Plasmodium vivax Genetic Diversity in Panama: Challenges for Malaria Elimination in Mesoamerica

Panama and all nations within the Mesoamerican region have committed to eliminate malaria within this decade. With more than 90% of the malaria cases in this region caused by Plasmodium vivax, an efficient national/regional elimination plan must include a comprehensive study of this parasite’s genetic diversity. Here, we retrospectively analyzed P. vivax genetic diversity in autochthonous and imported field isolates collected in different endemic regions in Panama from 2007 to 2020, using highly polymorphic markers (csp, msp-1, and msp-3α). We did the analysis using molecular techniques that are cost-effective for malaria molecular surveillance within Mesoamerica. Thus, we used molecular analyses that are feasible for malaria molecular surveillance within the region, and that can provide useful information for policy and decision making about malaria elimination. We also evaluated if haplotypes established by combining the genotypes found in these genes were associated with relevant epidemiological variables and showed structure across the transmission foci that have been observed in Panama. Ten different haplotypes were identified, some of them strongly associated with geographical origin, age, and collection year. Phylogenetic analysis of csp (central repeat domain) revealed that both major variant types (vk210 and vk247) were circulating in Panama. Variant vk247 was restricted to the eastern endemic regions, while vk210 was predominant (77.3%) and widespread, displaying higher diversity (14 alleles) and geographically biased alleles. The regional implications of these molecular findings for the control of P. vivax malaria to achieve elimination across Mesoamerica are discussed.

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Evolution of Plasmodium vivax populations in border areas of the Greater Mekong Subregion during malaria elimination

10.21203/rs.2.22640/v1 ◽

2020 ◽

Author(s):

Yuling Li ◽

Yubing Hu ◽

Yan Zhao ◽

Qinghui Wang ◽

Huguette Gaelle Ngassa Mbenda ◽

...

Keyword(s):

Genetic Diversity ◽

Plasmodium Vivax ◽

Malaria Elimination ◽

Scale Up ◽

Transmission Dynamics ◽

Clinical Samples ◽

Effective Population ◽

Border Areas ◽

Greater Mekong Subregion ◽

International Border

Abstract Background: Countries within the Greater Mekong Subregion (GMS) of Southeast Asia have committed to eliminating malaria by 2030. Although malaria situation has greatly improved, Plasmodium vivax remains at international border regions. Therefore, to gain a better understanding of transmission dynamics, knowledge on the evolution of P. vivax populations after the scale-up of control interventions will guide more effective targeted control efforts. Methods: We investigated genetic diversity and population structures in 206 longitudinally collected P. vivax clinical samples in two international border areas at the China-Myanmar border (CMB, n=50 in 2004 and n=52 in 2016) and western Thailand border (n=50 in 2012 and n=54 in 2015). Parasites were genotyped using 10 microsatellite markers. Results: Despite intensified control efforts, genetic diversity in the four populations remained high (HE = 0.66-0.86). The proportions of polyclonal infections showed substantial decreases to 23.7 and 30.7% in the CMB and western Thailand, respectively, with corresponding decreases in the multiplicity of infection. Consistent with the shrinking map of malaria transmission in the GMS over time, there were also increases in multilocus linkage disequilibrium, suggesting of more fragmented and increasingly inbred parasite populations. There were considerable genetic differentiation and subdivision with the four tested populations. Various degrees of clustering were evident between the older parasite samples collected in 2004 at the CMB with the 2016 CMB and 2012 Thailand populations, suggesting some of these parasites had shared ancestry. In contrast, the 2015 Thailand population was genetically distinctive, which may reflect a process of population replacement. The moderately large effective population sizes and proportions of polyclonal infections highlight the necessity of further coordinated and integrated control efforts on both sides of the borders in the pursuit of malaria elimination. Conclusions: With enhanced control efforts on malaria elimination, P. vivax population in the GMS has fragmented into a limited number of clustered foci, but the presence of large P. vivax reservoirs still sustains genetic diversity and transmission. These findings provide new insights into P. vivax transmission dynamics and population structure in this area.

