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Štrkolcová G, Fiľakovská Bobáková D, Kaduková M, Schreiberová A, Klein D, Halán M, Urbančíková I. Intestinal parasitic infections in children from marginalised Roma communities: prevalence and risk factors. BMC Infect Dis 2024; 24:596. [PMID: 38890608 PMCID: PMC11184866 DOI: 10.1186/s12879-024-09500-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2024] [Accepted: 06/12/2024] [Indexed: 06/20/2024] Open
Abstract
BACKGROUND Intestinal parasitic infections remain a significant global health issue, particularly affecting poor and marginalised populations. These infections significantly contribute to children's diseases, malnutrition, poor school performance, cognitive disorders, and future economic losses. This study aimed to explore and compare the occurrence of intestinal parasites in early childhood among the group of infants from the Slovak majority population and from marginalised Roma communities (MRCs). Furthermore, it aimed to explore the health complaints of children with and without intestinal parasitic infection in the past month and assess the effect of various risk factors on the occurrence of intestinal parasitic infection in infants from MRCs. METHODS We obtained cross-sectional data from mothers and stool samples of their children aged 13-21 months using the first wave of the longitudinal RomaREACH study. A total of 181 stools from infants were analysed: 105 infants from the Slovak majority population and 76 from MRCs. RESULTS Infants from MRCs are significantly more often infected by Ascaris lumbricoides, Trichuris trichiura and Giardia duodenalis than their better-off peers from the majority population. Infection rates are 30% in infants from MRCs vs. 0% in the majority population (p < 0.001). Single and mixed infections were observed in children from MRCs. Infants with intestinal parasitic infections suffer significantly more often from various health complaints, particularly cough, stomach ache, irritability, and diarrhoea. Within MRCs, the risk of parasitic infections in infants is significantly increased by risk factors such as the absence of flushing toilets in households (OR = 4.17, p < 0.05) and contact with un-dewormed animals (OR = 3.61, p < 0.05). Together with the absence of running water in the household, these three factors combined increase the risk more than ten times (p < 0.01). CONCLUSION Maintaining hygienic standards in conditions of socioeconomic deprivation in MRCs without running water and sewage in the presence of un-dewormed animals is problematic. These living conditions contribute to the higher prevalence of parasitic infections in children from MRCs, causing various health complaints and thus threatening their health and healthy development.
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Affiliation(s)
- Gabriela Štrkolcová
- Department of Epizootiology and Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in Košice, Košice, Slovakia
| | - Daniela Fiľakovská Bobáková
- Department of Health Psychology and Research Methodology, Faculty of Medicine, PJ Safarik University, Kosice, Slovakia.
- Olomouc University Social Health Institute, Palacky University in Olomouc, Olomouc, Czechia, Czechia.
| | - Michaela Kaduková
- Department of Epizootiology and Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in Košice, Košice, Slovakia
| | - Andrea Schreiberová
- Department of Epizootiology and Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in Košice, Košice, Slovakia
| | - Daniel Klein
- Institute of Mathematics, Faculty of Natural Sciences, PJ Safarik University, Kosice, Slovakia
| | - Miloš Halán
- Department of Epizootiology and Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in Košice, Košice, Slovakia
| | - Ingrid Urbančíková
- Department of Epidemiology, Faculty of Medicine, PJ Safarik University, Kosice, Slovakia
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Ionică AM, Ieremia A, Kalmár Z, Lupșe M, Flonta M, Muntean M, Cismaru C, Horvat M, Rădulescu A, Topan A, Jianu C, Deak G, Briciu V. Prevalence and Genotyping of Water- and Food-Borne Parasitic Protozoans ( Giardia duodenalis and Cryptosporidium spp.) in Hospitalized Patients from Northwestern Romania. Microorganisms 2024; 12:762. [PMID: 38674706 PMCID: PMC11052221 DOI: 10.3390/microorganisms12040762] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2024] [Revised: 04/07/2024] [Accepted: 04/09/2024] [Indexed: 04/28/2024] Open
Abstract
Giardia duodenalis and Cryptosporidium spp. are important zoonotic protozoan pathogens that infect the gastro-intestinal tract of numerous vertebrates, including humans, and both parasites are responsible for water- or food-borne outbreaks of disease worldwide. Although, globally, both parasites are highly prevalent, particularly in developing countries, epidemiological data from Romania are scarce, and genotyping has rarely been performed. The aims of the present study were to investigate the occurrence and genetic diversity of G. duodenalis and Cryptosporidium spp. in patients hospitalized in Northwestern Romania in relation to clinical and paraclinical presentation and to identify the relative frequency of non-specific symptoms and potential risk factors. Between June 2022 and January 2024, 426 fecal samples were screened for gastro-intestinal parasites by rapid tests and microscopical examination, further confirmed by PCR and sequencing. Giardia duodenalis was detected and characterized in 12 samples (2.82%), while Cryptosporidium parvum was confirmed in four samples (0.94%). A majority of positive patients were symptomatic and reported nausea and vomiting with a significantly higher frequency compared to negative ones. This study provides new insights into the epidemiological status and clinical implications of gastro-intestinal parasite species and genospecies in Romania that are necessary for an in-depth understanding of the potential zoonotic transmission and improvement of patient care.
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Affiliation(s)
- Angela Monica Ionică
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
| | - Anca Ieremia
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
| | - Zsuzsa Kalmár
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
- Department of Microbiology, Immunology, and Epidemiology, University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca, 400372 Cluj-Napoca, Romania
| | - Mihaela Lupșe
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Mirela Flonta
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
| | - Monica Muntean
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Cristina Cismaru
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Melinda Horvat
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Amanda Rădulescu
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Adriana Topan
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
| | - Cristian Jianu
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
| | - Georgiana Deak
- Department of Parasitology and Parasitic Diseases, University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca, 400372 Cluj-Napoca, Romania;
| | - Violeta Briciu
- Clinical Hospital of Infectious Diseases of Cluj-Napoca, 400348 Cluj-Napoca, Romania (M.L.); (M.F.); (M.M.); (C.J.)
- Department of Infectious Diseases, “Iuliu Haţieganu” University of Medicine and Pharmacy, 4000348 Cluj-Napoca, Romania
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Ghosal A, Sardar SK, Haldar T, Maruf M, Saito-Nakano Y, Dutta S, Nozaki T, Ganguly S. Genotyping and epidemiological distribution of diarrhea-causing isolates of Giardia duodenalis in southeastern part of West Bengal, India. Parasitol Res 2023; 122:2567-2584. [PMID: 37682345 DOI: 10.1007/s00436-023-07956-7] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2023] [Accepted: 08/24/2023] [Indexed: 09/09/2023]
Abstract
The prevalence and genetic diversity of the protozoan pathogen Giardia duodenalis have been extensively studied worldwide. There is currently a lack of data regarding the genetic variability of the organism in eastern India. Understanding the circulating genotypes and associated risk factors is crucial for effective planning and implementing control measures. Therefore, the objective of the study was to conduct an epidemiological study to determine the prevalence and identify the various genotypes present. This survey adds to our knowledge on the occurrence and distribution of Giardia genotypes in the studied region. The overall prevalence was found to be 6.8%. This parasitic infection was significantly associated with two age groups, i.e., >0-5 years and >5-12 years. Using a multilocus genotyping method, we genotyped 52 human Giardia isolates that were obtained from diarrheal patients. Two distinct assemblages were found in the population-30.8% belonged to assemblage A; 63.5% belonged to assemblage B, prevalent in the population; and 5.7% belonged to a combined assemblage A+B. Sub-assemblage AII was found in 17.3% of the cases, followed by sub-assemblage AI (13.5%). High levels of genetic diversity were found within the population of assemblage B undergoing balancing selection. Overall, the high prevalence of the parasite observed, particularly among children, raises a major concern and necessitates implementation of robust control measures. Furthermore, we report the presence of numerous unique genotypes, circulating in this limited geographical boundary, which can be useful dataset for future studies.
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Affiliation(s)
- Ajanta Ghosal
- Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India
| | - Sanjib K Sardar
- Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India
| | - Tapas Haldar
- Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India
| | - Maimoon Maruf
- Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India
| | - Yumiko Saito-Nakano
- Department of Parasitology, National Institute of Infectious Diseases (NIID), Tokyo, Japan
| | - Shanta Dutta
- Division of Bacteriology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India
| | - Tomoyoshi Nozaki
- Department of Biomedical Chemistry, School of International Health, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
| | - Sandipan Ganguly
- Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India.
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Prabakaran M, Weible LJ, Champlain JD, Jiang RY, Biondi K, Weil AA, Van Voorhis WC, Ojo KK. The Gut-Wrenching Effects of Cryptosporidiosis and Giardiasis in Children. Microorganisms 2023; 11:2323. [PMID: 37764167 PMCID: PMC10538111 DOI: 10.3390/microorganisms11092323] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/23/2023] [Revised: 09/04/2023] [Accepted: 09/12/2023] [Indexed: 09/29/2023] Open
Abstract
Cryptosporidium species and Giardia duodenalis are infectious intestinal protozoan pathogens that cause alarming rates of morbidity and mortality worldwide. Children are more likely to have clinical symptoms due to their less developed immune systems and factors such as undernutrition, especially in low- and middle-income countries. The severity of the symptoms and clinical manifestations in children may vary from asymptomatic to life-threatening depending on the Cryptosporidium species/G. duodenalis strains and the resulting complex stepwise interactions between the parasite, the host nutritional and immunologic status, and the gut microbiome profile. Structural damages inflicted by both parasites to epithelial cells in the large and small intestines could severely impair children's gut health, including the ability to absorb nutrients, resulting in stunted growth, diminished neurocognitive development, and other long-term effects. Clinically approved cryptosporidiosis and giardiasis drugs have broad antimicrobial effects that have incomprehensible impacts on growing children's gut health.
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Affiliation(s)
- Mayuri Prabakaran
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Lyssa J. Weible
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Joshua D. Champlain
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Ryan Ye Jiang
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Katalina Biondi
- Human Center for Artificial Intelligence, Department of Computer Science, University of Central Florida, Orlando, FL 32816, USA;
| | - Ana A. Weil
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Wesley C. Van Voorhis
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
| | - Kayode K. Ojo
- Center for Emerging and Reemerging Infectious Diseases (CERID), Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98109, USA; (M.P.); (L.J.W.); (J.D.C.); (R.Y.J.); (A.A.W.); (W.C.V.V.)
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Natali L, Luna Pizarro G, Moyano S, de la Cruz-Thea B, Musso J, Rópolo AS, Eichner N, Meister G, Musri MM, Feliziani C, Touz MC. The Exosome-like Vesicles of Giardia Assemblages A, B, and E Are Involved in the Delivering of Distinct Small RNA from Parasite to Parasite. Int J Mol Sci 2023; 24:ijms24119559. [PMID: 37298511 DOI: 10.3390/ijms24119559] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2023] [Revised: 03/27/2023] [Accepted: 04/03/2023] [Indexed: 06/12/2023] Open
Abstract
The genetically related assemblages of the intestinal protozoa parasite Giardia lamblia are morphologically indistinguishable and are often derived from specific hosts. The Giardia assemblages are separated by large genetic distances, which might account for their relevant biological and pathogenic differences. In this work, we analyzed the RNAs cargo released into exosomal-like vesicles (ElVs) by the assemblages A and B, which differentially infect humans, and the assemblage E, which infects hoofed animals. The RNA sequencing analysis revealed that the ElVs of each assemblage contained distinct small RNA (sRNA) biotypes, suggesting a preference for specific packaging in each assemblage. These sRNAs were classified into three categories, ribosomal-small RNAs (rsRNAs), messenger-small RNAs (msRNAs), and transfer-small RNAs (tsRNAs), which may play a regulatory role in parasite communication and contribute to host-specificity and pathogenesis. Uptake experiments showed, for the first time, that ElVs were successfully internalized by the parasite trophozoites. Furthermore, we observed that the sRNAs contained inside these ElVs were first located below the plasma membrane but then distributed along the cytoplasm. Overall, the study provides new insights into the molecular mechanisms underlying the host-specificity and pathogenesis of G. lamblia and highlights the potential role of sRNAs in parasite communication and regulation.
