scholarly journals The Phlebotomus Papatasi Transcriptomic Response to Trypanosomatid-contaminated Blood is Robust but non-specific

2020 ◽  
Author(s):  
Megan A. Sloan ◽  
Jovana Sadlova ◽  
Tereza Lestinova ◽  
Mandy J. Sanders ◽  
James A. Cotton ◽  
...  
Keyword(s):  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Sand Fly ◽  
Phlebotomus Papatasi ◽  
Transcriptomic Response ◽  
Infected Blood ◽  
New Thinking ◽  
Contaminated Blood ◽  
Generation Sequencing

Abstract Background Leishmaniasis, caused by parasites of the genus Leishmania, is a disease that effects up to 8 million people worldwide. Parasites are transmitted to human and animal hosts through the bite of an infected sand fly. Novel strategies for disease control, require a better understanding of the key step for transmission namely, the establishment of infection inside the fly.Methods In this work we wanted to identify fly transcriptomic signatures associated with infected blood meal with non-infected blood meal as our baseline. We used next generation sequencing to describe the transcriptome of the sand fly Phlebotomus papatasi when fed with blood alone or with blood containing one of three trypanosomatids: Leishmania major, Leishmania donovani and Herpetomonas muscarum: a parasite not transmitted to humans.Results Of these, only L. major was able to successfully establish an infection in P. papatasi. However, the transcriptional signatures observed were not specific to success or failure of infection but a generalised response to the blood meal.Conclusions This implies that sand flies perceive Leishmania as just one feature of their microbiome landscape and that any strategy to tackle transmission should focus on the response towards the blood meal rather than parasite establishment. This result will generate new thinking around the concept of stopping transmission by controlling the parasite inside the insect.

scholarly journals The Phlebotomus papatasi systemic transcriptional response to trypanosomatid-contaminated blood does not differ from the non-infected blood meal

2020 ◽  
Author(s):  
Megan A. Sloan ◽  
Jovana Sadlova ◽  
Tereza Lestinova ◽  
Mandy J. Sanders ◽  
James A. Cotton ◽  
...  
Keyword(s):  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Transcriptional Response ◽  
Sand Fly ◽  
Host Responses ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
New Thinking ◽  
Contaminated Blood

Abstract Background Leishmaniasis, caused by parasites of the genus Leishmania, is a disease that effects up to 8 million people worldwide. Parasites are transmitted to human and animal hosts through the bite of an infected sand fly. Novel strategies for disease control, require a better understanding of the key step for transmission namely, the establishment of infection inside the fly. Methods In this work we wanted to identify fly systemic transcriptomic signatures associated with Leishmania infection. We used next generation sequencing to describe the transcriptome of whole Phlebotomus papatasi sand flies when fed with blood alone (control) or with blood containing one of three trypanosomatids: Leishmania major, Leishmania donovani and Herpetomonas muscarum: a parasite not transmitted to humans. Results Of these, only L. major is able to successfully establish an infection in P. papatasi. However, the transcriptional signatures observed after each parasite-contaminated blood meal were not specific to success or failure of a specific infection and were not different from each other. They were also indistinguishable from non-contaminated blood. Conclusions This implies that sand flies perceive Leishmania as just one feature of their microbiome landscape and that any strategy to tackle transmission should focus on the response towards the blood meal rather than parasite establishment. Alternatively, Leishmania could suppress host responses. These results will generate new thinking around the concept of stopping transmission by controlling the parasite inside the insect.

scholarly journals The Phlebotomus papatasi systemic transcriptional response to trypanosomatid-contaminated blood does not differ from the non-infected blood meal

2020 ◽  
Author(s):  
Megan A. Sloan ◽  
Jovana Sadlova ◽  
Tereza Lestinova ◽  
Mandy J. Sanders ◽  
James A. Cotton ◽  
...  
Keyword(s):  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Transcriptional Response ◽  
Sand Fly ◽  
Host Responses ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
New Thinking ◽  
Contaminated Blood

