Molecular variation and phylogeny of members of the Minimus Group of Anopheles subgenus Cellia (Diptera: Culicidae)

Intra‐ and interspecific molecular variation were investigated in four members of the Minimus Group of Anopheles subgenus Cellia: An. aconitus, An. varuna, An. minimus A and An. minimus C. DNA sequence divergence between these species at a mitochondrial locus (cytochrome oxidase II) and at three nuclear loci (ITS2 and D3 regions of rDNA and guanylate cyclase) is reported. The data confirm the presence of two cryptic species, A and C, within An. minimus and provide evidence for the existence of a third species. Anopheles minimus A and C are estimated to have diverged 0.57–1.5 million years ago. The discrepancy observed using the guanylate cyclase intron, which is the fastest evolving region known in the Gambiae Complex but is relatively slowly evolving in the Minimus Group, is discussed. The long‐term effective population sizes of An. minimus A and C are estimated to be in their millions, with that of species A being approximately twice the size of species C. This implies that An. minimus C has a much wider distribution than currently recognized, with possible widespread implications for vector control. No evidence was found for population structuring in either species A or C: there was greater variation of mitochondrial haplotypes within than among localities. The phylogenetic relationships of Oriental members of the Myzomyia Series are reconstructed.     

Molecular phylogenetics of the Oriental members of the Myzomyia Series of Anopheles subgenus Cellia (Diptera: Culicidae) inferred from nuclear and mitochondrial DNA sequences

Abstract. The phylogenetic relationships of fifteen Oriental and two Afrotropical taxa of the Myzomyia Series of Anopheles subgenus Cellia and two outgroup species, An. maculatus (Neocellia Series) and An. dirus A (Neomyzomyia Series), were inferred from nucleotide sequences of the entire 685 bp of the mitochondrial cytochrome oxidase subunit II locus (COII) and 372 bp of the third domain (D3) of the 28S rDNA locus, both separately and together. Alignment of the D3 sequences was achieved with the aid of secondary structure comparisons, and the pattern of nucleotide substitution was best explained by the GTR + I + G model for either separate or combined datasets. Maximum likelihood and maximum parsimony analyses robustly identified five monophylies: An. fluviatilis U and T; An. fluviatilis U and T + An. minimus A, C, E and #157 + An. leesoni; An. filipinae + An. mangyanus; An. filipinae + An. mangyanus + An. aconitus; and An. culicifacies A and B. The results confirm the specific status of An. flavirostris, the close relationship of An. leesoni with the Minimus Complex, and the exclusion of An. jeyporiensis, An. culicifacies s.l and An. funestus from the Minimus Group. All of the species classified as members of the Minimus Group on morphological grounds formed a single clade, which comprised two subgroups: the Minimus Subgroup, including An. minimus s.l., An. fluviatilis s.l., An. leesoni and An. flavirostris, and the Aconitus Subgroup, including An. filipinae, An. mangyanus, An. aconitus, An. pampanai and An. varuna. However, these clades are only weakly supported by the present dataset.     

Salivary polytene chromosome mapping of Anopheles (Cellia) subpictus Grassi (Culicidae: Diptera).

With the introduction of molecular taxonomy of mosquitoes, polytene chromosome maps have become indispensable as standard references for locating genes, puffs, and inversion breakpoints of unique DNA sequences. We present a line map and a photomap of the salivary polytene chromosomes of Anopheles (Cellia) subpictus Grassi, an important emerging vector of malaria in India. In addition, we discuss the nature of this species complex consisting of sibling species A, B, C, and D. The comparative study is in relevance to the X chromosome heterozygous inversion differences between 2 allopatric populations of the species and the recognition of 4 X-chromosome inversion genotypes viz: species A-X(+a+b), B-X(ab), C-X(a+b) and D-X(+ab).     

Cladistic analysis of the subgenus Anopheles (Anopheles) Meigen (Diptera: Culicidae) based on morphological characters

In the present study, we used morphological characters to estimate phylogenetic relationships among members of the subgenus Anopheles Meigen. Phylogenetic analyses were carried out for 36 species of Anopheles (Anopheles). An. (Stethomyia) kompi Edwards, An. (Lophopodomyia) gilesi (Peryassú), Bironella hollandi Taylor, An. (Nyssorhynchus) oswaldoi (Peryassú) and An. (Cellia) maculatus Theobald were employed as outgroups. One hundred one characters of the external morphology of the adult male, adult female, fourth-instar larva, and pupa were scored and analyzed under the parsimony criterion in PAUP. Phylogenetic relationships among the series and several species informal groups of Anopheles (Anopheles) were hypothesized. The results suggest that Anopheles (Anopheles) is monophyletic. Additionally, most species groups included in the analysis were demonstrated to be monophyletic.     