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Affiliation(s)
- Lautaro Natali
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Gabriel Luna Pizarro
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Sofía Moyano
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Benjamin de la Cruz-Thea
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Juliana Musso
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Andrea S Rópolo
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Norbert Eichner
- Regensburg Center for Biochemistry (RCB), Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany
| | - Gunter Meister
- Regensburg Center for Biochemistry (RCB), Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany
| | - Melina M Musri
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - Constanza Feliziani
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
| | - María C Touz
- Instituto de Investigación Médica Mercedes y Martín Ferreyra, Consejo Nacional de Investigaciones Científicas y Técnicas (INIMEC-CONICET), Universidad Nacional de Córdoba, Córdoba 5016, Argentina
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The Investigation of Giardiasis (Foodborne and Waterborne Diseases) in Buffaloes in Van Region, Türkiye: First Molecular Report of Giardia duodenalis Assemblage B from Buffaloes. Pathogens 2023; 12:pathogens12010106. [PMID: 36678454 PMCID: PMC9863494 DOI: 10.3390/pathogens12010106] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2022] [Revised: 01/03/2023] [Accepted: 01/06/2023] [Indexed: 01/11/2023] Open
Abstract
Giardia duodenalis (G. duodenalis) is an important zoonotic protozoan agent that causes foodborne and waterborne diarrhea in humans and other mammals. Molecular-based tests are critical in diagnosing giardiasis in humans and animals, identifying species, understanding the zoonotic potential and transmission routes, and evaluating taxonomy. Therefore, this study aimed to investigate the molecular characterization of G. duodenalis in buffaloes in the Van region in Türkiye. Buffaloes are a species that has been poorly studied in this regard. For this purpose, 100 fecal samples were collected from buffaloes in the Van region. The DNA extraction was performed using the GeneMATRIX STOOL DNA Purification Kit from stool samples. The nested PCR test was performed with the appropriate primers from the obtained DNA samples. The obtained bands suitable for sequencing were sent for sequence analysis, and the sequence results were aligned bidirectionally and compared with the database of GenBank by BLAST. As a result of the study, an 11% positivity rate for G. duodenalis was found in buffaloes, and assemblage E and assemblage B were isolated. To our knowledge, assemblage B in buffaloes was reported for the first time in this study. As a result, it was concluded that buffaloes are an important reservoir for waterborne and foodborne giardiasis.
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Solaymani-Mohammadi S. Mucosal Defense Against Giardia at the Intestinal Epithelial Cell Interface. Front Immunol 2022; 13:817468. [PMID: 35250996 PMCID: PMC8891505 DOI: 10.3389/fimmu.2022.817468] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2021] [Accepted: 01/31/2022] [Indexed: 02/05/2023] Open
Abstract
Human giardiasis, caused by the protozoan parasite Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis, Lamblia intestinalis), is one of the most commonly-identified parasitic diseases worldwide. Chronic G. duodenalis infections cause a malabsorption syndrome that may lead to failure to thrive and/or stunted growth, especially in children in developing countries. Understanding the parasite/epithelial cell crosstalk at the mucosal surfaces of the small intestine during human giardiasis may provide novel insights into the mechanisms underlying the parasite-induced immunopathology and epithelial tissue damage, leading to malnutrition. Efforts to identify new targets for intervening in the development of intestinal immunopathology and the progression to malnutrition are critical. Translating these findings into a clinical setting will require analysis of these pathways in cells and tissues from humans and clinical trials could be devised to determine whether interfering with unwanted mucosal immune responses developed during human giardiasis provide better therapeutic benefits and clinical outcomes for G. duodenalis infections in humans.
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Affiliation(s)
- Shahram Solaymani-Mohammadi
- Laboratory of Mucosal Immunology, Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND, United States
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Zajaczkowski P, Lee R, Fletcher-Lartey SM, Alexander K, Mahimbo A, Stark D, Ellis JT. The controversies surrounding Giardia intestinalis assemblages A and B. CURRENT RESEARCH IN PARASITOLOGY & VECTOR-BORNE DISEASES 2022; 1:100055. [PMID: 35284870 PMCID: PMC8906113 DOI: 10.1016/j.crpvbd.2021.100055] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 09/17/2021] [Accepted: 10/09/2021] [Indexed: 11/27/2022]
Abstract
Giardia intestinalis continues to be one of the most encountered parasitic diseases around the world. Although more frequently detected in developing countries, Giardia infections nonetheless pose significant public health problems in developed countries as well. Molecular characterisation of Giardia isolates from humans and animals reveals that there are two genetically different assemblages (known as assemblage A and B) that cause human infections. However, the current molecular assays used to genotype G. intestinalis isolates are quite controversial. This is in part due to a complex phenomenon where assemblages are incorrectly typed and underreported depending on which targeted locus is sequenced. In this review, we outline current knowledge based on molecular epidemiological studies and raise questions as to the reliability of current genotyping assays and a lack of a globally accepted method. Additionally, we discuss the clinical symptoms caused by G. intestinalis infection and how these symptoms vary depending on the assemblage infecting an individual. We also introduce the host-parasite factors that play a role in the subsequent clinical presentation of an infected person, and explore which assemblages are most seen globally.
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Affiliation(s)
- Patricia Zajaczkowski
- Faculty of Science, School of Life Sciences, University of Technology Sydney, Australia
| | - Rogan Lee
- Centre for Infectious Diseases and Microbiology Laboratory Services, ICPMR, Westmead Hospital, Westmead, New South Wales, Australia.,Westmead Clinical School, Faculty of Medicine and Health, The University of Sydney, Westmead Hospital, Westmead, NSW, Australia
| | | | - Kate Alexander
- Public Health Unit, South Western Sydney Local Health District, Liverpool, Australia
| | - Abela Mahimbo
- Faculty of Health, School of Public Health, University of Technology Sydney, Australia
| | - Damien Stark
- Department of Microbiology, St Vincent's Hospital Sydney, Darlinghurst, New South Wales, Australia
| | - John T Ellis
- Faculty of Science, School of Life Sciences, University of Technology Sydney, Australia
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Dhodapkar R, Langbang D, Parija S, Premarajan KC, Rajkumari N. Molecular characterization of Giardia intestinalis assemblages in children among the rural and urban population of Pondicherry, India. Trop Parasitol 2022; 12:8-14. [PMID: 35923262 PMCID: PMC9341134 DOI: 10.4103/tp.tp_52_20] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/05/2020] [Revised: 06/19/2020] [Accepted: 07/21/2020] [Indexed: 11/05/2022] Open
Abstract
Introduction: Giardiasis is one of the greatest public parasitic infections causing diarrheal and also known to be associated with high morbidity and mortality, among the children's particularly in developing countries with less cleanliness practices. Thus, studying genomic variety of Giardia intestinalis aids to improve our perspective related to the variability in the genome of the parasite. Materials and Methods: In this cross-sectional study, 1006 stool samples were collected from the rural (n = 500) and urban settings (n = 506) from the children (<15 years) with and without symptoms and were screened for the presence of G. intestinalis by polymerase chain reaction (PCR) targeting triosephosphate isomerase gene. Further, all PCR-positive amplicons were subjected to restriction fragment length polymorphism using RsaI restriction enzyme. Results: Of the total 1006 stool samples, 500 samples from rural screened by PCR 108 (21%) were found to be positive for assemblage A, 116 (23.2%) belong to assemblage B, and 5 (1%) were mixed assemblages (A + B). Whereas in urban, of the 506 samples screened by PCR, 92 (18.1%) were found to be positive for assemblage A, 93 (18.3%) assemblage B, and 10 (1.9%) were mixed assemblages (A + B). No significant difference was found between the G. intestinalis assemblages with clinical details of symptomatic and asymptomatic in children. Conclusions: This signifies the first study inspection in our location to shed lights and delivers some preliminary data on assemblages and subassemblages. The results suggest that anthroponotic transmission could be a foremost transmission path for giardiasis among the study population.
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Scorza AV, Buch J, Franco P, McDonald C, Chandrashekar R, Lappin MR. Evaluation for associations amongst Giardia duodenalis assemblages and diarrhea in dogs. Vet Parasitol 2021; 300:109581. [PMID: 34735843 DOI: 10.1016/j.vetpar.2021.109581] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/11/2021] [Revised: 09/11/2021] [Accepted: 09/18/2021] [Indexed: 01/25/2023]
Abstract
Giardia duodenalis is a species complex comprising at least eight assemblages. Most dogs harbor the host-adapted assemblages C and D and approximately 30 % harbor the zoonotic assemblages. Humans and dogs with giardiosis can exhibit a variety of clinical manifestations ranging from the absence of clinical signs to acute or chronic diarrhea. Human studies report conflicting results concerning associations between clinical signs and assemblage type. The objective of this study was to use results of molecular and phylogenetic analyses to evaluate associations between G. duodenalis assemblages and diarrhea in client-owned dogs from the United States. Fecal samples that were positive for Giardia cysts were classified as normal or diarrheal. Samples were analyzed by PCR assays of the beta-giardin (bg), glutamate dehydrogenase (gdh), and triose phosphate isomerase (tpi) genes. Sequences of the three genes were analyzed by BLAST analysis and phylogenetic analysis was performed by Neighbor-Joining analysis. Two hundred and eighty-eight Giardia-positive fecal samples were evaluated by the three PCRs. One or more genes were amplified from 95 normal samples and 93 diarrheal samples, 27 samples were positive for one or more genes but could not be sequenced due to low quality DNA, and 73 samples tested negative. Ninety seven percent of the samples (182/188) in both the diarrheal and normal groups typed as dog-specific assemblages (D or C) by at least one gene. Phylogenetic analysis of the three genes placed the isolates from assemblages A, B, C and D separated from each other with strong bootstrap support. Diarrhea was not associated with the Giardia assemblage or other parasitic co-infection in this sample set. Other factors, such as the role of gut microbiota in giardiosis should be considered in future studies.
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Affiliation(s)
- Andrea V Scorza
- Center for Companion Animal Studies, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, 80523, USA.
| | - Jesse Buch
- IDEXX Laboratories Inc., Westbrook, ME, 04092, USA
| | - Patricia Franco
- Center for Companion Animal Studies, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, 80523, USA
| | - Cassandra McDonald
- Center for Companion Animal Studies, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, 80523, USA
| | | | - Michael R Lappin
- Center for Companion Animal Studies, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, 80523, USA
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Detection of Giardia intestinalis assemblages A and B among children from three villages in the West Delta region, Egypt using assemblage specific primers. J Parasit Dis 2021; 45:655-663. [PMID: 34475646 DOI: 10.1007/s12639-020-01338-x] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2020] [Accepted: 12/09/2020] [Indexed: 10/22/2022] Open
Abstract
Giardia intestinalis is a common diarrheagenic parasite infecting children globally. It has been classified into eight morphologically identical but genetically distinct genotypes. Human infection is mainly associated with A and B assemblages with variable geographical distribution. The present work aimed to study the epidemiology of assemblages A and B in children inhabiting different areas in Lower Egypt. Stool samples were collected from 315 children and examined microscopically for parasitic infections. Giardia positive samples were genotyped using tpi assemblage specific primers. The prevalence of Giardia was 18.1% among the examined children. Mixed assemblages A and B was more common (47.4%) than single assemblage B (36.8%) or A (15.8%). The distribution of different genotypes was significantly associated with the residence area, animal contact, and handwashing habits. A non-significant association was observed between Giardia assemblages and the clinical manifestations. Assemblage B is the predominant genotype among Egyptian children. The distribution of different Giardia assemblages is strongly associated with the studied area and the habits of its people.
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Multilocus Genotyping of Giardia duodenalis in Mostly Asymptomatic Indigenous People from the Tapirapé Tribe, Brazilian Amazon. Pathogens 2021; 10:pathogens10020206. [PMID: 33672794 PMCID: PMC7917967 DOI: 10.3390/pathogens10020206] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2020] [Revised: 02/01/2021] [Accepted: 02/08/2021] [Indexed: 02/06/2023] Open
Abstract
Little information is available on the occurrence and genetic variability of the diarrhoea-causing enteric protozoan parasite Giardia duodenalis in indigenous communities in Brazil. This cross-sectional epidemiological survey describes the frequency, genotypes, and risk associations for this pathogen in Tapirapé people (Brazilian Amazon) at four sampling campaigns during 2008–2009. Microscopy was used as a screening test, and molecular (PCR and Sanger sequencing) assays targeting the small subunit ribosomal RNA, the glutamate dehydrogenase, the beta-giardin, and the triosephosphate isomerase genes as confirmatory/genotyping methods. Associations between G. duodenalis and sociodemographic and clinical variables were investigated using Chi-squared test and univariable/multivariable logistic regression models. Overall, 574 individuals belonging to six tribes participated in the study, with G. duodenalis prevalence rates varying from 13.5–21.7%. The infection was positively linked to younger age and tribe. Infected children <15 years old reported more frequent gastrointestinal symptoms compared to adults. Assemblage B accounted for three out of four G. duodenalis infections and showed a high genetic diversity. No association between assemblage and age or occurrence of diarrhoea was demonstrated. These data indicate that the most likely source of infection was anthropic and that different pathways (e.g., drinking water) may be involved in the transmission of the parasite.
