, is inhibited by digitonin and the CS inhibitors, polyoxin D and Nikkomycin, but not by tunicamycin. The product is digested by chitinase. Ei enzyme does not require a reducing agent and is stimulated by 1 mg/ml digitonin, but inhibited by higher concentrations. These studies demonstrate CS enzymes which may play important roles in encystation of Gl and Ei.

  5. Decontamination of radioactive materials using a household water purifier

    International Nuclear Information System (INIS)

    Yanaga, Makoto; Oishi, Ayumi; Wada, Hideki

    2013-01-01

    A home water purifier was used to remove radioactive iodine and cesium from rainwater contaminated with the radioactive materials released by the Fukushima Daiichi Nuclear Power Plant accident following the earthquake and tsunami in 2011. To prevent internal radiation exposure, the rainwater was satisfactorily decontaminated by a simple method using the pitcher type water purifier, which is widely available to the general public. However, the results indicate that the ability for purification by the home water purifier will be reduced significantly if the water is not filtered by some method before the water purifier is used, especially in the case of natural water containing suspension matter. (author)

  6. A systematic review of the effects of Iranian pharmaceutical plant extracts on Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Hajar Ziaei Hezarjaribi

    2015-12-01

    Full Text Available This study aimed to provide a systematic review regarding anti-Giardia effect of different Iranian plant extracts used in vivo and in vitro on cysts and trophozoites. Many reports indicated that most of plant extracts used as anti-Giardia were obtained from Liliaceae, Apiaceae, Asteraceae, and Myrtaceae. These extracts included different fractions such as aqueous, alcoholic and chloroform extracts as well as Soxhlet extraction of juice or essence. The findings of this review showed that hydroalcoholic extract of asafoetida, Chenopodium botrys, and chloroformic extract of feverfew (Tanacetum parthenium have the maximum effect (100% efficacy on in vitro application against Giardia. However, the highest in vivo effect of 100% therapeutic significance was recorded for the extract of Allium sativum at 80 mg/mL concentration. Given the plant species richness of Iran in terms of herbal medicines with fewer side effects, it can be a good alternative to chemical drugs used to treat giardiasis.

  7. Gene expression changes during Giardia-host cell interactions in serum-free medium.

    Science.gov (United States)

    Ferella, Marcela; Davids, Barbara J; Cipriano, Michael J; Birkeland, Shanda R; Palm, Daniel; Gillin, Frances D; McArthur, Andrew G; Svärd, Staffan

    2014-10-01

    Serial Analysis of Gene Expression (SAGE) was used to quantify transcriptional changes in Giardia intestinalis during its interaction with human intestinal epithelial cells (IECs, HT-29) in serum free M199 medium. Transcriptional changes were compared to those in trophozoites alone in M199 and in TYI-S-33 Giardia growth medium. In total, 90 genes were differentially expressed, mainly those involved in cellular redox homeostasis, metabolism and small molecule transport but also cysteine proteases and structural proteins of the giardin family. Only 29 genes changed their expression due to IEC interaction and the rest were due to M199 medium. Although our findings generated a small dataset, it was consistent with our earlier microarray studies performed under different interaction conditions. This study has confined the number of genes in Giardia to a small subset that specifically change their expression due to interaction with IECs. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Administration of kefir-fermented milk protects mice against Giardia intestinalis infection.

    Science.gov (United States)

    Franco, Mariana Correa; Golowczyc, Marina A; De Antoni, Graciela L; Pérez, Pablo F; Humen, Martín; Serradell, María de los Angeles

    2013-12-01

    Giardiasis, caused by the protozoan Giardia intestinalis, is one of the most common intestinal diseases worldwide and constitutes an important problem for the public health systems of various countries. Kefir is a probiotic drink obtained by fermenting milk with 'kefir grains', which consist mainly of bacteria and yeasts that coexist in a complex symbiotic association. In this work, we studied the ability of kefir to protect mice from G. intestinalis infection, and characterized the host immune response to this probiotic in the context of the intestinal infection. Six- to 8-week-old C75BL/6 mice were separated into four groups: controls, kefir mice (receiving 1 : 100 dilution of kefir in drinking water for 14 days), Giardia mice (infected orally with 4×10(7) trophozoites of G. intestinalis at day 7) and Giardia-kefir mice (kefir-treated G. intestinalis-infected mice), and killed at 2 or 7 days post-infection. Kefir administration was able to significantly reduce the intensity of Giardia infection at 7 days post-infection. An increase in the percentage of CD4(+) T cells at 2 days post-infection was observed in the Peyer's patches (PP) of mice belonging to the Giardia group compared with the control and kefir groups, while the percentage of CD4(+) T cells in PP in the Giardia-kefir group was similar to that of controls. At 2 days post-infection, a reduction in the percentage of B220-positive major histocompatibility complex class II medium cells in PP was observed in infected mice compared with the other groups. At 7 days post-infection, Giardia-infected mice showed a reduction in RcFcε-positive cells compared with the control group, suggesting a downregulation of the inflammatory response. However, the percentages of RcFcε-positive cells did not differ from controls in the kefir and Giardia-kefir groups. An increase in IgA-positive cells was observed in the lamina propria of the kefir group compared with controls at 2 days post-infection. Interestingly, the

  9. Detection of anti-Giardia lamblia serum antibody among children of day care centers

    Directory of Open Access Journals (Sweden)

    Guimarães Semíramis

    2002-01-01

    Full Text Available OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF and enzyme-linked immunosorbent assay (ELISA for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3% showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3% and 100 (68% samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39% than IIF (70%. IIF also showed a higher concordance with microscopic examination than ELISA.

  10. Detection of small number of Giardia in biological materials prepared from stray dogs.

    Science.gov (United States)

    Esmailikia, Leila; Ebrahimzade, Elahe; Shayan, Parviz; Amininia, Narges

    2017-12-20

    Giardia lamblia is an intestinal protozoa with intermittent and low shedding especially in dogs, and the detection of Giardia is accompanied with problems such as sampling and diagnostic method. The objective of this study was to detection of Giardia in biological materials with low number of parasite using parasitological and molecular methods, and also to determine whether the examined stray dogs harbor known zoonotic genotype of Giardia. For this aim 85 fecal and duodenal samples were studied from which 1 was positive by Trichrome staining of stool, 4 were positive by staining of duodenal samples. The nested PCR analysis with primers derived from 18 SrRNA showed that the specific PCR product could be amplified in 4 stool and 4 duodenal samples. All positive samples in staining analysis were also positive in nested PCR. No amplification could be observed by nested PCR with primers derived from β giardin gene due to the single copy of gene. Interestingly, the extracted DNA from old fixed stained Giardia positive smears could be also amplified with primers derived from 18SrRNA gene. The sequence analysis of nested PCR products showed that they belong to the genotype D. In conclusion, it is to denote that the Trichrome or Giemsa methods were not suitable for the detection of small number of this parasite in stool and the nested PCR with primers derived from 18S rRNA gene can replace the traditional methods successfully. For detection of Giardia in stool, primers derived from β giardin will not be recommended.

  11. Giardia and other intestinal parasites in different dog populations in Northern Belgium.

    Science.gov (United States)

    Claerebout, E; Casaert, S; Dalemans, A-C; De Wilde, N; Levecke, B; Vercruysse, J; Geurden, T

    2009-04-06

    The objectives of this study were to obtain data on the prevalence of intestinal parasites in different dog populations in northern Belgium, to estimate the zoonotic risk associated with these infections and to identify potential risk factors. Between 2004 and 2007 a total of 1159 faecal samples were collected from 451 household dogs, 357 dogs from breeding kennels and 351 dogs with gastrointestinal disorders. The samples from dogs with gastrointestinal disorders were sent to the diagnostic Laboratory for Parasitology at Ghent University by veterinary practitioners. In household dogs the prevalence of intestinal parasites was relatively low. Giardia was the most commonly found parasite (9.3%, CI 5.5-13.1), followed by Toxocara canis (4.4%, CI 2.7-6.8). Much higher infection rates were observed in kennel dogs, especially for Giardia spp. (43.9%, CI 37.8-50.0); T. canis (26.3%, CI 21.8-31.2) and Cystoisospora spp. (26.3%, CI 21.8-31.2). Also in dogs with gastrointestinal problems, Giardia spp. (18.1%, CI 13.1-23.1), Cystoisospora spp. (8.8%, CI 6.1-12.3) and T. canis (7.4%, CI 4.9-10.7) were the most frequently detected parasites. In all dog populations pups were more frequently infected with Cystoisospora (PGiardia (PGiardia and T. canis (PGiardia spp. Breed and gender did not affect the risk of an infection in any of the study populations. Toxocara and Giardia present a zoonotic risk, especially in household dogs, where the majority of Giardia positive samples (80%) belonged to the zoonotic assemblage A. In kennel dogs and clinically affected dogs the host-specific Giardia assemblages C and D were most prevalent (94% and 80%, respectively).

  12. Determination of the recovery efficiency of cryptosporidium oocysts and giardia cysts from seeded bivalve mollusks.

    Science.gov (United States)

    Schets, Franciska M; van den Berg, Harold H J L; de Roda Husman, Ana Maria

    2013-01-01

    The intestinal parasites Cryptosporidium and Giardia are transmitted by water and food and cause human gastroenteritis. Filter-feeding bivalve mollusks, such as oysters and mussels, filter large volumes of water and thus concentrate such pathogens, which makes these bivalves potential vectors of disease. To assess the risk of infection from consumption of contaminated bivalves, parasite numbers and parasite recovery data are required. A modified immunomagnetic separation (IMS) procedure was used to determine Cryptosporidium oocyst and Giardia cyst numbers in individually homogenized oysters (Crassostrea gigas) and mussels (Mytilus edulis). About 12% of the commercial bivalves were positive, with low (oo)cyst numbers per specimen. The recovery efficiency of the IMS procedure was systematically evaluated. Experiments included seeding of homogenized bivalves and whole animals with 100 to 1,000 (oo)cysts. Both seeding procedures yielded highly variable recovery rates. Median Cryptosporidium recoveries were 7.9 to 21% in oysters and 62% in mussels. Median Giardia recoveries were 10 to 25% in oysters and 110% in mussels. Giardia recovery was significantly higher than Cryptosporidium recovery. (Oo)cysts were less efficiently recovered from seeded whole animals than from seeded homogenates, with median Cryptosporidium recoveries of 5.3% in oysters and 45% in mussels and median Giardia recoveries of 4.0% in oysters and 82% in mussels. Both bivalve homogenate seeding and whole animal seeding yielded higher (oo)cyst recovery in mussels than in oysters, likely because of the presence of less shellfish tissue in IMS when analyzing the smaller mussels compared with the larger oysters, resulting in more efficient (oo)cyst extraction. The data generated in this study may be used in the quantitative assessment of the risk of infection with Cryptosporidium or Giardia associated with the consumption of raw bivalve mollusks. This information may be used for making risk management

  13. Cryptosporidium spp. and Giardia sp. in aquatic mammals in northern and northeastern Brazil.

    Science.gov (United States)

    Borges, João Carlos; Lima, Danielle Dos; da Silva, Edson Moura; Moreira, André Lucas de Oliveira; Marmontel, Miriam; Carvalho, Vitor Luz; Amaral, Rodrigo de; Lazzarini, Stella Maris; Alves, Leucio Câmara

    2017-09-20

    Cryptosporidium and Giardia are protozoans that can infect humans and wild and domestic animals. Due to the growing importance of diseases caused by protozoan parasites in aquatic species, we aimed to evaluate the frequency of infection by Cryptosporidium spp. and Giardia sp. in aquatic and marine mammals in the northern and northeastern regions of Brazil. We collected 553 fecal samples from 15 species of wild-ranging and captive aquatic mammals in northern and northeastern Brazil. All samples were analyzed by the Kinyoun technique for identification of Cryptosporidium spp. oocysts. Giardia sp. cysts were identified by means of the centrifugal-flotation technique in zinc sulfate solution. Subsequently, all samples were submitted for direct immunofluorescence testing. The overall frequency of infection was 15.55% (86/553) for Cryptosporidium spp. and 9.04% (50/553) for Giardia sp. The presence of Cryptosporidium spp. was detected in samples from 5 species: neotropical river otter Lontra longicaudis (15.28%), giant otter Pteronura brasiliensis (41.66%), Guiana dolphin Sotalia guianensis (9.67%), Amazonian manatee Trichechus inunguis (16.03%), and Antillean manatee T. manatus (13.79%). Giardia sp. was identified in L. longicaudis (9.23%), P. brasiliensis (29.16%), pygmy sperm whale Kogia breviceps (100%), dwarf sperm whale K. sima (25%), S. guianensis (9.67%), T. inunguis (3.81%), and T. manatus (10.34%). This is the first report of Cryptosporidium spp. in L. longicaudis, P. brasiliensis, and S. guianensis, while the occurrence of Giardia sp., in addition to the 2 otter species, was also identified in manatees, thus extending the number of hosts susceptible to these parasitic agents.

  14. CONTRIBUTIONS TO THE KNOWLEDGE OF GIARDIA INTESTINALIS PARASITE IN THE ARGES POPULATION

    Directory of Open Access Journals (Sweden)

    Daniela Bărbuceanu

    2013-07-01

    Full Text Available The present study, conducted in January-December 2007-2010 to the Argeş population releaved a high incidence of parazitization with Giardia intestinalis to the preschool children: 38.9%. There are not significant differences related to gender distribution of parasitization with G. intestinalis, and most cases come from urban areas. This study seeks to highlight the socio-medical importance of this pathology to Arges population, because infestation with Giardia intestinalis is the most common cause of acute and chronic diarrhea in children and in adults, too.

  15. Review of cryptosporidium and giardia in the eastern part of Europe, 2016

    DEFF Research Database (Denmark)

    Plutzer, Judit; Lassen, Brian; Jokelainen, Pikka

    2018-01-01

    Introduction: This paper reviews the current knowledge and understanding of Cryptosporidium spp. an d Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia...... and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results: Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices...

  16. New filtration system for efficient recovery of waterborne Cryptosporidium oocysts and Giardia cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, J. A.; Riber, Ulla

    2015-01-01

    AimsTo develop a filtration unit for efficient recovery of waterborne Cryptosporidium oocysts and Giardia cysts ((oo-)cysts) in drinking water.Methods and ResultsThis unit utilizes a metallic filter and an ultrasound transducer for eluting (oo-)cysts, with a fixed retentate backwash volume; approx...... was assessed by seeding ColorSeed (10 replicates) into the filter unit followed by air backwash to a glass slide and counting of (oo-)cysts by epifluorescent microscopy. High recovery rates (meanSD) were found: 849%48 for Giardia cysts and 70%+/- 65 for Cryptosporidium oocysts. DNA of seeded wild type (oo...

  17. Assay of partially purified glutamate dehydrogenase isolated from ...

    African Journals Online (AJOL)

    Glutamate dehydrogenase (E C 1.4.1.1) isolated from the seeds of asparagus beans was partially purified to a factor of 22 by dialysis after fractional precipitation with solid ammonium sulphate at 40 and 60% saturation. A specific activity of 11.78μmol min-1 mg-1 protein was calculated for the partially purified enzyme when ...

  18. Isoforms of purified methyltransferase from human blood platelets ...

    African Journals Online (AJOL)

    ... purification from normal human blood platelets have not been investigated, hence, the aim of this study was to purify, characterise the enzyme from human blood platelets and determine its possible role in phospholipid transmethylation. The plasma membranes were purified by velocity and sucrose gradient centrifugation ...

  19. Glucoamylase isoform (GAII) purified from a thermophilic fungus ...

    African Journals Online (AJOL)

    Scytalidium thermophilum 15.8 produced two extracellular glucoamylases. Using a DEAE-Cellulose chromatographic column glucoamylases form II (GAII) was separated and purified from glucoamylases form I (GAI) that was previously purified and characterised (Cereia et al., 2000) when the filtrate of the culture medium ...

  20. Reproducible in vitro regeneration system for purifying sugarcane ...

    African Journals Online (AJOL)

    This procedure may be considered as one of the best ever published report on regeneration from in vitro grown plants to purify clones without subjecting the plants to field conditions and harvesting the mature cane. This technique was used to purify transgenic sugarcane plants carrying Bacillus thuringiensis gene.

  1. Respirators: Air Purifying, Self-Study, Course 40723

    Energy Technology Data Exchange (ETDEWEB)

    Chochoms, Michael [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-12-21

    Respirators: Air Purifying Self-Study (COURSE 40723) is designed for Los Alamos National Laboratory (LANL) workers, support services subcontractors, and other LANL subcontractors who work under the LANL Respiratory Protection Program (RPP). This course also meets the air-purifying respirators (APRs) retraining requirement.

  2. Partially purified polygalacturonase from Aspergillus niger (SA6 ...

    African Journals Online (AJOL)

    Polygalacturonase (PG) was isolated from Aspergillus niger (A. niger) (SA6), partially purified and characterized. The PG showed two bands on SDS-PAGE suggesting an “endo and exo PG with apparent molecular weights of 35 and 40 KDa, respectively. It was purified 9-fold with a yield of 0.18% and specific activity of 246 ...

  3. Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs

    DEFF Research Database (Denmark)

    Rimhanen-Finne, R.; Enemark, Heidi L.; Kolehmainen, J.

    2007-01-01

    The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5) oocysts/g, while the detection limit for Giardia ELISA was 10(4) cysts/g. The Cryptosporidium ELISA showed 94% specificity...... zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs...

  4. Phosphonoxins: Rational design and discovery of a potent nucleotide anti-Giardia agent

    Czech Academy of Sciences Publication Activity Database

    Suk, D. H.; Rejman, Dominik; Dykstra, Ch.; Pohl, Radek; Pankiewicz, K. W.; Patterson, S. E.

    2007-01-01

    Roč. 17, č. 10 (2007), s. 2811-2816 ISSN 0960-894X Institutional research plan: CEZ:AV0Z40550506 Keywords : Giardia * encystation * cyst wall synthase * nucleotide Subject RIV: CC - Organic Chemistry Impact factor: 2.604, year: 2007

  5. Batch solar disinfection inactivates oocysts of Cryptosporidium parvum and cysts of Giardia muris in drinking water.

    Science.gov (United States)

    McGuigan, K G; Méndez-Hermida, F; Castro-Hermida, J A; Ares-Mazás, E; Kehoe, S C; Boyle, M; Sichel, C; Fernández-Ibáñez, P; Meyer, B P; Ramalingham, S; Meyer, E A

    2006-08-01

    To determine whether batch solar disinfection (SODIS) can be used to inactivate oocysts of Cryptosporidium parvum and cysts of Giardia muris in experimentally contaminated water. Suspensions of oocysts and cysts were exposed to simulated global solar irradiation of 830 W m(-2) for different exposure times at a constant temperature of 40 degrees C. Infectivity tests were carried out using CD-1 suckling mice in the Cryptosporidium experiments and newly weaned CD-1 mice in the Giardia experiments. Exposure times of > or =10 h (total optical dose c. 30 kJ) rendered C. parvum oocysts noninfective. Giardia muris cysts were rendered completely noninfective within 4 h (total optical dose >12 kJ). Scanning electron microscopy and viability (4',6-diamidino-2-phenylindole/propidium iodide fluorogenic dyes and excystation) studies on oocysts of C. parvum suggest that inactivation is caused by damage to the oocyst wall. Results show that cysts of G. muris and oocysts of C. parvum are rendered completely noninfective after batch SODIS exposures of 4 and 10 h (respectively) and is also likely to be effective against waterborne cysts of Giardia lamblia. These results demonstrate that SODIS is an appropriate household water treatment technology for use as an emergency intervention in aftermath of natural or man-made disasters against not only bacterial but also protozoan pathogens.