Abstract Background Leishmaniasis, caused by parasites of the genus Leishmania, is a disease that effects up to 8 million people worldwide. Parasites are transmitted to human and animal hosts through the bite of an infected sand fly. Novel strategies for disease control, require a better understanding of the key step for transmission namely, the establishment of infection inside the fly. Methods In this work we wanted to identify fly systemic transcriptomic signatures associated with Leishmania infection. We used next generation sequencing to describe the transcriptome of whole Phlebotomus papatasi sand flies when fed with blood alone (control) or with blood containing one of three trypanosomatids: Leishmania major, Leishmania donovani and Herpetomonas muscarum: a parasite not transmitted to humans. Results Of these, only L. major is able to successfully establish an infection in P. papatasi. However, the transcriptional signatures observed after each parasite-contaminated blood meal were not specific to success or failure of a specific infection and were not different from each other. They were also indistinguishable from non-contaminated blood. Conclusions This implies that sand flies perceive Leishmania as just one feature of their microbiome landscape and that any strategy to tackle transmission should focus on the response towards the blood meal rather than parasite establishment. Alternatively, Leishmania could suppress host responses. These results will generate new thinking around the concept of stopping transmission by controlling the parasite inside the insect.

scholarly journals The Phlebotomus papatasi systemic transcriptional response to trypanosomatid-contaminated blood does not differ from the non-infected blood meal

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Megan A. Sloan ◽  
Jovana Sadlova ◽  
Tereza Lestinova ◽  
Mandy J. Sanders ◽  
James A. Cotton ◽  
...  
Keyword(s):  
Blood Meal ◽  
Leishmania Major ◽  
Transcriptional Response ◽  
Sand Fly ◽  
Host Responses ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Infected Blood ◽  
New Thinking ◽  
Contaminated Blood

Abstract Background Leishmaniasis, caused by parasites of the genus Leishmania, is a disease that affects up to 8 million people worldwide. Parasites are transmitted to human and animal hosts through the bite of an infected sand fly. Novel strategies for disease control require a better understanding of the key step for transmission, namely the establishment of infection inside the fly. Methods The aim of this work was to identify sand fly systemic transcriptomic signatures associated with Leishmania infection. We used next generation sequencing to describe the transcriptome of whole Phlebotomus papatasi sand flies when fed with blood alone (control) or with blood containing one of three trypanosomatids: Leishmania major, L. donovani and Herpetomonas muscarum, the latter being a parasite not transmitted to humans. Results Of the trypanosomatids studied, only L. major was able to successfully establish an infection in the host P. papatasi. However, the transcriptional signatures observed after each parasite-contaminated blood meal were not specific to success or failure of a specific infection and they did not differ from each other. The transcriptional signatures were also indistinguishable after a non-contaminated blood meal. Conclusions The results imply that sand flies perceive Leishmania as just one feature of their microbiome landscape and that any strategy to tackle transmission should focus on the response towards the blood meal rather than parasite establishment. Alternatively, Leishmania could suppress host responses. These results will generate new thinking around the concept of stopping transmission by controlling the parasite inside the insect.

scholarly journals The Phlebotomus papatasi transcriptomic response to trypanosomatid-contaminated blood is robust but non-specific

2019 ◽  
Author(s):  
Megan A. Sloan ◽  
Jovana Sadlova ◽  
Tereza Lestinova ◽  
Mandy J. Sanders ◽  
James A. Cotton ◽  
...  
Keyword(s):  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Blood Feeding ◽  
Sand Fly ◽  
Insect Vector ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Transcriptomic Response ◽  
Gene Expression Response

AbstractLeishmaniasis, caused by parasites of the genus Leishmania, is a disease that effects up to 8 million people worldwide. Parasites are transmitted to human and animal hosts through the bite of an infected sand fly. Novel strategies for disease control, require a better understanding of the key step for transmission namely, the establishment of infection inside the fly. In this work we wanted to identify fly transcriptomic signatures associated with infection success or failure. We used next generation sequencing to describe the transcriptome of the sand fly Phlebotomus papatasi when fed with blood alone or with blood containing one of three trypanosomatids: Leishmania major, Leishmania donovani and Herpetomonas muscarum: a parasite not transmitted to humans. Of these, only L. major was able to successfully establish an infection in P. papatasi. However, the transcriptional signatures observed were not specific to success or failure of infection but a generalised response to the blood meal. This implies that sand flies perceive Leishmania as just a feature of their microbiome landscape and that any strategy to tackle transmission should focus on the response towards the blood meal rather than parasite establishment.Authors summaryLeishmania are parasites that cause leishmaniasis, a group of serious diseases that affect millions of people, mainly across the subtropics and tropics. They are transmitted to humans by phlebotomine sand flies. However, despite establishment in the insect’s midgut being key to transmission, early infection events inside the insect are still unclear. Here, we study the gene expression response of the insect vector to a Leishmania parasite that is able to establish infection (L. major) one that is unable to do so (L. donovani) as well as one that is not a natural parasite of sand flies (Herpetomonas muscarum). We found that responses following any of the infected blood meals was very similar to uninfected blood meal. However, changes post-blood meal from day 1 to day 9 were dramatic. As a blood feeding insect can accumulate three times its weight in one blood meal, this seems to be the most important physiological change rather than the presence of the parasite. The latter might be just one in a number of microbes the insect encounters. This result will generate new thinking around the concept of stopping transmission by controlling the parasite inside the insect.