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Messa A, Köster PC, Garrine M, Gilchrist C, Bartelt LA, Nhampossa T, Massora S, Kotloff K, Levine MM, Alonso PL, Carmena D, Mandomando I. Molecular diversity of Giardia duodenalis in children under 5 years from the Manhiça district, Southern Mozambique enrolled in a matched case-control study on the aetiology of diarrhoea. PLoS Negl Trop Dis 2021; 15:e0008987. [PMID: 33465074 PMCID: PMC7846004 DOI: 10.1371/journal.pntd.0008987] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2020] [Revised: 01/29/2021] [Accepted: 11/18/2020] [Indexed: 12/23/2022] Open
Abstract
Giardia duodenalis is an enteric parasite commonly detected in children. Exposure to this organism may lead to asymptomatic or symptomatic infection. Additionally, early-life infections by this protozoan have been associated with impaired growth and cognitive function in poor resource settings. The Global Enteric Multicenter Study (GEMS) in Mozambique demonstrated that G. duodenalis was more frequent among controls than in diarrhoeal cases (≥3 loosing stools in the previous 24 hours). However, no molecular investigation was conducted to ascertain the molecular variability of the parasite. Therefore, we describe here the frequency and genetic diversity of G. duodenalis infections in children younger than five years of age with and without diarrhoea from the Manhiça district in southern Mozambique enrolled in the context of GEMS. Genomic DNA from 757 G. duodenalis-positive stool samples by immunoassay collected between 2007-2012, were reanalysed by multiplex PCR targeting the E1-HP and C1-P21 genes for the differentiation of assemblages A and B. Overall, 47% (353) of the samples were successfully amplified in at least one locus. Assemblage B accounted for 90% (319/353) of all positives, followed by assemblage A (8%, 29/353) and mixed A+B infections (1%, 5/353). No association between the presence of a given assemblage and the occurrence of diarrhoea could be demonstrated. A total of 351 samples were further analysed by a multi-locus sequence genotyping (MLSG) approach at the glutamate dehydrogenase (gdh), ß-giardin (bg) and triose phosphate isomerase (tpi) genes. Overall, 63% (222/351) of samples were genotyped and/or sub-genotyped in at least one of the three markers. Sequence analysis revealed the presence of assemblages A (10%; 23/222) and B (90%; 199/222) with high molecular diversity at the nucleotide level within the latter; no mixed infections were identified under the MLSG scheme. Assemblage A sequences were assigned to sub-assemblages AI (0.5%, 1/222), AII (7%, 15/222) or ambiguous AII/AIII (3%, 7/222). Within assemblage B, sequences were assigned to sub-assemblages BIII (13%, 28/222), BIV (14%, 31/222) and ambiguous BIII/BIV (59%, 132/222). BIII/BIV sequences accumulated the majority of the single nucleotide polymorphisms detected, particularly in the form of double peaks at chromatogram inspection. This study demonstrated that the occurrence of gastrointestinal illness (diarrhoea) was not associated to a given genotype of G. duodenalis in Mozambican children younger than five years of age. The assemblage B of the parasite was responsible for nine out of ten infections detected in this paediatric population. The extremely high genetic diversity observed within assemblage B isolates was compatible with an hyperendemic epidemiological scenario where infections and reinfections were common. The obtained molecular data may be indicative of high coinfection rates by different G. duodenalis assemblages/sub-assemblages and/or genetic recombination events, although the exact contribution of both mechanisms to the genetic diversity of the parasite remains unknown.
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Affiliation(s)
- Augusto Messa
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
| | - Pamela C. Köster
- Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, Majadahonda, Madrid, Spain
| | - Marcelino Garrine
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
- Global Health and Tropical Medicine (GHTM), Instituto de Higiene e Medicina Tropical (IHMT), Universidade Nova de Lisboa (UNL), Lisbon, Portugal
| | - Carol Gilchrist
- University of Virginia, Charlottesville, Virginia, United States of America
| | - Luther A. Bartelt
- Division of Infectious Diseases, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America
| | - Tacilta Nhampossa
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
- Instituto Nacional de Saúde (INS), Ministério da Saúde, Maputo, Mozambique
| | - Sérgio Massora
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
| | - Karen Kotloff
- Center for Vaccine Development (CVD), University of Maryland School of Medicine, Baltimore, Maryland, United States of America
| | - Myron M. Levine
- Center for Vaccine Development (CVD), University of Maryland School of Medicine, Baltimore, Maryland, United States of America
| | - Pedro L. Alonso
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
- ISGlobal, Hospital Clínic-Universitat de Barcelona, Barcelona, Spain
| | - David Carmena
- Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, Majadahonda, Madrid, Spain
| | - Inácio Mandomando
- Centro de Investigação em Saúde de Manhiça (CISM), Maputo, Mozambique
- Instituto Nacional de Saúde (INS), Ministério da Saúde, Maputo, Mozambique
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Giardiasis in symptomatic children from Sharkia, Egypt: genetic assemblages and associated risk factors. J Parasit Dis 2020; 44:719-724. [PMID: 33184538 DOI: 10.1007/s12639-020-01254-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/04/2020] [Accepted: 07/22/2020] [Indexed: 10/23/2022] Open
Abstract
Giardia intestinalis (G. intestinalis) is a common enteric protozoan parasite worldwide and in Egypt. Identification of true prevailing Giardia assemblages helps in identification of the sources of infection. The study's aim was to determine the true prevalence of Giardia assemblages in Egyptian children from Sharkia governorate presenting with gastrointestinal symptoms and to investigate their association with molecularly detected Giardia. A total of 617 stool specimens were collected from children presenting with gastrointestinal symptoms in Alquraeen, Sharkia governorate, Egypt for 17 months. All stool specimens were microscopically examined by wet mount smear before and after stool concentration to recover parasitic stages. Giardia copro-DNA was amplified from microscopically detected stool specimens using Copro-nPCR targeting the tpi gene for Giardia, followed by sequencing products of nPCR. The molecular prevalence of Giardia among symptomatic children was 9.88%, 83% of which were assemblage B and 17% were assemblages A. Giardia affected both sexes and all ages and was most prevalent in preschool children. Abdominal pain was the most common GIT symptom followed by diarrhoea. However, none of the patients' demographic variables (sex, age, weight and height) nor clinical symptoms showed significant association with molecular detection of Giardia. Giardia was common among symptomatic children from Sharkia, Egypt, with the predominance of assemblage B, which suggests the possibility of sharing common transmission source and route. Giardia had age, sex and clinical symptom distributions without statistical significance. The results necessitate further genomic studies targeting multiple gene targets for a better understanding of the ecology, dynamics of transmission, pathogenicity and clinical impact of Giardia infection, to improve its management and strategic control.
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Ahmad AA, El-Kady AM, Hassan TM. Genotyping of Giardia duodenalis in children in upper Egypt using assemblage- specific PCR technique. PLoS One 2020; 15:e0240119. [PMID: 33002078 PMCID: PMC7529291 DOI: 10.1371/journal.pone.0240119] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/26/2020] [Accepted: 09/18/2020] [Indexed: 01/25/2023] Open
Abstract
Giardia duodenalis is a common gastrointestinal protozoan parasite, causing diarrheal illness in humans worldwide. Yet, the distribution of G. duodenalis genotypes among human patients and their clinical relevance remains controversial. This study aimed to detect G. duodenalis in children in Upper Egypt and identify causative genotypes and elucidate a possible correlation between genotype and clinical presentation. One hundred sixty-five children, regardless of symptoms, were tested for giardiasis. Giardia positive stool samples (40/165) were subjected to PCR amplification targeting the tpi gene with positive PCR results in only 35 cases (87.5%). Assemblage-specific amplification of genotypes (A, B, and the zoonotic E strains) revealed predominantly G. duodenalis Assemblage A (45.7%). Assemblage B and mixed A and B infections were detected in 31.4% and 22.8% of children, respectively. Assemblage E was not detected. G. duodenalis assemblage A was dominant in children who complained of diarrhea and abdominal cramps. In contrast, asymptomatic children with positive stool samples display a higher frequency of assemblage B and mixed infections. The study highlights the predominance of Giardia Assemblage A in our study locality. This study is the first for this endemic area to use the copro-PCR technique for diagnosis and genotyping of giardiasis. Study results show the value of simple species-specific primers for genotyping in communities with little access to laboratory resources. Further genetic studies are needed to clarify the association between parasite genetic diversity and patient symptomatology.
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Affiliation(s)
| | - Asmaa M. El-Kady
- Department of Medical Parasitology, Faculty of Medicine, South Valley University, Qena, Egypt
| | - Tasneem M. Hassan
- Department of Medical Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt
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Natural Infection with Giardia Is Associated with Altered Community Structure of the Human and Canine Gut Microbiome. mSphere 2020; 5:5/4/e00670-20. [PMID: 32759335 PMCID: PMC7407069 DOI: 10.1128/msphere.00670-20] [Citation(s) in RCA: 28] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
While enteric parasitic infections are among the most important infections in lower- and middle-income countries, their impact on gut microbiota is poorly understood. We reasoned that clinical symptoms associated with these infections may be influenced by alterations of the microbiome that occur during infection. To explore this notion, we took a two-pronged approach. First, we studied a cohort of dogs naturally infected with various enteric parasites and found a strong association between parasite infection and altered gut microbiota composition. Giardia, one of the most prevalent parasite infections globally, had a particularly large impact on the microbiome. Second, we took a database-driven strategy to integrate microbiome data with clinical data from large human field studies and found that Giardia infection is also associated with marked alteration of the gut microbiome of children, suggesting a possible explanation for why Giardia has been reported to be associated with protection from moderate to severe diarrhea. Enteric parasitic infections are among the most prevalent infections in lower- and middle-income countries (LMICs) and have a profound impact on global public health. While the microbiome is increasingly recognized as a key determinant of gut health and human development, the impact of naturally acquired parasite infections on microbial community structure in the gut, and the extent to which parasite-induced changes in the microbiome may contribute to gastrointestinal symptoms, is poorly understood. Enteric parasites are routinely identified in companion animals in the United States, presenting a unique opportunity to leverage this animal model to investigate the impact of naturally acquired parasite infections on the microbiome. Clinical, parasitological, and microbiome profiling of a cohort of 258 dogs revealed a significant correlation between parasite infection and composition of the bacterial community in the gut. Relative to other enteric parasites, Giardia was associated with a more pronounced perturbation of the microbiome. To compare our findings to large-scale epidemiological studies of enteric diseases in humans, a database mining approach was employed to integrate clinical and microbiome data. Substantial and consistent alterations to microbiome structure were observed in Giardia-infected children. Importantly, infection was associated with a reduction in the relative abundance of potential pathobionts, including Gammaproteobacteria, and an increase in Prevotella—a profile often associated with gut health. Taken together, these data show that widespread Giardia infection in young animals and humans is associated with significant remodeling of the gut microbiome and provide a possible explanation for the high prevalence of asymptomatic Giardia infections observed across host species. IMPORTANCE While enteric parasitic infections are among the most important infections in lower- and middle-income countries, their impact on gut microbiota is poorly understood. We reasoned that clinical symptoms associated with these infections may be influenced by alterations of the microbiome that occur during infection. To explore this notion, we took a two-pronged approach. First, we studied a cohort of dogs naturally infected with various enteric parasites and found a strong association between parasite infection and altered gut microbiota composition. Giardia, one of the most prevalent parasite infections globally, had a particularly large impact on the microbiome. Second, we took a database-driven strategy to integrate microbiome data with clinical data from large human field studies and found that Giardia infection is also associated with marked alteration of the gut microbiome of children, suggesting a possible explanation for why Giardia has been reported to be associated with protection from moderate to severe diarrhea.