  6. Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in brazilian captive birds

    Science.gov (United States)

    A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, 3 birds were found positive f...

  7. Het rendement van de detectiemethode voor Cryptosporidium en Giardia in water

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Schijven JF; MGB

    2004-01-01

    Nederlandse waterleidingbedrijven zijn verplicht om te berekenen of als gevolg van consumptie van drinkwater infectie met Cryptosporidium of Giardia kan optreden. De kans hierop moet kleiner dan een infectie per 10000 personen per jaar zijn. De berekening (risicoanalyse) wordt gebaseerd op de

  8. Modellering van de verwijdering van Cryptosporidium parvum,Giardia lamblia en enterovirussen in spaarbekkens

    NARCIS (Netherlands)

    Ratsak CH; MGB

    1996-01-01

    De protozoa Cryptosporidium en Giardia hebben de laatste decennia in de Verenigde Staten en in Groot-Brittannie een aanzienlijk aantal grote en kleine epidemieen van maag-darminfecties via drinkwater veroorzaakt. Wiskundige modellen kunnen worden gebruikt om de verwijdering van pathogene

  9. Curcumin alters the cytoskeleton and microtubule organization on trophozoites of Giardia lamblia.

    Science.gov (United States)

    Gutiérrez-Gutiérrez, Filiberto; Palomo-Ligas, Lissethe; Hernández-Hernández, José Manuel; Pérez-Rangel, Armando; Aguayo-Ortiz, Rodrigo; Hernández-Campos, Alicia; Castillo, Rafael; González-Pozos, Sirenia; Cortés-Zárate, Rafael; Ramírez-Herrera, Mario Alberto; Mendoza-Magaña, María Luisa; Castillo-Romero, Araceli

    2017-08-01

    Giardia lamblia is a worldwide protozoan responsible for a significant number of intestinal infections. There are several drugs for the treatment of giardiasis, but they often cause side effects. Curcumin, a component of turmeric, has antigiardial activity; however, the molecular target and mechanism of antiproliferative activity are not clear. The effects of curcumin on cellular microtubules have been widely investigated. Since tubulin is the most abundant protein in the cytoskeleton of Giardia, to elucidate whether curcumin has activity against the microtubules of this parasite, we treated trophozoites with curcumin and the cells were analyzed by scanning electron microscopy and confocal microscopy. Curcumin inhibited Giardia proliferation and adhesion in a time-concentration-dependent mode. The higher inhibitory concentrations of curcumin (3 and 15μM) disrupted the cytoskeletal structures of trophozoites; the damage was evident on the ventral disk, flagella and in the caudal region, also the membrane was affected. The immunofluorescence images showed altered distribution of tubulin staining on ventral disk and flagella. Additionally, we found that curcumin caused a clear reduction of tubulin expression. By docking analysis and molecular dynamics we showed that curcumin has a high probability to bind at the interface of the tubulin dimer close to the vinblastine binding site. All the data presented indicate that curcumin may inhibit Giardia proliferation by perturbing microtubules. Copyright © 2017. Published by Elsevier B.V.

  10. Prevalence and Associated Risk Factors of Giardia Infection among Indigenous Communities in Rural Malaysia

    Science.gov (United States)

    Choy, Seow Huey; Al-Mekhlafi, Hesham M.; Mahdy, Mohammed A. K.; Nasr, Nabil N.; Sulaiman, Maria; Lim, Yvonne A. L.; Surin, Johari

    2014-01-01

    This study was carried out to investigate the prevalence and risk factors of Giardia infection among indigenous people in rural Malaysia. Faecal samples were collected from 1,330 participants from seven states of Malaysia and examined by wet mount and formalin-ether sedimentation methods while demographic, socioeconomic and environmental information was collected using a pre-tested questionnaire. The overall prevalence of Giardia infection was 11.6% and was significantly higher among those aged ≤ 12 years compared to their older counterparts. Multivariate logistic regression identified age of ≤12 years, lacking of toilet at household, not washing hands before eating, not washing hands after playing with animals, not boiling water before consumption, bathing in the river, and not wearing shoes when outside as the significant risk factors of Giardia infection among these communities. Based on a multilocus genotyping approach (including tpi, gdh and bg gene sequences), 69 isolates were identified as assemblage A, and 69 as assemblage B. No association between the assemblages and presence of symptoms was found. Providing proper sanitation, as well as provision of clean drinking water and proper health education regarding good personal hygiene practices will help significantly in reducing the prevalence and burden of Giardia infection in these communities. PMID:25366301

  11. Viability staining of Cryptosporidium oocysts and Giardia cysts combined with flow cytometry

    NARCIS (Netherlands)

    Schets FM; Medema GJ; MGB

    1998-01-01

    The incorporation of flow cytometry as an additional purification step has improved the detection method for Cryptosporidium oocysts and Giardia cysts in water. Flow cytometry allows separation of (oo)cysts from interfering debris particles present in water concentrates and thus enables the

  12. Het rendement van de detectiemethode voor Cryptosporidium en Giardia in water

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Schijven JF; MGB

    2004-01-01

    Dutch drinking water legislation requires drinking water companies to perform a quantitative risk assessment for Cryptosporidium and Giardia. The risk of infection through consumption of drinking water should be below one infection in 10,000 persons per year. Risk assessment is based on the number

  13. Small-intestinal factors promote encystation of Giardia lamblia in vitro.

    OpenAIRE

    Gillin, F D; Reiner, D S; Boucher, S E

    1988-01-01

    Bile salts and fatty acids stimulated differentiation of cultured Giardia lamblia trophozoites into water-resistant cysts at the slightly alkaline pH of the small intestinal lumen. Maximum encystation occurred at pH 7.8. Thus, specific small-intestinal factors may influence encystation in vivo as well as in vitro.

  14. Cytology Preparations of Formalin Fixative Aid Detection of Giardia in Duodenal Biopsy Samples.

    Science.gov (United States)

    Panarelli, Nicole C; Gobara, Nariman; Hoda, Rana S; Chaump, Michael; Jessurun, Jose; Yantiss, Rhonda K

    2017-04-01

    Giardiasis is the most common intestinal parasitic infection in the United States. The organism elicits no, or minimal, inflammatory changes in duodenal biopsy samples, so it can be easily overlooked. We performed this study to determine whether Giardia could be isolated from the formalin fixative of biopsy samples, and to evaluate the value of fluid analysis in the assessment for potential infection. We prospectively evaluated duodenal biopsy samples from 92 patients with a clinical suspicion of giardiasis or symptoms compatible with that diagnosis (ie, diarrhea, bloating, or abdominal pain) Biopsy samples were routinely processed and stained with hematoxylin and eosin. Histologic diagnoses included giardiasis (5 cases, 4%), normal findings (64 cases, 70%), peptic injury/active duodenitis (12 cases, 13%), and intraepithelial lymphocytosis with villous blunting (10 cases, 12%). Fifteen cases (13%) showed detached degenerated epithelial cells or mucus droplets in the intervillous space that resembled Giardia. Cytology slides were prepared from formalin in the biopsy container using the standard Cytospin protocol and reviewed by a cytopathologist blinded to the biopsy findings. Cytologic evaluation revealed Giardia spp. in all 5 biopsy-proven cases, and identified an additional case that was not detected by biopsy analysis. Organisms were significantly more numerous (mean: 400 trophozoites; range, 120 to 810) and showed better morphologic features in cytology preparations compared with tissue sections (mean: 129 trophozoites; range, 37 to 253 organisms; P=0.05). Our findings suggest that cytology preparations from formalin fixative can resolve diagnostically challenging cases and even enhance Giardia detection in some cases.

  15. Structure of the prolyl-tRNA synthetase from the eukaryotic pathogen Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Larson, Eric T.; Kim, Jessica E.; Napuli, Alberto J.; Verlinde, Christophe L. M. J.; Fan, Erkang; Zucker, Frank H.; Van Voorhis, Wesley C.; Buckner, Frederick S.; Hol, Wim G. J.; Merritt, Ethan A., E-mail: merritt@u.washington.edu [Medical Structural Genomics of Pathogenic Protozoa, (United States); University of Washington, Seattle, WA 98195 (United States)

    2012-09-01

    The structure of Giardia prolyl-tRNA synthetase cocrystallized with proline and ATP shows evidence for half-of-the-sites activity, leading to a corresponding mixture of reaction substrates and product (prolyl-AMP) in the two active sites of the dimer. The genome of the human intestinal parasite Giardia lamblia contains only a single aminoacyl-tRNA synthetase gene for each amino acid. The Giardia prolyl-tRNA synthetase gene product was originally misidentified as a dual-specificity Pro/Cys enzyme, in part owing to its unexpectedly high off-target activation of cysteine, but is now believed to be a normal representative of the class of archaeal/eukaryotic prolyl-tRNA synthetases. The 2.2 Å resolution crystal structure of the G. lamblia enzyme presented here is thus the first structure determination of a prolyl-tRNA synthetase from a eukaryote. The relative occupancies of substrate (proline) and product (prolyl-AMP) in the active site are consistent with half-of-the-sites reactivity, as is the observed biphasic thermal denaturation curve for the protein in the presence of proline and MgATP. However, no corresponding induced asymmetry is evident in the structure of the protein. No thermal stabilization is observed in the presence of cysteine and ATP. The implied low affinity for the off-target activation product cysteinyl-AMP suggests that translational fidelity in Giardia is aided by the rapid release of misactivated cysteine.

  16. MATRIX ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY BASED ANALYSIS OF GIARDIA LAMBLIA

    Science.gov (United States)

    Giardia lamblia is a zoonotic protozoan parasite that is a leading cause of drinking water related gastro-intestinal disease outbreaks worldwide. Due to the genotypic complexity and high prevalence of this parasite in the environment, numerous research studies are being done to ...

  17. Adaptive Immunity-Dependent Intestinal Hypermotility Contributes to Host Defense against Giardia spp.

    OpenAIRE

    Andersen, Yolanda S.; Gillin, Frances D.; Eckmann, Lars

    2006-01-01

    Humans infected with Giardia exhibit intestinal hypermotility, but the underlying mechanisms and functional significance are uncertain. Here we show in murine models of giardiasis that small-intestinal hypermotility occurs in a delayed fashion relative to peak parasite burden, is dependent on adaptive immune defenses, and contributes to giardial clearance.

  18. Structure of the prolyl-tRNA synthetase from the eukaryotic pathogen Giardia lamblia

    International Nuclear Information System (INIS)

    Larson, Eric T.; Kim, Jessica E.; Napuli, Alberto J.; Verlinde, Christophe L. M. J.; Fan, Erkang; Zucker, Frank H.; Van Voorhis, Wesley C.; Buckner, Frederick S.; Hol, Wim G. J.; Merritt, Ethan A.

    2012-01-01

    The structure of Giardia prolyl-tRNA synthetase cocrystallized with proline and ATP shows evidence for half-of-the-sites activity, leading to a corresponding mixture of reaction substrates and product (prolyl-AMP) in the two active sites of the dimer. The genome of the human intestinal parasite Giardia lamblia contains only a single aminoacyl-tRNA synthetase gene for each amino acid. The Giardia prolyl-tRNA synthetase gene product was originally misidentified as a dual-specificity Pro/Cys enzyme, in part owing to its unexpectedly high off-target activation of cysteine, but is now believed to be a normal representative of the class of archaeal/eukaryotic prolyl-tRNA synthetases. The 2.2 Å resolution crystal structure of the G. lamblia enzyme presented here is thus the first structure determination of a prolyl-tRNA synthetase from a eukaryote. The relative occupancies of substrate (proline) and product (prolyl-AMP) in the active site are consistent with half-of-the-sites reactivity, as is the observed biphasic thermal denaturation curve for the protein in the presence of proline and MgATP. However, no corresponding induced asymmetry is evident in the structure of the protein. No thermal stabilization is observed in the presence of cysteine and ATP. The implied low affinity for the off-target activation product cysteinyl-AMP suggests that translational fidelity in Giardia is aided by the rapid release of misactivated cysteine

  19. Human Secretory Immune Response to Fatty Acid-Binding Protein Fraction from Giardia lamblia

    Science.gov (United States)

    Hasan, S. M. T.; Maachee, M.; Córdova, O. M.; Diaz de la Guardia, R.; Martins, M.; Osuna, A.

    2002-01-01

    The secretory immune response in humans infected with Giardia lamblia was studied by using saliva samples and an 8-kDa antigen capable of binding fatty acids. This antigen was not recognized by saliva samples from healthy individuals. The antigen may be useful in diagnostic studies of G. lamblia infection. PMID:11895992

  20. Anti-giardia activity of hexane extract of Citrus aurantifolia (Christim ...

    African Journals Online (AJOL)

    Background: Giardia lamblia is a parasite that causes giardiasis in humans and other mammals. The common treatment includes different drugs, which were described to produce unpleasant side effects. Citrus aurantifolia, popularly known as “lima”, is a plant used in traditional medicine to treat gastrointestinal symptoms.

  1. Human Secretory Immune Response to Fatty Acid-Binding Protein Fraction from Giardia lamblia

    OpenAIRE

    Hasan, S. M. T.; Maachee, M.; Córdova, O. M.; Diaz de la Guardia, R.; Martins, M.; Osuna, A.

    2002-01-01

    The secretory immune response in humans infected with Giardia lamblia was studied by using saliva samples and an 8-kDa antigen capable of binding fatty acids. This antigen was not recognized by saliva samples from healthy individuals. The antigen may be useful in diagnostic studies of G. lamblia infection.

  2. Consistency of direct microscopic examination and ELISA in detection of Giardia in stool specimen among children

    Directory of Open Access Journals (Sweden)

    Zohreh Torabi

    2014-09-01

    Full Text Available Objective: To investigate the consistency of direct microscopic examination and ELISA for determination of Giadia in stool specimen. Method: Study population consisted of children with any clinical symptoms of Giardia infestation since last two weeks. Fresh stool specimen was collected from each child. The stools specimens were assessed by two methods of direct microscopic examination and ELISA.The degree of agreement between direct stool exam and ELISA was calculated by Cohen's kappa coefficient. Results: In this study, 124 children with age range 2-12 years were investigated. A total of 64 (61.7% and 79 (65.7% of children had Giardia by direct stool exam and ELISA test respectively. There was association between frequency of constipation and Giardia infection (P=0.036. The Cohen's kappa coefficient calculated for degree of agreement between direct stool exam and ELISA showed κ=0.756 (P<0.001. Conclusions: The frequency of Giardia infection in symptomatic children was high and there was high agreement rate between ELISA and direct stool smear.

  3. Giardia in symptomatic dogs and cats in Europe--results of a European study.

    Science.gov (United States)

    Epe, C; Rehkter, G; Schnieder, T; Lorentzen, L; Kreienbrock, L

    2010-10-11

    The percentage of Giardia infection in dogs and cats with gastrointestinal signs presenting to clinics was examined across Europe using the IDEXX SNAP Giardia Test (IDEXX Laboratories). Veterinary practices were asked to identify diarrheic and/or vomiting dogs and cats and to use the test on fecal samples from these animals. A selection for "asymptomatic" testing was also available on the data collection sheet for testing that occurred outside the target "symptomatic" population. Results from 8685 dogs and 4214 cats were submitted during the sampling period from 2005 to 2006. Analysis of the data showed 24.78% of the samples positive for Giardia among tested dogs and 20.3% among tested cats. The younger the dog or the cat, the higher the risk of being positive, peaking in the category below 6 months. The main clinical symptom, diarrhoea, also showed a higher risk of being positive, OR's of acute diarrhoea were almost double of those of the category "asymptomatic" for dogs or cats, the same range was calculated for chronic diarrhoea. Differences in participating countries were also seen. The risk of being positive was half or less in UK, Spain, Netherlands, Italy compared to Germany, and only in Belgium it showed to be higher. The results of this study show that Giardia is a common enteric agent among dogs and cats with gastrointestinal signs in Europe. Copyright 2010 Elsevier B.V. All rights reserved.

  4. Evaluation of an inhouse rapid ELISA test for detection of giardia in domestic sheep (Ovis aries).

    Science.gov (United States)

    Wilson, Jolaine M; Hankenson, F Claire

    2010-11-01

    Sheep (Ovis aries) are increasingly used at our institution as models of human disease. Within the research environment, routine husbandry and handling of sheep has potential for transmission of zoonotic agents, including Giardia. The prevalence of Giardia in sheep may approach 68%. Classic diagnostic testing involves microscopic examination for fecal cysts or trophozoites; however, limitations of microscopy include time, labor, and potential false-negative results due to intermittent shedding. We wished to determine whether a commercial rapid ELISA used for Giardia detection in dogs and cats could be used in sheep. Fecal samples collected from sheep (n = 93) were tested with a combination of 6 methods: reference laboratory fecal flotation, reference laboratory ELISA, inhouse fecal flotation, and commercially available tests (enzyme immunoassay, direct fluorescence antibody assay, and rapid ELISA). Prevalence of Giardia infection in facility sheep was 11.8% (11 of 93 animals). Of the 11 samples considered positive, 3 were confirmed by multiple testing methods, and 5 were positive by microscopy alone. Inhouse fecal flotation for 8 samples was positive on only 1 of 2 consecutive testing days. The rapid ELISA test exhibited 0% sensitivity for sheep giardiasis. Overall, the examined methods had low sensitivities and low positive predictive values. Despite limitations, microscopic analysis of repeat fecal samples remained the most accurate diagnostic method for ovine giardiasis among the methods tested.

  5. Prevalence and associated risk factors of Giardia infection among indigenous communities in rural Malaysia.

    Science.gov (United States)

    Choy, Seow Huey; Al-Mekhlafi, Hesham M; Mahdy, Mohammed A K; Nasr, Nabil N; Sulaiman, Maria; Lim, Yvonne A L; Surin, Johari

    2014-11-04

    This study was carried out to investigate the prevalence and risk factors of Giardia infection among indigenous people in rural Malaysia. Faecal samples were collected from 1,330 participants from seven states of Malaysia and examined by wet mount and formalin-ether sedimentation methods while demographic, socioeconomic and environmental information was collected using a pre-tested questionnaire. The overall prevalence of Giardia infection was 11.6% and was significantly higher among those aged ≤ 12 years compared to their older counterparts. Multivariate logistic regression identified age of ≤ 12 years, lacking of toilet at household, not washing hands before eating, not washing hands after playing with animals, not boiling water before consumption, bathing in the river, and not wearing shoes when outside as the significant risk factors of Giardia infection among these communities. Based on a multilocus genotyping approach (including tpi, gdh and bg gene sequences), 69 isolates were identified as assemblage A, and 69 as assemblage B. No association between the assemblages and presence of symptoms was found. Providing proper sanitation, as well as provision of clean drinking water and proper health education regarding good personal hygiene practices will help significantly in reducing the prevalence and burden of Giardia infection in these communities.