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Evolution of Plasmodium vivax populations in border areas of the Greater Mekong Subregion during malaria elimination

10.21203/rs.2.19786/v1 ◽

2019 ◽

Author(s):

Yuling Li ◽

Yubing Hu ◽

Yan Zhao ◽

Qinghui Wang ◽

Huguette Gaelle Ngassa Mbenda ◽

...

Keyword(s):

Genetic Diversity ◽

Plasmodium Vivax ◽

Malaria Elimination ◽

Scale Up ◽

Transmission Dynamics ◽

Clinical Samples ◽

Effective Population ◽

Border Areas ◽

Greater Mekong Subregion ◽

International Border

Abstract BackgroundCountries within the Greater Mekong Subregion (GMS) of Southeast Asia have committed to eliminating malaria by 2030. Although malaria situation has greatly improved, Plasmodium vivax remains at international border regions. Therefore, to gain a better understanding of transmission dynamics, knowledge on the evolution of P. vivax populations after the scale-up of control interventions will guide more effective targeted control efforts. MethodsWe investigated genetic diversity and population structures in 206 longitudinally collected P. vivax clinical samples in two international border areas at the China-Myanmar border (CMB, n=50 in 2004 and n=52 in 2016) and western Thailand border (n=50 in 2012 and n=54 in 2015). Parasites were genotyped using 10 microsatellite markers. ResultsDespite intensified control efforts, genetic diversity in the four populations remained high (HE = 0.66-0.86). The proportions of polyclonal infections showed substantial decreases to 23.7 and 30.7% in the CMB and western Thailand, respectively, with corresponding decreases in the multiplicity of infection. Consistent with the shrinking map of malaria transmission in the GMS over time, there were also increases in multilocus linkage disequilibrium, suggesting of more fragmented and increasingly inbred parasite populations. There were considerable genetic differentiation and subdivision with the four tested populations. Various degrees of clustering were evident between the older parasite samples collected in 2004 at the CMB with the 2016 CMB and 2012 Thailand populations, suggesting some of these parasites had shared ancestry. In contrast, the 2015 Thailand population was genetically distinctive, which may reflect a process of population replacement. The moderately large effective population sizes and proportions of polyclonal infections highlight the necessity of further coordinated and integrated control efforts on both sides of the borders in the pursuit of malaria elimination. ConclusionsWith enhanced control efforts on malaria elimination, P. vivax population in the GMS has fragmented into a limited number of clustered foci, but the presence of large P. vivax reservoirs still sustains genetic diversity and transmission. These findings provide new insights into P. vivax transmission dynamics and population structure in this area.

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Plasmodium falciparum Genetic Diversity in Panamá Based on glurp, msp-1 and msp-2 Genes: Implications for Malaria Elimination in Mesoamerica

Life ◽

10.3390/life10120319 ◽

2020 ◽

Vol 10 (12) ◽

pp. 319

Author(s):

Ana María Santamaría ◽

Vanessa Vásquez ◽

Chystrie Rigg ◽

Dianik Moreno ◽

Luis Romero ◽

...

Keyword(s):