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Costache C, Kalmár Z, Colosi HA, Baciu AM, Opriş RV, Györke A, Colosi IA. First multilocus sequence typing (MLST) of Giardia duodenalis isolates from humans in Romania. Parasit Vectors 2020; 13:387. [PMID: 32736595 PMCID: PMC7393877 DOI: 10.1186/s13071-020-04248-2] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2020] [Accepted: 07/20/2020] [Indexed: 12/26/2022] Open
Abstract
Background Giardia duodenalis is one of the most prevalent and highly diverse human parasites, encompassing a complex of eight genetically distinct assemblages, each further divided into sub-assemblages. While in recent years, G. duodenalis genotype distribution patterns in humans have been intensely studied, there is still very little information available on the diversity of Giardia genotypes and sub-assemblages infecting people in Romania. In the present study, we investigated the genetic diversity of Giardia duodenalis in asymptomatic patients from Romania. Methods Over an 11-month period, human feces from 7805 healthy adults were screened by microscopic analysis for G. duodenalis cysts during their obligatory periodic check-ups. DNA extraction was performed from microscopic-positive fecal samples, followed by multilocus sequence typing of four genetic loci of the ITS region, gdh, tpi and bg genes, followed by DNA sequencing and phylogenetic analysis. Statistical analysis was performed using EpiInfo 2000 software. Results The prevalence of giardiasis in the present study was 0.42% (33/7805). Twenty-three samples (76.67%) were successfully genotyped at each locus. The bg and tpi genes had the highest typing success rate (100%). The identified assemblages were assemblage A in 27 cases (subtypes A2 and A3), and B in 3 cases. Conclusions To our knowledge, the present study is the first report of multilocus sequence typing of G. duodenalis isolated from humans in Romania. The present results may shed light on G. duodenalis infection in humans at a regional and national level, thus increasing awareness against this parasitic infection. ![]()
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Affiliation(s)
- Carmen Costache
- Department of Molecular Sciences, Discipline of Microbiology, Iuliu Hațieganu University of Medicine and Pharmacy, 6 Louis Pasteur Street, 400349, Cluj-Napoca, Romania
| | - Zsuzsa Kalmár
- Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, 3-5 Calea Mănăştur, 400372, Cluj-Napoca, Romania.
| | - Horațiu Alexandru Colosi
- Department of Medical Education, Discipline of Medical Informatics and Biostatistics, Iuliu Hațieganu University of Medicine and Pharmacy, 6 Louis Pasteur Street, 400349, Cluj-Napoca, Romania
| | - Alina Mihaela Baciu
- Department of Molecular Sciences, Discipline of Microbiology, Iuliu Hațieganu University of Medicine and Pharmacy, 6 Louis Pasteur Street, 400349, Cluj-Napoca, Romania
| | - Răzvan Vlad Opriş
- Department of Molecular Sciences, Discipline of Microbiology, Iuliu Hațieganu University of Medicine and Pharmacy, 6 Louis Pasteur Street, 400349, Cluj-Napoca, Romania
| | - Adriana Györke
- Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, 3-5 Calea Mănăştur, 400372, Cluj-Napoca, Romania
| | - Ioana Alina Colosi
- Department of Molecular Sciences, Discipline of Microbiology, Iuliu Hațieganu University of Medicine and Pharmacy, 6 Louis Pasteur Street, 400349, Cluj-Napoca, Romania
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Frequency and Molecular Detection of Giardia intestinalis in Children Attending Pediatrics of Punjab, Pakistan. Jundishapur J Microbiol 2020. [DOI: 10.5812/jjm.97080] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
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Samie A, Tanih NF, Seisa I, Seheri M, Mphahlele J, ElBakri A, Mbati P. Prevalence and genetic characterization of Giardia lamblia in relation to diarrhea in Limpopo and Gauteng provinces, South Africa. Parasite Epidemiol Control 2020; 9:e00140. [PMID: 32083192 PMCID: PMC7016452 DOI: 10.1016/j.parepi.2020.e00140] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2019] [Revised: 01/28/2020] [Accepted: 01/28/2020] [Indexed: 11/28/2022] Open
Abstract
Background Very few studies have determined the prevalence and assemblage distribution of Giardia lamblia in South Africa. The present study aimed to ascertain the prevalence of G. lamblia infection and the spread of the various assemblages in two communities in South Africa - Giyani, Limpopo province (rural community) and Pretoria Guateng province (urban community). Methods Prevalence was determined by immunological and molecular methods analyzing a total of 516 stool samples collected from patients visiting different health centres in Giyani and Pretoria. For immunological assays, samples were screened by ELISA to detect G. lamblia antigen. Furthermore, a semi nested PCR amplifying the triose phosphate isomerase (tpi) gene was used to differentiate between the two most common human assemblages (A and B). Findings Of the 516 participants, 40 (7.75%) were identified as positive by ELISA. A statistically significant correlation was observed between the stool texture and Giardia infection (ᵡ2 = 10.533; p = .005). G. lamblia was significantly associated with watery stool types in females p = .008. Furthermore, a significant association was also noticed between the origin of samples (ᵡ2 = 9.725; p = .002). No significant correlation between age and gender was noted. Regarding the age groups, most people who were infected were between 3 and 20 years. A statistically significant association was seen (p = .001) with the distribution of the pathogen with the stool type. The prevalence of Giardia infection was higher in watery stool samples (71.4%) in Giyani region (rural) whereas in Pretoria, high prevalence was found in loose stool samples (6.2%). Generally, the distribution was statistically significant in the stool type collected for the study (p = .005). Genotyping revealed more G. lamblia assemblage B (17.8%) than assemblage A (1.7%). Furthermore, 21.0% of the samples exhibited single infection while 4.2% had mixed infections. Assemblage B was more common in Giyani than in urban Pretoria. Conclusions The study confirms Giardia as an important cause of diarrhea in the concerned communities with people in rural areas more at risk compared to those in urban areas with higher prevalence among younger patients. Therefore, health education campaigns should target young age groups.
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Affiliation(s)
- Amidou Samie
- Molecular Parasitology and opportunistic infections program, Department of Microbiology, University of Venda, Private Bag X5050, Thohoyandou, Limpopo, South Africa
| | | | - Itumeleng Seisa
- Molecular Parasitology and opportunistic infections program, Department of Microbiology, University of Venda, Private Bag X5050, Thohoyandou, Limpopo, South Africa
| | - Mapaseka Seheri
- South African Medical Research Council Diarrhoeal Pathogens Research Unit, Department of Medical Virology, Sefako Makgatho Health Sciences University, Medunsa, 0204, Pretoria, South Africa
| | - Jeffrey Mphahlele
- South African Medical Research Council, 1 Soutpansberg Road, Pretoria, Gauteng, South Africa
| | - Ali ElBakri
- Department of Medical Laboratory Sciences,College of Health Sciences, University of Sharjah, PO Box 27272, Sharjah, United Arab Emirates
| | - Peter Mbati
- Deputy Pro Vice Chancellor Academics and Quality, Botho University, Botho Education Park, Kgale, Gaborone, Botswana
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Ma DW, Lee MR, Hong SH, Cho SH, Lee SE. Molecular Prevalence and Genotypes of Cryptosporidium parvum and Giardia duodenalis in Patients with Acute Diarrhea in Korea, 2013-2016. THE KOREAN JOURNAL OF PARASITOLOGY 2019; 57:531-536. [PMID: 31715696 PMCID: PMC6851252 DOI: 10.3347/kjp.2019.57.5.531] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 05/31/2019] [Accepted: 09/09/2019] [Indexed: 01/24/2023]
Abstract
Cryptosporidium parvum and Giardia duodenalis are the main diarrhea-causing parasitic pathogens; however, their prevalence in Korea is unknown. Here, we conducted a survey to determine the prevalence and genotype distribution of these 2 pathogens causing acute diarrhea in 8,571 patients hospitalized in 17 Regional Institute of Health Environment sites in Korea, during 2013–2016. C. parvum and G. duodenalis were detected and genotyped by nested PCR, and the isolate were molecularly characterized by sequencing the glycoprotein 60 (Gp60) and β-giardin genes, respectively. The overall prevalence of C. parvum and G. duodenalis was 0.37% (n=32) and 0.55% (n=47), respectively, and both pathogens were more prevalent in children under 9 years old. Molecular epidemiological analysis showed that the C. parvum isolates belonged to the IIa family and were subtyped as IIaA13G2R1, IIaA14G2R1, IIaA15G2R1, and IIaA18G3R1. Analysis of the β-giardin gene fragment from G. duodenalis showed that all positive strains belong to assemblage A. This is the first report on the molecular epidemiology and subtyping of C. parvum and G. duodenalis in such a large number of diarrheal patients in Korea. These results highlight the need for continuous monitoring of these zoonotic pathogens and provide a basis for implementing control and prevention strategies. Further, the results might be useful for epidemiological investigation of the source of outbreak.
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Affiliation(s)
- Da-Won Ma
- Division of Vectors and Parasitic Diseases, Centers for Disease Control and Prevention, Osong 28159, Korea
| | - Myoung-Ro Lee
- Division of Vectors and Parasitic Diseases, Centers for Disease Control and Prevention, Osong 28159, Korea
| | - Sung-Hee Hong
- Division of Vectors and Parasitic Diseases, Centers for Disease Control and Prevention, Osong 28159, Korea
| | - Shin-Hyeong Cho
- Division of Vectors and Parasitic Diseases, Centers for Disease Control and Prevention, Osong 28159, Korea
| | - Sang-Eun Lee
- Division of Vectors and Parasitic Diseases, Centers for Disease Control and Prevention, Osong 28159, Korea
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Wang Y, Gonzalez-Moreno O, Roellig DM, Oliver L, Huguet J, Guo Y, Feng Y, Xiao L. Epidemiological distribution of genotypes of Giardia duodenalis in humans in Spain. Parasit Vectors 2019; 12:432. [PMID: 31492183 PMCID: PMC6728964 DOI: 10.1186/s13071-019-3692-4] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2019] [Accepted: 08/28/2019] [Indexed: 12/13/2022] Open
Abstract
BACKGROUND Although the distribution of Giardia duodenalis genotypes in humans has been increasingly reported in recent years, data on possible differences in pathogen transmission between age groups and virulence between genotypes are scarce. The purpose of this study is to investigate the genetic diversity of G. duodenalis in humans in Spain and compare the distribution of G. duodenalis assemblages A and B between children and adults and clinical presentations between the two genotypes. METHODS In the present study, 125 microscopy-positive fecal samples were collected from humans in Spain over a 7-year period. PCR and sequence analyses of the triosephosphate isomerase, β-giardin and glutamate dehydrogenase genes were used to identify the multilocus genotypes of G. duodenalis. RESULTS Sequence analysis of three genetic loci identified both G. duodenalis assemblages A (29) and B (66), with co-infections of the two in two patients. Among the sequences obtained in this study, four multilocus genotypes (MLGs) of the sub-assemblage AII were observed within assemblage A. In contrast, 19 MLGs were detected within assemblage B due to the high sequence diversity at each locus. One MLG, however, was found in 51.9% (27/52) of assemblage B samples. Children were more commonly infected by assemblage B (44/53 or 83%) than adults (22/42 or 52.4%; χ2 = 10.371, df = 1, P = 0.001). Asymptomatic infection was more common in patients with assemblage A (4/29 or 13.8%) than in those with assemblage B (1/66 or 1.5%; χ2 = 6.091, df = 1, P = 0.029), and the frequency of abdominal pain occurrence was higher in assemblage B patients (65/66 or 98.5%) than assemblage A patients (25/29 or 86.2%; χ2 = 6.091, df = 1, P = 0.029). CONCLUSIONS These results illustrate the existence of differences in genotype distribution between children and adults and clinical presentations between G. duodenalis genotypes. They are useful in understanding the transmission of G. duodenalis in humans in Spain.