  6. NIGERIAN VETERINARY JOURNAL

    African Journals Online (AJOL)

    ADEYEYE

    Giardiasis is a common intestinal infection caused by single-celled flagellate protozoan,. Giardia duodenalis (syn. ... common cause of foodborne and waterborne gastroenteritis (Castro-Hermida et al.,. 2007). Giardia duodenalis ... prophylaxis with anthelmintic, antibiotic, anti-protozoan and acaricidal drugs were performed.

  7. Current status and future trends in Cryptosporidium and Giardia epidemiology in Malaysia.

    Science.gov (United States)

    Lim, Y A L; Ahmad, R A; Smith, H V

    2008-06-01

    Cryptosporidium and Giardia are major causes of diarrhoeal diseases of humans worldwide, and are included in the World Health Organisation's 'Neglected Diseases Initiative'. Cryptosporidium and Giardia occur commonly in Malaysian human and non-human populations, but their impact on disease, morbidity and cost of illness is not known. The commonness of contributions from human (STW effluents, indiscriminate defaecation) and non-human (calving, lambing, muck spreading, slurry spraying, pasturing/grazing of domestic animals, infected wild animals) hosts indicate that many Malaysian environments, particularly water and soil, are sufficiently contaminated to act as potential vehicles for the transmission of disease. To gain insight into the morbidity and mortality caused by human cryptosporidiosis and giardiasis, they should be included into differential diagnoses, and routine laboratory testing should be performed and (as for many infectious diseases) reported to a centralised public health agency. To understand transmission routes and the significance of environmental contamination better will require further multidisciplinary approaches and shared resources, including raising national perceptions of the parasitological quality of drinking water. Here, the detection of Cryptosporidium and Giardia should be an integral part of the water quality requirement. A multidisciplinary approach among public health professionals in the water industry and other relevant health- and environment-associated agencies is also required in order to determine the significance of Cryptosporidium and Giardia contamination of Malaysian drinking water. Lastly, adoption of validated methods to determine the species, genotype and subgenotype of Cryptosporidium and Giardia present in Malaysia will assist in developing effective risk assessment, management and communication models.

  8. Polymeric immunoglobulin receptor in intestinal immune defense against the lumen-dwelling protozoan parasite Giardia.

    Science.gov (United States)

    Davids, Barbara J; Palm, J E Daniel; Housley, Michael P; Smith, Jennifer R; Andersen, Yolanda S; Martin, Martin G; Hendrickson, Barbara A; Johansen, Finn-Eirik; Svärd, Staffan G; Gillin, Frances D; Eckmann, Lars

    2006-11-01

    The polymeric Ig receptor (pIgR) is conserved in mammals and has an avian homologue, suggesting evolutionarily important functions in vertebrates. It transports multimeric IgA and IgM across polarized epithelia and is highly expressed in the intestine, yet little direct evidence exists for its importance in defense against common enteric pathogens. In this study, we demonstrate that pIgR can play a critical role in intestinal defense against the lumen-dwelling protozoan parasite Giardia, a leading cause of diarrheal disease. The receptor was essential for the eradication of Giardia when high luminal IgA levels were required. Clearance of Giardia muris, in which IgA plays a dominant role, was severely compromised in pIgR-deficient mice despite significant fecal IgA output at 10% of normal levels. In contrast, eradication of the human strain Giardia lamblia GS/M, for which adaptive immunity is less IgA dependent in mice, was unaffected by pIgR deficiency, indicating that pIgR had no physiologic role when lower luminal IgA levels were sufficient for parasite elimination. Immune IgA was greatly increased in the serum of pIgR-deficient mice, conferred passive protection against Giardia, and recognized several conserved giardial Ags, including ornithine carbamoyltransferase, arginine deiminase, alpha-enolase, and alpha- and beta-giardins, that are also detected in human giardiasis. Corroborative observations were made in mice lacking the J chain, which is required for pIgR-dependent transepithelial IgA transport. These results, together with prior data on pIgR-mediated immune neutralization of luminal cholera toxin, suggest that pIgR is essential in intestinal defense against pathogenic microbes with high-level and persistent luminal presence.

  9. Molecular seasonality of Giardia lamblia in a cohort of Egyptian children: a circannual pattern.

    Science.gov (United States)

    Ismail, Mousa A M; El-Akkad, Dina M H; Rizk, Enas M A; El-Askary, Hala M; El-Badry, Ayman A

    2016-11-01

    Giardia lamblia (G. lamblia) is the most worldwide prevailing intestinal parasite, notorious for its broad range of seasonal and age-related prevalence. The potentially lethal nature of giardiasis makes it essential that the seasonality, the groups at risk, and other potential risk factors are identified. The present molecular epidemiological study was designed to determine the genetic diversity of G. lamblia infection, taking into account seasonal peaks, age distribution, and associated symptoms in a cohort of Egyptian diarrheic patients. Stool samples were collected from 1187 diarrheic patients attending outpatient clinics of Cairo University hospitals, of all age groups over a 12-month period. The patients were examined microscopically for fecal G. lamblia cysts, and/or trophozoites, and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays targeting beta giardin gene. PCR-positive samples were characterized molecularly by nPCR restriction fragment length polymorphism (RFLP) to determine Giardia assemblages. The findings revealed circannual prevalence of Giardia, with a seasonal pattern peaking in mid-summer and late winter, with the summer peak preceded by a peak in temperature. Infection was prevailing in 224 (18.9 %) cases, mainly assemblage B (81.2 %) followed by assemblage A (18.8 %). There were statistically significant associations between the detection of Giardia and flatulence, persistent diarrhea, vomiting, and abdominal pain, while gender and intermittent diarrhea showed no association. The pre-school age group was the most vulnerable. This is the first study of molecular characterization of Giardia to determine its circannual prevalence in Egypt, a finding which carries promising potential for the diagnosis, treatment, and elimination of the disease.

  10. Immunodiffusion Studies of Purified Equine Infectious Anemia Virus

    Science.gov (United States)

    Nakajima, Hideo; Ushimi, Chuzo

    1971-01-01

    Antigenicity of purified equine infectious anemia (EIA) virus was examined by immunodiffusion against sera obtained from horses experimentally infected with EIA virus. The purified virus reacted with the infected horse serum, and virus-specific precipitating antibody was demonstrated. Furthermore, it was found that purified EIA virus reacted against the serum of horses infected with all strains of EIA virus which were antigenically different from one another. From the result, group-specific components of the virus rather than strain-specific ones were considered to be involved in the reaction. Serological reactivity was lost by adding antiserum from the infected horse to the antigen. The precipitating antibody usually appeared in the serum 1 to 2 weeks after the first febrile attack of EIA and remained for a longer period. Some characteristics of the purified antigen and specificity of the reaction for EIA are described. Images PMID:16557982

  11. Detection of Cryptosporidium oocysts and Giardia cysts in water samples with a Becton Dickinson FACSort flow cytometer

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Boschman GD; LWL; Becton Dickinson Europe, Aalst, Belgie

    1995-01-01

    Current detection techniques for Cryptosporidium oocysts and Giardia cysts in water samples combine filtration of large volumes of water, concentration by centrifugation and flotation and immunofluorescense microscopy. The techniques are extremely labour-intensive and inefficient. The various steps

  12. A light microscopic study of the course of cellular division of Giardia intestinalis trophozoites grown in vitro.

    Science.gov (United States)

    Cerva, L; Nohýnková, E

    1992-01-01

    The cellular division of trophozoites of Giardia intestinalis group was followed microscopically in stained preparations from axenic cultures. Eleven successive stages were described, documented by photomicrographs and their survival times calculated.

  13. First report of Giardia sp. and Cryptosporidium sp. in fecal samples in blue macaw (Anodorhynchus hyacinthinus) in southern Brazil

    OpenAIRE

    Matheus Hillard Farret; Vinicius da Rosa Fanfa; Luisa Ragagnin; Aleksandro Schafer da Silva; Silvia Gonzalez Monteiro

    2010-01-01

    This study reports the gastrointestinal parasitism by Giardia sp. and Cryptosporidium sp. in blue macaw (Anodorhynchus hyacinthinus) in the southern region of Brazil. Fecal samples of two species kept in captivity in the state of Rio Grande do Sul were analyzed by the direct smear method, centrifugal flotation technique with zinc sulfate and Kinyoun staining technique for research of parasites. Mixed infection by eggs of Capillaria, cysts of Giardia sp. and oocysts of Cryptosporidium sp. was ...

  14. Modeling long-term host cell-Giardia lamblia interactions in an in vitro co-culture system.

    Science.gov (United States)

    Fisher, Bridget S; Estraño, Carlos E; Cole, Judith A

    2013-01-01

    Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite, Giardia lamblia, is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium, Giardia induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of Giardia pathology is the lack of a functionally relevant model for the long-term study of the parasite in vitro. Here we report on the development of an in vitro co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of Giardia trophozoites. Using the developed model, we show that Giardia trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and Giardia medium. Giardia induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by Giardia indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

  15. Determination of the level of parasitic infection (Cryptosporidium and Giardia of the vegetables marketed in Ilam city

    Directory of Open Access Journals (Sweden)

    Moyad Avazpoor

    2015-01-01

    Full Text Available Background: Infected with intestinal parasites is one of the most important health and economical problems, which could have different effects, such as diarrheal diseases or death associated. The purpose of this study was to determine the level of prevalence of Cryptosporidium and Giardia parasites in the vegetable marketed in Ilam city. Methods: This study was performed on 280 samples of fresh vegetables and lettuce in Ilam. The samples were taken at the level of 500 grams from the places where vegetables and lettuce are sold. Micro liters of each sample was placed on the slide using automatic micropipette, and Logel and Zyl-Nelson stainings were performed in order to identify Cryptosporidium and Giardia. Results: From 200 samples, 54 samples were contaminated to Cryptosporidium oocyte and 13 samples to Giardia cysts. From 80 lettuce samples also 32 samples were contaminated to Cryptosporidium oocyte, and 6 samples contaminated to Giardia cysts. The results showed that the overall infection was 37%. Infection with Giardia cysts was 6.8% and infection with Cryptosporidium oocyte was 30.7%, and Cryptosporidium infection rates in vegetables and lettuce were different. This difference was statistically significant (P<0.05. Conclusion: As a result of this research it is determined that the prevalence of Giardia and Cryptosporidium in Ilam vegetables is significantly higher, and the contamination of lettuce is far greater. Therefore, authorities should be more attentive to the field of education and the control of parasitic diseases.

  16. The hematological status, plasma vitamin B12 and folic acid levels, and intestinal pathology in rats infected with Giardia lamblia.

    Science.gov (United States)

    Cheeramakara, Cheeraratana; Nontprasert, Apichart; Siripanth, Chutatip; Tanomsak, Wanyarat; Chularerk, Udomporn; Sucharit, Pramualmal; Areekul, Suvit

    2004-12-01

    The purpose of our study was to investigate the hematological status, vitamin B12 and folic acid absorption and intestinal pathology after Giardia lamblia infection in a rat model. Adult Wistar rats were assigned randomly to receive human giardia cysts orally in the amount of 5 x 10(5) or 1.0 x 10(6) cysts, or none in the controls. The results showed that all the rats injected with giardia cysts became infected. The cyst output in the infected rats varied considerably. In rats infected with 5.0 x 10(5) giardia cysts, the incubation period until cyst output was 10 days compared with 4 days in rats infected with the higher amount of 1.0 x 106 giardia cysts. The highest peaks for cysts output in these 2 groups were on days 4-33, which decreased gradually to days 40-58. The hematocrit and hemoglobin levels in the infected rats were statistically significantly lower than in the controls on days 16, 22, 33, and 37 post-infection (p giardia cyst output and hemoglobin concentration was found in the infected rats (p = 0.05). There were no significant differences in plasma vitamin B12 and folic acid levels between the infected rats and the control rats. No pathological changes were found in the small intestine of infected rats. These findings suggest that giardiasis did not affect the absorption of plasma vitamin B12 and folic acid but caused anemia in a rat model.

  17. Robust sparse image reconstruction of radio interferometric observations with PURIFY

    Science.gov (United States)

    Pratley, Luke; McEwen, Jason D.; d'Avezac, Mayeul; Carrillo, Rafael E.; Onose, Alexandru; Wiaux, Yves

    2018-01-01

    Next-generation radio interferometers, such as the Square Kilometre Array, will revolutionize our understanding of the Universe through their unprecedented sensitivity and resolution. However, to realize these goals significant challenges in image and data processing need to be overcome. The standard methods in radio interferometry for reconstructing images, such as CLEAN, have served the community well over the last few decades and have survived largely because they are pragmatic. However, they produce reconstructed interferometric images that are limited in quality and scalability for big data. In this work, we apply and evaluate alternative interferometric reconstruction methods that make use of state-of-the-art sparse image reconstruction algorithms motivated by compressive sensing, which have been implemented in the PURIFY software package. In particular, we implement and apply the proximal alternating direction method of multipliers algorithm presented in a recent article. First, we assess the impact of the interpolation kernel used to perform gridding and degridding on sparse image reconstruction. We find that the Kaiser-Bessel interpolation kernel performs as well as prolate spheroidal wave functions while providing a computational saving and an analytic form. Secondly, we apply PURIFY to real interferometric observations from the Very Large Array and the Australia Telescope Compact Array and find that images recovered by PURIFY are of higher quality than those recovered by CLEAN. Thirdly, we discuss how PURIFY reconstructions exhibit additional advantages over those recovered by CLEAN. The latest version of PURIFY, with developments presented in this work, is made publicly available.

  18. A single-stranded RNA copy of the Giardia lamblia virus double-stranded RNA genome is present in the infected Giardia lamblia.

    OpenAIRE

    Furfine, E S; White, T C; Wang, A L; Wang, C C

    1989-01-01

    An isolate of Giardia lamblia infected with the double-stranded RNA virus (GLV) has two major species of RNA that are not present in an uninfected isolate. One of these species is the previously characterized double-stranded RNA genome of GLV (1). The second species of RNA appears to be a full length copy of one strand of the double-stranded RNA genome. This full length single-stranded RNA is not present in viral particles isolated from the growth medium. The cellular concentration of the sin...

  19. High occurrence of Blastocystis sp. subtypes 1-3 and Giardia intestinalis assemblage B among patients in Zanzibar, Tanzania.

    Science.gov (United States)

    Forsell, Joakim; Granlund, Margareta; Samuelsson, Linn; Koskiniemi, Satu; Edebro, Helén; Evengård, Birgitta

    2016-06-29

    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. 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. 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. Carriage of intestinal parasites was very

  20. Lectin Activation in Giardia lamblia by Host Protease: A Novel Host-Parasite Interaction

    Science.gov (United States)

    Lev, Boaz; Ward, Honorine; Keusch, Gerald T.; Pereira, Miercio E. A.

    1986-04-01

    A lectin in Giardia lamblia was activated by secretions from the human duodenum, the environment where the parasite lives. Incubation of the secretions with trypsin inhibitors prevented the appearance of lectin activity, implicating proteases as the activating agent. Accordingly, lectin activation was also produced by crystalline trypsin and Pronase; other proteases tested were ineffective. When activated, the lectin agglutinated intestinal cells to which the parasite adheres in vivo. The lectin was most specific to mannose-6-phosphate and apparently was bound to the plasma membrane. Activation of a parasite lectin by a host protease represents a novel mechanism of hostparasite interaction and may contribute to the affinity of Giardia lamblia to the infection site.

  1. Development of a biogas purifier for rural areas in Japan

    Energy Technology Data Exchange (ETDEWEB)

    Kimura, Y.; Hinata, T. [Hokkaido Central Agricultural Experiment Station, Hokkaido (Japan); Yasui, S. [Zukosha Co. Ltd., Obihiro, Hokkaido (Japan); Noguchi, N. [Hokkaido Univ., Sapporo, Hokkaido (Japan); Tsukamoto, T. [IHI Shibaura. Co. Ltd., Obihiro, Hokkaido (Japan); Imai, T. [Green Plan Co. Ltd., Sapporo, Hokkaido (Japan); Kanai, M. [Air Water Co. Ltd, Sakai, Osaka (Japan); Matsuda, Z. [Hokuren Agricultural Research Center, Sapporo, Hokkaido (Japan)

    2010-07-01

    Although the biogas that is currently produced for dairy farms in Japan is a carbon-neutral energy, its use is restricted to farming areas only because there is no effective method of transporting unused biogas. There is a need for establishing practical methods for biogas removal from operating systems. In this study, a gas separation membrane was used in order to modify biogas to city gas 12A specifications, and to develop a biogas purifier equipped with a device to fill high pressure purified gas into cylinders to be taken outside the farming area. The objective was to expand the use of biogas produced from stand-alone gas plants. The amount of purified gas produced at a newly created refining-compression-filling (RCF) facility was approximately 97.0 Nm{sup 3}/day, for a raw material amount of about 216.0 Nm{sup 3}/day. The heat quantity of the purified gas was 38.9 MJ/Nm{sup 3}, which was within city gas 12A specifications. A total of 14.3 cylinders were filled each day with the manufactured purified gas. Test calculations along with a simulation exercise revealed that it would be possible to provide purified gas to approximately 6 per cent of common residences in a town in northern Japan. It was concluded that the newly created RCF facility allowed the modification of carbon-neutral biogas to conform to city gas 12A specifications, and allowed the transport of this gas out of the farming area.

  2. Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices

    OpenAIRE

    Moss, Joseph A.; Gordy, John; Snyder, Richard A.

    2014-01-01

    Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Metho...

