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      Molecular Epidemiology and Assemblage Typing of Giardia duodenalis in School-Age Children Situated along the Southern Shoreline of Lake Malawi, Malawi

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          ABSTRACT.

          Almost all human giardiasis infections are caused by Giardia duodenalis assemblages A and B. Differentiation between human infections with these assemblages, as well as between single-assemblage (A or B) and mixed-assemblage (A and B) infections, is therefore needed to better understand the pathological impact of infection with either, or both, assemblages. We assessed the prevalence of G. duodenalis assemblages A and B using 305 fecal samples provided by school-age children situated along the southern shoreline of Lake Malawi. Concurrently, intestinal pathology data were also collected to test for association(s) between assemblage infection status and intestinal health. Prevalence of G. duodenalis infection was 39.3% by real-time polymerase chain reaction. Of all identified infections, 32% were single G. duodenalis assemblage A and 32% were single G. duodenalis assemblage B, whereas 33% were mixed-assemblage infections. Fifteen unique G. duodenalis assemblage A and 13 unique G. duodenalis assemblage B β-giardin haplotypes were identified. There was a positive association between single infection with G. duodenalis assemblage B and both self-reporting of abdominal pain (odds ratio [OR]: 3.05, P = 0.004) and self-reporting of diarrhea (OR: 3.1, P = 0.003). No association between single infection with assemblage A and any form of intestinal pathology was found. Additionally, there was a positive association between mixed-assemblage infections and self-reporting of abdominal pain (OR: 3.1, P = 0.002). Our study highlights the importance G. duodenalis assemblage typing and reaffirms the need for improved access to water, sanitation and hygiene infrastructure in rural areas of low- and middle-income countries.

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          popart: full-feature software for haplotype network construction

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            A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping and DNA sequence data. III. Cladogram estimation.

            We previously developed a cladistic approach to identify subsets of haplotypes defined by restriction endonuclease mapping or DNA sequencing that are associated with significant phenotypic deviations. Our approach was limited to segments of DNA in which little recombination occurs. In such cases, a cladogram can be constructed from the restriction site or sequence data that represents the evolutionary steps that interrelate the observed haplotypes. The cladogram is used to define a nested statistical design to identify mutational steps associated with significant phenotypic deviations. The central assumption behind this strategy is that any undetected mutation causing a phenotypic effect is embedded within the same evolutionary history that is represented by the cladogram. The power of this approach depends upon the confidence one has in the particular cladogram used to draw inferences. In this paper, we present a strategy for estimating the set of cladograms that are consistent with a particular sample of either restriction site or nucleotide sequence data and that includes the possibility of recombination. We first evaluate the limits of parsimony in constructing cladograms. Once these limits have been determined, we construct the set of parsimonious and nonparsimonious cladograms that is consistent with these limits. Our estimation procedure also identifies haplotypes that are candidates for being products of recombination. If recombination is extensive, our algorithm subdivides the DNA region into two or more subsections, each having little or no internal recombination. We apply this estimation procedure to three data sets to illustrate varying degrees of cladogram ambiguity and recombination.
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              Zoonotic potential of Giardia.

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

                Journal
                Am J Trop Med Hyg
                Am J Trop Med Hyg
                tpmd
                tropmed
                The American Journal of Tropical Medicine and Hygiene
                The American Society of Tropical Medicine and Hygiene
                0002-9637
                1476-1645
                7 August 2023
                September 2023
                7 August 2023
                : 109
                : 3
                : 626-639
                Affiliations
                [ 1 ]Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom;
                [ 2 ]Institute of Medical Microbiology, Semmelweis University, Budapest, Hungary;
                [ 3 ]Laboratory Department, Mangochi District Hospital, Mangochi, Malawi;
                [ 4 ]Malawi Liverpool Wellcome Trust Programme of Clinical Tropical Research, Queen Elizabeth Central Hospital, Blantyre, Malawi;
                [ 5 ]Department of Pathology, School of Medicine and Oral Health, Kamuzu University of Health Sciences, Blantyre, Malawi;
                [ 6 ]MASM Medi Clinics Limited, Medical Aid Society of Malawi, Area 12 Medi Clinic and Head Office, Lilongwe, Malawi
                Author notes
                [* ]Address correspondence to John Archer, Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, United Kingdom. E-mail: john.archer@ 123456lstmed.ac.uk

                Financial support: J. A. is supported by a Medical Research Council Doctoral Training Partnership (MRC-DTP) studentship.

                Authors’ addresses: John Archer, Lucas J. Cunningham, Sam Jones, Ffion Doull, James E. LaCourse, and J. Russell Stothard, Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom, E-mails: john.archer@ 123456lstmed.ac.uk , lucas.cunningham@ 123456lstmed.ac.uk , sam.jones@ 123456lstmed.ac.uk , ffiondoull@ 123456icloud.com , james.lacourse@ 123456lstmed.ac.uk , and russell.stothard@ 123456lstmed.ac.uk . Alexandra Juhàsz, Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom, and Institute of Medical Microbiology, Semmelweis University, Budapest, Hungary, E-mail: alexandra.juhasz@ 123456lstmed.ac.uk . Bright Mainga, Laboratory Department, Mangochi District Hospital, Mangochi, Malawi, and Malawi Liverpool Wellcome Trust Programme of Clinical Tropical Research, Queen Elizabeth Central Hospital, Blantyre, Malawi, E-mail: brichicco@ 123456gmail.com . Peter Makaula, Malawi Liverpool Wellcome Trust Programme of Clinical Tropical Research, Queen Elizabeth Central Hospital, Blantyre, Malawi, E-mail: pmakaula@ 123456mlw.mw . Sekeleghe A. Kayuni, Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom, Malawi Liverpool Wellcome Trust Programme of Clinical Tropical Research, Queen Elizabeth Central Hospital, Blantyre, Malawi, Department of Pathology, School of Medicine and Oral Health, Kamuzu University of Health Sciences, Blantyre, Malawi, and MASM Medi Clinics Limited, Medical Aid Society of Malawi, Area 12 Medi Clinic and Head Office, Lilongwe, Malawi, E-mail: skayuni@ 123456mlw.mw . Janelisa Musaya, Malawi Liverpool Wellcome Trust Programme of Clinical Tropical Research, Queen Elizabeth Central Hospital, Blantyre, Malawi, and Department of Pathology, School of Medicine and Oral Health, Kamuzu University of Health Sciences, Blantyre, Malawi, E-mail: jmusaya@ 123456mlw.mw .

                Article
                tpmd230156
                10.4269/ajtmh.23-0156
                10484258
                37549892
                715921f9-483a-41ec-8f04-769c6fc95e50
                © The author(s)

                This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 13 March 2023
                : 16 June 2023
                Page count
                Pages: 14
                Categories
                Research Article

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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