Population genetic structure of Plasmodium falciparum in the two main African vectors, Anopheles gambiae and Anopheles funestus

We investigated patterns of genetic diversity of Plasmodium falciparum associated with its two main African vectors: Anopheles gambiae and Anopheles funestus. We dissected 10,296 wild-caught mosquitoes from three tropical sites, two in Cameroon (Simbock and Tibati, separated by 320 km) and one in Kenya (Rota, >2,000 km from the other two sites). We assayed seven microsatellite loci in 746 oocysts from 183 infected mosquito guts. Genetic polymorphism was very high in parasites isolated from both vector species. The expected heterozygosity (HE) was 0.79 in both species; the observed heterozygosities (HO) were 0.32 in A. funestus and 0.42 in A. gambiae, indicating considerable inbreeding within both vector species. Mean selfing (s) between genetically identical gametes was s = 0.33. Differences in the rate of inbreeding were statistically insignificant among sites and between the two vector species. As expected, because of the high rate of inbreeding, linkage disequilibrium was very high; it was significant for all 21 loci pairs in A. gambiae and for 15 of 21 pairs in A. funestus, although only two pairwise comparisons were between loci on the same chromosome. Overall, the genetic population structure of P. falciparum, as evaluated by F statistics, was predominantly clonal rather than panmictic, a population structure that facilitates the spread of antimalarial drug and vaccine resistance and thus may impair the effectiveness of malaria control efforts.

[1]  E. Winzeler,et al.  The paradoxical population genetics of Plasmodium falciparum. , 2002, Trends in parasitology.

[2]  S. Nee,et al.  Gametocyte sex ratios as indirect measures of outcrossing rates in malaria , 1992, Parasitology.

[3]  B. Sharp,et al.  Reduced variation around drug-resistant dhfr alleles in African Plasmodium falciparum. , 2005, Molecular biology and evolution.

[4]  C. Dye,et al.  World Malaria Report, 2008. , 2008 .

[5]  C. Rogier,et al.  High annual and seasonal variations in malaria transmission by anophelines and vector species composition in Dielmo, a holoendemic area in Senegal. , 1997, The American journal of tropical medicine and hygiene.

[6]  D. Conway,et al.  Extreme geographical fixation of variation in the Plasmodium falciparum gamete surface protein gene Pfs48/45 compared with microsatellite loci. , 2001, Molecular and biochemical parasitology.

[7]  F. James Rohlf,et al.  Biometry: The Principles and Practice of Statistics in Biological Research , 1969 .

[8]  D. Conway,et al.  High recombination rate in natural populations of Plasmodium falciparum. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[9]  J. Goudet FSTAT (Version 1.2): A Computer Program to Calculate F-Statistics , 1995 .

[10]  F. Simard,et al.  Unravelling complexities in human malaria transmission dynamics in Africa through a comprehensive knowledge of vector populations. , 2004, Comparative immunology, microbiology and infectious diseases.

[11]  R. Sinden,et al.  Heterogeneity in patterns of malarial oocyst infections in the mosquito vector , 1993, Parasitology.

[12]  A. Githeko,et al.  Population Dynamics of Malaria Vectors in Western Kenya Highlands , 2006, Journal of medical entomology.

[13]  D. M. Power Biometry. The Principles and Practice of Statistics in Biological Research; Statistical Tables , 1970 .

[14]  M. Coluzzi,et al.  The clay feet of the malaria giant and its African roots: hypotheses and inferences about origin, spread and control of Plasmodium falciparum. , 1999, Parassitologia.

[15]  R. Hunt,et al.  A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group. , 2002, The American journal of tropical medicine and hygiene.

[16]  G. Killeen,et al.  Short report: entomologic inoculation rates and Plasmodium falciparum malaria prevalence in Africa. , 1999, The American journal of tropical medicine and hygiene.

[17]  F J Ayala,et al.  A clonal theory of parasitic protozoa: The population structures of Entamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, and Trypanosoma and their medical and taxonomical consequences , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[18]  F. Ayala,et al.  Plasmodium falciparum antigenic diversity: evidence of clonal population structure. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[19]  J. Olivry Fleuves et rivières du Cameroun , 1986 .

[20]  Sokal Rr,et al.  Biometry: the principles and practice of statistics in biological research 2nd edition. , 1981 .

[21]  F. Collins,et al.  Identification of single specimens of the Anopheles gambiae complex by the polymerase chain reaction. , 1993, The American journal of tropical medicine and hygiene.

