Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]

Background: The leishmaniases are neglected diseases that affect some of the most vulnerable populations in the tropical and sub-tropical world. The parasites are transmitted by sand flies and novel strategies to control this neglected vector-borne disease are needed. Blocking transmission by targeting the parasite inside the phlebotomine vector offers potential in this regard. Some experimental approaches can be best performed by longitudinal study of parasites within flies, for which non-destructive methods to identify infected flies and to follow parasite population changes are required. Methods: Lutzomyia longipalpis were reared under standard insectary conditions at the Wellcome Centre for Molecular Parasitology. Flies were artificially infected with L. tarentolae expressing green fluorescent protein (GFP. Parasite counts were carried out 5 days post-infection and the percentage of infected flies and survival of infected females was established up to days 5 post-infection. Whole living females were visualised using an epifluorescence inverted microscope to detect the presence parasites inferred by a localised green fluorescent region in the upper thorax. Confirmation of infection was performed by localised-fluorescence of dissected flies and estimates of the parasite population.

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Bibliographic Details
Main Authors: Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173, Regnault, Clément autor, Alpízar Sosa, Edubiel Arturo autor, McGuinness, Dagmara autora, Barrett, Michael P. autor, Dillon, Rod J. autor
Format: Texto biblioteca
Language:eng
Subjects:Leishmania tarentolae, Leishmaniasis, Relaciones huésped-patógeno, Lutzomyia longipalpis, Control de insectos,
Online Access:https://wellcomeopenresearch.org/articles/3-160
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id KOHA-OAI-ECOSUR:60441
record_format koha
institution ECOSUR
collection Koha
country México
countrycode MX
component Bibliográfico
access En linea
En linea
databasecode cat-ecosur
tag biblioteca
region America del Norte
libraryname Sistema de Información Bibliotecario de ECOSUR (SIBE)
language eng
topic Leishmania tarentolae
Leishmaniasis
Relaciones huésped-patógeno
Lutzomyia longipalpis
Control de insectos
Leishmania tarentolae
Leishmaniasis
Relaciones huésped-patógeno
Lutzomyia longipalpis
Control de insectos
spellingShingle Leishmania tarentolae
Leishmaniasis
Relaciones huésped-patógeno
Lutzomyia longipalpis
Control de insectos
Leishmania tarentolae
Leishmaniasis
Relaciones huésped-patógeno
Lutzomyia longipalpis
Control de insectos
Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173
Regnault, Clément autor
Alpízar Sosa, Edubiel Arturo autor
McGuinness, Dagmara autora
Barrett, Michael P. autor
Dillon, Rod J. autor
Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
description Background: The leishmaniases are neglected diseases that affect some of the most vulnerable populations in the tropical and sub-tropical world. The parasites are transmitted by sand flies and novel strategies to control this neglected vector-borne disease are needed. Blocking transmission by targeting the parasite inside the phlebotomine vector offers potential in this regard. Some experimental approaches can be best performed by longitudinal study of parasites within flies, for which non-destructive methods to identify infected flies and to follow parasite population changes are required. Methods: Lutzomyia longipalpis were reared under standard insectary conditions at the Wellcome Centre for Molecular Parasitology. Flies were artificially infected with L. tarentolae expressing green fluorescent protein (GFP. Parasite counts were carried out 5 days post-infection and the percentage of infected flies and survival of infected females was established up to days 5 post-infection. Whole living females were visualised using an epifluorescence inverted microscope to detect the presence parasites inferred by a localised green fluorescent region in the upper thorax. Confirmation of infection was performed by localised-fluorescence of dissected flies and estimates of the parasite population.
