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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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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Leishmania tarentolae Leishmaniasis Relaciones huésped-patógeno Lutzomyia longipalpis Control de insectos Leishmania tarentolae Leishmaniasis Relaciones huésped-patógeno Lutzomyia longipalpis Control de insectos |
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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. |
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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 |
work_keys_str_mv |
AT diazalbiterhectormanueldoctor19772021autor21173 noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations AT regnaultclementautor noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations AT alpizarsosaedubielarturoautor noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations AT mcguinnessdagmaraautora noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations AT barrettmichaelpautor noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations AT dillonrodjautor noninvasivevisualisationandidentificationoffluorescentleishmaniatarentolaeininfectedsandfliesversion1peerreview1approved2approvedwithreservations |
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1794792074812325888 |
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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 |