[22]  C.J.F. ter Braak,et al.  CANOCO - a FORTRAN program for canonical community ordination by [partial] [etrended] [canonical] correspondence analysis, principal components analysis and redundancy analysis (version 2.1) , 1988 .

[23]  J. Goudet HIERFSTAT , a package for R to compute and test hierarchical F -statistics , 2005 .

[24]  R. Paru,et al.  Mating patterns in malaria parasite populations of Papua New Guinea. , 1995, Science.

[25]  R. Poulin The disparity between observed and uniform distributions: a new look at parasite aggregation. , 1993, International journal for parasitology.

[26]  F. Simard,et al.  Complexity of the Malaria Vectorial System in Cameroon: Contribution of Secondary Vectors to Malaria Transmission , 2006, Journal of medical entomology.

[27]  I. Hastings,et al.  Complex dynamics and stability of resistance to antimalarial drugs , 2006, Parasitology.

[28]  X. Su,et al.  Toward a high-resolution Plasmodium falciparum linkage map: polymorphic markers from hundreds of simple sequence repeats. , 1996, Genomics.

[29]  M. Nei,et al.  Estimation of average heterozygosity and genetic distance from a small number of individuals. , 1978, Genetics.

[30]  John C. Wootton,et al.  Genetic diversity and chloroquine selective sweeps in Plasmodium falciparum , 2002, Nature.

[31]  J. T. Williams,et al.  Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum. , 2000, Molecular biology and evolution.

[32]  Z. Tu,et al.  On the distribution and genetic differentiation of Anopheles gambiae s.s. molecular forms. , 2005, Insect biochemistry and molecular biology.

[33]  D. Fontenille,et al.  Bionomics of Anopheles gambiae Giles, An. arabiensis Patton, An. funestus Giles and An. nili (Theobald) (Diptera: Culicidae) and Transmission of Plasmodium falciparum in a Sudano-Guinean Zone (Ngari, Senegal) , 2003, Journal of medical entomology.

[34]  J. Dushoff,et al.  The entomological inoculation rate and Plasmodium falciparum infection in African children , 2005, Nature.

[35]  C. Rogier,et al.  Genetic diversity and structure of African Plasmodium falciparum populations in urban and rural areas. , 2006, The American journal of tropical medicine and hygiene.

[36]  F. Ayala,et al.  "Clonal" population structure of the malaria agent Plasmodium falciparum in high-infection regions. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[37]  P. Awadalla,et al.  Local differentiation in Plasmodium falciparum drug resistance genes in Sudan , 2003, Parasitology.

[38]  C. Braak Canonical Correspondence Analysis: A New Eigenvector Technique for Multivariate Direct Gradient Analysis , 1986 .

[39]  B G Williams,et al.  Multigenic drug resistance among inbred malaria parasites , 1997, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[40]  M. Nei Molecular Evolutionary Genetics , 1987 .

[41]  Yingyao Zhou,et al.  A Systematic Map of Genetic Variation in Plasmodium falciparum , 2006 .

[42]  E. Feil,et al.  Population structure and evolutionary dynamics of pathogenic bacteria , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[43]  F. Ayala,et al.  Plasmodium falciparum: population genetic analysis by multilocus enzyme electrophoresis and other molecular markers. , 1999, Experimental parasitology.

[44]  C. Antonio-Nkondjio,et al.  High Malaria Transmission Intensity in a Village Close to Yaounde, the Capital City of Cameroon , 2002, Journal of medical entomology.

[45]  M. Tibayrenc,et al.  Evidence for clonal propagation in natural isolates of Plasmodium falciparum from Venezuela , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[46]  L. Ranford-Cartwright,et al.  Estimation of inbreeding coefficients from genotypic data on multiple alleles, and application to estimation of clonality in malaria parasites. , 1995, Genetical research.

[47]  G. Yan,et al.  The Effects of Climatic Factors on the Distribution and Abundance of Malaria Vectors in Kenya , 2002, Journal of medical entomology.

[48]  Y. Michalakis,et al.  Significant linkage disequilibrium and high genetic diversity in a population of Plasmodium falciparum from an area (Republic of the Congo) highly endemic for malaria. , 2003, The American journal of tropical medicine and hygiene.

[49]  J. Charlwood,et al.  Random mating in a natural population of the malaria parasite Plasmodium falciparum , 1994, Parasitology.

[50]  Lajos Rózsa,et al.  QUANTIFYING PARASITES IN SAMPLES OF HOSTS , 2000, The Journal of parasitology.

[51]  Kateryna D. Makova,et al.  Chromosome-wide SNPs reveal an ancient origin for Plasmodium falciparum , 2002, Nature.