  3. Genetic variation in potential Giardia vaccine candidates cyst wall protein 2 and α1-giardin.

    Science.gov (United States)

    Radunovic, Matej; Klotz, Christian; Saghaug, Christina Skår; Brattbakk, Hans-Richard; Aebischer, Toni; Langeland, Nina; Hanevik, Kurt

    2017-08-01

    Giardia is a prevalent intestinal parasitic infection. The trophozoite structural protein a1-giardin (a1-g) and the cyst protein cyst wall protein 2 (CWP2) have shown promise as Giardia vaccine antigen candidates in murine models. The present study assesses the genetic diversity of a1-g and CWP2 between and within assemblages A and B in human clinical isolates. a1-g and CWP2 sequences were acquired from 15 Norwegian isolates by PCR amplification and 20 sequences from German cultured isolates by whole genome sequencing. Sequences were aligned to reference genomes from assemblage A2 and B to identify genetic variance. Genetic diversity was found between assemblage A and B reference sequences for both a1-g (90.8% nucleotide identity) and CWP2 (82.5% nucleotide identity). However, for a1-g, this translated into only 3 amino acid (aa) substitutions, while for CWP2 there were 41 aa substitutions, and also one aa deletion. Genetic diversity within assemblage B was larger; nucleotide identity 92.0% for a1-g and 94.3% for CWP2, than within assemblage A (nucleotide identity 99.0% for a1-g and 99.7% for CWP2). For CWP2, the diversity on both nucleotide and protein level was higher in the C-terminal end. Predicted antigenic epitopes were not affected for a1-g, but partially for CWP2. Despite genetic diversity in a1-g, we found aa sequence, characteristics, and antigenicity to be well preserved. CWP2 showed more aa variance and potential antigenic differences. Several CWP2 antigens might be necessary in a future Giardia vaccine to provide cross protection against both Giardia assemblages infecting humans.

  4. Transcriptomic Profiling of High-Density Giardia Foci Encysting in the Murine Proximal Intestine.

    Science.gov (United States)

    Pham, Jonathan K; Nosala, Christopher; Scott, Erica Y; Nguyen, Kristofer F; Hagen, Kari D; Starcevich, Hannah N; Dawson, Scott C

    2017-01-01

    Giardia is a highly prevalent, understudied protistan parasite causing significant diarrheal disease worldwide. Its life cycle consists of two stages: infectious cysts ingested from contaminated food or water sources, and motile trophozoites that colonize and attach to the gut epithelium, later encysting to form new cysts that are excreted into the environment. Current understanding of parasite physiology in the host is largely inferred from transcriptomic studies using Giardia grown axenically or in co-culture with mammalian cell lines. The dearth of information about the diversity of host-parasite interactions occurring within distinct regions of the gastrointestinal tract has been exacerbated by a lack of methods to directly and non-invasively interrogate disease progression and parasite physiology in live animal hosts. By visualizing Giardia infections in the mouse gastrointestinal tract using bioluminescent imaging (BLI) of tagged parasites, we recently showed that parasites colonize the gut in high-density foci. Encystation is initiated in these foci throughout the entire course of infection, yet how the physiology of parasites within high-density foci in the host gut differs from that of cells in laboratory culture is unclear. Here we use BLI to precisely select parasite samples from high-density foci in the proximal intestine to interrogate in vivo Giardia gene expression in the host. Relative to axenic culture, we noted significantly higher expression (>10-fold) of oxidative stress, membrane transporter, and metabolic and structural genes associated with encystation in the high-density foci. These differences in gene expression within parasite foci in the host may reflect physiological changes associated with high-density growth in localized regions of the gut. We also identified and verified six novel cyst-specific proteins, including new components of the cyst wall that were highly expressed in these foci. Our in vivo transcriptome data support an

  5. Field portable mobile phone based fluorescence microscopy for detection of Giardia lamblia cysts in water samples

    Science.gov (United States)

    Ceylan Koydemir, Hatice; Gorocs, Zoltan; McLeod, Euan; Tseng, Derek; Ozcan, Aydogan

    2015-03-01

    Giardia lamblia is a waterborne parasite that causes an intestinal infection, known as giardiasis, and it is found not only in countries with inadequate sanitation and unsafe water but also streams and lakes of developed countries. Simple, sensitive, and rapid detection of this pathogen is important for monitoring of drinking water. Here we present a cost-effective and field portable mobile-phone based fluorescence microscopy platform designed for automated detection of Giardia lamblia cysts in large volume water samples (i.e., 10 ml) to be used in low-resource field settings. This fluorescence microscope is integrated with a disposable water-sampling cassette, which is based on a flow-through porous polycarbonate membrane and provides a wide surface area for fluorescence imaging and enumeration of the captured Giardia cysts on the membrane. Water sample of interest, containing fluorescently labeled Giardia cysts, is introduced into the absorbent pads that are in contact with the membrane in the cassette by capillary action, which eliminates the need for electrically driven flow for sample processing. Our fluorescence microscope weighs ~170 grams in total and has all the components of a regular microscope, capable of detecting individual fluorescently labeled cysts under light-emitting-diode (LED) based excitation. Including all the sample preparation, labeling and imaging steps, the entire measurement takes less than one hour for a sample volume of 10 ml. This mobile phone based compact and cost-effective fluorescent imaging platform together with its machine learning based cyst counting interface is easy to use and can even work in resource limited and field settings for spatio-temporal monitoring of water quality.

  6. Persistent gut barrier damage and commensal bacterial influx following eradication of Giardia infection in mice

    Science.gov (United States)

    2013-01-01

    Background Recent studies of Giardia lamblia outbreaks have indicated that 40–80% of infected patients experience long-lasting functional gastrointestinal disorders after parasitic clearance. Our aim was to assess changes in the intestinal barrier and spatial distribution of commensal bacteria in the post-clearance phase of Giardia infection. Methods Mice were orogastrically inoculated with G. lamblia trophozoites (strain GS/M) or pair-fed with saline and were sacrificed on post-infective (PI) days 7 (colonization phase) and 35 (post-clearance phase). Gut epithelial barrier function was assessed by Western blotting for occludin cleavage and luminal-to-serosal macromolecular permeability. Gut-associated, superficial adherent, and mucosal endocytosed bacteria were measured by agar culturing and were examined by fluorescence in situ hybridization. Intracellular bacteria cultured from isolated mucosal cells were characterized by 16S rDNA sequencing. Neutrophil-specific esterase staining, a myeloperoxidase activity assay, and enzyme-linked immunosorbent assays for cytokine concentrations were used to verify intestinal tissue inflammation. Results Tight junctional damage was detected in the intestinal mucosa of Giardia-infected mice on PI days 7 and 35. Although intestinal bacterial overgrowth was evident only during parasite colonization (PI day 7), enhanced mucosal adherence and endocytosis of bacteria were observed on PI days 7 and 35. Multiple bacterial strains, including Bacillus, Lactobacillus, Staphylococcus, and Phenylobacterium, penetrated the gut mucosa in the post-infective phase. The mucosal influx of bacteria coincided with increases in neutrophil infiltration and myeloperoxidase activity on PI days 7 and 35. Elevated intestinal IFNγ, TNFα, and IL-1β levels also were detected on PI day 35. Conclusions Giardia-infected mice showed persistent tight junctional damage and bacterial penetration, accompanied by mucosal inflammation, after parasite clearance

  7. Occurrence of Giardia cysts and Cryptosporidium oocysts in the Temuan Orang Asli (aborigine) River System.

    Science.gov (United States)

    Lim, Y A L; Aahmad, R A

    2004-12-01

    A survey of the river water frequently used by the Temuan Orang Asli (aborigine) indicated that 66.7% of the river water samples were Giardia cyst positive and 5.6% were Cryptosporidium oocyst positive. Although Giardia cysts were detected in samples from all the sites (e.g. upstream, midstream, and downstream), Cryptosporidium was only present in one river water sample taken from downstream from a village. The only sample of upstream water which contained Giardia cysts had a concentration of 0.7 cysts/l. All samples taken from midstream contained cysts with a mean concentration of 9.8 +/- 6.6 cysts/l (range = 1-20 cysts/l). Eighty-three point three percent of the samples collected from downstream contained cysts and 16.7% had oocysts. The average concentration of cysts was 12.9 +/- 16.4 cysts/l (range = 0-44 cysts/l), whereas the oocyst concentration was 0.4 oocysts/l. All river samples tested positive for the presence of E. coli, indicating fecal contamination. The results of this study imply that the river system is contaminated with fecal-oral transmitted parasites. The river water, used by the Orang Asli, is a probable route for Giardia and Cryptosporidium transmission in this community. Long term strategies, incorporating health education regarding personal hygiene, and provision of toilets and the importance of their proper usage, need to be embraced by this community in order to control the spread of these parasites.

  8. Local and global genetic diversity of protozoan parasites: Spatial distribution of Cryptosporidium and Giardia genotypes.

    Science.gov (United States)

    Garcia-R, Juan C; French, Nigel; Pita, Anthony; Velathanthiri, Niluka; Shrestha, Rima; Hayman, David

    2017-07-01

    Cryptosporidiosis and giardiasis are recognized as significant enteric diseases due to their long-term health effects in humans and their economic impact in agriculture and medical care. Molecular analysis is essential to identify species and genotypes causing these infectious diseases and provides a potential tool for monitoring. This study uses information on species and genetic variants to gain insights into the geographical distribution and spatial patterns of Cryptosporidium and Giardia parasites. Here, we describe the population heterogeneity of genotypic groups within Cryptosporidium and Giardia present in New Zealand using gp60 and gdh markers to compare the observed variation with other countries around the globe. Four species of Cryptosporidium (C. hominis, C. parvum, C. cuniculus and C. erinacei) and one species of Giardia (G. intestinalis) were identified. These species have been reported worldwide and there are not unique Cryptosporidium gp60 subtype families and Giardia gdh assemblages in New Zealand, most likely due to high gene flow of historical and current human activity (travel and trade) and persistence of large host population sizes. The global analysis revealed that genetic variants of these pathogens are widely distributed. However, genetic variation is underestimated by data biases (e.g. neglected submission of sequences to genetic databases) and low sampling. New genotypes are likely to be discovered as sampling efforts increase according to accumulation prediction analyses, especially for C. parvum. Our study highlights the need for greater sampling and archiving of genotypes globally to allow comparative analyses that help understand the population dynamics of these protozoan parasites. Overall our study represents a comprehensive overview for exploring local and global protozoan genotype diversity and advances our understanding of the importance for surveillance and potential risk associated with these infectious diseases.

  9. A detailed, hierarchical study of Giardia lamblia's ventral disc reveals novel microtubule-associated protein complexes.

    Directory of Open Access Journals (Sweden)

    Cindi L Schwartz

    Full Text Available Giardia lamblia is a flagellated, unicellular parasite of mammals infecting over one billion people worldwide. Giardia's two-stage life cycle includes a motile trophozoite stage that colonizes the host small intestine and an infectious cyst form that can persist in the environment. Similar to many eukaryotic cells, Giardia contains several complex microtubule arrays that are involved in motility, chromosome segregation, organelle transport, maintenance of cell shape and transformation between the two life cycle stages. Giardia trophozoites also possess a unique spiral microtubule array, the ventral disc, made of approximately 50 parallel microtubules and associated microribbons, as well as a variety of associated proteins. The ventral disc maintains trophozoite attachment to the host intestinal epithelium. With the help of a combined SEM/microtome based slice and view method called 3View® (Gatan Inc., Pleasanton, CA, we present an entire trophozoite cell reconstruction and describe the arrangement of the major cytoskeletal elements. To aid in future analyses of disc-mediated attachment, we used electron-tomography of freeze-substituted, plastic-embedded trophozoites to explore the detailed architecture of ventral disc microtubules and their associated components. Lastly, we examined the disc microtubule array in three dimensions in unprecedented detail using cryo-electron tomography combined with internal sub-tomogram volume averaging of repetitive domains. We discovered details of protein complexes stabilizing microtubules by attachment to their inner and outer wall. A unique tri-laminar microribbon structure is attached vertically to the disc microtubules and is connected to neighboring microribbons via crossbridges. This work provides novel insight into the structure of the ventral disc microtubules, microribbons and associated proteins. Knowledge of the components comprising these structures and their three-dimensional organization is

  10. Local and global genetic diversity of protozoan parasites: Spatial distribution of Cryptosporidium and Giardia genotypes.

    Directory of Open Access Journals (Sweden)

    Juan C Garcia-R

    2017-07-01

    Full Text Available Cryptosporidiosis and giardiasis are recognized as significant enteric diseases due to their long-term health effects in humans and their economic impact in agriculture and medical care. Molecular analysis is essential to identify species and genotypes causing these infectious diseases and provides a potential tool for monitoring. This study uses information on species and genetic variants to gain insights into the geographical distribution and spatial patterns of Cryptosporidium and Giardia parasites. Here, we describe the population heterogeneity of genotypic groups within Cryptosporidium and Giardia present in New Zealand using gp60 and gdh markers to compare the observed variation with other countries around the globe. Four species of Cryptosporidium (C. hominis, C. parvum, C. cuniculus and C. erinacei and one species of Giardia (G. intestinalis were identified. These species have been reported worldwide and there are not unique Cryptosporidium gp60 subtype families and Giardia gdh assemblages in New Zealand, most likely due to high gene flow of historical and current human activity (travel and trade and persistence of large host population sizes. The global analysis revealed that genetic variants of these pathogens are widely distributed. However, genetic variation is underestimated by data biases (e.g. neglected submission of sequences to genetic databases and low sampling. New genotypes are likely to be discovered as sampling efforts increase according to accumulation prediction analyses, especially for C. parvum. Our study highlights the need for greater sampling and archiving of genotypes globally to allow comparative analyses that help understand the population dynamics of these protozoan parasites. Overall our study represents a comprehensive overview for exploring local and global protozoan genotype diversity and advances our understanding of the importance for surveillance and potential risk associated with these infectious diseases.

  11. Characterization of RNase MRP RNA and novel snoRNAs from Giardia intestinalis and Trichomonas vaginalis

    Directory of Open Access Journals (Sweden)

    Chen Xiaowei S

    2011-11-01

    Full Text Available Abstract Background Eukaryotic cells possess a complex network of RNA machineries which function in RNA-processing and cellular regulation which includes transcription, translation, silencing, editing and epigenetic control. Studies of model organisms have shown that many ncRNAs of the RNA-infrastructure are highly conserved, but little is known from non-model protists. In this study we have conducted a genome-scale survey of medium-length ncRNAs from the protozoan parasites Giardia intestinalis and Trichomonas vaginalis. Results We have identified the previously 'missing' Giardia RNase MRP RNA, which is a key ribozyme involved in pre-rRNA processing. We have also uncovered 18 new H/ACA box snoRNAs, expanding our knowledge of the H/ACA family of snoRNAs. Conclusions Results indicate that Giardia intestinalis and Trichomonas vaginalis, like their distant multicellular relatives, contain a rich infrastructure of RNA-based processing. From here we can investigate the evolution of RNA processing networks in eukaryotes.

  12. The concentration of Cryptosporidium and Giardia in water--the role and importance of recovery efficiency.

    Science.gov (United States)

    Ongerth, Jerry E

    2013-05-01

    The concentration of Cryptosporidium and of Giardia in surface water is a subject of importance to public health and public water supply. The term concentration is a fundamental property of any water quality parameter having a classical definition as used in chemistry and biology. Analytical methods for measuring the occurrence of Cryptosporidium and Giardia in water find only a fraction of the organisms actually present. This paper collects recently available data to examine the role and importance of recovery efficiency measurement to description of the concentrations of these organisms. Data from Australian sources graphically illustrate the variability of recovery efficiency at individual sites over relatively short time scales. Additional data on replicated recovery measurements establish their reproducibility. The recently released USEPA LT2 data along with those from Australia illustrate the independent variation of Cryptosporidium and Giardia occurrence and recovery efficiency at individual sampling locations. Calculation of concentration from paired raw numbers and recovery efficiency measurements clearly shows the magnitude and importance of taking recovery into account in expressing the concentration of these organisms. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. A study on protozoan infections (Giardia, Entamoeba, Isoapora and Cryptosporidium in stray dogs in Ilam province

    Directory of Open Access Journals (Sweden)

    S Kakekhani

    2011-11-01

    Full Text Available Giardia, Entamoeba, Isospora and Cryptosporidium are important protozoan parastites that caused diarrhea in human and animals. In the present study, fecal samples were collected fresh, directly from the rectum of 112 stray dogs in Ilam province. Giardia and Entamoeba were concentrated by using the formalin ether sedimentation method followed by the trichrome and iodine staining technique andCryptosporidium  oocysts  were  concentrated  by  using  the  formalin  ether  sedimentation  method  followed by the modified Ziehl-Neelsen staining technique. Of 112 stray dogs, protozoan infections were detected from feces of 46 dogs (41.07% that Giardia infection was detected from feces of 21 dogs (18.75%, Isospora 17 (15.17%, Cryptosporidium 8 (7.14% and synchronization infection to 2 protozoan in 9 dogs (8.03% and to 3 protozoan in 3 (2.67%. In the present study not observed to Entamoeba. No statistically significant differences in prevalence of protozoan parasites occurred between female (34.21 % and male (55.5 % stray dogs (p>0/05. But statistically significant differences in prevalence occurred between 1≥0 and 0 ≥1 stray dogs (p>0/05. So that stray dogs of Ilam province can cause infection of human water and food sources.

  14. Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices

    Directory of Open Access Journals (Sweden)

    Joseph A. Moss

    2014-01-01

    Full Text Available Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Method 1623. Results were equivalent to immunomagnetic procedures for Cryptosporidium, and Giardia. Overall percent recovery for Cryptosporidium with Method 1623 averaged 26.89% (std 21.44%; min = 0%; max = 73% and was similar but less variable for qPCR method at an estimated average of 27.67 (std 17.65%; min = 5%; max = 63%. For Giardia, Method 1623 had an overall average recovery of 27.11% (std 17.98%; min = 1%; max = 58%, while multistage purification and qPCR had an estimated lower overall recovery at 18.58% (std 13.95%; min = 0%; max = 35%. Microsporidia were also readily detected with an estimated recovery of 46.81% overall (std 17.66%; min = 18%; max = 70% for E. intestinalis and 38.90% (std 14.36%; min = 13%; max = 62% for E. bieneusi.

  15. Syrian hamsters (Mesocricetus auratus) with simultaneous intestinal Giardia sp., Spironucleus sp., and trichomonad infections.

    Science.gov (United States)

    Sheppard, Barbara J; Stockdale Walden, Heather D; Kondo, Hirotaka

    2013-11-01

    A commercial facility producing hamsters with a history of infection by dwarf tapeworm (Hymenolepis nana) submitted 15 animals for necropsy and postmortem parasitological and microscopic examination. No tapeworms were detected grossly or microscopically. Fecal examination including gastrointestinal mucosal smears demonstrated mixed intestinal bacteria and low numbers of Giardia sp. Histologic examination of small intestine demonstrated filling of the small intestinal crypts by large numbers of 7-9 µm × 3 µm, rod to crescent or teardrop-shaped flagellates consistent with Spironucleus sp. These organisms had two 1-µm, basophilic, oval nuclei and multiple superficial flagella-like structures. Much larger 10-15 µm × 8-10 µm, oval to pear-shaped organisms were also present in lower numbers and usually located with the crypts. These larger flagellates had multiple flagella and a basophilic rod-shaped nucleus. The larger flagellates included Giardia sp., which had an intimate interface with the surface of the mucosal epithelium, bilaterally symmetry, and binucleation. Lower numbers of trichomonads were also present and were distinguished by an undulating surface membrane and a single nucleus. The mucosa was hyperplastic and moderately inflamed. Although the tapeworm infection was resolved, diagnosis of multiple intestinal flagellates by fecal examination is complicated by the varying sensitivity and diagnostic accuracy of different types of fecal analysis for different flagellate types. Key differences in the morphology and location of the different types of flagellates as observed by histology of intestinal tissues provide important additional diagnostic information to distinguish trichomonads, Spironucleus sp., and Giardia sp.

