Influence of <i>Leishmania</i> products on midgut trypsin production.

<p>Midguts were exposed for 2 hours to <i>Leishmania infantum</i> products and to amino acids at a final concentration of 30 mM, and the trypsin activity was measured in the supernatant; no products from <i>Leishmania</i> were included the control group. Each dot represents the activity normalized to one midgut of 11 independent experiments. There was no difference in the quantity of trypsin produced (<i>p</i> = 0.7421).</p

Evaluation of the trypsin activity in infected insects.

<p>Sand flies were dissected at 48 hours after feeding on an uninfected dog (n = 26) or a naturally infected dog (n = 29); the trypsin activity was measured using the synthetic substrate L-BApNA and was expressed as enzymatic units (U), normalized to the quantity of homogenate equivalent to one midgut (U.midgut⁻¹). Each dot represents the activity obtained from an individual insect, and the horizontal line represents the mean value. The trypsin activity was significantly lower in the infected sand flies compared to the uninfected insects (<i>p</i> = 0.0128).</p

Delay in blood digestion in <i>Lutzomyia longipalpis</i> females infected with <i>Leishmania infantum</i>.

<p>Fisher's exact test was used to compare the presence of digesting blood inside uninfected and infected females (*significant difference).</p><p>Delay in blood digestion in <i>Lutzomyia longipalpis</i> females infected with <i>Leishmania infantum</i>.</p

pH in the abdominal midgut of <i>Lutzomyia longipalpis</i> females fed 199 medium plus 100 mM amino acids with or without lipophosphoglycan (LPG) purified from <i>Leishmania infantum</i>.

<p>Fisher's exact test was used to compare the treatments.</p><p>pH in the abdominal midgut of <i>Lutzomyia longipalpis</i> females fed 199 medium plus 100 mM amino acids with or without lipophosphoglycan (LPG) purified from <i>Leishmania infantum</i>.</p

pH in the abdominal midgut of infected and uninfected <i>Lutzomyia longipalpis</i> females during and after blood digestion.

<p>Students' T test was used to compare the intestinal pH measured in uninfected and infected females (*significant difference).</p><p>pH in the abdominal midgut of infected and uninfected <i>Lutzomyia longipalpis</i> females during and after blood digestion.</p

Activity of trypsin at different pH values.

<p>At 24 hours after a blood meal using hamsters, the midguts from 12 sand flies were dissected and pooled. The supernatant was incubated with L-BApNA at different pH values (7.0–8.5). The experiment was performed in duplicate and repeated independently three times. The graphic represents the mean values ± standard deviation.</p

<i>Phlebotomus papatasi</i> circadian rhythm pathway annotation.

Phlebotomus papatasi circadian rhythm pathway annotation.</p

Chitinase family annotation.

Phlebotomine sand flies are of global significance as important vectors of human disease, transmitting bacterial, viral, and protozoan pathogens, including the kinetoplastid parasites of the genus Leishmania, the causative agents of devastating diseases collectively termed leishmaniasis. More than 40 pathogenic Leishmania species are transmitted to humans by approximately 35 sand fly species in 98 countries with hundreds of millions of people at risk around the world. No approved efficacious vaccine exists for leishmaniasis and available therapeutic drugs are either toxic and/or expensive, or the parasites are becoming resistant to the more recently developed drugs. Therefore, sand fly and/or reservoir control are currently the most effective strategies to break transmission. To better understand the biology of sand flies, including the mechanisms involved in their vectorial capacity, insecticide resistance, and population structures we sequenced the genomes of two geographically widespread and important sand fly vector species: Phlebotomus papatasi, a vector of Leishmania parasites that cause cutaneous leishmaniasis, (distributed in Europe, the Middle East and North Africa) and Lutzomyia longipalpis, a vector of Leishmania parasites that cause visceral leishmaniasis (distributed across Central and South America). We categorized and curated genes involved in processes important to their roles as disease vectors, including chemosensation, blood feeding, circadian rhythm, immunity, and detoxification, as well as mobile genetic elements. We also defined gene orthology and observed micro-synteny among the genomes. Finally, we present the genetic diversity and population structure of these species in their respective geographical areas. These genomes will be a foundation on which to base future efforts to prevent vector-borne transmission of Leishmania parasites.</div

Molecular phylogenetic analysis of <i>Lu</i>. <i>longipalpis</i>, <i>P</i>. <i>papatasi</i> and <i>D</i>. <i>melanogaster</i> TRP channel sequences.

The different TRP subfamilies are displayed on the right. The evolutionary history was inferred by using the Maximum Likelihood method based on the Whelan and Goldman +Freq. model with 1000 bootstrap replicates. (TIF)</p

Glycosidase Hydrolase family 13 annotation.

Phlebotomine sand flies are of global significance as important vectors of human disease, transmitting bacterial, viral, and protozoan pathogens, including the kinetoplastid parasites of the genus Leishmania, the causative agents of devastating diseases collectively termed leishmaniasis. More than 40 pathogenic Leishmania species are transmitted to humans by approximately 35 sand fly species in 98 countries with hundreds of millions of people at risk around the world. No approved efficacious vaccine exists for leishmaniasis and available therapeutic drugs are either toxic and/or expensive, or the parasites are becoming resistant to the more recently developed drugs. Therefore, sand fly and/or reservoir control are currently the most effective strategies to break transmission. To better understand the biology of sand flies, including the mechanisms involved in their vectorial capacity, insecticide resistance, and population structures we sequenced the genomes of two geographically widespread and important sand fly vector species: Phlebotomus papatasi, a vector of Leishmania parasites that cause cutaneous leishmaniasis, (distributed in Europe, the Middle East and North Africa) and Lutzomyia longipalpis, a vector of Leishmania parasites that cause visceral leishmaniasis (distributed across Central and South America). We categorized and curated genes involved in processes important to their roles as disease vectors, including chemosensation, blood feeding, circadian rhythm, immunity, and detoxification, as well as mobile genetic elements. We also defined gene orthology and observed micro-synteny among the genomes. Finally, we present the genetic diversity and population structure of these species in their respective geographical areas. These genomes will be a foundation on which to base future efforts to prevent vector-borne transmission of Leishmania parasites.</div