Population analysis using the nuclear white gene detects Pliocene/Pleistocene lineage divergence within Anopheles nuneztovari in South America

Anopheles (Nyssorhynchus) nuneztovari Gabaldón (Diptera: Culicidae), a locally important malaria vector in some regions of South America, has been hypothesized to consist of at least two cryptic incipient species. We investigated its phylogeographic structure in several South American localities to determine the number of lineages and levels of divergence using the nuclear white gene, a marker that detected two recently diverged genotypes in the primary Neotropical malaria vector Anopheles darlingi Root. In An. nuneztovari, five distinct lineages (1-5) were elucidated: (1) populations from northeastern and central Amazonia; (2) populations from Venezuela east and west of the Andes; (3) populations from Colombia and Venezuela west of the Andes; (4) southeastern and western Amazonian Brazil populations, and (5) southeastern and western Amazonian Brazil and Bolivian populations. There was a large amount of genetic differentiation among these lineages. The deepest and earliest divergence was found between lineage 3 and lineages 1, 2 and 4, which probably accounts for the detection of lineage 3 in some earlier studies. The multiple lineages within Amazonia are partially congruent with previous mtDNA and ITS2 data, but were undetected in many earlier studies, probably because of their recent (Pleistocene) divergence and the differential mutation rates of the markers. The estimates for the five lineages, interpreted as recently evolved or incipient species, date to the Pleistocene and Pliocene. We hypothesize that the diversification in An. nuneztovari is the result of an interaction between the Miocene/Pliocene marine incursion and Pleistocene climatic changes leading to refugial isolation. The identification of cryptic lineages in An. nuneztovari could have a significant impact on local vector control measures.     

中国甜菜夜蛾地理种群的遗传分化与基因流

【目的】为了明确我国甜菜夜蛾Spodoptera exigua地理种群间的遗传分化及基因流,阐明该种害虫在我国的种群历史动态。【方法】本研究对采自我国20个地理种群的529头甜菜夜蛾样品进行线粒体COI基因序列测定与分析,利用DnaSP 5.0和Arlequin 3.11软件分析种群间遗传多样性、遗传分化、基因流水平及分子变异,构建了单倍型系统发育树与单倍型网络图。【结果】在所分析的所有529个序列样本中,共检测出10个单倍型,其中Hap_1为所有种群所共享。总群体遗传多样性较低（Hd=0.257±0.025,Pi=0.0007±0.0001,Kxy=0.323）,群体间遗传分化较小（F_ST=0.211）,基因流水平较高（N_m=1.870）。AMOVA分析表明,甜菜夜蛾遗传变异主要来自种群内,种群间变异水平较低。各种群间遗传分化程度与地理距离无显著相关性（R²=0.005,P>0.05）。各单倍型相互散布在不同种群中,未形成明显系统地理结构。中性检验（Tajima’s D=-2.177, P<0.05; Fu’s F_S=-8.629, P<0.05)与错配分布分析表明,我国甜菜夜蛾种群曾经历种群近期扩张。【结论】研究结果揭示,甜菜夜蛾各种群间的基因交流并未受到地理距离的影响,验证了甜菜夜蛾具有高度的迁飞能力。     

Molecular comparison of topotypic specimens confirms Anopheles (Nyssorhynchus) dunhami Causey (Diptera: Culicidae) in the Colombian Amazon

The presence of Anopheles (Nyssorhynchus) dunhami Causey in Colombia (Department of Amazonas) is confirmed for the first time through direct comparison of mtDNA cytochrome c oxidase I (COI) barcodes and nuclear rDNA second internal transcribed spacer (ITS2) sequences with topotypic specimens of An. dunhami from Tefé, Brazil. An. dunhami was identified through retrospective correlation of DNA sequences following misidentification as Anopheles nuneztovari s.l. using available morphological keys for Colombian mosquitoes. That An. dunhami occurs in Colombia and also possibly throughout the Amazon Basin, is of importance to vector control programs, as this non-vector species is morphologically similar to known malaria vectors including An. nuneztovari, Anopheles oswaldoi and Anopheles trinkae. Species identification of An. dunhami and differentiation from these closely related species are highly robust using either DNA ITS2 sequences or COI DNA barcode. DNA methods are advocated for future differentiation of these often sympatric taxa in South America.     