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Affiliation(s)
- Yuanfei Wang
- State Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai, 200237 China
| | - Olga Gonzalez-Moreno
- Laboratory of Microbiology and Parasitology, SYNLAB, 08950 Barcelona, Spain
- Laboratory of Parasitology, Department of Biology, Healthcare and Environment, Faculty of Pharmacy, University of Barcelona, 08028 Barcelona, Spain
| | - Dawn M. Roellig
- Division of Foodborne, Waterborne, and Environmental Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333 USA
| | - Laura Oliver
- Laboratory of Microbiology and Parasitology, SYNLAB, 08950 Barcelona, Spain
| | - Jordi Huguet
- Laboratory of Microbiology and Parasitology, SYNLAB, 08950 Barcelona, Spain
| | - Yaqiong Guo
- Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642 China
| | - Yaoyu Feng
- State Key Laboratory of Bioreactor Engineering, School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai, 200237 China
- Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642 China
| | - Lihua Xiao
- Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642 China
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Molecular Genotyping of Giardia duodenalis in Humans in the Andimeshk County, Southwestern Iran. Acta Parasitol 2019; 64:376-383. [PMID: 30968348 DOI: 10.2478/s11686-019-00051-x] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2018] [Accepted: 03/29/2019] [Indexed: 01/10/2023]
Abstract
INTRODUCTION Giardia duodenalis, one of the most common intestinal protozoa, infects a wide range of vertebrates, including humans and animals. MATERIALS AND METHODS In this study, 84 Giardia duodenalis positive stool samples were collected from 3580 patients attending the Imam Ali Hospital and two public health centers in Andimeshk County, southwestern Iran. Stool samples were examined initially by microscopy, and then G. duodenalis was confirmed by SSU rRNA gene and genotypes were determined by amplification of the gdh and β-giardin genes. RESULTS The SSU rRNA, gdh, and β-giardin genes were successfully amplified in 89.3%, 58.3%, and 51.2% samples, respectively. Of the positive samples for gdh and β-giardin, 40 isolates were successfully sequenced. Twenty-three isolates belonged to assemblage A, sub-assemblage AII, and 17 belonged to assemblage B. Of the 24 successfully amplified asymptomatic cases, 12 belonged to assemblage A and 12 belonged to assemblage B. CONCLUSION The current study found that 64.3% of the patients were asymptomatic. From an epidemiological point of view, the high percentage of asymptomatic patients is important because of their role in the transmission of Giardia. The predominant assemblage was assemblage A, sub-assemblage AII. In general, therefore, it seems that most infections are probably transmitted by anthroponotic pathways in the region.
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Gasparinho C, Ferreira FS, Mayer AC, Mirante MC, Vaz Nery S, Santos-Reis A, Portugal-Calisto D, Brito M. Molecular characterization of Giardia lamblia in children less than 5 years of age with diarrhoea attending the Bengo General Hospital, Angola. Trans R Soc Trop Med Hyg 2019; 111:497-503. [PMID: 29438541 PMCID: PMC5914390 DOI: 10.1093/trstmh/try004] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2017] [Accepted: 01/16/2018] [Indexed: 11/12/2022] Open
Abstract
Background Giardia lamblia is a pathogenic intestinal protozoan with high prevalence in developing countries, especially among children. Molecular characterization has revealed the existence of eight assemblages, with A and B being more commonly described in human infections. Despite its importance, to our knowledge this is the first published molecular analysis of G. lamblia assemblages in Angola. Methods The present study aimed to identify the assemblages of G. lamblia in children with acute diarrhoea presenting at the Bengo General Hospital, Angola. A stool sample was collected and microscopy and immunochromatographic tests were used. DNA was extracted and assemblage determination was performed through amplification of the gene fragment ssu-rRNA (175 bp) and β-giardin (511 bp) through polymerase chain reaction and DNA sequencing. Results Of the 16 stool samples screened, 12 were successfully sequenced. Eleven isolates were assigned to assemblage B and one to assemblage A. Subassemblage determination was not possible for assemblage B, while the single isolate assigned to assemblage A was identified as belonging to subassemblage A3. Conclusion This study provides information about G. lamblia assemblages in Bengo Province, Angola and may contribute as a first step in understanding the molecular epidemiology of this protozoan in the country. GenBank accession numbers for the ssur-RNA gene: MF479750, MF479751, MF479752, MF479753, MF479754, MF479755, MF479756, MF479757, MF479758, MF479759, MF479760, MF479761. GenBank accession numbers for the β-giardin gene: MF565378, MF565379, MF565380, MF565381.
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Affiliation(s)
- Carolina Gasparinho
- Clinical Research, Centro de Investigação em Saúde de Angola, Caxito, Angola.,Unidade de Saúde Pública Internacional e Bioestatística, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, 1349-008 Lisboa, Portugal
| | - Filipa S Ferreira
- Global Health and Tropical Medicine, Unidade de Clínica Tropical, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal
| | | | - Maria Clara Mirante
- Laboratory, Centro de Investigação em Saúde de Angola, Caxito, Angola.,Serviço de Patologia Clínica, Centro Hospitalar Lisboa Ocidental E.P.E., 1449-005 Lisboa, Portugal
| | - Susana Vaz Nery
- Department of Global Health, Research School of Population Health, Australian National University, Canberra, Australian Capital Territory, Australia
| | - Ana Santos-Reis
- Laboratory, Centro de Investigação em Saúde de Angola, Caxito, Angola.,Global Health and Tropical Medicine, Unidade de Clínica Tropical, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, 1349-008 Lisboa, Portugal
| | - Daniela Portugal-Calisto
- Global Health and Tropical Medicine, Unidade de Clínica Tropical, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal.,Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland
| | - Miguel Brito
- Clinical Research, Centro de Investigação em Saúde de Angola, Caxito, Angola.,Lisbon School of Health Technology, Lisboa, Portugal
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Villamizar X, Higuera A, Herrera G, Vasquez-A LR, Buitron L, Muñoz LM, Gonzalez-C FE, Lopez MC, Giraldo JC, Ramírez JD. Molecular and descriptive epidemiology of intestinal protozoan parasites of children and their pets in Cauca, Colombia: a cross-sectional study. BMC Infect Dis 2019; 19:190. [PMID: 30808303 PMCID: PMC6390308 DOI: 10.1186/s12879-019-3810-0] [Citation(s) in RCA: 52] [Impact Index Per Article: 8.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2018] [Accepted: 02/12/2019] [Indexed: 01/18/2023] Open
Abstract
BACKGROUND Parasitic infections, particularly those caused by protozoa, represent a considerable public health problem in developing countries. Blastocystis, Giardia duodenalis, Cryptosporidium spp. and the Entamoeba complex (Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii) are the most common etiological causes of intestinal parasitic infections. METHODS We carried out a descriptive cross-sectional study in school-age children attending a daycare institution in commune eight of Popayán, Cauca (Southwest Colombia). A total of 266 fecal samples were collected (258 from children and eight from pets). Blastocystis, G. duodenalis, Cryptosporidium spp. and the Entamoeba complex were identified by microscopy, quantitative real-time PCR (qPCR) and conventional PCR. The concordance of qPCR and microscopy was assessed using the Kappa index. Molecular characterization was conducted to identify Blastocystis subtypes (18S), G. duodenalis assemblages (tpi and gdh) and Cryptosporidium species/subtypes (18S and GP60). Potential associations between intestinal parasitism and sociodemographic factors were examined using bivariate analyses. RESULTS A total of 258 fecal samples from children were analyzed by microscopy and 255 samples were analyzed by qPCR. The prevalence of Blastocystis was between 25.19% (microscopy) and 39.22% (qPCR), that of G. duodenalis was between 8.14% (microscopy) and 10.59% (qPCR), that of Cryptosporidium spp. was estimated at 9.8% (qPCR), and that of the Entamoeba complex was between 0.39% (conventional PCR) and 0.78% (microscopy). The concordance between microscopy and qPCR was very low. Blastocystis ST1 (alleles 4, 8, and 80), ST2 (alleles 11, 12, and 15), ST3 (alleles 31, 34, 36, 38,57, and 151), and ST4 (alleles 42 and 91), G. duodenalis assemblages AII, BIII, BIV and D, C. parvum subtype IIa and C. hominis subtype IbA9G3R2 were identified. The only identified member of the Entamoeba complex corresponded to E. histolytica. No statistically significant association was identified between parasitic infection and any sociodemographic variable. CONCLUSION This study revealed the usefulness of molecular methods to depict the transmission dynamics of parasitic protozoa in southwest Colombia. The presence of some of these protozoa in domestic animals may be involved in their transmission.
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Affiliation(s)
- Ximena Villamizar
- Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia
| | - Adriana Higuera
- Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia
| | - Giovanny Herrera
- Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia
| | - Luis Reinel Vasquez-A
- Centro de estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, Universidad del Cauca, Popayán, Colombia
| | - Lorena Buitron
- Centro de estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, Universidad del Cauca, Popayán, Colombia
| | - Lina Maria Muñoz
- Centro de estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, Universidad del Cauca, Popayán, Colombia
| | - Fabiola E. Gonzalez-C
- Centro de estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, Universidad del Cauca, Popayán, Colombia
| | - Myriam Consuelo Lopez
- Departamento de Salud Pública, Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia
| | | | - Juan David Ramírez
- Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia
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25
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DNA profiling reveals Neobenedenia girellae as the primary parasitic monogenean in global fisheries and aquaculture. Mol Phylogenet Evol 2018; 129:130-137. [DOI: 10.1016/j.ympev.2018.05.012] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/13/2017] [Revised: 01/23/2018] [Accepted: 05/11/2018] [Indexed: 11/19/2022]
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Kasaei R, Carmena D, Jelowdar A, Beiromvand M. Molecular genotyping of Giardia duodenalis in children from Behbahan, southwestern Iran. Parasitol Res 2018. [PMID: 29541855 DOI: 10.1007/s00436-018-5826-6] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/07/2022]
Abstract
Giardia duodenalis is an intestinal flagellated protozoan that infects humans and several animal species. Giardiasis causing more than 200 million symptomatic infections globally is one of the most common causes of diarrhea in developing countries. Based on molecular studies mainly targeting the small-subunit (SSU) rRNA gene locus of the parasite, eight assemblages (A to H) have been identified in humans and other animal species. The aim of the current study was to evaluate the frequency and molecular diversity of G. duodenalis in children from rural and urban day care centers from Behbahan, southwestern Iran. This cross-sectional study was based on a concentration method for the microscopic detection of G. duodenalis in stool samples of 450 children, aged 1-7 years, in Behbahan, southwestern Iran. The survey was conducted from December 2015 to May 2016. PCR methods targeting the SSU rRNA and triose phosphate isomerase (TPI) genes of G. duodenalis were used for the identification and genotyping of the parasite isolates. Based on sucrose flotation and microscopy techniques, 2.7% (12/450) of children were infected with G. duodenalis, of which six (50.0%) were males and the other six (50.0%) were females. Overall, 91.7% (11/12) of the infections were detected in children from rural areas. The SSU rRNA and TPI genes were amplified successfully in nine and eight, respectively, of the Giardia-positive samples at microscopy. Among the eight TPI sequences, assemblage A, sub-assemblage AII, was identified in five of the isolates. The sequences of the three remaining samples were untypable. Although no significantly statistical difference between genotype and clinical symptoms was found, five out of the eight isolates identified as assemblage A were obtained in asymptomatic children. Giardia duodenalis infections were more prevalent in children from rural day care schools, and the predominant assemblage was A, sub-assemblage AII. The higher prevalence of giardiasis in rural areas might be related to differences in personal hygiene habits, parents' education level, source of drinking water, and inadequate hygienic toilet facilities in rural areas.
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Affiliation(s)
- Raziyeh Kasaei
- Infectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, P.O. Box 61357-15794, Ahvaz, Iran.,Department of Parasitology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - David Carmena
- Parasitology Reference and Research Laboratory, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
| | - Ali Jelowdar
- Infectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, P.O. Box 61357-15794, Ahvaz, Iran.,Department of Parasitology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Molouk Beiromvand
- Infectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, P.O. Box 61357-15794, Ahvaz, Iran. .,Department of Parasitology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
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Cacciò SM, Lalle M, Svärd SG. Host specificity in the Giardia duodenalis species complex. INFECTION GENETICS AND EVOLUTION 2017; 66:335-345. [PMID: 29225147 DOI: 10.1016/j.meegid.2017.12.001] [Citation(s) in RCA: 133] [Impact Index Per Article: 16.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/03/2017] [Revised: 12/01/2017] [Accepted: 12/02/2017] [Indexed: 12/15/2022]
Abstract
Giardia duodenalis is a unicellular flagellated parasite that infects the gastrointestinal tract of a wide range of mammalian species, including humans. Investigations of protein and DNA polymorphisms revealed that G. duodenalis should be considered as a species complex, whose members, despite being morphologically indistinguishable, can be classified into eight groups, or Assemblages, separated by large genetic distances. Assemblages display various degree of host specificity, with Assemblages A and B occurring in humans and many other hosts, Assemblage C and D in canids, Assemblage E in hoofed animals, Assemblage F in cats, Assemblage G in rodents, and Assemblage H in pinnipeds. The factors determining host specificity are only partially understood, and clearly involve both the host and the parasite. Here, we review the results of in vitro and in vivo experiments, and clinical observations to highlight relevant biological and genetic differences between Assemblages, with a focus on human infection.