  16. Characterization of RNase MRP RNA and novel snoRNAs from Giardia intestinalis and Trichomonas vaginalis

    Science.gov (United States)

    2011-01-01

    Background Eukaryotic cells possess a complex network of RNA machineries which function in RNA-processing and cellular regulation which includes transcription, translation, silencing, editing and epigenetic control. Studies of model organisms have shown that many ncRNAs of the RNA-infrastructure are highly conserved, but little is known from non-model protists. In this study we have conducted a genome-scale survey of medium-length ncRNAs from the protozoan parasites Giardia intestinalis and Trichomonas vaginalis. Results We have identified the previously 'missing' Giardia RNase MRP RNA, which is a key ribozyme involved in pre-rRNA processing. We have also uncovered 18 new H/ACA box snoRNAs, expanding our knowledge of the H/ACA family of snoRNAs. Conclusions Results indicate that Giardia intestinalis and Trichomonas vaginalis, like their distant multicellular relatives, contain a rich infrastructure of RNA-based processing. From here we can investigate the evolution of RNA processing networks in eukaryotes. PMID:22053856

  17. The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target.

    Science.gov (United States)

    Kunz, Stefan; Balmer, Vreni; Sterk, Geert Jan; Pollastri, Michael P; Leurs, Rob; Müller, Norbert; Hemphill, Andrew; Spycher, Cornelia

    2017-09-01

    Giardiasis is an intestinal infection correlated with poverty and poor drinking water quality, and treatment options are limited. According to the Center for Disease Control and Prevention, Giardia infections afflict nearly 33% of people in developing countries, and 2% of the adult population in the developed world. This study describes the single cyclic nucleotide-specific phosphodiesterase (PDE) of G. lamblia and assesses PDE inhibitors as a new generation of anti-giardial drugs. An extensive search of the Giardia genome database identified a single gene coding for a class I PDE, GlPDE. The predicted protein sequence was analyzed in-silico to characterize its domain structure and catalytic domain. Enzymatic activity of GlPDE was established by complementation of a PDE-deficient Saccharomyces cerevisiae strain, and enzyme kinetics were characterized in soluble yeast lysates. The potency of known PDE inhibitors was tested against the activity of recombinant GlPDE expressed in yeast and against proliferating Giardia trophozoites. Finally, the localization of epitope-tagged and ectopically expressed GlPDE in Giardia cells was investigated. Giardia encodes a class I PDE. Catalytically important residues are fully conserved between GlPDE and human PDEs, but sequence differences between their catalytic domains suggest that designing Giardia-specific inhibitors is feasible. Recombinant GlPDE hydrolyzes cAMP with a Km of 408 μM, and cGMP is not accepted as a substrate. A number of drugs exhibit a high degree of correlation between their potency against the recombinant enzyme and their inhibition of trophozoite proliferation in culture. Epitope-tagged GlPDE localizes as dots in a pattern reminiscent of mitosomes and to the perinuclear region in Giardia. Our data strongly suggest that inhibition of G. lamblia PDE activity leads to a profound inhibition of parasite proliferation and that GlPDE is a promising target for developing novel anti-giardial drugs.

  18. Characterization of the Giardia intestinalis secretome during interaction with human intestinal epithelial cells: The impact on host cells.

    Science.gov (United States)

    Ma'ayeh, Showgy Y; Liu, Jingyi; Peirasmaki, Dimitra; Hörnaeus, Katarina; Bergström Lind, Sara; Grabherr, Manfred; Bergquist, Jonas; Svärd, Staffan G

    2017-12-01

    Giardia intestinalis is a non-invasive protozoan parasite that causes giardiasis in humans, the most common form of parasite-induced diarrhea. Disease mechanisms are not completely defined and very few virulence factors are known. To identify putative virulence factors and elucidate mechanistic pathways leading to disease, we have used proteomics to identify the major excretory-secretory products (ESPs) when Giardia trophozoites of WB and GS isolates (assemblages A and B, respectively) interact with intestinal epithelial cells (IECs) in vitro. The main parts of the IEC and parasite secretomes are constitutively released proteins, the majority of which are associated with metabolism but several proteins are released in response to their interaction (87 and 41 WB and GS proteins, respectively, 76 and 45 human proteins in response to the respective isolates). In parasitized IECs, the secretome profile indicated effects on the cell actin cytoskeleton and the induction of immune responses whereas that of Giardia showed anti-oxidation, proteolysis (protease-associated) and induction of encystation responses. The Giardia secretome also contained immunodominant and glycosylated proteins as well as new candidate virulence factors and assemblage-specific differences were identified. A minor part of Giardia ESPs had signal peptides (29% for both isolates) and extracellular vesicles were detected in the ESPs fractions, suggesting alternative secretory pathways. Microscopic analyses showed ESPs binding to IECs and partial internalization. Parasite ESPs reduced ERK1/2 and P38 phosphorylation and NF-κB nuclear translocation. Giardia ESPs altered gene expression in IECs, with a transcriptional profile indicating recruitment of immune cells via chemokines, disturbances in glucose homeostasis, cholesterol and lipid metabolism, cell cycle and induction of apoptosis. This is the first study identifying Giardia ESPs and evaluating their effects on IECs. It highlights the importance of

  19. Characterization of the Giardia intestinalis secretome during interaction with human intestinal epithelial cells: The impact on host cells.

    Directory of Open Access Journals (Sweden)

    Showgy Y Ma'ayeh

    2017-12-01

    Full Text Available Giardia intestinalis is a non-invasive protozoan parasite that causes giardiasis in humans, the most common form of parasite-induced diarrhea. Disease mechanisms are not completely defined and very few virulence factors are known.To identify putative virulence factors and elucidate mechanistic pathways leading to disease, we have used proteomics to identify the major excretory-secretory products (ESPs when Giardia trophozoites of WB and GS isolates (assemblages A and B, respectively interact with intestinal epithelial cells (IECs in vitro.The main parts of the IEC and parasite secretomes are constitutively released proteins, the majority of which are associated with metabolism but several proteins are released in response to their interaction (87 and 41 WB and GS proteins, respectively, 76 and 45 human proteins in response to the respective isolates. In parasitized IECs, the secretome profile indicated effects on the cell actin cytoskeleton and the induction of immune responses whereas that of Giardia showed anti-oxidation, proteolysis (protease-associated and induction of encystation responses. The Giardia secretome also contained immunodominant and glycosylated proteins as well as new candidate virulence factors and assemblage-specific differences were identified. A minor part of Giardia ESPs had signal peptides (29% for both isolates and extracellular vesicles were detected in the ESPs fractions, suggesting alternative secretory pathways. Microscopic analyses showed ESPs binding to IECs and partial internalization. Parasite ESPs reduced ERK1/2 and P38 phosphorylation and NF-κB nuclear translocation. Giardia ESPs altered gene expression in IECs, with a transcriptional profile indicating recruitment of immune cells via chemokines, disturbances in glucose homeostasis, cholesterol and lipid metabolism, cell cycle and induction of apoptosis.This is the first study identifying Giardia ESPs and evaluating their effects on IECs. It highlights the

  20. Air purification by cementitious materials : Evaluation of air purifying properties

    NARCIS (Netherlands)

    Hüsken, G.; Brouwers, H.J.H.; Al-Mattarneh, H.; Mustapha, K.N.; Nuruddin, M.F.

    2008-01-01

    This paper addresses the evaluation of the photocatalytic properties of concrete containing titanium dioxide (TiO2). Here, the assessment of the air purifying abilities of the hardened concrete regarding the degradation of nitric oxide (NO) is of major interest. A setup for measuring the performance

  1. Effect of partially purified angiotensin converting enzyme inhibitory ...

    African Journals Online (AJOL)

    This study evaluated the effect of partially-purified angiotensin converting enzyme (ACE) inhibitory proteins obtained from the leaves of Moringa oleifera on blood glucose, serum ACE activity and lipid profile of alloxaninduced diabetic rats. Twenty-five apparently healthy male albino rats were divided into five groups of five ...

  2. Predicting group of metabolites available in partially purified tomato ...

    African Journals Online (AJOL)

    GREGORY

    2011-12-16

    Dec 16, 2011 ... biochemistry and analytical chemistry to identify, quantify and purify the individual components of the mixture .... chemistry, organic chemistry, physical chemistry and biochemistry. Normally, isomers of a .... ced analytic capabilities and reduced analyzing time. When combined with infrared spectroscopy, the ...

  3. Delivery of metronidazole from purified Nigerian sheabutter in ...

    African Journals Online (AJOL)

    The drug delivery propensities of 1 %w/w metronidazole (MTZ) ointment formulations in purified Nigerian sheabutter base have been compared to those of similar formulations produced with standard or modified bases, with a view to determining bases capable of ensuring rapid release of MTZ from ointment formulations ...

  4. Influence of a highly purified senna extract on colonic epithelium

    NARCIS (Netherlands)

    van Gorkom, B A; Karrenbeld, A; van Der Sluis, T; Koudstaal, J; de Vries, E G; Kleibeuker, J H

    2000-01-01

    BACKGROUND: Chronic use of sennoside laxatives often causes pseudomelanosis coli. A recent study suggested that pseudomelanosis coli is associated with an increased colorectal cancer risk. A single high dose of highly purified senna extract increased proliferation rate and reduced crypt length in

  5. Method of purifying phosphoric acid after solvent extraction

    International Nuclear Information System (INIS)

    Kouloheris, A.P.; Lefever, J.A.

    1979-01-01

    A method of purifying phosphoric acid after solvent extraction is described. The phosphoric acid is contacted with a sorbent which sorbs or takes up the residual amount of organic carrier and the phosphoric acid separated from the organic carrier-laden sorbent. The method is especially suitable for removing residual organic carrier from phosphoric acid after solvent extraction uranium recovery. (author)

  6. Occurrence of Giardia cysts and Cryptosporidium oocysts in sewage and their reduction in treatment process; Gesuichu no Giardia shisuto oyobi Cryptosporidium oshisuto noudo to gesui syori ni okeru jyokyosei

    Energy Technology Data Exchange (ETDEWEB)

    Hashimoto, A.; Hirata, T. [Azabu University, Kanagawa (Japan); Magara, Y. [National Institute of Public Health, Tokyo (Japan); Ogaki, S. [The University of Tokyo, Tokyo (Japan)

    1997-06-10

    Recently, the water system infection owing to protozoan, virus and so on which is different from the conventional infection like represented by bacteria, is reported. There are few examples for the infection conditional study and epidemiological investigation of these protozoan and virus in Japan. Authors carried out a field survey at nine sewage treatment plants, in order to investigate the occurrence of Giardia cysts and Cryptosporidium oocysts in raw sewage and the removal in sewage treatment processes. As a result, in all plants, Giardia cysts were detected in raw sewage and effluent of primary sedimentation. Cryptosporidium oocysts were detected from only one raw sewage sample. Indicator bacteria counts tested, C.perfringens spores, coliforms and E.coli, were statistically correlated with Giardia cyst densities and the most significant relationship was observed between C.perfringens spore counts and Giardia cyst densities. It was found that turbidity was the most appropriate indicator for Giardia cyst removal in activated sludge treatment process. 18 refs., 6 figs., 2 tabs.

  7. Thermal inactivation kinetics of partially purified mango pectin methylesterase

    Directory of Open Access Journals (Sweden)

    Claudio Alonso DÍAZ-CRUZ

    2016-01-01

    Full Text Available Abstract Kinetic parameters of thermal inactivation of pectin methylesterase (PME in a partially purified mango enzyme extract were determined. The PME of mango partially purified by salting out showed different patterns of thermal inactivation, indicating the presence of a thermostable fraction at 70 °C and a thermolabile fraction at lower temperatures. The inactivation of the thermostable fraction exhibited a linear behavior that yielded a z-value of 9.44 °C and an activation energy (Ea of 245.6 kJ mol-1 K-1 using the Arrhenius model. The thermostable mango PME fraction represented 17% of total crude enzyme extract, which emphasizes the importance of residual enzyme activity after heat treatment.

  8. Ultrasonic-resonator-combined apparatus for purifying nuclear aerosol particles

    Energy Technology Data Exchange (ETDEWEB)

    Hou, Suxia; Zhang, Quanhu; Li, Sufen; Chen, Chen; Su, Xianghua [Xi' an Hi-Tech Institute, Xi' an (China)

    2017-12-15

    The radiation hazards of radionuclides in the air arising from the storage room of nuclear devices to the operators cannot be ignored. A new ultrasonic-resonator-combined method for purifying nuclear aerosol particles is introduced. To remove particles with diameters smaller than 0.3 μm, an ultrasonic chamber is induced to agglomerate these submicron particles. An apparatus which is used to purify the nuclear aerosol particles is described in the article. The apparatus consists of four main parts: two filtering systems, an ultrasonic chamber and a high-pressure electrostatic precipitator system. Finally, experimental results demonstrated the effectiveness of the implementation of the ultrasonic resonators. The feasibility of the method is proven by its application to the data analysis of the experiments.

  9. Effect of Amphiphilic Alkyl Chain Length Upon Purified LATEX Stability

    International Nuclear Information System (INIS)

    Amira Amir Hassan; Amir Hashim Mohd Yatim

    2015-01-01

    Rubber particles in purified latex (PL) are stabilized by a film of protein and fatty acid soap (surfactant). Saturated straight-chain fatty acid soaps can assist an enhancement of latex stability. However, whether the alkyl chain length plays an important role in increasing the stability is still an issue. The aim of this study is to investigate the effect of alkyl chain length of anionic surfactant on the stability of purified latex. The fatty acid soap of decanoate (9), laurate (11), sodium dodecyl sulphate (SDS) (12) and palmitate (15) were used. The numbers in parentheses indicating the number of carbon present in alkyl chain of the soap. The results showed that the impact of alkyl chain length on the stability of latex is in the order of laurate > decanoate > SDS > palmitate > purified latex accordingly. The alkyl chain length does giving a significant effect on latex stability after longer stirring time. The particle size of latex with the presence of surfactant is greater compare to a single particle itself due to extension of particles diameter. Thus suitable interaction of the nonpolar tail of surfactant with the hydrophobic regions of latex surface played a major role in maintaining a stable latex system. (author)

  10. Proteomic analysis of purified coronavirus infectious bronchitis virus particles

    Directory of Open Access Journals (Sweden)

    Shu Dingming

    2010-06-01

    Full Text Available Abstract Background Infectious bronchitis virus (IBV is the coronavirus of domestic chickens causing major economic losses to the poultry industry. Because of the complexity of the IBV life cycle and the small number of viral structural proteins, important virus-host relationships likely remain to be discovered. Toward this goal, we performed two-dimensional gel electrophoresis fractionation coupled to mass spectrometry identification approaches to perform a comprehensive proteomic analysis of purified IBV particles. Results Apart from the virus-encoded structural proteins, we detected 60 host proteins in the purified virions which can be grouped into several functional categories including intracellular trafficking proteins (20%, molecular chaperone (18%, macromolcular biosynthesis proteins (17%, cytoskeletal proteins (15%, signal transport proteins (15%, protein degradation (8%, chromosome associated proteins (2%, ribosomal proteins (2%, and other function proteins (3%. Interestingly, 21 of the total host proteins have not been reported to be present in virions of other virus families, such as major vault protein, TENP protein, ovalbumin, and scavenger receptor protein. Following identification of the host proteins by proteomic methods, the presence of 4 proteins in the purified IBV preparation was verified by western blotting and immunogold labeling detection. Conclusions The results present the first standard proteomic profile of IBV and may facilitate the understanding of the pathogenic mechanisms.

  11. Physicochemical and kinetic properties of purified sheep's milk xanthine oxidoreductase.

    Science.gov (United States)

    Benboubetra, Mustapha; Baghiani, Abderahmene; Atmani, Djebbar; Harrison, Roger

    2004-06-01

    Xanthine oxidoreductase (XOR) was purified for the first time from sheep's milk. The ultraviolet-visible absorption spectrum was essentially identical to those of the corresponding bovine, human, and goats' milk enzymes and showed an A280/A450 ratio of 5.35 +/- 0.24, indicating a high degree of purity. Like milk XOR from other species, sheep's milk enzyme showed a single band on SDS-PAGE corresponding to a subunit with approximate Mr 150,000. Xanthine oxidase activity of purified sheep's milk XOR (0.69 +/- 0.04 micromole urate min(-1) mg(-1)) was low relative to that of the bovine milk enzyme (1.83 +/- 0.02 micromole urate min(-1) mg(-1)), but higher than those of human or goats' milk XOR. As in the latter 2 cases, the low activity of sheep's milk XOR can be attributed to its relatively low molybdenum content (0.18 atoms per subunit), compared with that of the bovine milk enzyme (0.56 atoms Mo per subunit). Consistent with this, NADH oxidase activity of sheep's milk XOR was similar to that of enzymes purified from bovine, human, or goats' milk. The presence of desulpho-enzyme in sheep's milk XOR was demonstrated by resulfuration experiments, whereby xanthine oxidase activity was increased by approximately 75%.

  12. Comparative reduction of Giardia cysts, F+ coliphages, sulphite reducing clostridia and fecal coliforms by wastewater treatment processes.

    Science.gov (United States)

    Nasser, Abidelfatah M; Benisti, Neta-Lee; Ofer, Naomi; Hovers, Sivan; Nitzan, Yeshayahu

    2017-01-28

    Advanced wastewater treatment processes are applied to prevent the environmental dissemination of pathogenic microorganisms. Giardia lamblia causes a severe disease called giardiasis, and is highly prevalent in untreated wastewater worldwide. Monitoring the microbial quality of wastewater effluents is usually based on testing for the levels of indicator microorganisms in the effluents. This study was conducted to compare the suitability of fecal coliforms, F+ coliphages and sulfide reducing clostridia (SRC) as indicators for the reduction of Giardia cysts in two full-scale wastewater treatment plants. The treatment process consists of activated sludge, coagulation, high rate filtration and either chlorine or UV disinfection. The results of the study demonstrated that Giardia cysts are highly prevalent in raw wastewater at an average concentration of 3600 cysts/L. Fecal coliforms, F+ coliphages and SRC were also detected at high concentrations in raw wastewater. Giardia cysts were efficiently removed (3.6 log 10 ) by the treatment train. The greatest reduction was observed for fecal coliforms (9.6 log 10 ) whereas the least reduction was observed for F+ coliphages (2.1 log 10 ) following chlorine disinfection. Similar reduction was observed for SRC by filtration and disinfection by either UV (3.6 log 10 ) or chlorine (3.3 log 10 ). Since F+ coliphage and SRC were found to be more resistant than fecal coliforms for the tertiary treatment processes, they may prove to be more suitable as indicators for Giardia. The results of this study demonstrated that advanced wastewater treatment may prove efficient for the removal of Giardia cysts and may prevent its transmission when treated effluents are applied for crop irrigation or streams restoration.