Genetic Diversity ◽

Malaria Elimination ◽

Multiple Correspondence Analysis ◽

Strong Association ◽

Parasite Load ◽

Molecular Surveillance ◽

Disease Propagation ◽

Low Genetic Diversity ◽

Imported Cases ◽

Endemic Areas

Panamá, together with all the nations in Mesoamerica, has committed to eliminate malaria from the region by 2020. As these countries approach malaria elimination and local transmission decreases, an active molecular surveillance to identify genotypes circulating along the border areas is particularly needed to accurately infer infection origin, drug resistance and disease propagation patterns in the region. This study evaluated the genetic diversity and allele frequencies of msp-1, msp-2 and glurp genes using different molecular analyses (nested PCR, PCR-restriction fragment length polymorphism (RFLP) and sequencing) from 106 autochthonous and imported P. falciparum isolates collected from different endemic areas in Panamá between 2003 and 2019. We also explored if P. falciparum genotypes assessed with these molecular markers were associated with relevant malaria epidemiological parameters using a multiple correspondence analysis. A strong association of certain local haplotypes with their geographic distribution in endemic areas, but also with parasite load and presence of gametocytes, was evidenced. Few multiclonal infections and low genetic diversity among locally transmitted P. falciparum samples were detected, consequent with the low transmission intensity of this parasite in Panamá, a pattern likely to be extended across Mesoamerica. In addition, several imported cases were genetically dissimilar to local infections and representative of more diverse extra-continental lineages.

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561 Using AFLPs to Examine Genetic Diversity in Umbrella Fern

HortScience ◽

10.21273/hortsci.34.3.543a ◽

1999 ◽

Vol 34 (3) ◽

pp. 543A-543

Author(s):

F.J. Keiper ◽

R. McConchie

Keyword(s):

Genetic Diversity ◽

Genetic Variation ◽

Molecular Markers ◽

Dna Markers ◽

Morphological Traits ◽

Natural Populations ◽

Cost Effective ◽

Molecular Techniques ◽

Breeding Systems ◽

Detailed Knowledge

Umbrella fern [Sticherus flabellatus (R. Br.) St John] is a successful Australian native foliage product. Currently, all umbrella fern sold on the market is bush-harvested. To meet the growing demand for this product on local and international markets, a commercially viable method for its production must be developed, with effective management of the germplasm resource in terms of conservation and exploitation. To manage this resource, breeders require a detailed knowledge of the amount and distribution of genetic variability within the species. Traditionally, plant breeders focus on a combination of agronomic and morphological traits (phenotype) to measure genetic diversity. In umbrella fern there are a limited number of morphological traits, and these are influenced by environmental factors and therefore do not reflect true genetic diversity. To overcome these problems, molecular techniques such as PCR-based DNA markers are used to complement traditional strategies for genotype assessment. DNA markers have the advantages of being independent of environmental effects, as well as being fast, cost-effective, reproducible, and largely accessible to the nonmolecular geneticist. Amplified fragment length polymorphisms (AFLPs) fulfil many of the desirable features of molecular markers, as well as requiring little knowledge of the genome to be investigated. AFLPs have been used widely in the analysis of breeding systems, ecogeographical variation, and genetic variation within and between natural populations. To date there are no published accounts of DNA molecular marker research on umbrella fern. A DNA extraction protocol has been developed for this species, and AFLP markers have been used to analyse genetic diversity within and between natural populations sampled in the Sydney Basin. A large number of polymorphic loci were revealed using 11 primer combinations. The genetic variation detected was partitioned between rather than within populations, suggesting that the mating system in Sticherus is primarily inbreeding. Data will be presented illustrating AFLPs as useful molecular markers for assessing genetic diversity within and between populations of umbrella fern and providing insight on the breeding system used by the species.

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Dynamics of Plasmodium vivax populations in border areas of the Greater Mekong Sub-region during malaria elimination

10.21203/rs.2.22640/v2 ◽

2020 ◽

Author(s):

Yuling Li ◽

Yubing Hu ◽

Yan Zhao ◽

Qinghui Wang ◽

Huguette Gaelle Ngassa Mbenda ◽

...

Keyword(s):