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Affiliation(s)
- Simone M Cacciò
- Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy.
| | - Marco Lalle
- Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy
| | - Staffan G Svärd
- Department of Cell and Molecular Biology, Uppsala University, Uppsala, Sweden
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Abraham RJ, O'Dea M, Rusdi B, Page SW, O'Handley R, Abraham S. Giardia duodenalis mouse model for the development of novel antigiardial agents. J Microbiol Methods 2017; 145:7-9. [PMID: 29198594 DOI: 10.1016/j.mimet.2017.11.025] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/18/2017] [Revised: 11/28/2017] [Accepted: 11/29/2017] [Indexed: 10/18/2022]
Abstract
This study describes a neonatal mouse model of Giardia infection for development of novel antigiardials. Mice were infected with the axenically cultured Assemblage A BAH2c2 strain, with 105 trophozoites per animal recovered. This model proved to be robust and consistent for use in preliminary drug efficacy trials and drug development.
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Affiliation(s)
- Rebecca J Abraham
- Murdoch University, 90 South Street, Murdoch, Western Australia 6150, Australia; The University of Adelaide School of Animal and Veterinary Science, Mudla Wirra Rd, Roseworthy, South Australia 5352, Australia.
| | - Mark O'Dea
- Murdoch University, 90 South Street, Murdoch, Western Australia 6150, Australia
| | - Bertha Rusdi
- Murdoch University, 90 South Street, Murdoch, Western Australia 6150, Australia
| | | | - Ryan O'Handley
- The University of Adelaide School of Animal and Veterinary Science, Mudla Wirra Rd, Roseworthy, South Australia 5352, Australia
| | - Sam Abraham
- Murdoch University, 90 South Street, Murdoch, Western Australia 6150, Australia
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Xiao L, Feng Y. Molecular epidemiologic tools for waterborne pathogens Cryptosporidium spp. and Giardia duodenalis. Food Waterborne Parasitol 2017; 8-9:14-32. [PMID: 32095639 PMCID: PMC7034008 DOI: 10.1016/j.fawpar.2017.09.002] [Citation(s) in RCA: 169] [Impact Index Per Article: 21.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/01/2017] [Revised: 09/19/2017] [Accepted: 09/19/2017] [Indexed: 01/26/2023] Open
Abstract
Molecular diagnostic tools have played an important role in improving our understanding of the transmission of Cryptosporidium spp. and Giardia duodenalis, which are two of the most important waterborne parasites in industrialized nations. Genotyping tools are frequently used in the identification of host-adapted Cryptosporidium species and G. duodenalis assemblages, allowing the assessment of infection sources in humans and public health potential of parasites found in animals and the environment. In contrast, subtyping tools are more often used in case linkages, advanced tracking of infections sources, and assessment of disease burdens attributable to anthroponotic and zoonotic transmission. More recently, multilocus typing tools have been developed for population genetic characterizations of transmission dynamics and delineation of mechanisms for the emergence of virulent subtypes. With the recent development in next generation sequencing techniques, whole genome sequencing and comparative genomic analysis are increasingly used in characterizing Cryptosporidium spp. and G. duodenalis. The use of these tools in epidemiologic studies has identified significant differences in the transmission of Cryptosporidium spp. in humans between developing countries and industrialized nations, especially the role of zoonotic transmission in human infection. Geographic differences are also present in the distribution of G. duodenalis assemblages A and B in humans. In contrast, there is little evidence for widespread zoonotic transmission of giardiasis in both developing and industrialized countries. Differences in virulence have been identified among Cryptosporidium species and subtypes, and possibly between G. duodenalis assemblages A and B, and genetic recombination has been identified as one mechanism for the emergence of virulent C. hominis subtypes. These recent advances are providing insight into the epidemiology of waterborne protozoan parasites in both developing and developed countries.
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Affiliation(s)
- Lihua Xiao
- Division of Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA
| | - Yaoyu Feng
- College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
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30
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Faria CP, Zanini GM, Dias GS, Sousa MDC. Associations of Giardia lamblia assemblages with HIV infections and symptomatology: HIV virus and assemblage B were they born to each other? Acta Trop 2017; 172:80-85. [PMID: 28456597 DOI: 10.1016/j.actatropica.2017.04.026] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2016] [Revised: 04/20/2017] [Accepted: 04/22/2017] [Indexed: 12/13/2022]
Abstract
Giardia lamblia is an intestinal parasite that has an extensive genetic variation among isolates. This species is divided into eight different assemblages (A-H), but only assemblages A and B have been associated with human infections. Studies on the associations of G. lamblia assemblages and symptoms have been done but were inconclusive. The aim of this study was to correlate G. lamblia assemblages with symptoms in patients with and without HIV/AIDS and its association with the CD4T cell count. The cross-sectional survey was conducted among patients attending the Evandro Chagas National Institute of Infectious Diseases (INI/FIOCRUZ) in Rio de Janeiro from January 2011 to February 2015. Thirty-eight of 65 microscopically positive stool samples for G. lamblia were from HIV positive patients and 27 were from HIV negative patients. Of the HIV infected patients, 19 (55.9%) were genotyped as assemblage B of which 9 (47.4%) had a CD4Tcell count below 200cells/mm3. In addition, we found a greater number of samples belonging to assemblage B in symptomatic cases (11 of 19; 57.9%). Our data suggest that assemblage B is very likely to be found in HIV infected patients and probably the lower CD4T count gives advantages for assemblage B replication. Furthermore, assemblage B seems to be associated with symptomatology, particularly abdominal pain, asthenia, diarrhea, fever, headache and myalgia. This study provides information on G. lamblia assemblages and symptoms in patients with and without HIV/AIDS virus and their association with CD4Tcell counts.
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Affiliation(s)
- Clarissa Perez Faria
- CNC - Center for Neurosciences and Cell Biology, University of Coimbra, 3030-548 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, 3030-548 Coimbra, Portugal; Laboratory of Parasitology, Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Rio de Janeiro, 21040-900, Brazil
| | - Graziela Maria Zanini
- Laboratory of Parasitology, Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Rio de Janeiro, 21040-900, Brazil
| | - Gisele Silva Dias
- Laboratory of Parasitology, Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Rio de Janeiro, 21040-900, Brazil
| | - Maria do Céu Sousa
- CNC - Center for Neurosciences and Cell Biology, University of Coimbra, 3030-548 Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, 3030-548 Coimbra, Portugal.
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Lass A, Karanis P, Korzeniewski K. First detection and genotyping of Giardia intestinalis in stool samples collected from children in Ghazni Province, eastern Afghanistan and evaluation of the PCR assay in formalin-fixed specimens. Parasitol Res 2017; 116:2255-2264. [PMID: 28612148 PMCID: PMC5529491 DOI: 10.1007/s00436-017-5529-4] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/08/2017] [Accepted: 06/01/2017] [Indexed: 02/07/2023]
Abstract
It is estimated that faecal-orally transmitted diseases are common in Afghanistan, as a consequence of poor hygienic standards of life and widespread contamination of water and food with both human and animal faeces. However, there is little information in the literature concerning infections caused by intestinal parasites in the Afghan population. In this study, we report the occurrence of Giardia intestinalis assemblages (A and B) in formalin-fixed stool samples collected from 245 Afghan schoolchildren living in Ghazni Province in eastern Afghanistan. Detection of the parasite’s DNA and genotyping was performed using real-time PCR, specific to the β-giardin gene of G. intestinalis. Positive results were recorded in 52 (21.2%) samples. Genotyping was successful in 39 faecal samples and showed the predominance of assemblage B of G. intestinalis in this population (15 assemblage A and 24 assemblage B). Co-infection with both genotypes A and B was detected in four samples. Additionally, we evaluated the effect of 10% buffered formalin fixative on the detection of G. intestinalis DNA using real-time PCR and nested PCR characterised by different lengths of PCR products (74 and 479 bp, respectively). The human faeces containing the Giardia cysts were tested for 16 weeks. Amplification of G. intestinalis DNA with real-time PCR was possible up to 6 weeks of preservation of stool sample in formalin, compared to only 2 weeks with nested PCR. This suggests that real-time PCR is a more suitable tool in cases where stool samples have to be kept in formalin for longer periods of time.
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Affiliation(s)
- Anna Lass
- Department of Tropical Parasitology, Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gdansk, 9b Powstania Styczniowego Str, 81-519, Gdynia, Poland.
| | - Panagiotis Karanis
- State Key Laboratory of Plateau Ecology and Agriculture, Center for Biomedicine and Infectious Disease, Qinghai Academy of Animal Sciences and Veterinary Medicine, Qinghai University, Xining, 1#Wei'er Road, Qinghai Biological Scientific Estate Garden, Xining, 810016, People's Republic of China
| | - Krzysztof Korzeniewski
- Epidemiology and Tropical Medicine Department in Gdynia, Military Institute of Medicine in Warsaw, Grudzinskiego St. 4, 81-103, Gdynia, Poland
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Hussein EM, Ismail OA, Mokhtar AB, Mohamed SE, Saad RM. Nested PCR targeting intergenic spacer (IGS) in genotyping of Giardia duodenalis isolated from symptomatic and asymptomatic infected Egyptian school children. Parasitol Res 2016; 116:763-771. [PMID: 27975120 DOI: 10.1007/s00436-016-5347-0] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2016] [Accepted: 12/07/2016] [Indexed: 11/30/2022]
Abstract
Distinct sequences of Giardia duodenalis assemblages raised the hypothesis that certain assemblages may contribute to its clinical outcome. However, sequences analysis is time consuming, expensive, and needs many manual operations. Nested PCR targeting intergenic spacer (IGS) region was applied successfully to genotype G. duodenalis. This study aimed to identify the prevalence of G. duodenalis assemblages among giardiasis school children and its relation to the presence of symptoms using nested IGS/PCR. Of 65 microscopically confirmed Giardia-positive samples, 65 samples were genotyped proving high sensitivity (92.3%) of IGS/PCR. Negative IGS/PCR samples were also negative for β-giardin gene. Subassemblage AI was the commonest with 66.6% (20/30) among asymptomatic children compared to 53.3% (16/30) of symptomatic, while assemblage B was found in 40% (12/30) of symptomatic compared to 20% (6/30) of asymptomatic. The difference was significant. AII was only found in asymptomatic with 13.4% (4/30), while mixed infections (AI&B) were recorded only in 6.6% (2/30) of symptomatic group. A significant relation was found between younger children susceptibility for AI and B infections as presented in 77.7 (12/16) and 83.3% (10/12) of symptomatic, respectively, and 80 (16/80) and 33.4% (2/4) of asymptomatic, respectively. Significant relations were found between AI with intermittent diarrhea and B with chronic. A significant relation was found between assemblage distributions and heavy infection intensity. In conclusion, higher incidence of assemblage B among symptomatic children compared to asymptomatic could denote its possible pathogenic potential.
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Affiliation(s)
- Eman M Hussein
- Medical Parasitology Department, Faculty of Medicine, Suez Canal University, Ismailia, 41522, Egypt.
| | - Ola A Ismail
- Medical Parasitology Department, Faculty of Medicine, Suez Canal University, Ismailia, 41522, Egypt
| | - Amira B Mokhtar
- Medical Parasitology Department, Faculty of Medicine, Suez Canal University, Ismailia, 41522, Egypt
| | - Samer E Mohamed
- Medical Parasitology Department, Faculty of Medicine, Suez Canal University, Ismailia, 41522, Egypt
| | - Rania M Saad
- Medical Parasitology Department, Faculty of Medicine, Suez Canal University, Ismailia, 41522, Egypt
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Ghoshal U, Shukla R, Pant P, Ghoshal UC. Frequency, diagnostic performance of coproantigen detection and genotyping of the Giardia among patients referred to a multi-level teaching hospital in northern India. Pathog Glob Health 2016; 110:316-320. [PMID: 27852150 PMCID: PMC5189869 DOI: 10.1080/20477724.2016.1254141] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023] Open
Abstract
Giardiasis, a common gastrointestinal parasitic infection in tropics, is diagnosed on stool microscopy (gold standard); however, its sensitivity is low due to intermittent fecal shedding. Coproantigen detection (ELISA) is useful but requires further evaluation. We aimed to study: (a) detection of Giardia by stool microscopy and/or coproantigen, (b) diagnostic performance of fecal antigen detection and microscopy, and c) genotypic characterization of G. lamblia using PCR specific for triose phosphate isomerase (tpi) gene. Stool samples from 2992 patients were examined by microscopy from March 2013 to March 2015 in a multi level teaching hospital in northern India. Giardia coproantigen detection was performed by ELISA in a subset of patients. Genetic characterization of G. lamblia was performed by PCR targeting tpi gene in a subset of microscopy positive stool samples. Of 2992 patients, 132 (4.4%) had Giardia by microscopy (cyst/trophozoite) and/or ELISA. ELISA was performed in 264 patients; of them, 127 were positive by microscopy. Sensitivity, specificity, positive and negative predictive values of ELISA were 91, 91, 94, and 91%, respectively, using microscopy as a gold standard. PCR was performed in 116 randomly selected samples having Giardia using tpi gene. Assemblages A and B were found among 44 (38%) and 72 (62%) patients, respectively. Assemblage B was more often associated with malnutrition and loss of appetite than A (48/72 [67%] vs. 21/44 [48%], P = 0.044 and 17/72 [24%] vs. 14/44 [32%], P = 0.019). We conclude that 4.4% of studied population had giardiasis. Fecal antigen is a useful method for diagnosis and assemblage B is the most common genotype.
