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      Mosquito Feeding Assays to Determine the Infectiousness of Naturally Infected Plasmodium falciparum Gametocyte Carriers

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          Abstract

          Introduction

          In the era of malaria elimination and eradication, drug-based and vaccine-based approaches to reduce malaria transmission are receiving greater attention. Such interventions require assays that reliably measure the transmission of Plasmodium from humans to Anopheles mosquitoes.

          Methods

          We compared two commonly used mosquito feeding assay procedures: direct skin feeding assays and membrane feeding assays. Three conditions under which membrane feeding assays are performed were examined: assays with i) whole blood, ii) blood pellets resuspended with autologous plasma of the gametocyte carrier, and iii) blood pellets resuspended with heterologous control serum.

          Results

          930 transmission experiments from Cameroon, The Gambia, Mali and Senegal were included in the analyses. Direct skin feeding assays resulted in higher mosquito infection rates compared to membrane feeding assays (odds ratio 2.39, 95% confidence interval 1.94–2.95) with evident heterogeneity between studies. Mosquito infection rates in membrane feeding assays and direct skin feeding assays were strongly correlated (p<0.0001). Replacing the plasma of the gametocyte donor with malaria naïve control serum resulted in higher mosquito infection rates compared to own plasma (OR 1.92, 95% CI 1.68–2.19) while the infectiousness of gametocytes may be reduced during the replacement procedure (OR 0.60, 95% CI 0.52–0.70).

          Conclusions

          Despite a higher efficiency of direct skin feeding assays, membrane feeding assays appear suitable tools to compare the infectiousness between individuals and to evaluate transmission-reducing interventions. Several aspects of membrane feeding procedures currently lack standardization; this variability makes comparisons between laboratories challenging and should be addressed to facilitate future testing of transmission-reducing interventions.

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          Most cited references55

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          Statistical methods for assessing agreement between two methods of clinical measurement.

          In clinical measurement comparison of a new measurement technique with an established one is often needed to see whether they agree sufficiently for the new to replace the old. Such investigations are often analysed inappropriately, notably by using correlation coefficients. The use of correlation is misleading. An alternative approach, based on graphical techniques and simple calculations, is described, together with the relation between this analysis and the assessment of repeatability.
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            Epidemiology and infectivity of Plasmodium falciparum and Plasmodium vivax gametocytes in relation to malaria control and elimination.

            Malaria remains a major cause of morbidity and mortality in the tropics, with Plasmodium falciparum responsible for the majority of the disease burden and P. vivax being the geographically most widely distributed cause of malaria. Gametocytes are the sexual-stage parasites that infect Anopheles mosquitoes and mediate the onward transmission of the disease. Gametocytes are poorly studied despite this crucial role, but with a recent resurgence of interest in malaria elimination, the study of gametocytes is in vogue. This review highlights the current state of knowledge with regard to the development and longevity of P. falciparum and P. vivax gametocytes in the human host and the factors influencing their distribution within endemic populations. The evidence for immune responses, antimalarial drugs, and drug resistance influencing infectiousness to mosquitoes is reviewed. We discuss how the application of molecular techniques has led to the identification of submicroscopic gametocyte carriage and to a reassessment of the human infectious reservoir. These components are drawn together to show how control measures that aim to reduce malaria transmission, such as mass drug administration and a transmission-blocking vaccine, might better be deployed.
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              Submicroscopic Plasmodium falciparum gametocyte densities frequently result in mosquito infection.

