- INTRODUCTION
- MATERIALS AND METHODS
- Study site and collection method.
- Laboratory rearing of wild-caught material.
- Molecular identification of field material.
- Sequencing and primer design for unidentified Malawi samples.
- Application of the MalaF-specific PCR primer.
- Cytogenetics.
- Cross-mating studies.
- Enzyme-linked immunosorbent assay for sporozoite detection.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Gillies MT, de Meillon B, 1968. The Anophelinae of Africa South of the Sahara. Publications of the South African Institute for Medical Research, no. 54. Johannesburg, South Africa.
-
2
Gillies MT, Coetzee M, 1987. A Supplement to the Anophelinae of Africa South of the Sahara (Afrotropical Region). Publications of the South African Institute for Medical Research, no 55. Johannesburg, South Africa.
-
3
Harbach RE, 2004. The classification of genus Anopheles (Diptera: Culicidae): a working hypothesis of phylogenetic relationships. Bull Entomol Res 94 :537–553.
-
4
Garros C, Harbach RE, Manguin S, 2005. Morphological assessment and molecular phylogenetics of the Funestus and Minimus Groups of Anopheles (Cellia). J Med Entomol 42 :522–536.
-
5
Hackett BJ, Gimnig J, Guelbeogo W, Costantini C, Koekemoer LL, Coetzee M, Collins FH, Besansky NJ, 2000. Ribosomal DNA internal transcribed spacer (ITS2) sequences differentiate Anopheles funestus and An. rivulorum, and uncover a cryptic taxon. Insect Mol Biol 9 :369–374.
-
6
Couhet A, Simard F, Toto J-C, Kengne P, Coetzee M, Fontenille D, 2003. Species identification within the Anopheles funestus group of malaria vectors in Cameroon and evidence for a new species. Am J Trop Med Hyg 69 :200–205.
-
7
Green CA, 1982. Cladistic analysis of chromosome data (Anopheles (Cellia) Myzomyia). J Hered 73 :2–11.
-
8
Wilkes TJ, Matola YG, Charlwood JD, 1996. Anopheles rivulorum, a vector of human malaria in Africa. Med Vet Entomol 10 :108–110.
-
9
De Meillon B, Van Eeden GJ, Coetzee L, Coetzee M, Meiswinkel R, Du Toit CL, Hansford CF, 1977. Observations on a species of the Anopheles funestus subgroup, a sustpected exophilic vector of malaria parasites in North Eastern Transvaal, South Africa. Mosq News 37 :657–661.
-
10
Evans AM, Symes CB, 1937. Anopheles funestus and its allies in Kenya. Ann Trop Med Parasitol 31 :105–111.
-
11
Evans AM, 1938. Mosquitoes of the Ethiopian Region. London: Bernard Quartich Ltd., Dulau and Co. Ltd., and The Oxford University Press.
-
12
De Meillon B, 1947. New records and species of biting insects from the Ethiopian Region II. J Entomol Soc S Afr 10 :110–124.
-
13
Green CA, Hunt RH, 1980. Interpretations of variation in ovarian polytene chromosomes of Anopheles funestus Giles, A. parensis Gillies and A. aruni? Genetica 51 :187–195.
-
14
Lochouarn L, Dia I, Boccolini D, Coluzzi M, Fontenille D, 1998. Bionomical and cytogenetic heterogeneities of Anopheles funestus in Senegal. Trans R Soc Trop Med Hyg 92 :607–612.
-
15
Costantini C, Sagnon, N’F, Ilboudo-Sanogo E, Coluzzi M, Boccolini D, 1999. Chromosomal and bionomic heterogeneities suggest incipient speciation in Anopheles funestus from Burkina Faso. Parassitologia 41 :595–611.
-
16
Koekemoer LL, Kamau L, Hunt RH, Coetzee M, 2002. A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group. Am J Trop Med Hyg 6 :804–811.
