Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia

Anopheles (Nyssorhynchus) benarrochi, An. (N.) oswaldoi, and An. (N.) rangeli are the most common anthropophilic mosquitoes in the southern Colombian state of Putumayo. Adult females are most commonly collected in epidemiological studies, and this stage poses significant problems for correct identif...

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Main Authors: Freddy Ruiz, Martha L Quiñones, Holmes F Erazo, David A Calle, Juan F Alzate, Yvonne-Marie Linton
Format: Article
Language:English
Published: Instituto Oswaldo Cruz, Ministério da Saúde 2005-04-01
Series:Memórias do Instituto Oswaldo Cruz.
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762005000200008
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spelling doaj-b1e07e19f1b34d0d94f4a78123d211e62020-11-24T23:47:59ZengInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz.0074-02761678-80602005-04-01100215516010.1590/S0074-02762005000200008Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern ColombiaFreddy RuizMartha L QuiñonesHolmes F ErazoDavid A CalleJuan F AlzateYvonne-Marie LintonAnopheles (Nyssorhynchus) benarrochi, An. (N.) oswaldoi, and An. (N.) rangeli are the most common anthropophilic mosquitoes in the southern Colombian state of Putumayo. Adult females are most commonly collected in epidemiological studies, and this stage poses significant problems for correct identification, due to overlapping inter-specific morphological characters. Although An. rangeli is easy to identify, the morphological variant of An. benarrochi found in the region and An. oswaldoi are not always easy to separate. Herein we provide a rapid molecular method to distinguish these two species in Southern Colombia. Sequence data for the second internal transcribed spacer (ITS2) region of rDNA was generated for link-reared progeny of An. benarrochi and An. oswaldoi, that had been identified using all life stages. ITS2 sequences were 540 bp in length in An. benarrochi (n = 9) and 531 bp in An. oswaldoi (n = 7). Sequences showed no intra-specific variation and ungapped inter-specific sequence divergence was 6.4%. Species diagnostic banding patterns were recovered following digestion of the ITS2 amplicons with the enzyme Hae III as follows: An. benarrochi (365, 137, and 38 bp) and An. oswaldoi (493 and 38 bp). This polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay provides rapid, accurate, and inexpensive species diagnosis of adult females. This will benefit future epidemiological studies and, as PCR amplification can be achieved using a single mosquito leg, the remaining specimen can be either retained as a morphological voucher or further used in vector incrimination studies. That An. benarrochi comprises a complex of at least two species across Latin America is discussed.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762005000200008Anopheles benarrochi BAn. oswaldoiNyssorhynchusinternal transcribed spacer 2diagnostic assayColombia
collection DOAJ
language English
format Article
sources DOAJ
author Freddy Ruiz
Martha L Quiñones
Holmes F Erazo
David A Calle
Juan F Alzate
Yvonne-Marie Linton
spellingShingle Freddy Ruiz
Martha L Quiñones
Holmes F Erazo
David A Calle
Juan F Alzate
Yvonne-Marie Linton
Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
Memórias do Instituto Oswaldo Cruz.
Anopheles benarrochi B
An. oswaldoi
Nyssorhynchus
internal transcribed spacer 2
diagnostic assay
Colombia
author_facet Freddy Ruiz
Martha L Quiñones
Holmes F Erazo
David A Calle
Juan F Alzate
Yvonne-Marie Linton
author_sort Freddy Ruiz
title Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
title_short Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
title_full Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
title_fullStr Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
title_full_unstemmed Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia
title_sort molecular differentiation of anopheles (nyssorhynchus) benarrochi and an. (n.) oswaldoi from southern colombia
publisher Instituto Oswaldo Cruz, Ministério da Saúde
series Memórias do Instituto Oswaldo Cruz.
issn 0074-0276
1678-8060
publishDate 2005-04-01
description Anopheles (Nyssorhynchus) benarrochi, An. (N.) oswaldoi, and An. (N.) rangeli are the most common anthropophilic mosquitoes in the southern Colombian state of Putumayo. Adult females are most commonly collected in epidemiological studies, and this stage poses significant problems for correct identification, due to overlapping inter-specific morphological characters. Although An. rangeli is easy to identify, the morphological variant of An. benarrochi found in the region and An. oswaldoi are not always easy to separate. Herein we provide a rapid molecular method to distinguish these two species in Southern Colombia. Sequence data for the second internal transcribed spacer (ITS2) region of rDNA was generated for link-reared progeny of An. benarrochi and An. oswaldoi, that had been identified using all life stages. ITS2 sequences were 540 bp in length in An. benarrochi (n = 9) and 531 bp in An. oswaldoi (n = 7). Sequences showed no intra-specific variation and ungapped inter-specific sequence divergence was 6.4%. Species diagnostic banding patterns were recovered following digestion of the ITS2 amplicons with the enzyme Hae III as follows: An. benarrochi (365, 137, and 38 bp) and An. oswaldoi (493 and 38 bp). This polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay provides rapid, accurate, and inexpensive species diagnosis of adult females. This will benefit future epidemiological studies and, as PCR amplification can be achieved using a single mosquito leg, the remaining specimen can be either retained as a morphological voucher or further used in vector incrimination studies. That An. benarrochi comprises a complex of at least two species across Latin America is discussed.
topic Anopheles benarrochi B
An. oswaldoi
Nyssorhynchus
internal transcribed spacer 2
diagnostic assay
Colombia
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762005000200008
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