Evidence for late Pleistocene population expansion of the malarial mosquitoes, Anopheles arabiensis and Anopheles gambiae in Nigeria.

Anopheles gambiae Giles s.s. and Anopheles arabiensis Patton (Diptera: Culicidae) are major vectors of malaria in Nigeria. We used 1115 bp of the mitochondrial COI gene to assess their population genetic structures based on samples from across Nigeria (n = 199). The mtDNA neighbour-joining tree, based on F(ST) estimates, separated An. gambiae M and S forms, except that samples of An. gambiae M from Calabar clustered with all the An. gambiae S form. Anopheles arabiensis and An. gambiae could be combined into a single star-shaped, parsimonious haplotype network, and shared three haplotypes. Haplotype diversity values were high in An. arabiensis and An. gambiae S, and intermediate in An. gambiae M; all nucleotide diversities were relatively low. Taken together, patterns of haplotype diversity, the star-like genealogy of haplotypes, five of seven significant neutrality tests, and the violation of the isolation-by-distance model indicate population expansion in An. arabiensis and An. gambiae S, but the signal was weak in An. gambiae M. Selection is supported as an important factor shaping genetic structure in An. gambiae in Nigeria. There were two geographical subdivisions in An. arabiensis: one included all southern localities and all but two central localities; the other included all northern and two central localities. Re-analysing an earlier microsatellite dataset of An. arabiensis using a Bayesian method determined that there were two distinctive clusters, northern and southern, that were fairly congruent with the mtDNA subdivisions. There was a trend towards decreasing genetic diversity in An. arabiensis from the northern savannah to the southern rainforest that corroborated previous data from microsatellites and polytene chromosomes.

Progress toward the development of a bacterial vaccine vector that induces high-titer long-lived broadly neutralizing antibodies against HIV-1.

Conformationally constrained HIV-1 Env and gp120 immunogens induce broadly cross-reactive neutralizing antibodies. Thus, it is now feasible to rationally design an HIV-1 vaccine that affords protection through humoral mechanisms. This paper reviews our progress toward the development of an oral bacterial vaccine vector that is capable of delivering an HIV-1 DNA vaccine to host lymphoid tissues and inducing broadly neutralizing antibodies to HIV-1 in the mucosal and systemic immune compartments.

Immunogenicity of DNA vaccines that direct the coincident expression of the 120 kDa glycoprotein of human immunodeficiency virus and the catalytic domain of cholera toxin.

Passive antibody studies unequivocally demonstrate that sterilizing immunity against lentiviruses is obtainable through humoral mechanisms. In this regard, DNA vaccines represent an inexpensive alternative to subunit vaccine for mass vaccination programs designed to induce such responses to human immunodeficiency virus type I (HIV-1). At present, however, this vaccine modality has proven relatively ineffective at inducing humoral responses. In this report, we describe the immunogenicity of DNA vaccines that direct the coincident expression of the cholera toxin catalytic domain (CTA1) with that of the human immunodeficiency virus type I gp120 through genes either encoded in individual plasmids or in a single dicistronic plasmid. In BALB/cJ mice, coincident expression of CTA1 in either a separate plasmid or in the dicistronic plasmid in the DNA vaccines induced serum IgG responses to gp120 that were at least 1000-fold greater, and remained elevated longer than, the analogous responses in mice vaccinated with a DNA vaccine that expressed gp120 alone. In addition, mice vaccinated with CTA1 and gp120 produced significantly more gp120-specific IFN-gamma ELISPOTs than mice vaccinated with the gp120 DNA vaccine. Combined, these data show that the adjuvant properties of cholera toxin can be harnessed in DNA vaccine modalities.

The distribution of M and S molecular forms of Anopheles gambiae in Nigeria.

The distribution of M and S molecular forms of Anopheles gambiae sensu stricto across Nigeria was determined. The molecular form of 40 to 45 specimens per locality from 9 localities was determined using mostly the same specimens from our recent study of genetic differentiation of A. gambiae across Nigeria (Onyabe & Conn, 2001). These samples were previously genotyped at 10 microsatellite loci, 5 located within chromosome inversions and 5 outside inversions. Both molecular forms occurred throughout the country, with no apparent relationship to the ecological transition from dry savannah in the north to humid forest in southern Nigeria. In all localities, however, 1 form or the other occurred virtually exclusively. No hybrids between forms were found. Across all loci, F(ST) values were as high within molecular forms as between forms. Regardless of molecular form, F(ST) values calculated across loci within inversions were much higher (range 0.0016 to 0.1988) than those calculated across loci outside inversions (range -0.0035 to 0.0260). Genetic distance was not significantly correlated with geographical distance within either form (P> 0.05). These observations suggest that, in addition to partial reproductive barriers between molecular forms, selection is a major factor shaping genetic differentiation of A. gambiae across Nigeria.

Genetic differentiation of the malaria vector Anopheles gambiae across Nigeria suggests that selection limits gene flow.