-
17
Collins FH, Mendez MA, Rasmussen MO, Meheffey PC, Besansky NJ, Finnerty V, 1987. A ribosomal RNA gene probes differentiates members of the Anopheles gambiae complex. Am J Trop Med Hyg 37 :37–41.
-
18
Litvaitis MK, Nunn G, Thomas WK, Kocher TD, 1994. A molecular approach for the identification of Meiofaunal turbellarians (Platyhelminthes: Turbellaria). Mar Biol 158 :17–35.
-
19
Hunt RH, 1973. A cytological technique for the study of Anopheles gambiae complex. Parassitologia 15 :137–139.
-
20
Davidson G, Paterson H, Coluzzi M, Mason G, Micks D, 1967. The Anopheles gambiae complex. Wright J, Pal R, eds. Genetics of Insect Vectors of Disease. London: Elsevier Publishing Company, 211–250.
-
21
Wirtz RA, Zavala F, Charoenvit Y, Cambell GH, Burkot TR, Schneider I, Esser KM, Beaudoin RL, Andre GR, 1987. Comparative testing of Plasmodium falciparum sporozoite monoclonal antibodies for ELISA development. Bull World Health Organ 65 :39–45.
-
22
Singh OP, Chandra D, Nanda N, Raghavendra K, Sunil S, Sharma SK, Dua VK, Subbarao SK, 2004. Differentiation of members of the Anopheles fluviatilis species complex by an allele-specific polymerase chain reaction based on 28S ribosomal DNA sequences. Am J Trop Med Hyg 70 :27–32.
-
23
Sharpe RG, Harbach RE, Butlin RK, 2000. Molecular variation and phylogeny of the members of the Minimus group of Anopheles subgenus Cellia (Diptera: Culicidae). Syst Entomol 25 :263–272.
-
24
Paskewitz SM, Wesson DM, Collins FH, 1993. The internal transcribed spacers of ribosomal DNA in five members of the Anopheles gambiae species complex. Insect Mol Biol 2 :247–257.
-
25
Frizzi G, 1947. Salivary gland chromosomes of Anopheles. Nature 160 :226–227.
-
26
Frizzi G, 1953. Étude cytog’enetique d’anopheles maculipennis en Italie. Extension des recherches ‘a d’autres esp’eces d’anoph’eles. Bull World Health Organ 9 :335–344.
-
27
Coluzzi M, Sabatini A, 1967. Cytogenetic observations on species A and B of the Anopheles gambiae complex. Parassitologia 9 :73–88.
-
28
Coluzzi M, Sabatini A, 1968. Cytogenetic observations on species C of the Anopheles gambiae complex. Parassitologia 10 :155–166.
-
29
Coluzzi M, Sabatini A, 1969. Cytogenetic observations on the salt water species Anopheles merus and Anopheles melas, of the Gambiae complex. Parassitologia 11 :155–166.
-
30
Thongwat D, Morgan K, O’Loughlin SM, Walton C, Choochote W, Somboon P, 2008. Crossing experiments supporting the specific status of Anopheles maculatus chromosomal form K. J Am Mosq Control Assoc 24 :194–202.
-
31
Coetzee M, Estrada-Franco JG, Wunderlich CA, Hunt RH, 1999. Cytogenetic evidence for a species complex within Anopheles pseudopunctipennis Theobald (Diptera: Culicidae). Am J Trop Med Hyg 60 :649–653.
-
32
Somboon P, Thongwat D, Choochote W, Walton C, Takagi M, 2005. Crossing experiments of Anopheles minimus species A and putative species E. J Am Mosq Control Assoc 21 :5–9.
-
33
Kaiser PE, 1988. Cytotaxonomy as a tool for identification of siblings of the Anopheles quadrimaculatus complex. Fla Entomol 71 :311–323.
-
34
Foley DH, Bryan JH, 1991. Anopheles annulipes Walker (Diptera: Culicidae) at Griffith New South Wales. 1. Two sibling species in sympatry. Aust J Entomol 30 :109–112.
