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Molecular detection of Leishmania in phlebotomine sand flies (Diptera: Psychodidae) from a cutaneous leishmaniasis focus atXakriabá Indigenous Reserve, Brazil.

Rêgo FD, Rugani JM, Shimabukuro PH, Tonelli GB, Quaresma PF, Gontijo CM - PLoS ONE (2015)

Bottom Line: The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one).The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively.The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.

View Article: PubMed Central - PubMed

Affiliation: Grupo de Estudos em Leishmanioses, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Av. Augusto de Lima, 1715 Barro Preto, CEP 30190-002, Belo Horizonte, Minas Gerais, Brasil.

ABSTRACT
Autochthonous cases of American cutaneous leishmaniasis (ACL) have been reported since 2001 in the Xakriabá Indigenous Reserve located in the municipality of São João das Missões in northern Minas Gerais state, Brazil. In order to study the presence of Leishmania DNA in phlebotomine sand flies, six entomological collections were carried out from July 2008 through July 2009, using 40 light traps placed in peridomicile areas of 20 randomly selected houses. From October 2011 through August 2012, another six collections were carried out with 20 light traps distributed among four trails (five traps per trail) selected for a previous study of wild and synanthropic hosts of Leishmania. A total of 4,760 phlebotomine specimens were collected belonging to ten genera and twenty-three species. Single female specimens or pools with up to ten specimens of the same locality, species and date, for Leishmania detection by molecular methods. Species identification of parasites was performed with ITS1 PCR-RFLP using HaeIII enzyme and genetic sequencing for SSU rRNA target. The presence of Leishmania DNA was detected in eleven samples from peridomicile areas: Lu. longipalpis (two), Nyssomyia intermedia (four), Lu. renei (two), Lu. ischnacantha, Micropygomyia goiana and Evandromyia lenti (one pool of each specie). The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one). The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively. The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.

No MeSH data available.


Related in: MedlinePlus

Detection of Leishmania in sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.Sand flies were pooled with up to ten specimens of the same locality, species and date, and ITS1 PCR was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 2% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of Le. amazonensis (IFLA/BR/67/PH8), Le. braziliensis (MHOM/BR/75/M2903), Le. infantum (MHOM/BR/74/PP75), Le. guyanensis (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools; NC, negative control.
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pone.0122038.g002: Detection of Leishmania in sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.Sand flies were pooled with up to ten specimens of the same locality, species and date, and ITS1 PCR was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 2% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of Le. amazonensis (IFLA/BR/67/PH8), Le. braziliensis (MHOM/BR/75/M2903), Le. infantum (MHOM/BR/74/PP75), Le. guyanensis (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools; NC, negative control.

Mentions: Out of the 1,026 pools from the trails twelve were ITS1-PCR positive (Table 3). The twelve positive pools were Mt. minasensis (five), Ny. intermedia (three), Mi. peresi (two) and one sample of each of the following species: Mi. capixaba and Ev. lenti (Fig 2). The PCR-RFLP technique identified to the species level the Leishmania in all samples (Fig 3). The most prevalent species was Leishmania braziliensis (41.6%), followed by Leishmania guyanensis and Leishmania infantum chagasi (25% of each species).


Molecular detection of Leishmania in phlebotomine sand flies (Diptera: Psychodidae) from a cutaneous leishmaniasis focus atXakriabá Indigenous Reserve, Brazil.

Rêgo FD, Rugani JM, Shimabukuro PH, Tonelli GB, Quaresma PF, Gontijo CM - PLoS ONE (2015)

Detection of Leishmania in sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.Sand flies were pooled with up to ten specimens of the same locality, species and date, and ITS1 PCR was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 2% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of Le. amazonensis (IFLA/BR/67/PH8), Le. braziliensis (MHOM/BR/75/M2903), Le. infantum (MHOM/BR/74/PP75), Le. guyanensis (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools; NC, negative control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390197&req=5

pone.0122038.g002: Detection of Leishmania in sand flies collected from trails in the Xakriabá Indigenous Reserve, Brazil, during the study period.Sand flies were pooled with up to ten specimens of the same locality, species and date, and ITS1 PCR was performed with total DNA extracted from these pools. The figure represents an ethidium bromide-stained 2% agarose gel in which the amplicons were submitted to electrophoresis. Lanes: MW, molecular weight marker—100 bp; lanes 1–4, positive controls of Le. amazonensis (IFLA/BR/67/PH8), Le. braziliensis (MHOM/BR/75/M2903), Le. infantum (MHOM/BR/74/PP75), Le. guyanensis (MHOM/BR/75/M4147) respectively; 5–14, phlebotomine positive pools; NC, negative control.
Mentions: Out of the 1,026 pools from the trails twelve were ITS1-PCR positive (Table 3). The twelve positive pools were Mt. minasensis (five), Ny. intermedia (three), Mi. peresi (two) and one sample of each of the following species: Mi. capixaba and Ev. lenti (Fig 2). The PCR-RFLP technique identified to the species level the Leishmania in all samples (Fig 3). The most prevalent species was Leishmania braziliensis (41.6%), followed by Leishmania guyanensis and Leishmania infantum chagasi (25% of each species).

Bottom Line: The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one).The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively.The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.

View Article: PubMed Central - PubMed

Affiliation: Grupo de Estudos em Leishmanioses, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Av. Augusto de Lima, 1715 Barro Preto, CEP 30190-002, Belo Horizonte, Minas Gerais, Brasil.

ABSTRACT
Autochthonous cases of American cutaneous leishmaniasis (ACL) have been reported since 2001 in the Xakriabá Indigenous Reserve located in the municipality of São João das Missões in northern Minas Gerais state, Brazil. In order to study the presence of Leishmania DNA in phlebotomine sand flies, six entomological collections were carried out from July 2008 through July 2009, using 40 light traps placed in peridomicile areas of 20 randomly selected houses. From October 2011 through August 2012, another six collections were carried out with 20 light traps distributed among four trails (five traps per trail) selected for a previous study of wild and synanthropic hosts of Leishmania. A total of 4,760 phlebotomine specimens were collected belonging to ten genera and twenty-three species. Single female specimens or pools with up to ten specimens of the same locality, species and date, for Leishmania detection by molecular methods. Species identification of parasites was performed with ITS1 PCR-RFLP using HaeIII enzyme and genetic sequencing for SSU rRNA target. The presence of Leishmania DNA was detected in eleven samples from peridomicile areas: Lu. longipalpis (two), Nyssomyia intermedia (four), Lu. renei (two), Lu. ischnacantha, Micropygomyia goiana and Evandromyia lenti (one pool of each specie). The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one). The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively. The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.

No MeSH data available.


Related in: MedlinePlus