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Affiliation(s)
- Ujjala Ghoshal
- Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - Ratnakar Shukla
- Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - Priyannk Pant
- Department of Microbiology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
| | - Uday C. Ghoshal
- Department of Gastroenterology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India
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Positive correlation of HIV infection with Giardia intestinalis assemblage B but not with assemblage A in asymptomatic Kenyan children. AIDS 2016; 30:2385-7. [PMID: 27478987 DOI: 10.1097/qad.0000000000001216] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/02/2023]
Abstract
A cross-sectional molecular epidemiological study of Giardia intestinalis infection was conducted among asymptomatic Kenyan children with (n = 123) and without (n = 111) HIV infection. G. intestinalis assemblage B infection was positively correlated with HIV infection [HIV (+), 18.7% vs. HIV (-), 11.7%; P = 0.013], whereas assemblage A infection was not [HIV (+), 4.1% vs. HIV (-), 6.3%; P = 0.510]. Thus, HIV infection is a risk factor for G. intestinalis assemblage B infection but not for assemblage A infection.
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Quadros RMD, Weiss PHE, Marques SMT, Miletti LC. POTENTIAL CROSS-CONTAMINATION OF SIMILAR Giardia duodenalis ASSEMBLAGE IN CHILDREN AND PET DOGS IN SOUTHERN BRAZIL, AS DETERMINED BY PCR-RFLP. Rev Inst Med Trop Sao Paulo 2016; 58:66. [PMID: 27680171 PMCID: PMC5048637 DOI: 10.1590/s1678-9946201658066] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2015] [Accepted: 05/10/2016] [Indexed: 11/21/2022] Open
Abstract
Giardia duodenalis is an enteric parasite that has distinct genetic
groups. Human infections are mainly caused by assemblages A and B, although sporadic
infections by assemblages C and D have also been reported. Animals can be infected by
a wide range of assemblages (A to H). The aim of this study is to identify the
assemblages and sub-assemblages of G. duodenalis with zoonotic
features in fecal samples of school-aged children, and in dogs that coexist in the
same households in Lages, Santa Catarina, Brazil. Fecal samples of
91 children and 108 dogs were obtained and G. duodenalis cysts were
detected in samples from 11 (12.08%) children and 10 (9.25%) dogs. DNA extracted from
the 21 positive samples was analyzed by PCR-RFLP, using the gdh
gene. Results showed the presence of sub-assemblages AI (2/11), AII (4/11), BIII
(2/11), and BIV(3/11) among children and AI (5/10) and BIV(3/10) in dogs, with
zoonotic characteristics, and the carnivore specific assemblage C (2/10). G.
duodenalis was found to infect both children and dogs living in the same
household, with the same sub-assemblage (BIV) indicating that pet dogs are a
potential risk of transmission of G. duodenalis to humans.
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Affiliation(s)
- Rosiléia Marinho de Quadros
- Universidade do Planalto Catarinense (UNIPLAC), Faculdade de Ciências Biológicas e Biomedicina. Lages, SC, Brazil
| | - Paulo Henrique Exterchoter Weiss
- Universidade do Estado de Santa Catarina (CAV/UDESC), Laboratório de Bioquímica de Hemoparasitas e Vetores do Centro de Ciências. Lages, SC, Brazil
| | - Sandra Marcia Tietz Marques
- Universidade Federal do Rio Grande do Sul (UFRGS), Departamento de Patologia Clínica Veterinária da Faculdade de Veterinária. Porto Alegre, RS, Brazil
| | - Luiz Claudio Miletti
- Universidade do Estado de Santa Catarina (CAV/UDESC), Laboratório de Bioquímica de Hemoparasitas e Vetores do Centro de Ciências. Lages, SC, Brazil
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Pijnacker R, Mughini-Gras L, Heusinkveld M, Roelfsema J, van Pelt W, Kortbeek T. Different risk factors for infection with Giardia lamblia assemblages A and B in children attending day-care centres. Eur J Clin Microbiol Infect Dis 2016; 35:2005-2013. [DOI: 10.1007/s10096-016-2753-2] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/07/2016] [Accepted: 08/08/2016] [Indexed: 11/30/2022]
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Epidemiology of Cryptosporidiosis and Giardiasis: What Pediatricians Need to Know. CURRENT TROPICAL MEDICINE REPORTS 2016. [DOI: 10.1007/s40475-016-0081-3] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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Emery SJ, Lacey E, Haynes PA. Quantitative proteomics in Giardia duodenalis —Achievements and challenges. Mol Biochem Parasitol 2016; 208:96-112. [DOI: 10.1016/j.molbiopara.2016.07.002] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2016] [Revised: 07/13/2016] [Accepted: 07/16/2016] [Indexed: 12/31/2022]
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Ismail MAM, El-Akkad DMH, Rizk EMA, El-Askary HM, El-Badry AA. Molecular seasonality of Giardia lamblia in a cohort of Egyptian children: a circannual pattern. Parasitol Res 2016; 115:4221-4227. [PMID: 27449642 DOI: 10.1007/s00436-016-5199-7] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/03/2016] [Accepted: 07/13/2016] [Indexed: 11/25/2022]
Abstract
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.
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Affiliation(s)
- Mousa A M Ismail
- Medical Parasitology Department, Kasr Al-Ainy Faculty of Medicine, Cairo University, El-Manial, Cairo, Egypt
| | - Dina M H El-Akkad
- Medical Parasitology Department, Kasr Al-Ainy Faculty of Medicine, Cairo University, El-Manial, Cairo, Egypt
| | - Enas M A Rizk
- Medical Parasitology Department, Kasr Al-Ainy Faculty of Medicine, Cairo University, El-Manial, Cairo, Egypt
| | - Hala M El-Askary
- Medical Parasitology Department, Faculty of Medicine, Beni-Suef University, Beni Suef, Egypt
| | - Ayman A El-Badry
- Medical Parasitology Department, Kasr Al-Ainy Faculty of Medicine, Cairo University, El-Manial, Cairo, Egypt.
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Erlandsen SL, Jarroll E, Wallis P, van Keulen H. Development of Species-specific rDNA Probes for Giardia by Multiple Fluorescent In Situ Hybridization Combined with Immunocytochemical Identification of Cyst Wall Antigens. J Histochem Cytochem 2016; 53:917-27. [PMID: 15879572 DOI: 10.1369/jhc.5c6656.2005] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17–22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin.
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Affiliation(s)
- Stanley L Erlandsen
- Department of Genetics, Cell Biology, and Development, 6-160 Jackson Hall, University of Minnesota Medical School, Minneapolis, MN 55455, USA.
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Wegayehu T, Karim MR, Li J, Adamu H, Erko B, Zhang L, Tilahun G. Multilocus genotyping of Giardia duodenalis isolates from children in Oromia Special Zone, central Ethiopia. BMC Microbiol 2016; 16:89. [PMID: 27209324 PMCID: PMC4875739 DOI: 10.1186/s12866-016-0706-7] [Citation(s) in RCA: 25] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2016] [Accepted: 05/12/2016] [Indexed: 01/05/2023] Open
Abstract
BACKGROUND Giardia duodenalis is the etiologic agent of giardiasis in humans and other mammals worldwide. The burden of disease is high among children in developing countries where sanitation is inadequate. However, the epidemiology and genetic diversity of this parasite is poorly understood in Ethiopia. This study aimed to determine the prevalence and genetic diversity of G. duodenalis in asymptomatic children in Oromia Special Zone, central Ethiopia. RESULTS A total of 286 fresh fecal specimens were collected from children and screened using microscopy and PCR. The prevalence of Giardia duodenalis was 10.8 % (31/286) and 16.8 % (48/286) as detected by microscopy and nested PCR, respectively. The infection rate by the study area, sex and age group difference was not significant (P > 0.05). Genotyping results showed that 22.9 % (11/48) of the isolates belonged to assemblage A while 77.1 % (37/48) belonged to assemblage B. Although double peaks were observed at the chromatogram level, no mixed assemblage or sub-assemblage infections were demonstrated. Isolates of assemblage A mostly belonged to the sub-assemblage AII and showed similarity with previously described isolates. However, there was great genetic variability within assemblage B that showed heterogeneous nucleotide positions. Fifteen of them were new genotypes: 5 at the triose phosphate isomerase (tpi), 2 at the β-giardin (bg), and 8 at the glutamate dehydrogenase (gdh) genes. CONCLUSIONS Giardia duodenalis mainly assemblage B infection was predominant among the asymptomatic children in the study area. The high polymorphism found in isolates of assemblage B warrants a more defining tool to discriminate assemblage B at the sub-assemblage level. The findings of the present study indicate that there is a need to carry out national screening programs aiming to detect asymptomatic infections to minimize the reservoir of the disease.
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Affiliation(s)
- Teklu Wegayehu
- Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia. .,College of Natural Sciences, Arba Minch University, Arba Minch, Ethiopia.
| | - Md Robiul Karim
- Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.,College of Animal Sciences and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, China
| | - Junqiang Li
- College of Animal Sciences and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, China
| | - Haileeyesus Adamu
- Institute of Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia
| | - Berhanu Erko
- Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia
| | - Longxian Zhang
- College of Animal Sciences and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, China
| | - Getachew Tilahun
- Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia
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42
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Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections. Clin Microbiol Rev 2016; 27:371-418. [PMID: 24696439 DOI: 10.1128/cmr.00122-13] [Citation(s) in RCA: 146] [Impact Index Per Article: 16.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
Over the past few decades, nucleic acid-based methods have been developed for the diagnosis of intestinal parasitic infections. Advantages of nucleic acid-based methods are numerous; typically, these include increased sensitivity and specificity and simpler standardization of diagnostic procedures. DNA samples can also be stored and used for genetic characterization and molecular typing, providing a valuable tool for surveys and surveillance studies. A variety of technologies have been applied, and some specific and general pitfalls and limitations have been identified. This review provides an overview of the multitude of methods that have been reported for the detection of intestinal parasites and offers some guidance in applying these methods in the clinical laboratory and in epidemiological studies.
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Hawash Y, Ghonaim MM, Al-Shehri SS. An Improved PCR-RFLP Assay for Detection and Genotyping of Asymptomatic Giardia lamblia Infection in a Resource-Poor Setting. THE KOREAN JOURNAL OF PARASITOLOGY 2016; 54:1-8. [PMID: 26951972 PMCID: PMC4792328 DOI: 10.3347/kjp.2016.54.1.1] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/21/2015] [Revised: 10/18/2015] [Accepted: 11/04/2015] [Indexed: 11/23/2022]
Abstract
Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, RIDA® Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.
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Affiliation(s)
- Yoursry Hawash
- Department of Medical Laboratory Science, College of Applied Medical Sciences, Taif University, Taif 21944, Saudi Arabia ; Department of Medical Parasitology, National Liver Institute, Menoufia University, Shebin El-Koom, Menoufia 23513, Egypt
| | - M M Ghonaim
- Department of Medical Laboratory Science, College of Applied Medical Sciences, Taif University, Taif 21944, Saudi Arabia ; Medical Microbiology and Immunology Department, Faculty of Medicine, Menoufia University, Shebin El-Koom, Menoufia 23513, Egypt
| | - S S Al-Shehri
- Department of Medical Laboratory Science, College of Applied Medical Sciences, Taif University, Taif 21944, Saudi Arabia
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El Basha NR, Zaki MM, Hassanin OM, Rehan MK, Omran D. GiardiaAssemblagesAandBin Diarrheic Patients: A Comparative Study in Egyptian Children and Adults. J Parasitol 2016; 102:69-74. [DOI: 10.1645/14-676] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023] Open
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Štrkolcová G, Goldová M, Maďar M, Čechová L, Halánová M, Mojžišová J. Giardia duodenalis and Giardia enterica in children: first evidence of assemblages A and B in Eastern Slovakia. Parasitol Res 2016; 115:1939-44. [PMID: 26818944 DOI: 10.1007/s00436-016-4935-3] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2015] [Accepted: 01/19/2016] [Indexed: 11/26/2022]
Abstract
For an overview on the occurrence of Giardia assemblages in children in Eastern Slovakia, we examined 259 faecal samples of children from the segregated settlement in Medzev, 30 samples of children from the orphanage in Medzev and 40 samples of children with autism from the Special Elementary School in Košice. Thirty-eight samples (14.67 %) from the segregated settlement, 19 samples (63.33 %) from the orphanage and two samples (5.0 %) from the Special Elementary School were positive for Giardia by flotation. The initial microscopic diagnostics were completed by the genotyping of the triosephosphate isomerase-gene loci (tpi genes) which revealed the existence of two Giardia assemblages in Slovak population, namely Giardia duodenalis (assemblage A) and Giardia enterica (assemblage B). These results represent the first evidence of A and B assemblages in children in Slovakia. Epidemiological significance and the impact on the public health of Giardia infection are highlighted.