基于线粒体COI基因序列的大豆食心虫中国东北地理种群遗传多样性分析

【目的】大豆食心虫Leguminivora glycinivorella (Matsumura)是一种危害大豆的主要害虫,在中国北方地区危害较重。本研究旨在探讨大豆食心虫在中国东北不同地理种群间的遗传变异。【方法】测定了10个不同地理种群153个个体的线粒体细胞色素氧化酶亚基Ⅰ (mtCOI)基因的657 bp序列,利用DnaSP 5. 0和Arlequin 3. 5. 1. 2等软件对大豆食心虫种群间的遗传多样性、基因流水平和分子变异进行分析。【结果】结果表明：10个地理种群间的COI基因共有36个变异位点和17个单倍型,其中1个单倍型为10个种群所共享。总种群的单倍型多样性指数Hd为0.456,各地理种群单倍型多样度范围在0~0.634之间。总群体的固定系数Fst为0.12545,遗传分化系数Gst为0.06326,总基因流Nm为3.49,且各种群间的基因流均大于1,种群间基因交流的水平较高。【结论】大豆食心虫种群内遗传多样性水平处于中低等水平。总群体和各种群的Tajima’s D检验结果皆不显著,说明中国东北地区大豆食心虫在较近的历史时期内没有出现种群扩张现象。AMOVA分子变异分析结果表明,大豆食心虫的遗传分化主要来自种群内部,而种群间未发生明显的遗传分化。各地理种群的单倍型在系统发育树上和中介网络图上散布在不同的分布群中,缺乏明显的地理分布格局。各种群的遗传距离与地理距离之间没有显著线性相关性,种群间的基因交流并未受到地理距离的影响。     

Genetic variation of Mytilus coruscus Gould (Bivalvia: Mytilidae) populations in the East China Sea inferred from mtDNA COI gene Sequence

In order to characterize the genetic relationship of six populations of Mytilus coruscus Gould in the East China Sea, a 681 bp region of mtDNA COI gene was sequenced and analyzed. Eighty four individuals in total were collected from three cultured populations and three wild populations from three localities of the coast of East China Sea. The sequences from these different populations identified 62 polymorphic sites, which included 41 singleton variable sites and 21 parsimony informative sites that defined 45 distinct haplotypes. Phylogenetic analysis showed that most haplotypes were highly interconnected with each other. Thirty seven of the 45 haplotypes were only found in their own populations, seven were found at two-four localities and only haplotype NO.2 was found in all six populations, indicating that most haplotypes were locally restricted. All haplotypes had shaped two similar branches, each including individuals from all six strains. The results of F_ST values indicated that the genetic distances between populations are not closely associated with their geographic distances.     

Genetic differentiation between populations of swimming crab <Emphasis Type="Italic">Portunus trituberculatus</Emphasis> along the coastal waters of the East China Sea

Portunus trituberculatus is a commercially important species widely spread in the East China Sea. Intraspecific variation of the mitochondrial DNA cytochrome oxidase subunit I (mtDNA COI) gene was investigated in 213 individuals from six localities (Changjiang Estuary, Shengsi Islands, Zhoushan Islands, Dongtou Islands, Dinghai Bay, and Quanzhou Bay) ranging from north (31°21′N) to south (24°55′N) coastal waters of the East China Sea. Overall, a total of 27 mtDNA haplotypes and 21 variable sites were detected in the 787 bp segment of COI gene. Analysis of mtDNA COI sequence data revealed that crabs from the six localities were characterized by moderately high haplotypic diversity (h = 0.787 ± 0.026), while sequence divergence values between haplotypes were relatively low (π = 0.00241 ± 0.00098). Each population was characterized by a single most frequent haplotype, shared among all six localities, and a small number of rare ones, typically present in only one or two individuals and representative of a specific population. However, neither the neighbor-joining tree nor the minimum spanning network (MSN) based on the haplotype data exhibited geographical patterns of the six populations. Mismatch distribution analysis of P. trituberculatus individuals sampled from the six localities suggested that sudden population expansion might have occurred in CJ and SS population that might be consistent with over-exploitation of the swimming crab. Analysis of molecular variance (AMOVA) and F _ST statistics showed that significant genetic differentiation existed among the SS, ZS, DT, DH, and QZ populations, suggesting that gene flow might be reduced, even between the geographically close sites, despite the high potential of dispersal. The possible causes of the observed genetic heterogeneity among the P. trituberculatus populations and the potential applications of the mtDNA COI marker in the artificial breeding and fisheries management are discussed. Handling editor: C. Sturmbauer     