Genetic Diversity ◽

Genetic Differentiation ◽

Plasmodium Vivax ◽

Malaria Elimination ◽

Malaria Incidence ◽

Scale Up ◽

Transmission Dynamics ◽

Clinical Samples ◽

Border Areas ◽

International Border

Abstract Background Countries within the Greater Mekong Sub-region (GMS) of Southeast Asia have committed to eliminating malaria by 2030. Although the malaria situation has greatly improved, malaria transmission remains at international border regions. In some areas, Plasmodium vivax has become the predominant parasite. To gain a better understanding of transmission dynamics, knowledge on the changes of P. vivax populations after the scale-up of control interventions will guide more effective targeted control efforts. Methods This study investigated genetic diversity and population structures in 206 P. vivax clinical samples collected at two time points in two international border areas: the China-Myanmar border (CMB) (n=50 in 2004 and n=52 in 2016) and Thailand-Myanmar border (TMB) (n=50 in 2012 and n=54 in 2015). Parasites were genotyped using 10 microsatellite markers. Results Despite intensified control efforts, genetic diversity remained high ( H E = 0.66-0.86) and was not significantly different among the four populations ( P >0.05). Specifically, H E slightly decreased from 0.76 in 2004 to 0.66 in 2016 at the CMB and increased from 0.80 in 2012 to 0.86 in 2015 at the TMB. The proportions of polyclonal infections varied significantly among the four populations ( P < 0.05), and showed substantial decreases from 48.0% in 2004 to 23.7 at the CMB and from 40.0% in 2012 to 30.7% in 2015 at the TMB, with corresponding decreases in the multiplicity of infection. Consistent with the continuous decline of malaria incidence in the GMS over time, there were also increases in multilocus linkage disequilibrium, suggesting more fragmented and increasingly inbred parasite populations. There were considerable genetic differentiation and sub-division among the four tested populations. Temporal genetic differentiation was observed at each site ( F ST = 0.081 at the CMB and F ST = 0.133 at the TMB). Various degrees of clustering were evident between the older parasite samples collected in 2004 at the CMB with the 2016 CMB and 2012 TMB populations, suggesting some of these parasites had shared ancestry. In contrast, the 2015 TMB population was genetically distinctive, which may reflect a process of population replacement. Whereas the effective population size ( N e ) at the CMB showed a decrease from 4979 in 2004 to 3052 in 2016 with the infinite allele model, the N e at the TMB experienced an increase from 6289 to 10259. Conclusions With enhanced control efforts on malaria, P. vivax at the TMB and CMB showed considerable spatial and temporal differentiation, but the presence of large P. vivax reservoirs still sustained genetic diversity and transmission. These findings provide new insights into P. vivax transmission dynamics and population structure in these border areas of the GMS. Coordinated and integrated control efforts on both sides of international borders are essential to reach the goal of regional malaria elimination.

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Current and Future Pathotyping Platforms for Plasmodiophora brassicae in Canada

Plants ◽

10.3390/plants10071446 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1446

Author(s):

Heather H. Tso ◽

Leonardo Galindo-González ◽

Stephen E. Strelkov

Keyword(s):

Crop Production ◽

Plasmodiophora Brassicae ◽

Cost Effective ◽

Turnaround Time ◽

Molecular Techniques ◽

Differential Set ◽

Management Plans ◽

Increased Sensitivity ◽

Phenotypic Methods ◽

Important Disease

Clubroot, caused by Plasmodiophora brassicae, is one of the most detrimental threats to crucifers worldwide and has emerged as an important disease of canola (Brassica napus) in Canada. At present, pathotypes are distinguished phenotypically by their virulence patterns on host differential sets, including the systems of Williams, Somé et al., the European Clubroot Differential set, and most recently the Canadian Clubroot Differential set and the Sinitic Clubroot Differential set. Although these are frequently used because of their simplicity of application, they are time-consuming, labor-intensive, and can lack sensitivity. Early, preventative pathotype detection is imperative to maximize productivity and promote sustainable crop production. The decreased turnaround time and increased sensitivity and specificity of genotypic pathotyping will be valuable for the development of integrated clubroot management plans, and interest in molecular techniques to complement phenotypic methods is increasing. This review provides a synopsis of current and future molecular pathotyping platforms for P. brassicae and aims to provide information on techniques that may be most suitable for the development of rapid, reliable, and cost-effective pathotyping assays.