scholarly journals Molecular detection of Leishmania donovani, Leishmania major, and Trypanosoma species in Sergentomyia squamipleuris sand flies from a visceral leishmaniasis focus in Merti sub-County, eastern Kenya

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Barrack O. Owino ◽  
Jackline Milkah Mwangi ◽  
Steve Kiplagat ◽  
Hannah Njiriku Mwangi ◽  
Johnstone M. Ingonga ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Mus Musculus ◽  
Blood Meal ◽  
Leishmania Donovani ◽  
Leishmania Major ◽  
Sand Fly ◽  
Health Concern ◽  
Sand Flies ◽  
Reservoir Hosts ◽  
Blood Meals

Abstract Background Visceral leishmaniasis (VL) and zoonotic cutaneous leishmaniasis (ZCL) are of public health concern in Merti sub-County, Kenya, but epidemiological data on transmission, vector abundance, distribution, and reservoir hosts remain limited. To better understand the disease and inform control measures to reduce transmission, we investigated the abundance and distribution of sand fly species responsible for Leishmania transmission in the sub-County and their blood-meal hosts. Methods We conducted an entomological survey in five villages with reported cases of VL in Merti sub-County, Kenya, using CDC miniature light traps and castor oil sticky papers. Sand flies were dissected and identified to the species level using standard taxonomic keys and PCR analysis of the cytochrome c oxidase subunit 1 (cox1) gene. Leishmania parasites were detected and identified by PCR and sequencing of internal transcribed spacer 1 (ITS1) genes. Blood-meal sources of engorged females were identified by high-resolution melting analysis of vertebrate cytochrome b (cyt-b) gene PCR products. Results We sampled 526 sand flies consisting of 8 species, Phlebotomus orientalis (1.52%; n = 8), and 7 Sergentomyia spp. Sergentomyia squamipleuris was the most abundant sand fly species (78.71%; n = 414) followed by Sergentomyia clydei (10.46%; n = 55). Leishmania major, Leishmania donovani, and Trypanosoma DNA were detected in S. squamipleuris specimens. Humans were the main sources of sand fly blood meals. However, we also detected mixed blood meals; one S. squamipleuris specimen had fed on both human and mouse (Mus musculus) blood, while two Ph. orientalis specimens fed on human, hyrax (Procavia capensis), and mouse (Mus musculus) blood. Conclusions Our findings implicate the potential involvement of S. squamipleuris in the transmission of Leishmania and question the dogma that human leishmaniases in the Old World are exclusively transmitted by sand flies of the Phlebotomus genus. The presence of Trypanosoma spp. may indicate mechanical transmission, whose efficiency should be investigated. Host preference analysis revealed the possibility of zoonotic transmission of leishmaniasis and other pathogens in the sub-County. Leishmania major and L. donovani are known to cause ZCL and VL, respectively. However, the reservoir status of the parasites is not uniform. Further studies are needed to determine the reservoir hosts of Leishmania spp. in the area.