  13. Identification and analysis of the RNA degrading complexes and machinery of Giardia lamblia using an in silico approach.

    Science.gov (United States)

    Williams, Christopher W; Elmendorf, Heidi G

    2011-11-29

    RNA degradation is critical to the survival of all cells. With increasing evidence for pervasive transcription in cells, RNA degradation has gained recognition as a means of regulating gene expression. Yet, RNA degradation machinery has been studied extensively in only a few eukaryotic organisms, including Saccharomyces cerevisiae and humans. Giardia lamblia is a parasitic protist with unusual genomic traits: it is binucleated and tetraploid, has a very compact genome, displays a theme of genomic minimalism with cellular machinery commonly comprised of a reduced number of protein components, and has a remarkably large population of long, stable, noncoding, antisense RNAs. Here we use in silico approaches to investigate the major RNA degradation machinery in Giardia lamblia and compare it to a broad array of other parasitic protists. We have found key constituents of the deadenylation and decapping machinery and of the 5'-3' RNA degradation pathway. We have similarly found that all of the major 3'-5' RNA degradation pathways are present in Giardia, including both exosome-dependent and exosome-independent machinery. However, we observe significant loss of RNA degradation machinery genes that will result in important differences in the protein composition, and potentially functionality, of the various RNA degradation pathways. This is most apparent in the exosome, the central mediator of 3'-5' degradation, which apparently contains an altered core configuration in both Giardia and Plasmodium, with only four, instead of the canonical six, distinct subunits. Additionally the exosome in Giardia is missing both the Rrp6, Nab3, and Nrd1 proteins, known to be key regulators of noncoding transcript stability in other cells. These findings suggest that although the full complement of the major RNA degradation mechanisms were present - and likely functional - early in eukaryotic evolution, the composition and function of the complexes is more variable than previously

  14. First report of Giardia sp. and Cryptosporidium sp. in fecal samples in blue macaw (Anodorhynchus hyacinthinus in southern Brazil

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    Matheus Hillard Farret

    2010-09-01

    Full Text Available This study reports the gastrointestinal parasitism by Giardia sp. and Cryptosporidium sp. in blue macaw (Anodorhynchus hyacinthinus in the southern region of Brazil. Fecal samples of two species kept in captivity in the state of Rio Grande do Sul were analyzed by the direct smear method, centrifugal flotation technique with zinc sulfate and Kinyoun staining technique for research of parasites. Mixed infection by eggs of Capillaria, cysts of Giardia sp. and oocysts of Cryptosporidium sp. was observed. This is the first report this protozoa in blue macaw.

  15. Estimating Cryptosporidium and Giardia disease burdens for children drinking untreated groundwater in a rural population in India.

    Science.gov (United States)

    Daniels, Miles E; Smith, Woutrina A; Jenkins, Marion W

    2018-01-01

    In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children < 5 in rural India. We studied a population of 3,385 children < 5 years of age in 100 communities of Puri District, Odisha, India. We developed a coupled quantitative microbial risk assessment (QMRA) and susceptible-infected-recovered (SIR) population model based on observed levels of Cryptosporidium and Giardia in improved groundwater sources used for drinking and compared the QMRA-SIR estimates with independently measured all-cause (i.e., all fecal-oral enteric pathogens and exposure pathways) child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013). We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06-1.53%), far exceeded the tolerable daily risk of infection from drinking water in the US (< 0.0001%). Depending on which protozoa species were present, median estimates of daily child diarrhea prevalence due to either Cryptosporidium or Giardia infection from drinking water was as high as 6.5% or as low as < 1% and accounted for at least 2.9% and as much as 65.8% of the all-cause diarrhea disease burden measured in children < 5 during the study period. Cryptosporidium tended to account for a greater share of estimated waterborne protozoa infections causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne Cryptosporidium

  16. Spatial and Molecular Epidemiology of Giardia intestinalis Deep in the Amazon, Brazil.

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    Beatriz Coronato Nunes

    Full Text Available Current control policies for intestinal parasitosis focuses on soil-transmitted helminths, being ineffective against Giardia intestinalis, a highly prevalent protozoon that impacts children's nutritional status in developing countries. The objective of this study was to explore spatial and molecular epidemiology of Giardia intestinalis in children of Amerindian descent in the Brazilian Amazon.A cross sectional survey was performed in the Brazilian Amazon with 433 children aged 1 to 14 years. Fecal samples were processed through parasitological techniques and molecular characterization. Prevalence of G. intestinalis infection was 16.9% (73/433, reaching 22.2% (35/158 among children aged 2-5 years, and a wide distribution throughout the city with some hot spots. Positivity-rate was similar among children living in distinct socioeconomic strata (48/280 [17.1%] and 19/116 [16.4%] below and above the poverty line, respectively. Sequencing of the β-giardin gene revealed 52.2% (n = 12 of assemblage A and 47.8% (n = 11 of assemblage B with high haplotype diversity for the latter. The isolates clustered into two well-supported G. intestinalis clades. A total of 38 haplotypes were obtained, with the following subassemblages distribution: 5.3% (n = 2 AII, 26.3% (n = 10 AIII, 7.9% (n = 3 BIII, and 60.5% (n = 23 new B genotypes not previously described.Giardia intestinalis infection presents a high prevalence rate among Amerindian descended children living in Santa Isabel do Rio Negro/Amazon. The wide distribution observed in a small city suggests the presence of multiple sources of infection, which could be related to environmental contamination with feces, possibly of human and animal origin, highlighting the need of improving sanitation, safe water supply and access to diagnosis and adequate treatment of infections.

  17. Differential gene expression in Giardia lamblia under oxidative stress: significance in eukaryotic evolution.

    Science.gov (United States)

    Raj, Dibyendu; Ghosh, Esha; Mukherjee, Avik K; Nozaki, Tomoyoshi; Ganguly, Sandipan

    2014-02-10

    Giardia lamblia is a unicellular, early branching eukaryote causing giardiasis, one of the most common human enteric diseases. Giardia, a microaerophilic protozoan parasite has to build up mechanisms to protect themselves against oxidative stress within the human gut (oxygen concentration 60 μM) to establish its pathogenesis. G. lamblia is devoid of the conventional mechanisms of the oxidative stress management system, including superoxide dismutase, catalase, peroxidase, and glutathione cycling, which are present in most eukaryotes. NADH oxidase is a major component of the electron transport chain of G. lamblia, which in concurrence with disulfide reductase, protects oxygen-labile proteins such as pyruvate: ferredoxin oxidoreductase against oxidative stress by sustaining a reduced intracellular environment. It also contains the arginine dihydrolase pathway, which occurs in a number of anaerobic prokaryotes, includes substrate level phosphorylation and adequately active to make a major contribution to ATP production. To study differential gene expression under three types of oxidative stress, a Giardia genomic DNA array was constructed and hybridized with labeled cDNA of cells with or without stress. The transcriptomic data has been analyzed and further validated using real time PCR. We identified that out of 9216 genes represented on the array, more than 200 genes encoded proteins with functions in metabolism, oxidative stress management, signaling, reproduction and cell division, programmed cell death and cytoskeleton. We recognized genes modulated by at least ≥ 2 fold at a significant time point in response to oxidative stress. The study has highlighted the genes that are differentially expressed during the three experimental conditions which regulate the stress management pathway differently to achieve redox homeostasis. Identification of some unique genes in oxidative stress regulation may help in new drug designing for this common enteric parasite prone to

  18. Prevalence and genetic characterization of Cryptosporidium, Enterocytozoon, Giardia and Cyclospora in diarrheal outpatients in China.

    Science.gov (United States)

    Liu, Hua; Shen, Yujuan; Yin, Jianhai; Yuan, Zhongying; Jiang, Yanyan; Xu, Yuxin; Pan, Wei; Hu, Yuan; Cao, Jianping

    2014-01-13

    Cryptosporidium spp., Enterocytozoon spp., Giardia spp. and Cyclospora spp. are important intestinal protozoan parasites causing diarrhea in humans, livestocks and wildlife and have a significant impact on public health. No reports exist about simultaneous prevalence rates or genotyping data of these four parasites in outpatients from China. Fecal specimens from 252 diarrhea patients in a pediatric clinic (n = 169) and an intestinal clinic (n = 83) of a hospital in Shanghai, China, were collected between October 2012 and March 2013. All samples were examined for the presence of the four parasites by using molecular methods. In total, 76/252 (30.16%) patients were positive for at least one intestinal parasite, of which Cryptosporidium spp., Enterocytozoon bieneusi and Giardia intestinalis were detected by nested PCR in 34 (13.49%), 34 (13.49%) and 17 (6.75%) of the fecal specimens, respectively. Sequence analysis showed that all Cryptosporidium-positive specimens were C. andersoni and that most G. intestinalis- positive patients were infected by assemblage C, which is usually found in canids, while only one sample was from assemblage B. Eight patients were co-infected with Cryptosporidium spp. and Enterocytozoon, while one was co-infected with Cryptosporidium and Giardia. The patients infected with Cryptosporidium and Enterocytozoon bieneusi had higher infection rates in winter than in spring in this area. Data indicated that C. andersoni is the fourth major Cryptosporidium species infecting humans in addition to C. hominis, C. parvum and C. meleagridis. Our study also revealed a short-term outbreak of cryptosporidiosis and microsporidiosis and sporadic cases of giardiasis that occurred among humans in Shanghai, China.

  19. Arginine consumption by the intestinal parasite Giardia intestinalis reduces proliferation of intestinal epithelial cells.

    Science.gov (United States)

    Stadelmann, Britta; Merino, María C; Persson, Lo; Svärd, Staffan G

    2012-01-01

    In the field of infectious diseases the multifaceted amino acid arginine has reached special attention as substrate for the hosts production of the antimicrobial agent nitric oxide (NO). A variety of infectious organisms interfere with this part of the host immune response by reducing the availability of arginine. This prompted us to further investigate additional roles of arginine during pathogen infections. As a model we used the intestinal parasite Giardia intestinalis that actively consumes arginine as main energy source and secretes an arginine-consuming enzyme, arginine deiminase (ADI). Reduced intestinal epithelial cell (IEC) proliferation is a common theme during bacterial and viral intestinal infections, but it has never been connected to arginine-consumption. Our specific question was thereby, whether the arginine-consumption by Giardia leads to reduced IEC proliferation, in addition to NO reduction. In vitro cultivation of human IEC lines in arginine-free or arginine/citrulline-complemented medium, as well as in interaction with different G. intestinalis isolates, were used to study effects on host cell replication by MTT assay. IEC proliferation was further analyzed by DNA content analysis, polyamine measurements and expressional analysis of cell cycle regulatory genes. IEC proliferation was reduced upon arginine-withdrawal and also in an arginine-dependent manner upon interaction with G. intestinalis or addition of Giardia ADI. We show that arginine-withdrawal by intestinal pathogens leads to a halt in the cell cycle in IECs through reduced polyamine levels and upregulated cell cycle inhibitory genes. This is of importance with regards to intestinal tissue homeostasis that is affected through reduced cell proliferation. Thus, the slower epithelial cell turnover helps the pathogen to maintain a more stable niche for colonization. This study also shows why supplementation therapy of diarrhea patients with arginine/citrulline is helpful and that

  20. Age and sex distribution of infections with Entamoeba histolytica and Giardia intestinalis in the Lagos population.

    Science.gov (United States)

    Oyerinde, J P; Ogunbi, O; Alonge, A A

    1977-09-01

    Stool samples were collected from 2,099 persons and examined for parasites: 9.5 percent were infected with Entamoeba histolytica, 7.9 percent were infected with Giardia intestinalis and 15.5 percent were infected with Iodamoeba butschlii. The infections with E. histolytica and I. butschlii occurred more frequently in females than in males and adults were also more frequently infected than children. Infection with G. intestinalis, on the other hand, was more prevalent in males than in females and children were more frequently infected than adults with the highest incidence of infection (13.8 percent) in the age-group 1-5 years.

  1. In vitro inhibition of Giardia lamblia and Trichomonas vaginalis growth by bithionol, dichlorophene, and hexachlorophene.

    OpenAIRE

    Takeuchi, T; Kobayashi, S; Tanabe, M; Fujiwara, T

    1985-01-01

    Bithionol, dichlorophene, and hexachlorophene, which are used in treating some helminthic infections, killed trophozoites of Giardia lamblia and Trichomonas vaginalis in modified BI-S-33 and Asami media, respectively. Virtually all G. lamblia and T. vaginalis cells were killed within 24 h with a 0.42 mM concentration of these compounds, except that 0.93 mM dichlorophene was required for sterilizing T. vaginalis in the same period. In modified BI-S-33 and Asami media from which bovine and huma...

  2. Phosphonoxins III: synthesis of α-aminophosphonate analogs of antifungal polyoxins with anti-Giardia activity.

    Science.gov (United States)

    Staake, Michael; Chauhan, Jay; Zhou, Ding; Shanker, Aaron; De Chatterjee, Atasi; Das, Siddhartha; Patterson, Steven E

    2010-10-15

    A synthesis of α-aminophosphonate analogs of polyoxins, termed phosphonoxin C1, C2, and C3, has been achieved. The key step was the addition of lithium dimethyl phosphite to the aldehyde of a protected threose derivative. α-Hydroxyphosphonate analogs C4 and C5 were also obtained by taking advantage of an unprecedented conversion of an azide to hydroxyl during treatment with hydrogen on palladium on carbon. The resulting phosphonoxin C5 inhibited the growth of an intestinal protozoan, Giardia lamblia, at low micromolar concentration.

  3. Imunodeficiência comum variável : manifestações gastrintestinais a Giardia lamblia

    OpenAIRE

    Carvalho, Maria Dinah Jacob da Silva de, 1967-

    2011-01-01

    Tese de mestrado, Microbiologia Clínica, Faculdade de Medicina, Universidade de Lisboa, 2012 A imunodeficiência comum variável (IDCV) é segunda imunodeficiência primária mais frequente a seguir à deficiência selectiva de IgA. As manifestações gastrintestinais na IDCV apresentam uma prevalência de 20% a 60% e a infecção por Giardia lamblia é a causa infecciosa mais frequentemente identificada de diarreia e mal-absorção, em cerca de 30% dos doentes. Escassos estudos documentam a ...

  4. Phosphonoxins III: Synthesis of α-Aminophosphonate Analogs of Antifungal Polyoxins with Anti-Giardia activity

    Science.gov (United States)

    Staake, Michael; Chauhan, Jay; Zhou, Ding; Shanker, Aaron; De Chatterjee, Atasi; Das, Siddhartha; Patterson, Steven E.

    2010-01-01

    A synthesis of α-aminophosphonate analogs of polyoxins, termed phosphonoxin C1, C2 and C3, has been achieved. The key step was the addition of lithium dimethyl phosphite to the aldehyde of a protected threose derivative. α-Hydroxyphosphonate analogs C4 and C5 were also obtained by taking advantage of an unprecedented conversion of an azide to hydroxyl during treatment with hydrogen on palladium on carbon. The resulting phosphonoxin C5 inhibited the growth of an intestinal protozoan, Giardia lamblia, at low micromolar concentration. PMID:20857975

  5. Cross-modulation of pathogen-specific pathways enhances malnutrition during enteric co-infection with Giardia lamblia and enteroaggregative Escherichia coli.

    Science.gov (United States)

    Bartelt, Luther A; Bolick, David T; Mayneris-Perxachs, Jordi; Kolling, Glynis L; Medlock, Gregory L; Zaenker, Edna I; Donowitz, Jeffery; Thomas-Beckett, Rose Viguna; Rogala, Allison; Carroll, Ian M; Singer, Steven M; Papin, Jason; Swann, Jonathan R; Guerrant, Richard L

    2017-07-01

    Diverse enteropathogen exposures associate with childhood malnutrition. To elucidate mechanistic pathways whereby enteric microbes interact during malnutrition, we used protein deficiency in mice to develop a new model of co-enteropathogen enteropathy. Focusing on common enteropathogens in malnourished children, Giardia lamblia and enteroaggregative Escherichia coli (EAEC), we provide new insights into intersecting pathogen-specific mechanisms that enhance malnutrition. We show for the first time that during protein malnutrition, the intestinal microbiota permits persistent Giardia colonization and simultaneously contributes to growth impairment. Despite signals of intestinal injury, such as IL1α, Giardia-infected mice lack pro-inflammatory intestinal responses, similar to endemic pediatric Giardia infections. Rather, Giardia perturbs microbial host co-metabolites of proteolysis during growth impairment, whereas host nicotinamide utilization adaptations that correspond with growth recovery increase. EAEC promotes intestinal inflammation and markers of myeloid cell activation. During co-infection, intestinal inflammatory signaling and cellular recruitment responses to EAEC are preserved together with a Giardia-mediated diminishment in myeloid cell activation. Conversely, EAEC extinguishes markers of host energy expenditure regulatory responses to Giardia, as host metabolic adaptations appear exhausted. Integrating immunologic and metabolic profiles during co-pathogen infection and malnutrition, we develop a working mechanistic model of how cumulative diet-induced and pathogen-triggered microbial perturbations result in an increasingly wasted host.

  6. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

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    Renata Angeli

    2009-01-01

    Full Text Available A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A and Cratylia mollis (Cramoll 1 and Cramoll 1,4 did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column.