Molecular Phylogeny of Neotropical Anopheles (Nyssorhynchus) Albitarsis Species Complex (Diptera: Culicidae)

A phylogeny was reconstructed for four species belonging to the Neotropical Anopheles (Nyssorhynchus) albitarsis complex using partial sequences from the mitochondrial cytochrome oxidase I (COI) and NADH dehydrogenase 4 (ND4) genes and the ribosomal DNA ITS2 and D2 expansion region of the 28S subunit. The basis for initial characterization of each member of the complex was by correlated random amplification of polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Analyses were carried out with and without an outgroup (An.(Nys.) argyritarsis Robineau-Desvoidy) by using maximum parsimony, maximum likelihood, and Bayesian methods. A total evidence approach without the outgroup, using separate models for "fast" (COI and ND4 position 3) and "slow" (rDNA ITS2 and D2, and COI and ND4 position 1) partitions, gave the best supported topology, showing close relationships of An. albitarsis Lynch-Arribálzaga to An. albitarsis B and An. marajoara Galv?o & Damasceno to An. deaneorum Rosa-Freitas. Analyses with the outgroup included showed poorer support, possibly because of a long branch attraction effect caused by a divergent outgroup, which caused one of the An. marajoara specimens to cluster with An. deaneorum in some analyses. The relationship of the above-mentioned result to a separately proposed hypothesis suggesting a fifth species in the complex is discussed.     

Description of a new species Anopheles (Cellia) kosiensis (Diptera: Culicidae) from Zululand, South Africa

ABSTRACT. The Anopheles marshallii group of mosquitoes consists of four chromosomally typed species of which three have already been described and named. The fourth chromosomal species, formally known as species E, is described here as Anopheles kosiensis from Kosi Bay, Zululand, South Africa. A photograph of the X chromosome is given. The taxonomic status of Anopheles pitchfordi , a synonym of marshallii , is discussed.     

Anopheles halophylus,a new species of the subgenus Nyssorhynchus (Diptera: Culicidae) from Brazil

Anopheles halophylus n.sp. is described from Central Brazil. It is distinguished from An. triannulatus (Neiva and Pinto) by morphological and morphometric characters, especially in the male genitalia, larva and egg. Illustrations of the male and female genitalia, egg, larva and pupa, and discussion of the status of names associated with An. triannulatus are provided.     

The subgenus Sabethinus of Sabethes (Diptera: Culicidae)

Abstract. The subgenus Sabethinus of Sabethes is redescribed and defined on the basis of shared morphological features observed in the adult, larval and pupal stages of four known species, two of which are described as new. Each species is described and illustrated in detail, and keys are provided for the identification of the adult and larval stages. The treatment of each species also includes an annotated summary of previous literature, a brief systematic discussion and information on bionomics and distribution. The species comprising the subgenus are Saintermedins, Samelcinonymphe, Saxhyphydes sp.n. and Sakliogenes sp.n.     

Phylogeny and classification of the Culicidae (Diptera)

The generic relationships and higher classification of the family Culicidae are examined on the basis of a phylogenetic analysis. New and traditional morphological characters studied and compared throughout the Culicidae resulted in the acquisition of character data relative to the majority of species within each genus. Polymorphisms and morphological observations are discussed and additional information and illustrations are provided for the majority of characters and their character states. The analysis of seventy-three adult, pupal and fourth-instar larval characters coded for the thirty-eight currently recognized genera of mosquitoes resulted in relationships and groupings which differ significantly from traditional hypotheses. The analysis supports the monophyly of the subfamily Anophelinae and the tribes Culicini and Sabethini. The Anophelinae form the most basal clade of the family. The results indicate that Aedini is a paraphyletic assemblage with respect to the Mansoniini, each of which is monophyletic in itself. The Aedini + Mansoniini form a sister group to the Culicini + Sabethini, with the Aedini and the Culicini placed in ancestral relationships to the Mansoniini and the Sabethini, respectively. Based on the topography of generic relationships among more 'generalized' mosquitoes, the boundaries and relationships of the tribes Aedeomyiini, Uranotaeniini, Ficalbiini, Hodgesiini, Orthopodomyiini and Culisetini appear to be problematic. Relationships between genera of the tribe Aedini are generally poorly resolved due to a significant amount of polymorphism, especially within the genus Aedes as currently defined. There is no support for separate subfamily recognition for the genus Toxorhynchites , which is downgraded to tribal status as a result of the analysis. The results are discussed in relation to previous hypotheses based on subjective inference and cladistic analyses.     

Anopheles (Cellia) epiroticus (Diptera: Culicidae), a new malaria vector species in the Southeast Asian Sundaicus Complex

Anopheles sundaicus species A of the Southeast Asian A. sundaicus complex is formally named Anopheles epiroticus Linton & Harbach based on DNA sequence differentiation of the whole nuclear ITS2 region and a portion of both the cytochrome b and cytochrome c oxidase I mitochondrial genes. Detailed comparative morphological studies of the adult, larval and pupal stages did not reveal any differential or diagnostic differences that reliably distinguish A. epiroticus from A. sundaicus s.s. Information is provided on the bionomics and systematics of the new species.     