-
35
Hunt RH, Coetzee M, Fettene M, 1998. The Anopheles gambiae complex: a new species from Ethiopia. Trans R Soc Trop Med Hyg 92 :231–235.
-
36
Garros C, Koekemoer LL, Kamau L, Awolola TS, Van Bortel W, Coetzee M, Coosemans M, Manguin S, 2004. Restriction fragment length polymorphism method for the identification of major African and Asian malaria vectors within the Anopheles funestus and An. minimus groups. Am J Trop Med Hyg 70 :260–265.
-
37
Michel AP, Ingracsi MJ, Schemerhorn BJ, Kern M, Le Goff G, Coetzee M, Elissa N, Fontenille D, Vulule J, Lehmann T, Sagnon N’F, Costantini C, Besansky NJ, 2005. Rangewide population genetics structure of the African malaria vector Anopheles funestus. Mol Ecol 14 :4235–4248.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 486 | 296 | 13 |
| Full Text Views | 384 | 11 | 0 |
| PDF Downloads | 113 | 8 | 0 |
A New Species Concealed by Anopheles funestus Giles, a Major Malaria Vector in Africa
Belinda L. Spillings
Belinda L. Spillings
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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Basil D. Brooke
Basil D. Brooke
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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Lizette L. Koekemoer
Lizette L. Koekemoer
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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John Chiphwanya
John Chiphwanya
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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Maureen Coetzee
Maureen Coetzee
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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Richard H. Hunt
Richard H. Hunt
Vector Control Reference Unit, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa; Division of Virology and Communicable Disease Surveillance, School of Pathology of the University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa; School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa; National Research Foundation Chair in Medical Entomology and Vector Control, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa; National Malaria Control Programme, Lilongwe, Malawi
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The major malaria vector Anopheles funestus belongs to a group of morphologically similar species that are commonly distinguished from one another through the use of chromosomal and molecular techniques. Indoor resting collections of mosquitoes from Malawi were initially identified as An. funestus by morphology, but failed to have this confirmed by the species-specific polymerase chain reaction assay. Sequence analysis of the internal transcribed spacer region 2 identified variations within the An. funestus-specific primer binding site and showed a sequence variation of 4.5% compared with An. funestus. Domain 3 analysis showed sequence variation of 1.5% from An. funestus. Cytogenetic analysis of the polytene chromosome banding arrangements showed that the specimens were homosequential with An. funestus, with fixed inverted arrangements of the 3a, 3b, and 5a inversions commonly polymorphic in An. funestus. The chromosomes of hybrid females showed levels of asynapsis typical of inter-species crosses. These molecular and cytogenetic observations support the conclusion that this Malawi population is a new species and it has provisionally been named An. funestus-like.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Study site and collection method.
- Laboratory rearing of wild-caught material.
- Molecular identification of field material.
- Sequencing and primer design for unidentified Malawi samples.
- Application of the MalaF-specific PCR primer.
- Cytogenetics.
- Cross-mating studies.
- Enzyme-linked immunosorbent assay for sporozoite detection.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Gillies MT, de Meillon B, 1968. The Anophelinae of Africa South of the Sahara. Publications of the South African Institute for Medical Research, no. 54. Johannesburg, South Africa.
-
2
Gillies MT, Coetzee M, 1987. A Supplement to the Anophelinae of Africa South of the Sahara (Afrotropical Region). Publications of the South African Institute for Medical Research, no 55. Johannesburg, South Africa.
-
3
Harbach RE, 2004. The classification of genus Anopheles (Diptera: Culicidae): a working hypothesis of phylogenetic relationships. Bull Entomol Res 94 :537–553.
-
4
Garros C, Harbach RE, Manguin S, 2005. Morphological assessment and molecular phylogenetics of the Funestus and Minimus Groups of Anopheles (Cellia). J Med Entomol 42 :522–536.
