Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border
Abstract Background Polyclonal blood-stage infections of Plasmodium vivax are frequent even in low transmission settings, allowing meiotic recombination between heterologous parasites. Empirical data on meiotic products are however lacking. This study examined microsatellites in oocysts derived by m...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2017-09-01
|
| Series: | Malaria Journal |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12936-017-2002-x |
| _version_ | 1819159040324796416 |
|---|---|
| author | Ingfar Soontarawirat Chiara Andolina Richard Paul Nicholas P. J. Day Francois Nosten Charles J. Woodrow Mallika Imwong |
| author_facet | Ingfar Soontarawirat Chiara Andolina Richard Paul Nicholas P. J. Day Francois Nosten Charles J. Woodrow Mallika Imwong |
| author_sort | Ingfar Soontarawirat |
| collection | DOAJ |
| description | Abstract Background Polyclonal blood-stage infections of Plasmodium vivax are frequent even in low transmission settings, allowing meiotic recombination between heterologous parasites. Empirical data on meiotic products are however lacking. This study examined microsatellites in oocysts derived by membrane feeding of mosquitoes from blood-stage P. vivax infections at the Thai–Myanmar border. Methods Blood samples from patients presenting with vivax malaria were fed to Anopheles cracens by membrane feeding and individual oocysts from midguts were obtained by dissection after 7 days. DNA was extracted from oocysts and parental blood samples and tested by microsatellite analysis. Results A focused study of eight microsatellite markers was undertaken for nine blood stage infections from 2013, for which derived oocysts were studied in six cases. One or more alleles were successfully amplified for 131 oocysts, revealing high levels of allelic diversity in both blood and oocyst stages. Based on standard criteria for defining minor alleles, there was evidence of clear deviation from random mating (inbreeding) with relatively few heterozygous oocysts compared to variance across the entire oocyst population (FIT = 0.89). The main explanation appeared to be natural compartmentalisation at mosquito (FSC = 0.27) and human stages (FCT = 0.68). One single human case produced a total of 431 successfully amplified loci (across 70 oocysts) that were homozygous and identical to parental alleles at all markers, indicating clonal infection and transmission. Heterozygous oocyst alleles were found at 15/176 (8.5%) successfully amplified loci in the other five cases. There was apparently reduced oocyst heterozygosity in individual oocysts compared to diversity within individual mosquitoes (FIS = 0.55), but this may simply reflect the difficulty of detecting minor alleles in oocysts, given the high rate of amplification failure. Inclusion of minor allele peaks (irrespective of height) when matching peaks were found in related blood or oocyst samples, added 11 minor alleles for 9 oocysts, increasing the number of heterozygous loci to 26/176 (14.8%; p = 0.096). Conclusion There was an apparently low level of heterozygous oocysts but this can be explained by a combination of factors: relatively low complexity of parental infection, natural compartmentalisation in humans and mosquitoes, and the methodological challenge of detecting minor alleles. |
| first_indexed | 2024-12-22T16:34:14Z |
| format | Article |
| id | doaj.art-3d4982071a724c60b3a9c34b3bec2448 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-22T16:34:14Z |
| publishDate | 2017-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-3d4982071a724c60b3a9c34b3bec24482022-12-21T18:19:59ZengBMCMalaria Journal1475-28752017-09-0116111110.1186/s12936-017-2002-xPlasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar borderIngfar Soontarawirat0Chiara Andolina1Richard Paul2Nicholas P. J. Day3Francois Nosten4Charles J. Woodrow5Mallika Imwong6Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol UniversityShoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol UniversityUnité de Génétique Fonctionnelle Des Maladies Infectieuses, Institut PasteurCentre for Tropical Medicine and Global Health, Nuffield Department of Medicine Research Building, University of OxfordShoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol UniversityCentre for Tropical Medicine and Global Health, Nuffield Department of Medicine Research Building, University of OxfordMahidol Oxford Research Unit, Faculty of Tropical Medicine, Mahidol UniversityAbstract Background Polyclonal blood-stage infections of Plasmodium vivax are frequent even in low transmission settings, allowing meiotic recombination between heterologous parasites. Empirical data on meiotic products are however lacking. This study examined microsatellites in oocysts derived by membrane feeding of mosquitoes from blood-stage P. vivax infections at the Thai–Myanmar border. Methods Blood samples from patients presenting with vivax malaria were fed to Anopheles cracens by membrane feeding and individual oocysts from midguts were obtained by dissection after 7 days. DNA was extracted from oocysts and parental blood samples and tested by microsatellite analysis. Results A focused study of eight microsatellite markers was undertaken for nine blood stage infections from 2013, for which derived oocysts were studied in six cases. One or more alleles were successfully amplified for 131 oocysts, revealing high levels of allelic diversity in both blood and oocyst stages. Based on standard criteria for defining minor alleles, there was evidence of clear deviation from random mating (inbreeding) with relatively few heterozygous oocysts compared to variance across the entire oocyst population (FIT = 0.89). The main explanation appeared to be natural compartmentalisation at mosquito (FSC = 0.27) and human stages (FCT = 0.68). One single human case produced a total of 431 successfully amplified loci (across 70 oocysts) that were homozygous and identical to parental alleles at all markers, indicating clonal infection and transmission. Heterozygous oocyst alleles were found at 15/176 (8.5%) successfully amplified loci in the other five cases. There was apparently reduced oocyst heterozygosity in individual oocysts compared to diversity within individual mosquitoes (FIS = 0.55), but this may simply reflect the difficulty of detecting minor alleles in oocysts, given the high rate of amplification failure. Inclusion of minor allele peaks (irrespective of height) when matching peaks were found in related blood or oocyst samples, added 11 minor alleles for 9 oocysts, increasing the number of heterozygous loci to 26/176 (14.8%; p = 0.096). Conclusion There was an apparently low level of heterozygous oocysts but this can be explained by a combination of factors: relatively low complexity of parental infection, natural compartmentalisation in humans and mosquitoes, and the methodological challenge of detecting minor alleles.http://link.springer.com/article/10.1186/s12936-017-2002-xRelapsePlasmodium vivaxOocystMeiosisGenetic diversity |
| spellingShingle | Ingfar Soontarawirat Chiara Andolina Richard Paul Nicholas P. J. Day Francois Nosten Charles J. Woodrow Mallika Imwong Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border Malaria Journal Relapse Plasmodium vivax Oocyst Meiosis Genetic diversity |
| title | Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border |
| title_full | Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border |
| title_fullStr | Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border |
| title_full_unstemmed | Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border |
| title_short | Plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the Thai–Myanmar border |
| title_sort | plasmodium vivax genetic diversity and heterozygosity in blood samples and resulting oocysts at the thai myanmar border |
| topic | Relapse Plasmodium vivax Oocyst Meiosis Genetic diversity |
| url | http://link.springer.com/article/10.1186/s12936-017-2002-x |
| work_keys_str_mv | AT ingfarsoontarawirat plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT chiaraandolina plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT richardpaul plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT nicholaspjday plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT francoisnosten plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT charlesjwoodrow plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder AT mallikaimwong plasmodiumvivaxgeneticdiversityandheterozygosityinbloodsamplesandresultingoocystsatthethaimyanmarborder |