              Submicroscopic Plasmodium falciparum gametocytemia (<5,000 gametocytes/mL) is common and may result in mosquito infection. We assessed the relation between gametocyte density and mosquito infection under experimental and field conditions using real-time quantitative nucleic acid sequence-based amplification (QT-NASBA) for gametocyte quantification. Serial dilutions of NF54 P. falciparum gametocytes showed a positive association between gametocyte density and the proportion of infected mosquitoes (beta=6.1; 95% confidence interval [CI], 2.7-9.6; P=0.001). Successful infection became unlikely below an estimated density of 250-300 gametocytes/mL. In the field, blood samples of 100 naturally infected children showed a positive association between gametocyte density and oocyst counts in mosquitoes (beta=0.38; 95% CI, 0.14-0.61; P=0.002). The relative contribution to malaria transmission was similar for carriers with submicroscopic and microscopic gametocytemia. Our results show that transmission occurs efficiently at submicroscopic gametocyte densities and that carriers harboring submicroscopic gametocytemia constitute a considerable proportion of the human infectious reservoir.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                22 August 2012
                : 7
                : 8
                : e42821
                Affiliations
                [1 ]Department of Immunity and Infection, London School of Hygiene & Tropical Medicine, London, United Kingdom
                [2 ]Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
                [3 ]W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health and Malaria Research Institute, Baltimore, Maryland, United States of America
                [4 ]Laboratoire de Recherche sur le Paludisme, Organisation de Coordination pour la lutte contre les Endémies en Afrique Centrale (OCEAC), Institut de Recherche pour le Développement, Yaoundé, Cameroun
                [5 ]Maladies Infectieuses et Vecteurs : Écologie, Génétique, Évolution et Contrôle (MIVEGEC), Institut de Recherche pour le Développement, Montpellier, France
                [6 ]Centre de Recherche et de Veille sur les Maladies Emergentes dans l'Océan Indien, La Réunion, France
                [7 ]USC INRA Bartonella-tiques, Agence National de Sécurité Sanitaire, Maisons Alfort, France
                [8 ]Malaria Research and Training Center, University of Sciences, Techniques and Technology, Bamako, Mali
                [9 ]Malaria Research Unit of the Laboratory of Applied Biology and Ecology, University of Dschang, Dschang, Cameroon
                [10 ]Department of Medical Microbiology, Microbiology Laboratory Twente, Enschede, The Netherlands
                [11 ]School of Public Health-Tropical Medicine and Rehabilitation Sciences, James Cook University, Cairns, Queensland, Australia
                [12 ]PATH Malaria Vaccine Initiative, Washington, D.C., United States of America
                [13 ]Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, Rockville, Maryland, United States of America
                [14 ]Infectious Disease Epidemiology-MRC Centre for Outbreak Analysis and Modelling, Imperial College London, London, United Kingdom
                Tulane University, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: TB IM LCG PHA-A SB MD MC TT BM GT CD CS VR OD YT PG WR RS YW. Performed the experiments: TB IM LCG PHA-A SB MD MC TT BM GT CD CS VR OD YT PG YW. Analyzed the data: TB RRD IM YW TSC TW. Wrote the paper: TB RRD IM AB EL MM YW TSC.

                Article
                PONE-D-12-12445
                10.1371/journal.pone.0042821
                3425579
                22936993
                fb26845c-c14c-4acc-af39-2c4c50d5af33
                Copyright @ 2012

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

                History
                : 2 May 2012
                : 11 July 2012
                Page count
                Pages: 13
                Funding
                The work of TB is supported by grants from the Bill & Melinda Gates Foundation and the European FP7 framework (REDMAL project, #242079) and the PATH Malaria Vaccine Initiative (MVI). RRD and TW were supported by MVI, the Bloomberg Family Foundation and the Johns Hopkins Malaria Research Institute. IM, LCG, PHAA, SB and VR were partly supported by the Institut de Recherche pour le Développement. The MRTC group in Mali is supported by extramural (TMRC) and intramural grants from NIAID/NIH. YW is supported by Division of Intramural Research, National Institute of Allergy and Infectious Diseases. TSC is supported by a Junior Research Fellowship from Imperial College London and an EC FP7 Collaborative project TransMalariaBloc (HEALTH-F3-2008-223736). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Microbiology
                Parasitology
                Population Biology
                Epidemiology
                Infectious Disease Epidemiology
                Medicine
                Epidemiology
                Infectious Disease Epidemiology
                Infectious Diseases
                Parasitic Diseases
                Malaria
                Plasmodium Falciparum

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                Uncategorized

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