-
5
Hackett BJ, Gimnig J, Guelbeogo W, Costantini C, Koekemoer LL, Coetzee M, Collins FH, Besansky NJ, 2000. Ribosomal DNA internal transcribed spacer (ITS2) sequences differentiate Anopheles funestus and An. rivulorum, and uncover a cryptic taxon. Insect Mol Biol 9 :369–374.
-
6
Couhet A, Simard F, Toto J-C, Kengne P, Coetzee M, Fontenille D, 2003. Species identification within the Anopheles funestus group of malaria vectors in Cameroon and evidence for a new species. Am J Trop Med Hyg 69 :200–205.
-
7
Green CA, 1982. Cladistic analysis of chromosome data (Anopheles (Cellia) Myzomyia). J Hered 73 :2–11.
-
8
Wilkes TJ, Matola YG, Charlwood JD, 1996. Anopheles rivulorum, a vector of human malaria in Africa. Med Vet Entomol 10 :108–110.
-
9
De Meillon B, Van Eeden GJ, Coetzee L, Coetzee M, Meiswinkel R, Du Toit CL, Hansford CF, 1977. Observations on a species of the Anopheles funestus subgroup, a sustpected exophilic vector of malaria parasites in North Eastern Transvaal, South Africa. Mosq News 37 :657–661.
-
10
Evans AM, Symes CB, 1937. Anopheles funestus and its allies in Kenya. Ann Trop Med Parasitol 31 :105–111.
-
11
Evans AM, 1938. Mosquitoes of the Ethiopian Region. London: Bernard Quartich Ltd., Dulau and Co. Ltd., and The Oxford University Press.
-
12
De Meillon B, 1947. New records and species of biting insects from the Ethiopian Region II. J Entomol Soc S Afr 10 :110–124.
-
13
Green CA, Hunt RH, 1980. Interpretations of variation in ovarian polytene chromosomes of Anopheles funestus Giles, A. parensis Gillies and A. aruni? Genetica 51 :187–195.
-
14
Lochouarn L, Dia I, Boccolini D, Coluzzi M, Fontenille D, 1998. Bionomical and cytogenetic heterogeneities of Anopheles funestus in Senegal. Trans R Soc Trop Med Hyg 92 :607–612.
-
15
Costantini C, Sagnon, N’F, Ilboudo-Sanogo E, Coluzzi M, Boccolini D, 1999. Chromosomal and bionomic heterogeneities suggest incipient speciation in Anopheles funestus from Burkina Faso. Parassitologia 41 :595–611.
-
16
Koekemoer LL, Kamau L, Hunt RH, Coetzee M, 2002. A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group. Am J Trop Med Hyg 6 :804–811.
-
17
Collins FH, Mendez MA, Rasmussen MO, Meheffey PC, Besansky NJ, Finnerty V, 1987. A ribosomal RNA gene probes differentiates members of the Anopheles gambiae complex. Am J Trop Med Hyg 37 :37–41.
-
18
Litvaitis MK, Nunn G, Thomas WK, Kocher TD, 1994. A molecular approach for the identification of Meiofaunal turbellarians (Platyhelminthes: Turbellaria). Mar Biol 158 :17–35.
-
19
Hunt RH, 1973. A cytological technique for the study of Anopheles gambiae complex. Parassitologia 15 :137–139.
-
20
Davidson G, Paterson H, Coluzzi M, Mason G, Micks D, 1967. The Anopheles gambiae complex. Wright J, Pal R, eds. Genetics of Insect Vectors of Disease. London: Elsevier Publishing Company, 211–250.
-
21
Wirtz RA, Zavala F, Charoenvit Y, Cambell GH, Burkot TR, Schneider I, Esser KM, Beaudoin RL, Andre GR, 1987. Comparative testing of Plasmodium falciparum sporozoite monoclonal antibodies for ELISA development. Bull World Health Organ 65 :39–45.