基于线粒体COI基因序列的黄胫小车蝗不同地理种群的遗传分化及基因流分析

黄胫小车蝗Oedaleus infernalis Saussure是一种在我国分布广泛、 对农牧业生产危害严重的经济害虫。本文对黄胫小车蝗10个地理种群的线粒体COI基因序列进行测序和分析, 利用DnaSP 5.0和Arlequin 3.5.1.2软件对该蝗虫种群间的遗传多样性、 遗传分化程度、 基因流水平及分子变异进行了分析, 建立了单倍型贝叶斯系统发育进化树和单倍型网络图。结果表明： 在所分析的144个序列样本中, 共检测到21种单倍型, 其中1种单倍型为10个地理种群所共享。总群体的单倍型多样性指数为0.653, 各地理种群单倍型多样度范围在0.423~0.790之间。总群体和各种群的Tajima’s D检验结果皆不显著, 说明该种害虫在较近的历史上未经历群体扩张。总群体的遗传分化系数Gst为0.04436, 固定系数Fst为0.05255, 基因流Nm为9.01。AMOVA分子方差分析结果表明, 黄胫小车蝗的遗传分化主要来自种群内部, 种群间的遗传变异水平较低。各地理种群的遗传距离的大小与其地理距离间没有显著的相关性。贝叶斯系统发育进化树与单倍型网络图显示, 黄胫小车蝗各地理种群中的单倍型散布在不同的分布群中, 分布格局较为混杂, 未形成明显的系统地理结构。研究结果揭示, 黄胫小车蝗各种群间的基因交流并未受到地理距离的影响。     

Cryptic divergence and strong population structure in the colonial invertebrate Pycnoclavella communis (Ascidiacea) inferred from molecular data

We studied sequence variation in the mitochondrial gene cytochrome c oxidase subunit I (COI) for 135 individuals from eight Mediterranean populations of the colonial ascidian Pycnoclavella communis across most of its presently known range of distribution in the Mediterranean. Three haplotypes from Atlantic locations were also included in the study. Phylogenetic, phylogeographic and population genetic analyses were used to unravel the genetic variability within and between populations. The study revealed 32 haplotypes for COI, 29 of them grouped within two Mediterranean lineages of P. communis (mean nucleotide divergence between lineages was 8.55%). Phylogenetic and network analyses suggest the possible existence of cryptic species corresponding to these two lineages. Population genetic analyses were restricted to the five populations belonging to the main genetic lineage, and for these localities we compared the information gleaned from COI sequence data and from eight microsatellite loci. A high genetic divergence between populations was substantiated using both kinds of markers (COI, global Fst=0.343; microsatellite loci, global Fst=0.362). There were high numbers of private haplotypes (COI) and alleles (microsatellites) in the populations studied. Restricted gene flow and inbreeding occur in the present range of distribution of the species. Microsatellite loci showed a strong incidence of failed amplifications, which we attribute to the marked intraspecies variability that hampered the application of these highly specific markers. Our results show important genetic variability at all levels studied, from within populations to between basins, possibly coupled to speciation processes. This variability is attributable to restricted gene flow among populations due to short-distance dispersal of the larvae.     

Malaria vectors in the municipality of Serra do Navio, State of Amapá, Amazon Region, Brazil

We conducted a survey to determine the vectors of malaria in six localities of Serra do Navio municipality, State of Amapá, from 1990 to 1991. Malaria infection rates of 29.3%, 6.2% and 20.4% were detected by human blood smears in Col?nia Agua Branca, Porto Terezinha and Arrependido, respectively. There was no malaria infection detected in Serra do Navio. Fifteen species were identified among 3,053 anopheline mosquitoes collected by human bait and 64.4% were identified as Anopheles albitarsis s.l., 16.7% An. braziliensis, 9.5% An. nuneztovari and 5.8% An. triannulatus. An. darlingi, the main vector of malaria in the Amazon region of Brazil, was scare. Using enzyme-linked immunosorbent assay (ELISA), a total positive rate of 0.8% (23/2876) was found for six species: fifteen An. albitarsis s.l., four An. nuneztovari, and one of each: An. braziliensis, An. triannulatus, An. oswaldoi and An. rangeli. Nine of 23 positive mosquitoes were infected with Plasmodium malariae, eight with P. vivax VK210, three with P. vivax VK247 and three with P. falciparum. Since An. albitarsis s.l. was collected feeding on humans, was present in the highest density and was positive by ELISA for malaria sporozoites, it probably plays an important role in malaria transmission in this area.     