  7. Identification and molecular characterization of Cryptosporidium and Giardia in children and cattle populations from the province of Alava, North of Spain

    Energy Technology Data Exchange (ETDEWEB)

    Cardona, Guillermo A. [Livestock Laboratory, Regional Government of Alava, Ctra. de Azua 4, 01520 Vitoria-Gasteiz (Spain); Carabin, Helene [Department of Biostatistics and Epidemiology, College of Public Health, Oklahoma University Health Sciences Center, 801 Northeast 13th Street, Room 309, Oklahoma City, OK 73104 (United States); Goni, Pilar [Department of Microbiology, Preventive Medicine and Public Health, Faculty of Medicine, University of Zaragoza, Domingo Miral s/n, 50009 Zaragoza (Spain); Arriola, Larraitz [Epidemiology Unit, Public Health Division of Guipuzcoa, Basque Government, Av. Navarra 4, 2013 San Sebastian (Spain); Robinson, Guy [UK Cryptosporidium Reference Unit, Public Health Wales, Microbiology ABM, Swansea, Singleton Hospital, Swansea SA2 8QA (United Kingdom); Fernandez-Crespo, Juan C. [Sub-direction of Public Health of Alava, Department of Health, Basque Government, Avda. Santiago 11, 01002 Vitoria-Gasteiz (Spain); Clavel, Antonio [Department of Microbiology, Preventive Medicine and Public Health, Faculty of Medicine, University of Zaragoza, Domingo Miral s/n, 50009 Zaragoza (Spain); Chalmers, Rachel M. [UK Cryptosporidium Reference Unit, Public Health Wales, Microbiology ABM, Swansea, Singleton Hospital, Swansea SA2 8QA (United Kingdom); Carmena, David, E-mail: d.carmena@imperial.ac.uk [MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN (United Kingdom)

    2011-12-15

    The prevalence of and factors associated with the protozoan enteropathogens Cryptosporidium and Giardia have been investigated in selected children and cattle populations from the province of Alava (Northern Spain). The presence of these organisms was detected in fecal samples using commercially available coproantigen-ELISA (CpAg-ELISA) and immunochromatographic (ICT) assays. A total of 327 caregivers of children participants were asked to answer questions on risk factors potentially associated to the prevalence of Cryptosporidium and Giardia, including water-use practices, water sports and contact with domestic or pet animals. Molecular analyses were conducted using a nested-PCR technique to amplify the small-subunit (SSU) rRNA gene of Cryptosporidium and the triosephosphate isomerase (tpi) gene of Giardia. Cryptosporidium oocysts and Giardia cysts were found in 3 and 16 samples using the CpAg-ELISA, and in 5 and 9 samples using the ICT test, respectively. Cryptosporidium and Giardia were also found in 7 and 17 samples by CpAg-ELISA, and 4 and 14 samples by ICT, respectively, of 227 cattle fecal samples. The overall Cryptosporidium and Giardia infection prevalences, based on a Bayesian approach accounting for the imperfect sensitivities and specificities of both diagnostic tests, were estimated to 1.0% (95% BCI: 0.2%-2.8%) and 3.1% (1.5%-5.3%) in children and 3.0% (0.5%-9.2%) and 1.4% (0.0%-6.4%) in cattle, respectively. In humans, a single Cryptosporidium isolate was characterized as C. hominis. Of seven Giardia isolates, four were identified as assemblage B, two as assemblage A-II and one was a mixed assemblage B + A-II infection. No Cryptosporidium or Giardia isolates could be obtained from cattle samples. Although limited, these results seem to suggest that cattle are unlikely to be an important reservoir of zoonotic Cryptosporidium and/or Giardia in the province of Alava.

  8. Identification of an immunogenic protein of Giardia lamblia using monoclonal antibodies generated from infected mice

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    Jael Quintero

    2013-08-01

    Full Text Available The humoral immune response plays an important role in the clearance of Giardia lamblia. However, our knowledge about the specific antigens of G. lamblia that induce a protective immune response is limited. The purpose of this study was to identify and characterise the immunogenic proteins of G. lamblia in a mouse model. We generated monoclonal antibodies (moAbs specific to G. lamblia (1B10, 2C9.D11, 3C10.E5, 3D10, 5G8.B5, 5F4, 4C7, 3C5 and 3C6 by fusing splenocytes derived from infected mice. Most of these moAbs recognised a band of ± 71 kDa (5G8 protein and this protein was also recognised by serum from the infected mice. We found that the moAbs recognised conformational epitopes of the 5G8 protein and that this antigen is expressed on the cell surface and inside trophozoites. Additionally, antibodies specific to the 5G8 protein induced strong agglutination (> 70-90% of trophozoites. We have thus identified a highly immunogenic antigen of G. lamblia that is recognised by the immune system of infected mice. In summary, this study describes the identification and partial characterisation of an immunogenic protein of G. lamblia. Additionally, we generated a panel of moAbs specific for this protein that will be useful for the biochemical and immunological characterisation of this immunologically interesting Giardia molecule.

  9. The presence of Giardia intestinalis in donkeys, Equus asinus, in China.

    Science.gov (United States)

    Zhang, Xiao-Xuan; Zhang, Fu-Kai; Li, Fa-Cai; Hou, Jun-Ling; Zheng, Wen-Bin; Du, Shuai-Zhi; Zhao, Quan; Zhu, Xing-Quan

    2017-01-03

    Giardia intestinalis is one of the most important zoonotic enteric parasites. As no information regarding prevalence and genotype of G. intestinalis in donkeys (Equus asinus) in China is available, 181 faecal samples from 48 donkeys from Jilin Province, from 104 from Shandong Province and from 29 from Liaoning Province were examined between May and December 2015. Twenty-eight (15.47%) out of 181 donkey samples were tested G. intestinalis-positive by nested amplification of the triosephosphate isomerase (tpi) gene. The prevalence in different regional groups varied from 10.42 to 18.27%. The prevalence in adult and young donkeys was 14.29 and 22.92%, respectively. Otherwise, the prevalence was 11.69% in summer and 18.27% in winter. However, no statistically significant differences were found in relation to region or age group. Sequence analysis of the tpi, glutamate dehydrogenase (gdh) and beta giardin (bg) loci identified 4, 1 and 3 subtypes of assemblage B, respectively. Moreover, four novel multilocus genotypes (MLGs novel-1 to novel-4) were identified in assemblage B. This first report of G. intestinalis in donkeys in China indicates that further studies of nation-wide molecular epidemiology and geographical distribution of Giardia in donkeys are warranted. Effective strategies should be implemented to control G. intestinalis infection in donkeys, other animals and humans.

  10. The Study of the Multilayer Filtration Process Efficiency on Giardia lamblia Elimination in Isfahan Water Treatment Plant

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    Zahra Mosaviyan

    2014-04-01

    Full Text Available Introduction: Giardia lamblia is a waterborne highly infectious protozoan parasite capable of causing gastrointestinal illness. The multilayer filtration is used in many applications such as turbidity removes from surface waters and Bacteria, Viruses and protozoan removes. This study was investigated in full-scale the efficiency of filtration process during water treatment that remove Giardia cyst in water at Isfahan province. Materials and methods: We used Information Collection Rule method (ICR. In this method the polypropylene yarn-wound cartridge filter for isolation of these parasites was examined and followed by elution, sample concentration, flotation by percoll-sucrose solution and immunofluorescence antibody (IFA staining to recognize them. Results: Result showed that three layer filtration had a maximum 2.3 log10 for remove Giardia cyst. Cysts removal in water filtration is likely to be comparable to the efficiency of turbidity. Efficiency removal was 99.5% for Giardia and 92.7% for turbidity in filtration stage. We detected 0.2 cyst per 100 liter and per 100 liter in filtered water. This observation is according to U.S.EPA standards. The number of cysts were more in high pH samples. Discussion and conclusion: Analysis of physical processes of treatment water in Isfahan plant configurations showed that granular filters (include sand, anthracite and garnet filter were more likely to have effluence in removal cysts than dual filters.

  11. Relationships between the occurrence of Giardia and Cryptosporidium and physicochemical properties of marine waters of the Pacific Coast of Mexico.

    Science.gov (United States)

    Magana-Ordorica, Dalia; Mena, Kristina; Valdez-Torres, Jose B; Soto-Beltran, Marcela; Leon-Felix, Josefina; Chaidez, Cristobal

    2010-12-01

    Untreated sewage has adversely affected the quality of marine recreational waters worldwide. Exposure to marine recreational water with poor microbial quality may pose a threat to bathers. The objectives of this study were to assess the effect of physicochemical parameters on Cryptosporidium and Giardia presence in marine recreational water of Sinaloa, Mexico, by Logistic Regression Analyses. Thirty-two 10-litre water samples were collected from two tourist beaches, Altata and Mazatlan, between November 2006 and May 2007. Water samples were processed by the EPA 1623 method and pH, temperature, salinity and turbidity were also determined. Cryptosporidium and Giardia were present in 71 and 57% of the samples collected from Altata, respectively. In Mazatlan, Cryptosporidium and Giardia were found in 83 and 72% of the samples, respectively. The overall concentration of Cryptosporidium ranged from 150 to 2,050 oocysts/10 L with an average of 581 oocysts/10 L and Giardia ranged from 10 to 300 cysts/10 L with an average of 73 cysts/10 L. The occurrence of both parasites increased in water with decreasing temperatures and increasing turbidity of the water.

  12. Investigation into Cryptosporidium and Giardia in bivalve mollusks farmed in Sardinia region and destined for human consumption

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    Tiziana Tedde

    2013-06-01

    Full Text Available Cryptosporidium and Giardia are protozoan parasites transmitted by fecal-oral ingestion of (oocysts, and are responsible for enteritis in several animal species and humans worldwide. These (oocysts can survive for over a year in aquatic environments and can accumulate in bivalve mollusks, which filter large volumes of water. The aim of this study is to evaluate the natural occurrence of Cryptosporidium and Giardia contamination in different specimens of edible bivalves mollusks from farming sites of the western and north-eastern coasts of Sardinia. From April 2011 to February 2012, 1095 specimens of Mytilus galloprovincialis and 240 of Crassostrea gigas were sampled from Olbia and Oristano gulf and San Teodoro pond. Hepatopancreas and gills, including the labial palp, were examined for oocysts and cysts after pooling and homogenisation using different techniques: i staining for light and fluorescence microscopy; ii direct immunofluorescence (IF Merifluor® test Cryptosporidium/ Giardia (Meridian Bioscience Inc., Cincinnati, OH, USA; and iii molecular procedures. However, in the context under study, all mollusks examined with the three main diagnostic techniques were negative for both parasites pointing out the hypothetically low zoonotic risk related to Cryptosporidium and Giardia in bivalves, especially Mytilus galloprovincialis and Crassostrea gigas.

  13. Detection of Cryptosporidium oocysts and Giardia cysts in water samples with a Becton Dickinson FACSort flow cytometer

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Boschman GD; LWL; Becton Dickinson Europe; Aalst; Belgie

    1995-01-01

    De huidige methode die in de Verenigde Staten, Groot- Brittannie en Nederland wordt gebruikt voor de detectie van Cryptosporidium en Giardia in water bestaat uit een combinatie van filtratie, concentratie door centrifugeren en flotatie en immuunfluorescentie microscopie. De methode is zeer

  14. Simultaneous detection of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum in fecal samples by using multiplex real-time PCR

    NARCIS (Netherlands)

    Verweij, Jaco J.; Blangé, Roy A.; Templeton, Kate; Schinkel, Janke; Brienen, Eric A. T.; van Rooyen, Marianne A. A.; van Lieshout, Lisette; Polderman, Anton M.

    2004-01-01

    Entamoeba histolytica, Giardia lamblia, and Cryptosporidium are three of the most important diarrhea-causing parasitic protozoa. For many years, microscopic examination of stool samples has been considered to be the "gold standard" for diagnosis of E. histolytica, G. lamblia, and C. parvum

  15. The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target

    NARCIS (Netherlands)

    Kunz, Stefan; Balmer, Vreni; Sterk, Geert Jan; Pollastri, Michael P.; Leurs, Rob; Müller, Norbert; Hemphill, Andrew; Spycher, Cornelia

    2017-01-01

    Background: Giardiasis is an intestinal infection correlated with poverty and poor drinking water quality, and treatment options are limited. According to the Center for Disease Control and Prevention, Giardia infections afflict nearly 33% of people in developing countries, and 2% of the adult

  16. Molecular epidemiology and multilocus sequence analysis of potentially zoonotic Giardia spp. from humans and dogs in Jamaica.

    Science.gov (United States)

    Lee, Mellesia F; Cadogan, Paul; Eytle, Sarah; Copeland, Sonia; Walochnik, Julia; Lindo, John F

    2017-01-01

    Giardia spp. are the causative agents of intestinal infections in a wide variety of mammals including humans and companion animals. Dogs may be reservoirs of zoonotic Giardia spp.; however, the potential for transmission between dogs and humans in Jamaica has not been studied. Conventional PCR was used to screen 285 human and 225 dog stool samples for Giardia targeting the SSU rDNA gene followed by multilocus sequencing of the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), and β-giardin (bg) genes. Prevalence of human infections based on PCR was 6.7 % (19/285) and canine infections 19.6 % (44/225). Nested PCR conducted on all 63 positive samples revealed the exclusive presence of assemblage A in both humans and dogs. Sub-assemblage A-II was responsible for 79.0 % (15/19) and 70.5 % (31/44) of the infections in humans and dogs, respectively, while sub-assemblage A-I was identified at a rate of 15.8 % (3/19) and 29.5 % (13/44) in humans and dogs, respectively. The predominance of a single circulating assemblage among both humans and dogs in Jamaica suggests possible zoonotic transmission of Giardia infections.

  17. Protection against diarrhea associated with Giardia intestinalis is lost with Multi-Nutrient Supplementation: A Study in Tanzanian Children

    NARCIS (Netherlands)

    Veenemans, J.; Mank, T.; Ottenhof, M.; Baidjoe, A.Y.; Mbugi, E.V.; Demir, A.Y.; Wielders, J.P.M.; Savelkoul, H.F.J.; Verhoef, J.C.M.

    2011-01-01

    Background - Asymptomatic carriage of Giardia intestinalis is highly prevalent among children in developing countries, and evidence regarding its role as a diarrhea-causing agent in these settings is controversial. Impaired linear growth and cognition have been associated with giardiasis, presumably

  18. Draft genome sequencing of giardia intestinalis assemblage B isolate GS: is human giardiasis caused by two different species?

    Directory of Open Access Journals (Sweden)

    Oscar Franzén

    2009-08-01

    Full Text Available Giardia intestinalis is a major cause of diarrheal disease worldwide and two major Giardia genotypes, assemblages A and B, infect humans. The genome of assemblage A parasite WB was recently sequenced, and the structurally compact 11.7 Mbp genome contains simplified basic cellular machineries and metabolism. We here performed 454 sequencing to 16x coverage of the assemblage B isolate GS, the only Giardia isolate successfully used to experimentally infect animals and humans. The two genomes show 77% nucleotide and 78% amino-acid identity in protein coding regions. Comparative analysis identified 28 unique GS and 3 unique WB protein coding genes, and the variable surface protein (VSP repertoires of the two isolates are completely different. The promoters of several enzymes involved in the synthesis of the cyst-wall lack binding sites for encystation-specific transcription factors in GS. Several synteny-breaks were detected and verified. The tetraploid GS genome shows higher levels of overall allelic sequence polymorphism (0.5 versus <0.01% in WB. The genomic differences between WB and GS may explain some of the observed biological and clinical differences between the two isolates, and it suggests that assemblage A and B Giardia can be two different species.

  19. A Purified Recombinant Lipopeptide as Adjuvant for Cancer Immunotherapy

    Directory of Open Access Journals (Sweden)

    Ying-Chyi Song

    2014-01-01

    Full Text Available Synthetic lipopeptides have been widely used as vaccine adjuvants to enhance immune responses. The present study demonstrated that the tryptic N-terminal fragment of the lipoprotein rlipo-D1E3 (lipo-Nter induces superior antitumor effects compared to a synthetic lipopeptide. The lipo-Nter was purified and formulated with protein or peptide vaccines to determine if lipo-Nter could be used as a novel adjuvant and could induce antitumor immunity in a cervical cancer model. Purified lipo-Nter activated the maturation of bone marrow-derived dendritic cells (BM-DCs, leading to the secretion of TNF-α through TLR2/6 but not TLR1/2. A recombinant mutant HPV16 E7 (rE7m protein was mixed with lipo-Nter to immunize the mice; the anti-E7 antibody titers were increased, and the T helper cells were skewed toward the Th1 fate (increased IL-2 and decreased IL-5 secretion. Single-dose injection of rE7m and lipo-Nter inhibited tumor growth, but the injection of rE7m alone did not. Accordingly, lipo-Nter also enhanced the antitumor immunity of the E7-derived peptide but not the synthetic lipopeptide (Pam3CSK4. We demonstrated that the lipo-Nter of a bacterial-derived recombinant lipoprotein is a novel adjuvant that could be used for the development of a new generation of vaccines.

  20. Tea derived galloylated polyphenols cross-link purified gastrointestinal mucins.

    Directory of Open Access Journals (Sweden)

    Pantelis Georgiades

    Full Text Available Polyphenols derived from tea are thought to be important for human health. We show using a combination of particle tracking microrheology and small-angle neutron scattering that polyphenols acts as cross-linkers for purified gastrointestinal mucin, derived from the stomach and the duodenum. Both naturally derived purified polyphenols, and green and black tea extracts are shown to act as cross-linkers. The main active cross-linking component is found to be the galloylated forms of catechins. The viscosity, elasticity and relaxation time of the mucin solutions experience an order of magnitude change in value upon addition of the polyphenol cross-linkers. Similarly small-angle neutron scattering experiments demonstrate a sol-gel transition with the addition of polyphenols, with a large increase in the scattering at low angles, which is attributed to the formation of large scale (>10 nm heterogeneities during gelation. Cross-linking of mucins by polyphenols is thus expected to have an impact on the physicochemical environment of both the stomach and duodenum; polyphenols are expected to modulate the barrier properties of mucus, nutrient absorption through mucus and the viscoelastic microenvironments of intestinal bacteria.

  1. Risk assessment of Giardia from a full scale MBR sewage treatment plant caused by membrane integrity failure.

    Science.gov (United States)

    Zhang, Yu; Chen, Zhimin; An, Wei; Xiao, Shumin; Yuan, Hongying; Zhang, Dongqing; Yang, Min

    2015-04-01

    Membrane bioreactors (MBR) are highly efficient at intercepting particles and microbes and have become an important technology for wastewater reclamation. However, many pathogens can accumulate in activated sludge due to the long residence time usually adopted in MBR, and thus may pose health risks when membrane integrity problems occur. This study presents data from a survey on the occurrence of water-borne Giardia pathogens in reclaimed water from a full-scale wastewater treatment plant with MBR experiencing membrane integrity failure, and assessed the associated risk for green space irrigation. Due to membrane integrity failure, the MBR effluent turbidity varied between 0.23 and 1.90 NTU over a period of eight months. Though this turbidity level still met reclaimed water quality standards (≤5 NTU), Giardia were detected at concentrations of 0.3 to 95 cysts/10 L, with a close correlation between effluent turbidity and Giardia concentration. All β-giardin gene sequences of Giardia in the WWTP influents were genotyped as Assemblages A and B, both of which are known to infect humans. An exponential dose-response model was applied to assess the risk of infection by Giardia. The risk in the MBR effluent with chlorination was 9.83×10(-3), higher than the acceptable annual risk of 1.0×10(-4). This study suggested that membrane integrity is very important for keeping a low pathogen level, and multiple barriers are needed to ensure the biological safety of MBR effluent. Copyright © 2015. Published by Elsevier B.V.