-
22
Singh OP, Chandra D, Nanda N, Raghavendra K, Sunil S, Sharma SK, Dua VK, Subbarao SK, 2004. Differentiation of members of the Anopheles fluviatilis species complex by an allele-specific polymerase chain reaction based on 28S ribosomal DNA sequences. Am J Trop Med Hyg 70 :27–32.
-
23
Sharpe RG, Harbach RE, Butlin RK, 2000. Molecular variation and phylogeny of the members of the Minimus group of Anopheles subgenus Cellia (Diptera: Culicidae). Syst Entomol 25 :263–272.
-
24
Paskewitz SM, Wesson DM, Collins FH, 1993. The internal transcribed spacers of ribosomal DNA in five members of the Anopheles gambiae species complex. Insect Mol Biol 2 :247–257.
-
25
Frizzi G, 1947. Salivary gland chromosomes of Anopheles. Nature 160 :226–227.
-
26
Frizzi G, 1953. Étude cytog’enetique d’anopheles maculipennis en Italie. Extension des recherches ‘a d’autres esp’eces d’anoph’eles. Bull World Health Organ 9 :335–344.
-
27
Coluzzi M, Sabatini A, 1967. Cytogenetic observations on species A and B of the Anopheles gambiae complex. Parassitologia 9 :73–88.
-
28
Coluzzi M, Sabatini A, 1968. Cytogenetic observations on species C of the Anopheles gambiae complex. Parassitologia 10 :155–166.
-
29
Coluzzi M, Sabatini A, 1969. Cytogenetic observations on the salt water species Anopheles merus and Anopheles melas, of the Gambiae complex. Parassitologia 11 :155–166.
-
30
Thongwat D, Morgan K, O’Loughlin SM, Walton C, Choochote W, Somboon P, 2008. Crossing experiments supporting the specific status of Anopheles maculatus chromosomal form K. J Am Mosq Control Assoc 24 :194–202.
-
31
Coetzee M, Estrada-Franco JG, Wunderlich CA, Hunt RH, 1999. Cytogenetic evidence for a species complex within Anopheles pseudopunctipennis Theobald (Diptera: Culicidae). Am J Trop Med Hyg 60 :649–653.
-
32
Somboon P, Thongwat D, Choochote W, Walton C, Takagi M, 2005. Crossing experiments of Anopheles minimus species A and putative species E. J Am Mosq Control Assoc 21 :5–9.
-
33
Kaiser PE, 1988. Cytotaxonomy as a tool for identification of siblings of the Anopheles quadrimaculatus complex. Fla Entomol 71 :311–323.
-
34
Foley DH, Bryan JH, 1991. Anopheles annulipes Walker (Diptera: Culicidae) at Griffith New South Wales. 1. Two sibling species in sympatry. Aust J Entomol 30 :109–112.
-
35
Hunt RH, Coetzee M, Fettene M, 1998. The Anopheles gambiae complex: a new species from Ethiopia. Trans R Soc Trop Med Hyg 92 :231–235.
-
36
Garros C, Koekemoer LL, Kamau L, Awolola TS, Van Bortel W, Coetzee M, Coosemans M, Manguin S, 2004. Restriction fragment length polymorphism method for the identification of major African and Asian malaria vectors within the Anopheles funestus and An. minimus groups. Am J Trop Med Hyg 70 :260–265.
-
37
Michel AP, Ingracsi MJ, Schemerhorn BJ, Kern M, Le Goff G, Coetzee M, Elissa N, Fontenille D, Vulule J, Lehmann T, Sagnon N’F, Costantini C, Besansky NJ, 2005. Rangewide population genetics structure of the African malaria vector Anopheles funestus. Mol Ecol 14 :4235–4248.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 486 | 296 | 13 |
| Full Text Views | 384 | 11 | 0 |
| PDF Downloads | 113 | 8 | 0 |