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Affiliation(s)
- G Štrkolcová
- Department of Epizootiology and Parasitology, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, Košice, 041 81, Slovak Republic
| | - M Goldová
- Department of Epizootiology and Parasitology, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, Košice, 041 81, Slovak Republic.
| | - M Maďar
- Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, Košice, 041 81, Slovak Republic
| | - L Čechová
- Department of Epidemiology, Faculty of Medicine, Pavol Jozef Šafárik University, Šrobárova 2, Košice, 041 80, Slovak Republic
| | - M Halánová
- Department of Epidemiology, Faculty of Medicine, Pavol Jozef Šafárik University, Šrobárova 2, Košice, 041 80, Slovak Republic
| | - J Mojžišová
- Department of Epizootiology and Parasitology, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, Košice, 041 81, Slovak Republic
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Predominance of Giardia lamblia assemblage A among iron deficiency anaemic pre-school Egyptian children. Parasitol Res 2016; 115:1537-45. [PMID: 26758448 DOI: 10.1007/s00436-015-4888-y] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/22/2015] [Accepted: 12/11/2015] [Indexed: 10/22/2022]
Abstract
Intestinal parasites and nutritional deficiency can coexist and influence each other. This study aimed to clarify the association between Giardia genotypes and presence of iron deficiency anaemia (IDA) among pre-school Egyptian children. Two groups (IDA and non-anaemic) of giardiasis children (44/group) were selected according to their recovery response after treatment of giardiasis. Each group included 24 and 20 gastrointestinal symptomatic and asymptomatic, respectively. Giardia human genotypes were performed by intergenic spacer (IGS) gene based polymerase chain reaction (PCR) with high-resolution melting curve (HRM). PCR/HRM proved that Tms of assemblage A and B ranged from 79.31 ± 0.29 to 84.77 ± 0.31. In IDA patients, assemblages A and B were found among 40/44 (90.9 %) and 4/44 (9.1 %), respectively, while in non-anaemic patients, assemblages A and B were found in 10/44 (22.7 %) and 32/44 (72.7 %), respectively, beside two (4.6 %) cases had mixed infection. The difference was statistically significant. No significant relation was found between symptomatic or asymptomatic assemblages and IDA as assemblage A was found in 21/24 (87.5 %) and 19/20 (95 %) of symptomatic and asymptomatic, respectively, while 3/24 (12.5 %) and 1/20 (5 %) of assemblage B were symptomatic was asymptomatic, respectively. A significant relation was found between assemblage A subtypes distribution among IDA patients as AI and AII were detected on 23 (52.3 %) and 16 (36.4 %) of patients, respectively, while one case (2.3 %) had mixed infection. In conclusion, assemblage A is predominant among IDA giardiasis children suggesting its role in enhancing the occurrence of IDA while B has a protective role.
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Nishida Y, Morimoto N, Korenaga M, Komatsu Y, Takeuchi H, Matsumura Y, Sugiura T. Genotyping <i>Giardia intestinalis</i> by Using DNA Extracted from Long-Term Preserved Human Specimens Stained with Chlorazol Black E. Jpn J Infect Dis 2016; 69:244-7. [DOI: 10.7883/yoken.jjid.2014.543] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Affiliation(s)
- Yoshie Nishida
- Department of Clinical Laboratory, Kochi Medical School Hospital
- Department of Clinical Laboratory Medicine, Kochi Medical School, Kochi University
| | | | | | - Yutaka Komatsu
- Department of Clinical Laboratory, Kochi Medical School Hospital
| | - Hiroaki Takeuchi
- Department of Clinical Laboratory Medicine, Kochi Medical School, Kochi University
| | - Yoshihisa Matsumura
- Department of Clinical Laboratory Medicine, Kochi Medical School, Kochi University
| | - Tetsuro Sugiura
- Department of Clinical Laboratory, Kochi Medical School Hospital
- Department of Clinical Laboratory Medicine, Kochi Medical School, Kochi University
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de Lucio A, Martínez-Ruiz R, Merino FJ, Bailo B, Aguilera M, Fuentes I, Carmena D. Molecular Genotyping of Giardia duodenalis Isolates from Symptomatic Individuals Attending Two Major Public Hospitals in Madrid, Spain. PLoS One 2015; 10:e0143981. [PMID: 26641082 PMCID: PMC4671680 DOI: 10.1371/journal.pone.0143981] [Citation(s) in RCA: 42] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/02/2015] [Accepted: 11/11/2015] [Indexed: 11/18/2022] Open
Abstract
BACKGROUND 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. METHODS 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. PRINCIPAL FINDINGS 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. BIV was the most prevalent genetic variant of G. duodenalis found in individuals with symptomatic giardiasis in the population under study. CONCLUSIONS Human giardiasis is an ongoing public health problem in Spain affecting primarily young children under four years of age but also individuals of all age groups. Our typing and sub-typing results demonstrate that assemblage B is the most prevalent G. duodenalis assemblage circulating in patients with clinical giardiasis in Central Spain. Our analyses also revealed a large genetic variability in assemblage B (but not assemblage A) isolates of the parasite, corroborating the information obtained in similar studies in other geographical regions. We believe that molecular data presented here provide epidemiological evidence at the population level in support of the existence of genetic exchange within assemblages of G. duodenalis.
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Affiliation(s)
- Aida de Lucio
- Parasitology Service, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
| | - Rocío Martínez-Ruiz
- Microbiology and Clinical Parasitology Service, University Hospital Puerta de Hierro Majadahonda, Majadahonda, Madrid, Spain
| | - Francisco J. Merino
- Microbiology Service, University Hospital Severo Ochoa, Leganés, Madrid, Spain
| | - Begoña Bailo
- Parasitology Service, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
| | - María Aguilera
- Parasitology Service, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
| | - Isabel Fuentes
- Parasitology Service, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
| | - David Carmena
- Parasitology Service, National Centre for Microbiology, Carlos III Health Institute, Majadahonda, Madrid, Spain
- * E-mail:
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Molecular Characterization and Risk Factors of Giardia duodenalis among School Children from La Habana, Cuba. J Parasitol Res 2015; 2015:378643. [PMID: 26693345 PMCID: PMC4677018 DOI: 10.1155/2015/378643] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2015] [Accepted: 10/29/2015] [Indexed: 11/25/2022] Open
Abstract
Giardia duodenalis is considered the most common protozoan infecting humans worldwide. Molecular characterization of G. duodenalis isolates has revealed the existence of eight groups (assemblages A to H) which differ in their host distribution. A cross-sectional study was conducted in 639 children from La Habana between January and December 2013. Two assemblage-specific PCRs were carried out for the molecular characterization. The overall prevalence of Giardia infection was 11.9%. DNA from 63 of 76 (82.9%) samples was successfully amplified by PCR-tpi, while 58 from 76 (76.3%) were detected by PCRE1-HF. Similar results by both PCRs were obtained in 54 from 76 samples (71%). According to these analyses, assemblage B and mixed assemblages A + B account for most of the Giardia infections in the cohort of children tested. Our current study identified assemblage B as predominant genotype in children infected with Giardia. Univariate analysis indicated that omission of washing hands before eating and keeping dogs at home were significant risk factors for a Giardia infection. In the future, novel molecular tools for a better discrimination of assemblages at the subassemblages level are needed to verify possible correlations between Giardia genotypes and symptomatology of giardiasis.
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Ankarklev J, Franzén O, Peirasmaki D, Jerlström-Hultqvist J, Lebbad M, Andersson J, Andersson B, Svärd SG. Comparative genomic analyses of freshly isolated Giardia intestinalis assemblage A isolates. BMC Genomics 2015; 16:697. [PMID: 26370391 PMCID: PMC4570179 DOI: 10.1186/s12864-015-1893-6] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2015] [Accepted: 09/01/2015] [Indexed: 12/31/2022] Open
Abstract
Background The diarrhea-causing protozoan Giardia intestinalis makes up a species complex of eight different assemblages (A-H), where assemblage A and B infect humans. Comparative whole-genome analyses of three of these assemblages have shown that there is significant divergence at the inter-assemblage level, however little is currently known regarding variation at the intra-assemblage level. We have performed whole genome sequencing of two sub-assemblage AII isolates, recently axenized from symptomatic human patients, to study the biological and genetic diversity within assemblage A isolates. Results Several biological differences between the new and earlier characterized assemblage A isolates were identified, including a difference in growth medium preference. The two AII isolates were of different sub-assemblage types (AII-1 [AS175] and AII-2 [AS98]) and showed size differences in the smallest chromosomes. The amount of genetic diversity was characterized in relation to the genome of the Giardia reference isolate WB, an assemblage AI isolate. Our analyses indicate that the divergence between AI and AII is approximately 1 %, represented by ~100,000 single nucleotide polymorphisms (SNP) distributed over the chromosomes with enrichment in variable genomic regions containing surface antigens. The level of allelic sequence heterozygosity (ASH) in the two AII isolates was found to be 0.25–0.35 %, which is 25–30 fold higher than in the WB isolate and 10 fold higher than the assemblage AII isolate DH (0.037 %). 35 protein-encoding genes, not found in the WB genome, were identified in the two AII genomes. The large gene families of variant-specific surface proteins (VSPs) and high cysteine membrane proteins (HCMPs) showed isolate-specific divergences of the gene repertoires. Certain genes, often in small gene families with 2 to 8 members, localize to the variable regions of the genomes and show high sequence diversity between the assemblage A isolates. One of the families, Bactericidal/Permeability Increasing-like protein (BPIL), with eight members was characterized further and the proteins were shown to localize to the ER in trophozoites. Conclusions Giardia genomes are modular with highly conserved core regions mixed up by variable regions containing high levels of ASH, SNPs and variable surface antigens. There are significant genomic variations in assemblage A isolates, in terms of chromosome size, gene content, surface protein repertoire and gene polymorphisms and these differences mainly localize to the variable regions of the genomes. The large genetic differences within one assemblage of G. intestinalis strengthen the argument that the assemblages represent different Giardia species. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-1893-6) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Johan Ankarklev
- Department of Cell and Molecular Biology, Science for Life Laboratory, BMC, Uppsala University, Box 596, SE-751 24, Uppsala, Sweden.
| | - Oscar Franzén
- Department of Cell and Molecular Biology, Karolinska Institutet, Box 285, SE-171 77, Stockholm, Sweden. .,Science for Life Laboratory, KISP, Tomtebodavägen 23A, 171 65, Solna, Sweden.
| | - Dimitra Peirasmaki
- Department of Cell and Molecular Biology, Science for Life Laboratory, BMC, Uppsala University, Box 596, SE-751 24, Uppsala, Sweden.
| | - Jon Jerlström-Hultqvist
- Department of Cell and Molecular Biology, Science for Life Laboratory, BMC, Uppsala University, Box 596, SE-751 24, Uppsala, Sweden.
| | - Marianne Lebbad
- Department of Microbiology, Public Health Agency of Sweden, SE-171 82, Solna, Sweden.
| | - Jan Andersson
- Department of Cell and Molecular Biology, Science for Life Laboratory, BMC, Uppsala University, Box 596, SE-751 24, Uppsala, Sweden.
| | - Björn Andersson
- Department of Cell and Molecular Biology, Karolinska Institutet, Box 285, SE-171 77, Stockholm, Sweden. .,Science for Life Laboratory, KISP, Tomtebodavägen 23A, 171 65, Solna, Sweden.
| | - Staffan G Svärd
- Department of Cell and Molecular Biology, Science for Life Laboratory, BMC, Uppsala University, Box 596, SE-751 24, Uppsala, Sweden.
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