Genetic diversity and population connectivity of the Asian green mussel Perna viridis in South China Sea,inferred from mitochondria DNA markers

The Asian green mussel Perna viridis is ecologically and economically important in the coastal regions of China. In order to characterize the genetic diversity and population connectivity of P. viridis in South China Sea, a 664 bp region of mitochondrial COI gene and a 293 bp region of 16S rRNA gene were sequenced and analyzed for 78 and 92 individuals from four populations in South China Sea, respectively. A total of 15 haplotypes were defined by 14 variable nucleotide sites in COI gene, and 7 haplotypes by 6 variable nucleotide sites in 16S rRNA gene. High haplotype diversity and low nucleotide diversity were observed in COI gene, while moderate haplotype diversity and low nucleotide diversity were observed in 16S rRNA gene. Pairwise F_ST values of COI gene were all negative and those of 16S rRNA gene ranged from −0.01409 to 0.10289. The results showed that no significant genetic divergence (or shallow genetic structure) and high levels of population connectivity among the four populations of P. viridis in South China Sea.     

Phylogeography of Hyalomma (Euhyalomma) lusitanicum (Acarina,Parasitiformes, Ixodidae) in Andalusia based on mitochondrial cytochrome oxidase I gene

The genetic population structure relationships of Hyalomma (Euhyalomma) lusitanicum in Andalusia (the south of the Iberian Peninsula) were examined using mtDNA sequence data from 887 bp of cytochrome oxidase subunit I (COI) gene. The sequence for the COI region was determined for 84 individuals collected in several localities of Andalusia, and 10 for other localities (i.e., five from Toledo, central Iberian Peninsula, four from Sicily (Italy) and one from Canary Island). Seventeen haplotypes were detected, including 27 polymorphic sites. The number of amino acid substitutions per site from mean diversity calculations for the entire population was 0.017. AMOVA analysis revealed a low gene flow that characterises the genetic population structure of this species in South Iberian Peninsula, with a haplotype diversity (h) value of 0.815. No geographically induced differentiation was observed, and separate evolutionary units were not detected. Our results indicate low genetic diversity across the geographical range of H. lusitanicum tick in Andalusia. Our data do not show any genetic discontinuity between the tick populations studied, including specimens from Canary Island and Sicily (Italy).

     

Molecular characterization of Gonatocerus tuberculifemur (Ogloblin) (Hymenoptera: Mymaridae), a prospective Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) biological control candidate agent from South America: divergent clades

We genetically characterized the prospective South American egg parasitoid candidate, Gonatocerus tuberculifemur, of the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, for a neoclassical biological control program in California. Two molecular methods, inter-simple sequence repeat-polymerase chain reaction DNA fingerprinting and a phylogeographic approach inferred from the mitochondrial cytochrome oxidase subunit I gene (COI), were utilized. Five geographic populations from South America were analyzed; in addition, a phylogenetic analysis was performed with several named and one unnamed Gonatocerus species using the COI gene. DNA fingerprinting demonstrated a fixed geographic banding pattern difference in the population from San Rafael, Mendoza Province, Argentina. The COI analysis uncovered haplotype or geographic structure in G. tuberculifemur. A neighbour-joining distance (NJ) and a single most parsimonious tree (MP) clustered the populations into two well-supported distinct clades with strong bootstrap values (97-99% and 92-99%, respectively) with populations from San Rafael clustering into clade 2 and the rest of the populations clustering into clade 1. No haplotype sharing was observed between individuals from the two clades. Phylogenetic analyses performed by NJ and MP methods with 15 Gonatocerus species confirmed species boundaries and again uncovered two distinct clades in G. tuberculifemur with strong bootstrap support (95-100% and 68-100%, respectively). However, the NJ tree supported the morphologically defined relationships better than the MP tree. The molecular evidence in the present study is suggestive of a species level divergence. Because G. tuberculifemur is under consideration as a potential biological control agent for GWSS in California, understanding cryptic variation in this species is critical.     