A new species of Anopheles (Anopheles) from Namibia (Diptera: Culicidae)

Abstract. The adult, pupa, larva and egg of Anopheles {Anopheles) namibien-sis sp.n. from the Kavango district, Namibia (South West Africa), are described and comparisons made with Anopheles (Anopheles) ziemanni Griinberg. A photomap of the polytene chromosomes from the salivary glands of the fourth stage larvae is presented.     

Systematic studies on Anopheles galvaoi Causey,Deane & Deane from the subgenus Nysssorhynchus blanchard (Diptera: Culicidae)

Anopheles galvaoi, a member of the subgenus Nyssorhynchus, is redescribed based on morphological characters of the adults male and female, fourth-instar larva and pupa. Female, male genitalia, larval and pupal stages are illustrated. Data about medical importance, bionomics, and distribution are given based on literature records. Adult female of An. galvaoi can be easily misidentified as An. benarrochi Gabald n and An. aquasalis Curry. A few characters are indicated for identifying female and immatures of An. galvaoi. Phylogenetic relationships among An. galvaoi and six other species of the Oswaldoi Subgroup are estimated using COII mtDNA and ITS2 rDNA gene sequences. Lectotype of An. galvaoi, an adult female from Rio Branco, State of Acre, is invalidated.     

Satellite DNA of Anopheles stephensi liston (Diptera: Culicidae)

Four satellite DNAs in the Anopheles stephensi genome have been defined on the basis of their banding properties in Hoechst 33258-CsCl density gradients. Two of these satellites, satellites I and II, are visible on neutral CsCl density gradients as a light density peak forming approximately 15% of total cellular DNA. Hoechst-CsCl density gradient profiles of DNA extracted from polytene tissues indicates that these satellites are underreplicated in larval salivary gland cells and adult female Malpighian tubules and possibly also in ovarian nurse cells. The chromosomal location of satellite I on mitotic and polytene chromosomes has been determined by in situ hybridisation. Sequences complementary to satellite I are present in approximately equal amounts on a heterochromatic arm of the X and Y chromosomes and are also present, in smaller amounts, at the centromere of chromosome 3. A quantitative analysis of the in situ hybridisation experiments indicates that sequences complementary to satellite I at these two sites differ in their replicative behaviour during polytenisation: heterosomal satellite I sequences are under-replicated relative to chromosome 3 sequences in polytene larval salivary gland and ovarian nurse cell nuclei.     

基于rDNA-ITS2序列的中国按蚊属塞蚊亚属部分种类的系统发育研究(双翅目:蚊科)

应用rDNA-ITS2序列,采用邻接法(NJ)、最大简约法(MP)和最大似然法(ML),以按蚊亚属Anopheles的中华按蚊An.(An.)sinensis和赫坎按蚊An.(An.)hyrcanus为外群,对采自中国的按蚊属Anopheles塞蚊亚属Cellia 21种蚊进行了系统发育分析.结果表明:rDNA-ITS...     

Karyotype of Brazilian Anopheles albitarsis sensu lato (Diptera:Culicidae)

Anopheles (Nyssorhynchus) albitarsis sensu lato is an important malaria vector in Brazil, especially in the Brazilian Amazon region. Chromosome preparations of fourth-instar larvae of A. albitarsis from Iranduba and Coari (AM) and Ilha Comprida (SP) were analyzed for karyotype determination and to improve cytogenetic identification of this species. Anopheles albitarsis possesses 2n = 6 chromosomes, with two pairs (submetacentric and metacentric) of autosomes and one pair of sex chromosomes, with X-Y dimorphism. The sex pair is homomorphic and acrocentric in females and heteromorphic in males, with a punctiform Y chromosome. Somatic pairing was detected in the prometaphase and metaphase chromosomes of the three A. albitarsis populations. Apparently, sex chromosome evolution in the Culicidae does not function as does evolution in the Culicidae, since it occurs in the subfamily Anophelinae, which possesses heteromorphic sex chromosomes and is regarded as primitive, based on several criteria. These karyotype data on the albitarsis complex reinforce the hypothesis that sex chromosome evolution in the subfamily Anophelinae is conserved, and the variation revealed in the mean size of chromosomes in three populations indicates that selective pressure in these populations is occurring only at a genetic level.     

Morphological and molecular identity of Anopheles (Cellia) sundaicus (Diptera: Culicidae), the nominotypical member of a malaria vector species complex in Southeast Asia

A neotype for Anopheles (Cellia) sundaicus (Rodenwaldt), the nominotypical member of a malaria vector species complex, is selected from the progeny of a female collected at Pandan Beach near Lundu, Sarawak, Malaysia. Siblings of the neotype and other topotypic specimens are used to characterize and fix the morphological and molecular identity of the species as a foundation for systematic studies of the species complex. The species is described and illustrated in the adult, pupal and larval stages, and DNA sequence data are included for the mitochondrial cytochrome c oxidase I (COI) gene and the nuclear ITS‐2 region.