format Texto
topic_facet Leishmania tarentolae
Leishmaniasis
Relaciones huésped-patógeno
Lutzomyia longipalpis
Control de insectos
author Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173
Regnault, Clément autor
Alpízar Sosa, Edubiel Arturo autor
McGuinness, Dagmara autora
Barrett, Michael P. autor
Dillon, Rod J. autor
author_facet Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173
Regnault, Clément autor
Alpízar Sosa, Edubiel Arturo autor
McGuinness, Dagmara autora
Barrett, Michael P. autor
Dillon, Rod J. autor
author_sort Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173
title Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
title_short Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
title_full Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
title_fullStr Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
title_full_unstemmed Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
title_sort non-invasive visualisation and identification of fluorescent leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations]
url https://wellcomeopenresearch.org/articles/3-160
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spelling KOHA-OAI-ECOSUR:604412024-03-11T15:27:41ZNon-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies [version 1; peer review: 1 approved, 2 approved with reservations] Díaz Albíter, Héctor Manuel Doctor 1977-2021 autor 21173 Regnault, Clément autor Alpízar Sosa, Edubiel Arturo autor McGuinness, Dagmara autora Barrett, Michael P. autor Dillon, Rod J. autor textengBackground: The leishmaniases are neglected diseases that affect some of the most vulnerable populations in the tropical and sub-tropical world. The parasites are transmitted by sand flies and novel strategies to control this neglected vector-borne disease are needed. Blocking transmission by targeting the parasite inside the phlebotomine vector offers potential in this regard. Some experimental approaches can be best performed by longitudinal study of parasites within flies, for which non-destructive methods to identify infected flies and to follow parasite population changes are required. Methods: Lutzomyia longipalpis were reared under standard insectary conditions at the Wellcome Centre for Molecular Parasitology. Flies were artificially infected with L. tarentolae expressing green fluorescent protein (GFP. Parasite counts were carried out 5 days post-infection and the percentage of infected flies and survival of infected females was established up to days 5 post-infection. Whole living females were visualised using an epifluorescence inverted microscope to detect the presence parasites inferred by a localised green fluorescent region in the upper thorax. Confirmation of infection was performed by localised-fluorescence of dissected flies and estimates of the parasite population.Results: Leishmania tarentolae was successfully transfected and expressed GFP in vitro. L. tarentolae-GFP Infected flies showed similar parasite populations when compared to non-transfected parasites (L. tarentolae-WT). Survival of non-infected females was higher than L. tarentolae-infected groups, (Log-rank (Mantel-Cox) test, p<0.05). L. tarentolae-GFP infected females displayed an intense localised fluorescence in the thorax while other specimens from the same infected group did not. Localised fluorescent flies were dissected and showed higher parasite populations compared to those that did not demonstrate high concentrations in this region (t-test, p<0.005). Conclusion: These results demonstrate the feasibility of establishing a safe non-human infectious fluorescent Leishmania-sand fly infection model by allowing non-destructive imaging to signal the establishment of Leishmania infections in living sand flies.Background: The leishmaniases are neglected diseases that affect some of the most vulnerable populations in the tropical and sub-tropical world. The parasites are transmitted by sand flies and novel strategies to control this neglected vector-borne disease are needed. Blocking transmission by targeting the parasite inside the phlebotomine vector offers potential in this regard. Some experimental approaches can be best performed by longitudinal study of parasites within flies, for which non-destructive methods to identify infected flies and to follow parasite population changes are required. Methods: Lutzomyia longipalpis were reared under standard insectary conditions at the Wellcome Centre for Molecular Parasitology. Flies were artificially infected with L. tarentolae expressing green fluorescent protein (GFP. Parasite counts were carried out 5 days post-infection and the percentage of infected flies and survival of infected females was established up to days 5 post-infection. Whole living females were visualised using an epifluorescence inverted microscope to detect the presence parasites inferred by a localised green fluorescent region in the upper thorax. Confirmation of infection was performed by localised-fluorescence of dissected flies and estimates of the parasite population.Results: Leishmania tarentolae was successfully transfected and expressed GFP in vitro. L. tarentolae-GFP Infected flies showed similar parasite populations when compared to non-transfected parasites (L. tarentolae-WT). Survival of non-infected females was higher than L. tarentolae-infected groups, (Log-rank (Mantel-Cox) test, p<0.05). L. tarentolae-GFP infected females displayed an intense localised fluorescence in the thorax while other specimens from the same infected group did not. Localised fluorescent flies were dissected and showed higher parasite populations compared to those that did not demonstrate high concentrations in this region (t-test, p<0.005). Conclusion: These results demonstrate the feasibility of establishing a safe non-human infectious fluorescent Leishmania-sand fly infection model by allowing non-destructive imaging to signal the establishment of Leishmania infections in living sand flies.Adobe Acrobat profesional 6.0 o superiorLeishmania tarentolaeLeishmaniasisRelaciones huésped-patógenoLutzomyia longipalpisControl de insectosDisponible en líneaWellcome Open Researchhttps://wellcomeopenresearch.org/articles/3-160Acceso en línea sin restricciones