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. 2020 Feb 27;10(1):3566.
doi: 10.1038/s41598-020-60600-7.

Experimental infections and co-infections with Leishmania braziliensis and Leishmania infantum in two sand fly species, Lutzomyia migonei and Lutzomyia longipalpis

Joanna Alexandre  1 , Jovana Sadlova  2 , Tereza Lestinova  2 , Barbora Vojtkova  2 , Magda Jancarova  2 , Lucie Podesvova  3 , Vyacheslav Yurchenko  3   4 , Filipe Dantas-Torres  1 , Sinval P Brandão-Filho  5 , Petr Volf  2
Affiliations

Experimental infections and co-infections with Leishmania braziliensis and Leishmania infantum in two sand fly species, Lutzomyia migonei and Lutzomyia longipalpis

Joanna Alexandre et al. Sci Rep. .

Abstract

Leishmaniases are neglected tropical diseases and Leishmania (Leishmania) infantum and Leishmania (Viannia) braziliensis are the most important causative agents of leishmaniases in the New World. These two parasite species may co-circulate in a given endemic area but their interactions in the vector have not been studied yet. We conducted experimental infections using both single infections and co-infections to compare the development of L. (L.) infantum (OGVL/mCherry) and L. (V.) braziliensis (XB29/GFP) in Lutzomyia longipalpis and Lutzomyia migonei. Parasite labelling by different fluorescein proteins enabled studying interspecific competition and localization of different parasite species during co-infections. Both Leishmania species completed their life cycle, producing infective forms in both sand fly species studied. The same happens in the co infections, demonstrating that the two parasites conclude their development and do not compete with each other. However, infections produced by L. (L.) infantum reached higher rates and grew more vigorously, as compared to L. (V.) braziliensis. In late-stage infections, L. (L.) infantum was present in all midgut regions, showing typical suprapylarian type of development, whereas L. (V.) braziliensis was concentrated in the hindgut and the abdominal midgut (peripylarian development). We concluded that both Lu. migonei and Lu. longipalpis are equally susceptible vectors for L. (L.) infantum, in laboratory colonies. In relation to L. (V.) braziliensis, Lu. migonei appears to be more susceptible to this parasite than Lu. longipalpis.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Rates and intensities (a,c) and localization of Leishmania spp. (b,d) in Lu. migonei (a,b) and Lu. longipalpis (c,d) evaluated by fluorescence microscopy. Intensities of infections were classified into three categories: light (<100 parasites/gut), moderate (100–1000 parasites/gut), or heavy (>1000 parasites/gut). PI = post infections, INF = L. infantum, BRA = L. braziliensis. Columns represent intensity of the Leishmania species either in single infections (INF single, BRA single) of in coinfection (INF co-inf, BRA co-inf). Numbers of dissected sand fly females are shown above the bars.
Figure 2
Figure 2
Fluorescence micrographs of thoracic midguts with cardia section and stomodeal valve of Lutzomyia spp. females on days 8 postinfection. Cardia of Lu. longipalpis (A–C) and Lu. migonei (D–F) coinfected by Leishmania (Viannia) braziliensis (green) and Leishmania (Leishmania) infantum (red). Control uninfected gut of Lu. longipalpis (G–I). (A,D,G) images from red fluorescence, (B,E,H) images from green fluorescence and (C,F,I) merged images, scale bar: 5 mm.
Figure 3
Figure 3
Morphological forms of Leishmania spp. during development in Lutzomyia spp. Morphological forms of Leishmania parasites in Lutzomyia migonei and Lutzomyia longipalpis were evaluated by light microscopy using oil-immersion objective on days 5 and 8 postinfection. MIG (Lu. migonei), BRA (L. (V.) braziliensis), LONG (Lu. longipalpis), INF (L. (L.) infantum), EN (elongated nectomonads), SN (short nectomonads).
Figure 4
Figure 4
Morphological forms of Leishmania occurring in sand fly midgut of Lu. migonei. Scale bar 10 μm. (A) Elongated nectomonad of L. (V.) braziliensis in Lu. migonei; (B) Short nectomonad of L. (L.) infantum in Lu. migonei (C) Metacyclic promastigote of L. (L.) infantum in Lu. migonei; (D) haptomonads of L. (L.) infantum in Lu. migonei.
Figure 5
Figure 5
The curve growth of Leishmania spp. cultures at days 0, 1, 2, 3, 4, 5 and 6. Two initial concentrations were used for each Leishmania species. (a) Initial dose (at day 0) of 104 promastigotes/ml; (b) Initial dose (at day 0) of 105 promastigotes/ml.
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