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Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies

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Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies. / Diaz-Albiter, Hector M; Regnault, Clément; Alpizar-Sosa, Edubiel A et al.

In: Wellcome Open Research, Vol. 3, 160, 13.12.2018.

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Diaz-Albiter HM, Regnault C, Alpizar-Sosa EA, McGuinness D, Barrett M, Dillon RJ. Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies. Wellcome Open Research. 2018 Dec 13;3:160. doi: 10.12688/wellcomeopenres.14910.1, 10.12688/wellcomeopenres.14910.1

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Diaz-Albiter, Hector M ; Regnault, Clément ; Alpizar-Sosa, Edubiel A et al. / Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies. In: Wellcome Open Research. 2018 ; Vol. 3.

Bibtex

@article{5e0e926cfe074acb84a388114d7e649b,
title = "Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies",
abstract = "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.",
author = "Diaz-Albiter, {Hector M} and Cl{\'e}ment Regnault and Alpizar-Sosa, {Edubiel A} and Dagmara McGuinness and Michael Barrett and Dillon, {Rod J}",
year = "2018",
month = dec,
day = "13",
doi = "10.12688/wellcomeopenres.14910.1",
language = "English",
volume = "3",
journal = "Wellcome Open Research",
issn = "2398-502X",
publisher = "F1000 Research Ltd.",

}

RIS

TY - JOUR

T1 - Non-invasive visualisation and identification of fluorescent Leishmania tarentolae in infected sand flies

AU - Diaz-Albiter, Hector M

AU - Regnault, Clément

AU - Alpizar-Sosa, Edubiel A

AU - McGuinness, Dagmara

AU - Barrett, Michael

AU - Dillon, Rod J

PY - 2018/12/13

Y1 - 2018/12/13

N2 - 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.

AB - 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.

U2 - 10.12688/wellcomeopenres.14910.1

DO - 10.12688/wellcomeopenres.14910.1

M3 - Journal article

C2 - 30756095

VL - 3

JO - Wellcome Open Research

JF - Wellcome Open Research

SN - 2398-502X

M1 - 160

ER -