Multiple origins of stone loach,Barbatula barbatula (Teleostei: Nemacheilidae), in Sweden based on mitochondrial DNA

Stone loach, Barbatula barbatula, occurs in scattered localities in Sweden. Some of the populations have usually been considered as feral descendants of escaped 18th Century pond stock, but historical documentation is inconclusive. Using the mitochondrial COI gene as a marker, we analyzed specimens from seven Swedish localities. One of the middle Swedish localities, in Stockholm, belongs to a haplotype found also in Poland and Lithuania. Two other samples, from near Nyköping and Lake Hjälmaren, belong to a haplotype found in northeastern Europe (Finland, Estonia, Latvia, Volga basin in Russia). Those two Swedish populations are probably descendants from at least two introductions, probably for pond rearing for human consumption. Samples from Skåne and Halland in southern Sweden belong to the haplotype found in Denmark, northern Germany and Poland; and whereas it remains possible that they also represent feral populations, they may be naturally occurring, having reached Sweden during the Ancylus period, about 8,000–10,000 years ago. A recently discovered population from the central South Swedish Highlands belongs to a mainly southeastern European haplotype. It probably represents a release of imported aquarium specimens or live bait carried by sport fishing tourists.     

淮河水系日本沼虾群体遗传结构和系统演化的线粒体COI序列分析

测定了淮河水系17个日本沼虾(Macrobrachium nipponense)野生群体共248个个体的线粒体细胞色素氧化酶亚基I(COI)部分序列,获得623 bp核苷酸片段,包括48个变异位点,定义了31个单倍型,共享单倍型有12个,整体单倍型多样性和平均核苷酸多样性均处于中间水平。AMOVA分析表明,17个群体间的遗传分化系数Fst=0.041 3(P0.05),群体间遗传分化较小。Kimura 2-paramter遗传距离在五河与焦岗湖、花家湖及瓦埠湖群体间最大,为0.014,在高邮和邵伯湖群体之间最小,为0.003。MP系统树与单倍型进化网络关系图具有较高的一致性,31个单倍型被分为3个进化枝,其中一个进化枝主要以下游群体为主,另外2个进化枝主要以中游群体为主。群体中性检验、错配分析表明,淮河日本沼虾近期曾经历过种群扩张。     

Two Corbicula (Corbiculidae: Bivalvia) mitochondrial lineages are widely distributed in Asian freshwater environment

We investigated the biogeography of Asian Corbicula using mitochondrial gene sequence variation for Corbicula members sampled from 24 localities of eight Asian regions. A total of 210 individuals were genetically characterized by examining sequence variations of a 614 bp fragment of the mitochondrial cytochrome oxidase I (COI) gene. Phylogenetic analyses of the COI dataset revealed that Corbicula members are subdivided into two well-supported clades: estuarine and freshwater. A robust dichotomy between the Japanese/Korean (Corbicula japonica) and Chinese (Corbicula fluminalis) estuarine forms was evident, suggesting that these two regional populations represent a deep phylogeographic split. Our mitochondrial gene tree showed that among the freshwater members, two Corbicula mitochondrial lineages are the most common, having attained extensive geographic distribution in the Asian freshwater environment. While the present study provides significant biogeographic information on Asian Corbicula, a comprehensive phylogenetic study by cross-referencing the mitochondrial-based Corbicula phylogeny with nuclear gene data is required to fully understand the evolutionary origin(s) of triploidy/clonality in this genus.     

Phylogenetic and phylogeographic inferences based on two DNA markers reveal geographic structure of the orangeclaw hermit crab Calcinus tibicen (Anomura: Diogenidae) in the western Atlantic

The hermit crab Calcinus tibicen has an extensive distribution along the coasts of the western Atlantic Ocean, from the USA to southern Brazil. The present study aimed to test the hypothesis of the presence of phylogeographic structure of C. tibicen throughout its distribution, evaluating the genetic and morphological variabilities of the species. The molecular data, genetic diversity and demographic history were inferred from 16 different localities for the mitochondrial genes cytochrome oxidase subunit I (COI) (75 specimens) and 16S rRNA (20). The morphological information was based on 99 individuals from the same 16 localities. Results were obtained by the construction of haplotype networks and a phylogenetic tree, an Analysis of Molecular Variance (AMOVA), Tajima’s D and Fu’s Fs tests, pairwise mismatch distributions under the sudden expansion model, and by estimating divergence times. The haplotype networks show the existence of two genetically well-defined groups that do not share haplotypes. The phylogenetic tree and the matrix of genetic distances suggest lack of gene between North and South Atlantic groups, also corroborated by AMOVA for the COI gene. The genetic distance can be attributed to at least two ecological factors: the existence of a physiological barrier caused by the Amazon River freshwater plume, and the effect of the bifurcation of the westerly flowing South Atlantic oceanic currents creating a gap along the Brazilian coastline. Demographic history analyses suggest that C. tibicen experienced population expansion, probably after the Amazon River changed its direction. Despite the molecular evidence for the existence of a phylogeographic structure, no morphological pattern for each genetic group was observed. Therefore, based on the described molecular differences and the ecological and historical factors suggested, the question arises whether a cryptic species should be recognized for some of the populations of C. tibicen.     

