scholarly journals Estimation of vector competence of Moroccan Sand fly Populations Using a Proteomic Approach

2020 ◽  
Author(s):  
Mohamed Daoudi ◽  
George Dong ◽  
Samia Boussaa ◽  
Caroline Martel ◽  
Mohamed Hafidi ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Leishmania Major ◽  
Vector Competence ◽  
Sand Fly ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Chromatography Tandem Mass Spectrometry ◽  
Proteomic Approach ◽  
Liquid Chromatography Tandem Mass ◽  
Leishmania Spp

Abstract Background Phlebotomine vector born disease are distributed throughout the world, and different pathogens are associated with varying degrees of disease severity. In Morocco, sand fly populations are incriminated in the transmission of several pathogens, such Leishmania spp. and phlebovirus, in particular the most relevant sand fly species as the case of Phlebotomus papatasi. This species is the main vector of Leishmania major in Morocco, and incriminated in phlebovirus transmission. Not to pass over without investigating its possible role on the transmission of entomopathogenic parasite already reported in literature. Methodology and finding The present study focused on proteomic analysis in Phlebotomus papatasi from four localities in Morocco. Proteomic analysis in female P. papatasi was performed with Liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 239 identified proteins for Leishmania spp., 26 identified proteins for phlebovirus and 1132 identified proteins for nematode were detected in the 20 pools of P. papatasi from investigated localities. The microscopic examination of 1752 sandflies collected showed, the infection of two specimens of female’s P. sergenti with Tetranematid Didilia spp. and potential Microfilariae in Imintanout locality (Z2).Conclusion This study presents the first report of nematodes in sand flies of Africa. Also, the first use of proteomic tools for estimation of vector competence of P. papatasi which could be a specific tool to a better understanding of the vectorial capacity of Moroccan sandflies population.

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.

scholarly journals Exploring Lutzomyia longipalpis Sand Fly Vector Competence for Leishmania major Parasites

2020 ◽  
Vol 222 (7) ◽  
pp. 1199-1203 ◽  
Author(s):  
Pedro Cecílio ◽  
Ana Clara A M Pires ◽  
Jesus G Valenzuela ◽  
Paulo F P Pimenta ◽  
Anabela Cordeiro-da-Silva ◽  
...  
Keyword(s):  
New World ◽  
Leishmania Major ◽  
Vector Competence ◽  
Experimental Studies ◽  
Leishmania Species ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Potential Vector ◽  
Natural Vector

Abstract Lutzomyia longipalpis sand flies are the major natural vector of Leishmania infantum parasites, responsible for transmission of visceral leishmaniasis in the New World. Several experimental studies have demonstrated the ability of Lu. longipalpis to sustain development of different Leishmania species. However, no study had explored in depth the potential vector competence of Lu. longipalpis for Leishmania species other than L. infantum. Here, we show that Lu. longipalpis is a competent vector of L. major parasites, being able to acquire parasites from active cutaneous leishmaniasis lesions, sustain mature infections, and transmit them to naive hosts, causing disease.

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 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 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.

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 Development of a Diagnostic Marker for Phlebotomus papatasi to Initiate a Potential Vector Surveillance Program in North America

Insects ◽  
2018 ◽  
Vol 9 (4) ◽  
pp. 162 ◽  
Author(s):  
Austin Merchant ◽  
Tian Yu ◽  
Jizhe Shi ◽  
Xuguo Zhou
Keyword(s):  
Rapid Identification ◽  
International Travel ◽  
Sand Fly ◽  
Surveillance Program ◽  
Sand Flies ◽  
Phlebotomus Papatasi ◽  
Diagnostic Assay ◽  
Vector Surveillance ◽  
Geographical Regions ◽  
Imminent Threat

Phlebotomus papatasi, an Old World sand fly species, is primarily responsible for the transmission of leishmaniasis, a highly infectious and potentially lethal disease. International travel, especially military rotations, between domestic locations and P. papatasi-prevalent regions in the Middle East poses an imminent threat to the public health of US citizens. Because of its small size and cryptic morphology, identification of P. papatasi is challenging and labor-intensive. Here, we developed a ribosomal DNA-polymerase chain reaction (PCR)-based diagnostic assay that is capable of detecting P. papatasi genomic DNA from mixed samples containing multiple sand flies native to the Americas. Serial dilution of P. papatasi samples demonstrated that this diagnostic assay could detect one P. papatasi from up to 255 non-target sand flies. Due to its simplicity, sensitivity and specificity, this rapid identification tool is suited for a long-term surveillance program to screen for the presence of P. papatasi in the continental United States and to reveal geographical regions potentially vulnerable to sand fly-borne diseases.

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.

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