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Genotyping Strategies Using ddRAD Sequencing in Farmed Arctic Charr (Salvelinus alpinus)

Animals ◽

10.3390/ani11030899 ◽

2021 ◽

Vol 11 (3) ◽

pp. 899

Author(s):

Fotis Pappas ◽

Christos Palaiokostas

Keyword(s):

Genetic Diversity ◽

Salvelinus Alpinus ◽

Arctic Charr ◽

Cost Effective ◽

Limiting Factors ◽

Phenotypic Traits ◽

Production Volume ◽

High Coverage ◽

Genomic Relationships ◽

Low Coverage

Incorporation of genomic technologies into fish breeding programs is a modern reality, promising substantial advances regarding the accuracy of selection, monitoring the genetic diversity and pedigree record verification. Single nucleotide polymorphism (SNP) arrays are the most commonly used genomic tool, but the investments required make them unsustainable for emerging species, such as Arctic charr (Salvelinus alpinus), where production volume is low. The requirement to genotype a large number of animals for breeding practices necessitates cost effective genotyping approaches. In the current study, we used double digest restriction site-associated DNA (ddRAD) sequencing of either high or low coverage to genotype Arctic charr from the Swedish national breeding program and performed analytical procedures to assess their utility in a range of tasks. SNPs were identified and used for deciphering the genetic structure of the studied population, estimating genomic relationships and implementing an association study for growth-related traits. Missing information and underestimation of heterozygosity in the low coverage set were limiting factors in genetic diversity and genomic relationship analyses, where high coverage performed notably better. On the other hand, the high coverage dataset proved to be valuable when it comes to identifying loci that are associated with phenotypic traits of interest. In general, both genotyping strategies offer sustainable alternatives to hybridization-based genotyping platforms and show potential for applications in aquaculture selective breeding.

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Molecular markers and antioxidant activity in berry crops: Genetic diversity analysis

Canadian Journal of Plant Science ◽

10.4141/cjps2011-240 ◽

2012 ◽

Vol 92 (6) ◽

pp. 1121-1133 ◽

Cited By ~ 9

Author(s):

S. C. Debnath ◽

Y. L. Siow ◽

J. Petkau ◽

D. An ◽

N. V. Bykova

Keyword(s):

Genetic Diversity ◽

Antioxidant Activity ◽

Molecular Markers ◽

Molecular Techniques ◽

Diversity Analysis ◽

Full Potential ◽

Genetic Diversity Analysis ◽

Wide Range ◽

Berry Crops ◽

Improvement Programs

Debnath, S. C., Siow, Y. L., Petkau, J., An, D. and Bykova, N. V. 2012. Molecular markers and antioxidant activity in berry crops: Genetic diversity analysis. Can. J. Plant Sci. 92: 1121–1133. An improved understanding of important roles of dietary fruits in maintaining human health has led to a dramatic increase of global berry crop production. Berry fruits contain relatively high levels of vitamin C, cellulose and pectin, and produce anthocyanins, which have important therapeutic values, including antitumor, antiulcer, antioxidant and anti-inflammatory activities. There is a need to develop reliable methods to identify berry germplasm and assess genetic diversity/relatedness for dietary properties in berry genotypes for practical breeding purposes through genotype selection in a breeding program for cultivar development, and proprietary-rights protection. The introduction of molecular biology techniques, such as DNA-based markers, allows direct comparison of different genetic materials independent of environmental influences. Significant progress has been made in diversity analysis of wild cranberry, lowbush blueberry, lingonberry and cloudberry germplasm, and in strawberry and raspberry cultivars and advanced breeding lines developed in Canada. Inter simple sequence repeat (ISSR) markers detected an adequate degree of polymorphism to differentiate among berry genotypes, making this technology valuable for cultivar identification and for the more efficient choice of parents in the current berry improvement programs. Although multiple factors affect antioxidant activity, a wide range of genetic diversity has been reported in wild and cultivated berry crops. Diversity analysis based on molecular markers did not agree with those from antioxidant activity. The paper also discusses the issues that still need to be addressed to utilize the full potential of molecular techniques including expressed sequence tag-polymerase chain reaction (EST-PCR) analysis to develop improved environment-friendly berry cultivars suited to the changing needs of growers and consumers.

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