Molecular population genetics of the malaria vector Anopheles darlingi in Central and South America

To analyze the genetic relatedness and phylogeographic structure of Anopheles darlingi from 19 localities throughout Central and South America, we used a minimum spanning network, diversity measures, differentiation, neutrality tests, and mismatch distribution with mitochondrial cytochrome oxidase subunit I (COI) sequences. All the Central American haplotypes were separated by seven mutational steps from the South American haplotypes and the FST distance-based neighbor-joining tree showed a primary division between Central and South America, evidence for a putative gene pool division. More ancestral and diverse haplotypes were found in Amazonian and southern Brazil populations, suggesting that Central American populations may have originated in South America. The patterns of the mtDNA haplotype diversity and five of six tests for equilibrium implicate demographic expansion in the South American populations as the historical structure, but mismatch distribution depicts populations at mutation drift equilibrium (MDE). In South America, the departure from equilibrium was consistent with an expansion that occurred during the Pleistocene.     

Biting and resting behaviour of anophelines in western Venezuela and implications for control of malaria transmission

A study was conducted in three villages in western Venezuela between February 1988 and October 1989 to determine the biting and resting activity of anophelines in relation to human habits, rainfall and fenitrothion spraying of houses. Mosquitoes were collected landing on humans inside and outside experimental huts throughout the night. Only three mosquitoes were found resting in the huts in the morning, but 2470 were collected resting on vegetation in the early morning. The collections yielded eleven species of anopheline, the most abundant being Anopheles nuneztovari, comprising over 75% of the total anophelines collected, followed by An.triannulatus, An.albitarsis s.l. and An.oswaldoi. The four most abundant species showed different diel patterns of biting. For An.nuneztovari the peak of activity was close to midnight indoors and outdoors, for An.triannulatus between 19.00 and 20.00 hours outdoors, for An.albitarsis mainly before midnight indoors and outdoors and for An.oswaldoi outdoors at 19.00 hours, with an additional smaller peak indoors at midnight. Most of the human population use bednets, go to bed before 22.00 hours and are therefore most exposed to mosquitoes that bite outdoors early in the night. Fenitrothion house-spraying failed to prevent large mosquito populations developing in the wet season, presumably because of their exophilic resting habits. The possible advantages of impregnation of existing bednets with pyrethroids, and provision of nets for people who do not have them, are discussed.     

Genetic variation and population structure of the carpet shell clam <Emphasis Type="Italic">Ruditapes decussatus</Emphasis> along the Tunisian coast inferred from mtDNA and ITS1 sequence analysis

Surveys of allozyme polymorphisms in the carpet shell clam Ruditapes decussatus have revealed sharp genetic differentiation of populations. Analysis of population structure in this species has now been extended to include nuclear and mitochondrial genes. A partial sequence of a mitochondrial COI gene and of the internal transcribed spacer region (ITS-1) were used to study haplotype distribution, the pattern of gene flow, and population genetic structure of R. decussatus. The samples were collected from twelve populations from the eastern and western Mediterranean coasts of Tunisia, one from Concarneau and one from Thau. A total of twenty and twenty-one haplotypes were detected in the examined COI and ITS1 regions respectively. The study revealed higher levels of genetic diversity for ITS1 compared to COI. The analysis of haplotype frequency distribution and molecular variation indicated that the majority of the genetic variation was distributed within populations (93% and 86% for COI and ITS1 respectively). No significant differentiation was found among eastern and western groups on either side of the Siculo-Tunisian strait. However, distinct and significant clinal changes in haplotypes frequencies between eastern and western samples were found at the most frequent COI haplotype and at three out of five major ITS1 haplotypes. These results suggest the relative importance of historical processes and contemporary hydrodynamic features on the observed patterns of genetic structure.     

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.