Gene flow was investigated in Anopheles gambiae from eight localities that span the ecological zones of Nigeria (arid savanna zones in the north gradually turn into humid forest zones in the south). Genetic differentiation was measured over 10 microsatellite loci and, to determine any effects of selection, five loci were located within chromosome inversions and the other five were outside inversions. Over all loci, the largest estimates of differentiation were in comparisons between localities in the savanna vs. forest zones (range FST 0.024-0.087, Nm 2.6-10.1; RST 0.014-0.100, Nm 2.2-16.4). However, three loci located within inversions on chromosome II, whose frequencies varied clinically from north to south, were responsible for virtually all of the differentiation. When the three loci were removed, genetic distances across the remaining seven loci were markedly reduced even between localities in the forest and savanna zones (range FST 0.001-0.019, Nm 12.7-226.1) or no longer significant (P > 0.05) in the case of RST. Although tests of isolation by distance gave seemingly equivocal results, geographical distance does not appear to limit gene flow. These observations suggest that gene flow is extensive across the country but that selection on genes located within some inversions on chromosome II counters the homogenizing effects of gene flow.

The distribution of two major malaria vectors, Anopheles gambiae and Anopheles arabiensis, in Nigeria.

The distribution of Anopheles gambiae and An. arabiensis across the ecological zones of Nigeria (arid savanna in the north gradually turns into humid forest in the south) was investigated. Results of the present study were compared to the distributions determined from samples of indoor-resting females reported by an earlier study over 20 years ago. Larvae were sampled in the rainy seasons of 1997 and 1999 from 24 localities, 10 of which were sampled in both years. Specimens were identified by the polymerase chain reaction method. Results showed that species composition changed significantly among the 10 localities in both years (chi2=13.62, P = 0.0002), but this change was significant in only four of the 10 localities. The identity of the prevalent (more abundant) species changed between 1997 and 1999 in only three of 10 localities. An. arabiensis was prevalent in several localities in the southern Guinea savanna, an area where it was virtually absent over 20 years ago. The data suggest that An. arabiensis has extend its range, although differences in sampling technique (larval sampling versus adult collection) can not be ruled out as a possible explanation.

Population genetic structure of the malaria mosquito Anopheles arabiensis across Nigeria suggests range expansion.

Ten microsatellite loci, four located within and six outside chromosome inversions, were employed to study the genetic structure of Anopheles arabiensis across the ecological zones of Nigeria (arid savannah in the north gradually turns into humid forest in the south). Regardless of location within or outside inversions, genetic variability at all loci was characterized by a reduction in both the number of alleles per locus and heterozygosity from savannah to forest. Across all loci, all but one allele in the forest also occurred in the savannah, whereas at least 78 alleles in the savannah were missing in the forest. Genetic differentiation increased with geographical distance; consequently, genetic distances between zones exceeded those within zones. The largest genetic distances were between localities at the extremes of the transect (range F(ST) = 0.196-0.258 and R(ST) = 0.183-0.468) and were as large as those between A. arabiensis and Anopheles gambiae s.s. Gene flow across the country was very low, so that Nm between the extremes of the transect was < 1. These data suggest that A. arabiensis has extended its range from the savannah into the forest during which it experienced a reduction in effective population size due to sequential founder effects. Gene flow post range expansion appears too restricted by geographical distance to homogenize the gene pool of A. arabiensis across Nigeria.

Intragenomic heterogeneity of a ribosomal DNA spacer (ITS2) varies regionally in the neotropical malaria vector Anopheles nuneztovari (Diptera:Culicidae).

We investigated intragenomic heterogeneity of ITS2 within twenty individual Anopheles nuneztovari (subgenus Nyssorhynchus) from five geographical localities in the neotropics (three from Brazil and one each from Colombia and Venezuela) by cloning and sequencing PCR-amplified copies of this spacer. Intragenomic heterogeneity was observed in thirteen of twenty mosquitoes of both sexes from all localities. As estimated by uncorrected P, however, mean sequence divergence was greater in mosquitoes from Brazil (PR = 0.0100, BL = 0.0196, AB = 0.0182) than in those from Venezuela (SO = 0.0026) or Colombia (SI = 0.0078). Sequence divergence per genome was significantly higher in mosquitoes from Brazil than in those from SO and SI. In fact, divergence among ITS2 variants within single mosquitoes from the Brazilian localities was often as great as that between localities. Similarly, the number of variants per mosquito was significantly greater in Brazil (maximum of six variants) than in both SO and SI (maximum of two variants). These results indicate that homogenization of ITS2 has proceeded to a greater extent in SO and SI than in the localities in Brazil. The differences in intragenomic heterogeneity between both SO and SI versus the Brazilian localities probably reflect either differences in population-level processes such as gene flow and genetic drift, or the fact that these localities may represent two or more cryptic species, as suggested by other studies.

Feeding and mating strategies in anopheles (Diptera: Culicidae): theoretical modeling approach.

The effects of various ecological factors, such as the probability of finding mates and hosts and of successfully obtaining a blood meal, on the mating and feeding strategies of domestic female anopheline mosquitoes was investigated using theoretical models. The models calculated the mean fitness of 1,000 nonblood-fed, anautogenous, virgin anophelines. One model simulated females that always mate before blood feeding, whereas another simulated females that are able to feed opportunistically if a host was detected before they mated. The models demonstrated highest fitness for mosquitoes capable of opportunistic feeding under nearly all simulated conditions. This advantage increased as the probability of finding hosts and mates decreased as a function of host and mate availability.