A novel role for the peritrophic matrix in protecting Leishmania from the hydrolytic activities of the sand fly midgut

Parasitology ◽  
1997 ◽  
Vol 115 (4) ◽  
pp. 359-369 ◽  
Author(s):  
P. F. P. PIMENTA ◽  
G. B. MODI ◽  
S. T. PEREIRA ◽  
M. SHAHABUDDIN ◽  
D. L. SACKS
Keyword(s):  
Digestive Enzymes ◽  
Leishmania Major ◽  
Specific Inhibitor ◽  
Sand Fly ◽  
Peritrophic Matrix ◽  
Phlebotomus Papatasi ◽  
Hydrolytic Activities ◽  
Natural Vector ◽  
Parasite Survival

The role of the peritrophic matrix (PM) in the development of Leishmania major infections in a natural vector, Phlebotomus papatasi, was investigated by addition of exogenous chitinase to the bloodmeal, which completely blocked PM formation. Surprisingly, the absence of the PM was associated with the loss of midgut infections. The chitinase was not directly toxic to the parasite, nor were midgut infections lost due to premature expulsion of the bloodmeal. Most parasites were killed in chitinase-treated flies within the first 4 h after feeding. Substantial early killing was also observed in control flies, suggesting that the lack of PM exacerbates lethal conditions which normally exist in the blood-fed midgut. Early parasite mortality was reversed by soybean trypsin inhibitor. Allosamadin, a specific inhibitor of chitinase, led to a thickening of the PM, and also prevented the early parasite mortality seen in infected flies. Susceptibility to gut proteases was extremely high in transitional-stage parasites, while amastigotes and fully transformed promastigotes were relatively resistant. A novel role for the PM in promoting parasite survival is suggested, in which the PM creates a barrier to the rapid diffusion of digestive enzymes, and limits the exposure of parasites to these enzymes during the time when they are especially vulnerable to proteolytic damage.

scholarly journals Phlebotomus papatasi Antimicrobial Peptides in Larvae and Females and a Gut-Specific Defensin Upregulated by Leishmania major Infection

2021 ◽  
Vol 9 (11) ◽  
pp. 2307
Author(s):  
Barbora Kykalová ◽  
Lucie Tichá ◽  
Petr Volf ◽  
Erich Loza Telleria
Keyword(s):  
Gene Expression ◽  
Antimicrobial Peptides ◽  
Leishmania Major ◽  
Larval Instar ◽  
Gut Bacteria ◽  
Sand Fly ◽  
Control Group ◽  
Phlebotomus Papatasi ◽  
Major Infection ◽  
Defensin Gene

Phlebotomus papatasi is the vector of Leishmania major, causing cutaneous leishmaniasis in the Old World. We investigated whether P. papatasi immunity genes were expressed toward L. major, commensal gut microbes, or a combination of both. We focused on sand fly transcription factors dorsal and relish and antimicrobial peptides (AMPs) attacin and defensin and assessed their relative gene expression by qPCR. Sand fly larvae were fed food with different bacterial loads. Relish and AMPs gene expressions were higher in L3 and early L4 larval instars, while bacteria 16S rRNA increased in late L4 larval instar, all fed rich-microbe food compared to the control group fed autoclaved food. Sand fly females were treated with an antibiotic cocktail to deplete gut bacteria and were experimentally infected by Leishmania. Compared to non-infected females, dorsal and defensin were upregulated at early and late infection stages, respectively. An earlier increase of defensin was observed in infected females when bacteria recolonized the gut after the removal of antibiotics. Interestingly, this defensin gene expression occurred specifically in midguts but not in other tissues of females and larvae. A gut-specific defensin gene upregulated by L. major infection, in combination with gut-bacteria, is a promising molecular target for parasite control strategies.

scholarly journals Effect of Beauveria bassiana and Metarhizium anisopliae on different stages of Phlebotomus papatasi (Diptera: Psychodidae)

2021 ◽  
Vol 90 (4 - Ahead of print) ◽  
pp. 175-193
Author(s):  
Maha Moustafa Ahmed ◽  
Heba Yehia Mady ◽  
Amira Hassan El Namaky
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Beauveria Bassiana ◽  
Metarhizium Anisopliae ◽  
Leishmania Major ◽  
The Body ◽  
Sand Fly ◽  
Life Stages ◽  
Phlebotomus Papatasi ◽  
Scanning Electron