  2. Association between Whipple's disease and Giardia lamblia infection Asociación de enfermedad de Whipple e infección por Giardia lamblia

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    J. A. Gil Ruiz

    2005-07-01

    Full Text Available Whipple'disease is mainly characterized by affecting the digestive system, although it can be a multisystemic process with different clinical symptoms. The bacillus causing the disease has been isolated and cultivated in 2000 and the genome sequence has been recently analyzed in 2003, which means new perspectives for its diagnosis and treatment. Giardiasis is an infestation caused by a protozoo and may cause a malabsorption syndrome or run in a subclinic way. The case of a middle-aged male is described, who after a three-year period of migratory arthralgias, showed weight loss, diarrheas and abdominal pain, being diagnosed of Giardiasis, and after the persistent symptoms and a number of studies, was diagnosed with Whipple disease. Nineteen cases of Giardia-Whipple coinfection have been described in the literature, but the reason of this association has not been found yet. The discussion on whether there is an alteration in the inmunitary system which facilitates infections or, the development of an infection lead to the other one, goes on.La enfermedad de Whipple se caracteriza por afectar principalmente al aparato digestivo, aunque puede ser un proceso multisistémico con variadas manifestaciones clínicas. En el año 2000 se ha conseguido aislar y cultivar el bacilo causante y recientemente, en el 2003, se ha analizado la secuencia completa del genoma, lo que supone nuevas perspectivas para el diagnóstico y el tratamiento. La giardiasis es una infección originada por un protozoo que puede evolucionar de forma subclínica o dar lugar a un síndrome de malabsorción. Describimos el caso de un paciente varón de mediana edad, que tras un periodo de tres años de artralgias migratorias en articulaciones periféricas, presentó un cuadro de pérdida de peso, con aumento del número de deposiciones y molestias abdominales siendo diagnosticado de Giardiasis y ante la persistencia de los síntomas y la posterior realización de diferentes estudios

  3. Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

    Directory of Open Access Journals (Sweden)

    Silvia Cristina Osaki

    2011-10-01

    Full Text Available INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC. Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

  4. Measurement of Ozone Emission and Particle Removal Rates from Portable Air Purifiers

    Science.gov (United States)

    Mang, Stephen A.; Walser, Maggie L.; Nizkorodov, Sergey A.; Laux, John M.

    2009-01-01

    Portable air purifiers are popular consumer items, especially in areas with poor air quality. Unfortunately, most users of these air purifiers have minimal understanding of the factors affecting their efficiency in typical indoor settings. Emission of the air pollutant ozone (O[subscript 3]) by certain air purifiers is of particular concern. In an…

  5. Axial dispersion in a Kureha Crystal Purifier (KCP)

    Science.gov (United States)

    Otawara, K.; Matsuoka, T.

    2002-04-01

    It is widely accepted that the degree of axial back-mixing in the equipment affects the performance of a column crystallizer. Specifically, small such mixing is inevitable for obtaining highly pure crystal. In fact, it is believed that scale-up of some column crystallizers has been terminated due to the large axial back-mixing or maldistribution. Thus, experiments have been performed for estimating axial dispersion coefficients of liquid phase representing axial back-mixing in the column of a Kureha Crystal Purifier (KCP). The effect of the column diameter on such coefficients has been investigated and it has become evident that the axial back-mixing in the column is more significant in the larger column. Nevertheless, the results have also indicated that the axial back-mixing in KCPs of industrial sizes can be substantially smaller than those in other types of column crystallizers.

  6. Altered glycosylation profile of purified plasma ACT from Alzheimer's disease.

    Science.gov (United States)

    Ianni, Manuela; Manerba, Marcella; Di Stefano, Giuseppina; Porcellini, Elisa; Chiappelli, Martina; Carbone, Ilaria; Licastro, Federico

    2010-12-16

    Alzheimer's disease (AD) is one of the most frequent cause of neurodegenerative disorder in the elderly. Inflammation has been implicated in brain degenerative processes and peripheral markers of brain AD related impairment would be useful. Plasma levels of alpha-1-antichymotrypsin (ACT), an acute phase protein and a secondary component of amyloid plaques, are often increased in AD patients and high blood ACT levels correlate with progressive cognitive deterioration. During inflammatory responses changes in the micro-heterogeneity of ACT sugar chains have been described. N-Glycanase digestion from Flavobacterium meningosepticum (PNGase F) was performed on both native and denatured purified ACT condition and resolved to Western blot with the purpose to revealed the ACT de-glycosylation pattern.Further characterization of the ACT glycan profile was obtained by a glycoarray; each lectin group in the assay specifically recognizes one or two glycans/epitopes. Lectin-bound ACT produced a glyco-fingerprint and mayor differences between AD and controls samples were assessed by a specific algorithms. Western blot analysis of purified ACT after PNGase F treatment and analysis of sugar composition of ACT showed significantly difference in "glyco-fingerprints" patterns from controls (CTR) and AD; ACT from AD showing significantly reduced levels of sialic acid. A difference in terminal GlcNac residues appeared to be related with progressive cognitive deterioration. Low content of terminal GlcNac and sialic acid in peripheral ACT in AD patients suggests that a different pattern of glycosylation might be a marker of brain inflammation. Moreover ACT glycosylation analysis could be used to predict AD clinical progression and used in clinical trials as surrogate marker of clinical efficacy.

  7. Magnetism for understanding catalyst analysis of purified carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Bellouard, Christine; Mercier, Guillaume; Cahen, Sébastien; Ghanbaja, Jaafar; Medjahdi, Ghouti [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Gleize, Jérôme [Laboratoire de Chimie Physique-Approche Multi-échelle de Milieux Complexes-Université de Lorraine, 1 Bd Arago, 57078 Metz (France); Lamura, Gianrico [CNR-SPIN – Dipartimento di Fisica, via Dodecaneso 33, 16146 Genova (Italy); Hérold, Claire [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France); Vigolo, Brigitte, E-mail: Brigitte.Vigolo@univ-lorraine.fr [Institut Jean Lamour, CNRS-Université de Lorraine, BP 70239, 54506 Vandoeuvre-lès-Nancy (France)

    2016-08-01

    The precise quantification of catalyst residues in purified carbon nanotubes is often a major issue in view of any fundamental and/or applicative studies. More importantly, since the best CNTs are successfully grown with magnetic catalysts, their quantification becomes strictly necessary to better understand intrinsic properties of CNT. For these reasons, we have deeply analyzed the catalyst content remained in nickel–yttrium arc-discharge single walled carbon nanotubes purified by both a chlorine-gas phase and a standard acid-based treatment. The study focuses on Ni analysis which has been investigated by transmission electron microscopy, X-ray diffraction, thermogravimetry analysis, and magnetic measurements. In the case of the acid-based treatment, all quantifications result in a decrease of the nanocrystallized Ni by a factor of two. In the case of the halogen gas treatment, analysis and quantification of Ni content is less straightforward: a huge difference appears between X-ray diffraction and thermogravimetry results. Thanks to magnetic measurements, this disagreement is explained by the presence of Ni{sup 2+} ions, belonging to NiCl{sub 2} formed during the Cl-based purification process. In particular, NiCl{sub 2} compound appears under different magnetic/crystalline phases: paramagnetic or diamagnetic, or well intercalated in between carbon sheets with an ordered magnetic phase at low temperature. - Highlights: • Cl-gas treatment of Ni catalyst of carbon nanotubes leads to NiCl{sub 2} residue. • Magnetic measurements show the transformation of Ni{sup 0} in Ni{sup 2+}through a purification process. • High temperature Cl treatment removes 75% of metallic impurities. • Cl-purification yields to an amount of metal of 1.5% in arc-discharge CNT samples.

  8. Prevalence of Giardia spp. in young dogs using a combination of two diagnostic methods.

    Science.gov (United States)

    Alves, João; Santos, Ana

    2016-03-01

    In this study, prevalence of the protozoan parasites from the genus Giardia spp, with zoonotic potential and worldwide dissemination, was accessed in young dogs, which are reported as having higher prevalence rates. With that purpose, 49 animals from the Grupo de Intervenção Cinotécnico of the Guarda Nacional Republicana (Portuguese Gendarmerie Canine Unit) were chosen. They were housed individually in areas with a high number of kennels (up to 100), with ages ragging from newborns to 10 years old. Dogs were divided in four groups, according their age: under 6 months (n = 16), 6-12 months (n = 6), 12-18 months (n = 13) and 18-24 months (n = 14), comprising 22 females and 27 males. Fecal samples were collected from every animal and all were submitted to two different diagnostic tests, a passive flotation technique with a ZnSO4 solution and a detection of fecal antigen using a commercially available ELISA test (Witness® Giardia - Zoetis). From the 49 samples, 5 (10.2%) were considered positive with ZnSO4 flotation technique and 6 (12.24%) with the Witness® Giardia test. When considering the combination of both tests, 5 animals (10.2%) were considered positive. Of these, 3 (60%) were from the group under 6 months old, 1 (20%) from the 6-12 months and 1 from the 18-24 (20%) months. Within each group, in the under 6 months group 18.75% (n = 16) were considered positive, 16.67% in the 6-12 month group (n = 6), 0% in the 12-18 month group (n=13) and 7.14% in 18-24 month group (n = 14). None of the animals had clinical signs and no significant differences were found when comparing prevalence according to age, breed or gender. A combination of fecal flotation and antigen ELISA tests have good sensitivity and are easy to perform in practice and, therefore, could be a good choice to perform a diagnostic and small animal veterinarians should have this possible diagnostic in mind when in the present of clinical signs, particularly in young dogs.

  9. Prevalence and molecular characterization of Cryptosporidum spp. and Giardia spp. in environmental samples in Hanam province, Vietnam

    DEFF Research Database (Denmark)

    Nguyen, Tram Thuy; Traub, Rebecca J.; Pham, Phuc Duc

    2016-01-01

    Cryptosporidium and Giardia are protozoan parasites that cause human diarrheal disease worldwide. This study was done to evaluate the prevalence and concentrations of these protozoa in environmental samples in Hanam, Vietnam and to assess potential contamination sources using molecular epidemiolo......Cryptosporidium and Giardia are protozoan parasites that cause human diarrheal disease worldwide. This study was done to evaluate the prevalence and concentrations of these protozoa in environmental samples in Hanam, Vietnam and to assess potential contamination sources using molecular...

  10. Influence of selected stool concentration techniques on the effectiveness of PCR examination in Giardia intestinalis diagnostics.

    Science.gov (United States)

    Stojecki, K; Sroka, J; Karamon, J; Kusyk, P; Cencek, T

    2014-01-01

    Giardia intestinalis is a widespread parasitic protozoa which has great significance as a public health threat. Molecular diagnostics of stool sample can be unreliable because of the presence of inhibitors of enzymatic reactions. The aim of this study was to determine the effectiveness of selected pre-treatment methods of fecal samples for further PCR-based diagnostics of G. intestinalis, and the effect of each component of pre-treatment solutions on PCR reactions. Seven stool concentration techniques were compared. The results showed that the most efficient concentration method for stool sample preparation for detection of G. intestinalis by PCR is centrifugal flotation with Percoll (with saturated NaNO3 as the flotation solution). This method is relatively inexpensive, less labor-intensive, and suitable for epidemiological monitoring and clinical investigations.

  11. Effects of Ultrasound on the Survival and Characteristics of Cryptosporidium Oocysts and Giardia Cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, J.; Klinting, M.

    2011-01-01

    Background: Sonication of fluids has been widely used for cell lyses and decontamination of water and food due to its three major physical and chemical effects: cavitation, implosion of water bubbles with subsequent increase in substrate temperature and liberation of free radicals such as O...... power and time durations on two of the most common waterborne protozoa Cryptosporidium and Giardia, and examined its effects on parasite characteristics and survival rate using immunofluorescence dyes; DAPI (4’,6-diamidino-2-phenylindol) staining/PI (propidium iodide), and analyzed by flow cytometry...... determine the effects of ultrasound on the parasite, including the sonication power of ultrasound as well as substrate temperature. Conclusions: Ultrasound is harmful for waterborne protozoa even when momentarily applied. However, a mode of operation may exist in which ultrasound can be used for collection...

  12. Prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugee population of Mianwali district, Pakistan.

    Science.gov (United States)

    Abrar Ul Haq, Khan; Gul, Naz Asma; Hammad, Hussain Muhammad; Bibi, Yasmeen; Bibi, Asma; Mohsan, Javed

    2015-06-01

    Present study aimed to investigate prevalence of Giardia intestinalis and Hymenolepis nana in Afghan refugees visiting Central Health Unit (CHU), Kot Chandana (Mianwali, Northern Punjab) during two years period (February 2007 to December 2009). A total of 687 stool samples were collected from different age groups of both genders. Samples were processed under sterile conditions after gross examination. Microscopic examination was done on same day along with eggs (H. nana), cyst and trophozoites (G. intestinalis) detection after staining. The prevalence of G. intestinalis was significantly higher (x2=59.54, pintestinal clinical symptoms observed in G. intestinalis. Whereas, bloody diarrhea (OR: 1.56, 95%CI=1.00-2.43) and rectal prolapse (OR: 5.79, 95%CI=1.87-17.91) were associated with H. nana infections. Intestinal parasitic infections are common among Afghan refugees and serious preventive measures should be implemented to promote the safety and healthy lifestyle of these people.

  13. Detection and molecular characterisation of Giardia lamblia genotypes in Sharjah, United Arab Emirates.

    Science.gov (United States)

    ElBakri, Ali; Samie, Amidou; Bessong, Pascal; Potgieter, Natasha; Odeh, Ra'ed Abu

    2014-08-01

    No data are available on Giardia lamblia genotypes in the United Arab Emirates (UAE). This study aimed to identify G. lamblia from DNA extracted from human stool samples to gain information on its prevalence and to perform molecular analysis on isolates collected from expatriates from different localities residing in Sharjah, UAE. In total, 111 healthy expatriates residing in Sharjah were screened for G. lamblia using nested PCR amplification of the small subunit ribosomal RNA (ssu-rRNA) gene. Positive samples were genotyped using a nested PCR amplifying the triosephosphate isomerase (tpi) gene to differentiate between the two human assemblages (A and B). A subset of the PCR products (n=23) were sequenced and their phylogenetic relationships were determined. Of the 111 participants, 67 (60.4%) were identified as positive for the ssu-rRNA gene. When genotyped for the tpi gene, 18.9% (21/111) were of assemblage A, 17.1% (19/111) belonged to assemblage B and 5.4% (6/111) showed patterns compatible with mixed infections. A strong correlation between the presence of diarrhoea and assemblage B was observed (χ(2)=10.553; p=0.001). Moreover, an association was also observed between mixed infections (A+B) and diarrhoea (χ(2)=8.899; p=0.003). No correlation between age, gender and geographic origin of the infected individual was noted. Phylogenetic analysis showed three clusters for the tpi gene. No relationship between the clusters and the origin of samples was noted. This study is the first to determine the infection rate and genotypic composition of Giardia in Sharjah, UAE. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. Actividad lactásica en niños con enfermedad diarreica por Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Arnaldo Izquierdo Estévez

    1998-03-01

    Full Text Available Se realizó un estudio prospectivo, de corte transversal y analítico, con el propósito de establecer posibles diferencias en el comportamiento de la actividad lactásica en niños menores de 2 años con enfermedad diarreica por Giardia lamblia tratados en el Servicio de Diarreas del Hospital Pediátrico Docente "Dr. Ángel A. Aballí", en el período de diciembre de 1994 a noviembre de 1995. Se determinaron los niveles de la actividad enzimática según diferentes categorías de variables de caracterización del niño y su enfermedad. Los resultados permitieron afirmar su reducción en estos niños, y que tal actividad tiene un comportamiento diferencial asociado con la edad, estado nutricional y grado de atrofia de la mucosa.A prospective analytical cross-sectional study was made with the objective of setting likely differences in Beta-galactosidase performance in under 2 years-old children admitted with diarrheal disease due to Giardia lamblia to «Ángel A. Aballí» Teaching Pediatric Hospital’s Diarrhea Service Unit from December 1994 to November 1995. Levels of the enzime activity were determined according to the several characterization variable categories of the child and his disease. The outcome showed the decline in diarrheal disease in thesechildren and the different performance patterns of this enzime depending on age, nutrition and mucosa atrophic level.

  15. A detailed look at the cytoskeletal architecture of the Giardia lamblia ventral disc.

    Science.gov (United States)

    Brown, Joanna R; Schwartz, Cindi L; Heumann, John M; Dawson, Scott C; Hoenger, Andreas

    2016-04-01

    Giardia lamblia is a protistan parasite that infects and colonizes the small intestine of mammals. It is widespread and particularly endemic in the developing world. Here we present a detailed structural study by 3-D negative staining and cryo-electron tomography of a unique Giardia organelle, the ventral disc. The disc is composed of a regular array of microtubules and associated sheets, called microribbons that form a large spiral, held together by a myriad of mostly unknown associated proteins. In a previous study we analyzed by cryo-electron tomography the central microtubule portion (here called disc body) of the ventral disc and found a large portion of microtubule associated inner (MIPs) and outer proteins (MAPs) that render these microtubules hyper-stable. With this follow-up study we expanded our 3-D analysis to different parts of the disc such as the ventral and dorsal areas of the overlap zone, as well as the outer disc margin. There are intrinsic location-specific characteristics in the composition of microtubule-associated proteins between these regions, as well as large differences between the overall architecture of microtubules and microribbons. The lateral packing of microtubule-microribbon complexes varies substantially, and closer packing often comes with contracted lateral tethers that seem to hold the disc together. It appears that the marginal microtubule-microribbon complexes function as outer, laterally contractible lids that may help the cell to clamp onto the intestinal microvilli. Furthermore, we analyzed length, quantity, curvature and distribution between different zones of the disc, which we found to differ from previous publications. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Intestinal parasites and genotypes of Giardia intestinalis in school children from Berisso, Argentina.

    Science.gov (United States)

    Molina, Nora; Pezzani, Betina; Ciarmela, Maria; Orden, Alicia; Rosa, Diana; Apezteguía, Maria; Basualdo, Juan; Minvielle, Marta

    2011-07-27

    Intestinal parasitic infections have been reported in different regions of Argentina. Giardia intestinalis is recognized as "the national parasite". The aim of this work was to determine the prevalence of both intestinal parasites and G. intestinalis genotypes, as well as to analyze the clinical and epidemiological characteristics in schoolchildren from a suburban community. Serial coproparasitological analysis and perianal swab method were performed in 244 schoolchildren. Demographic, sociocultural and environmental variables were registered. The presence of signs/symptoms and risk behaviours were also recorded. Stools with G. intestinalis were selected for genotyping. Out of 244 schoolchildren, 179/244 (73.4%) were infected with intestinal parasites. The presence of intestinal parasitosis was associated only with house flooding. Multivariate analysis identified that use of a latrine is significantly correlated with G. intestinalis and age six to 11 years with E. vermicularis. Signs and symptoms were recorded in 62% of infected children and in 57.9% of those not infected. Genomic amplification was revealed that 65.7% (46/70) of Giardia positive samples corresponded to genotype B, 31.4% (22/70) to genotype AII, and two samples (2.8%) had mixed infection (AII + B). This study shows a high percentage of infected children living in a suburban community in poor sanitary conditions, and not visiting the doctor in spite of evident signs and symptoms associated a digestive pathology. This situation supports the need for continuing the development of community programs allowing the improvement of quality of life and control of parasitosis in deprived populations.

  17. Selective condensation drives partitioning and sequential secretion of cyst wall proteins in differentiating Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Christian Konrad

    2010-04-01

    Full Text Available Controlled secretion of a protective extracellular matrix is