The sand fly, Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae), is the main vector of Leishmania major Yakimoff and Schokhor, 1914, the causative agent of zoonotic cutaneous leishmaniasis North Africa, the Middle East, and North Sinai. The purpose of this study was to determine the effect of fungi on P. papatasi larvae, pupae, and adults using light microscopic analysis, scanning electron microscope (SEM), and histopathological studies. Specifically, larvae, pupae, and adult P. papatasi were infected with the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae. Scanning electron microscope and histopathological methods were used to investigate the destructive impact of the fungi on the external and internal structures of P. papatasi. The results revealed propagation of the conidia on the cuticles of all P. papatasi life stages, including on the compound eyes, leg setae, thorax, wings, and abdomen of the adults. Histological sections of the control and treated larvae, pupae, and adults showed many alterations and malformations in the body and tissues of all life stages after 72 h. These results demonstrated that B. bassiana was more effective than M. anisopliae as a biological control of phlebotomine sand flies. Further studies to determine the best methods for delivery and application in the diverse ecological settings of the various leishmaniasis vectors are recommended.

Salivary glands of the sand fly Phlebotomus papatasi contain pharmacologically active amounts of adenosine and 5′-AMP

1999 ◽  
Vol 202 (11) ◽  
pp. 1551-1559 ◽  
Author(s):  
J.M. Ribeiro ◽  
O. Katz ◽  
L.K. Pannell ◽  
J. Waitumbi ◽  
A. Warburg
Keyword(s):  
Mass Spectrometry ◽  
Salivary Gland ◽  
Salivary Glands ◽  
Blood Meal ◽  
Protein Phosphatase ◽  
Sand Fly ◽  
Phlebotomus Papatasi ◽  
Pharmacologically Active ◽  
Phosphatase Substrate ◽  
Allosteric Modification

Salivary gland homogenates of the sand fly Phlebotomus papatasi contain large amounts of adenosine and 5′-AMP, of the order of 1 nmol per pair of glands, as demonstrated by liquid chromatography, ultraviolet spectrometry, mass spectrometry and bioassays. These purines, 75–80 % of which are secreted from the glands following a blood meal, have vasodilatory and anti-platelet activities and probably help the fly to obtain a blood meal. Salivary 5′-AMP is also responsible for the previously reported protein phosphatase inhibitor in the salivary glands of P. papatasi, which is shown to be artifactual in nature as a result of allosteric modification by AMP of the phosphatase substrate used (phosphorylase a).

scholarly journals CRISPR/Cas9 Mutagenesis in Phlebotomus papatasi: the Immune Deficiency Pathway Impacts Vector Competence for Leishmania major

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Isabelle Louradour ◽  
Kashinath Ghosh ◽  
Ehud Inbar ◽  
David L. Sacks
Keyword(s):  
Immune Deficiency ◽  
Immune Response ◽  
Leishmania Major ◽  
Vector Competence ◽  
Targeted Mutagenesis ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Content Type ◽  
Imd Pathway

ABSTRACT Sand flies are the natural vectors for the Leishmania species that produce a spectrum of diseases in their mammalian hosts, including humans. Studies of sand fly/Leishmania interactions have been limited by the absence of genome editing techniques applicable to these insects. In this report, we adapted CRISPR (clustered regularly interspaced palindromic repeat)/Cas9 (CRISPR-associated protein 9) technology to the Phlebotomus papatasi sand fly, a natural vector for Leishmania major, targeting the sand fly immune deficiency (IMD) pathway in order to decipher its contribution to vector competence. We established a protocol for transformation in P. papatasi and were able to generate transmissible null mutant alleles for Relish (Rel), the only transcription factor of the IMD pathway. While the maintenance of a homozygous mutant stock was severely compromised, we were able to establish in an early generation their greater susceptibility to infection with L. major. Flies carrying different heterozygous mutant alleles variably displayed a more permissive phenotype, presenting higher loads of parasites or greater numbers of infective-stage promastigotes. Together, our data show (i) the successful adaptation of the CRISPR/Cas9 technology to sand flies and (ii) the impact of the sand fly immune response on vector competence for Leishmania parasites. IMPORTANCE Sand flies are the natural vectors of Leishmania parasites. Studies of sand fly/Leishmania interactions have been limited by the lack of successful genomic manipulation of these insects. This paper shows the first example of successful targeted mutagenesis in sand flies via adaptation of the CRISPR/Cas9 editing technique. We generated transmissible null mutant alleles of relish, a gene known to be essential for the control of immune response in other insects. In addition to the expected higher level of susceptibility to bacteria, the mutant flies presented higher loads of parasites when infected with L. major, showing that the sand fly immune response impacts its vector competence for this pathogen.

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