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Michalis Kotsyfakis - One of the best experts on this subject based on the ideXlab platform.
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plasmodium falciparum infection induces expression of a mosquito salivary protein agaphelin that targets neutrophil function and inhibits thrombosis without impairing hemostasis
PLOS Pathogens, 2014Co-Authors: Daniella M Mizurini, Michael Waisberg, Alvaro Molinacruz, Nidhi Gera, Beatriz Coutinho De Sousa, Ana C Leal, Taina Gomes, Michalis KotsyfakisAbstract:Background Invasion of mosquito salivary glands (SGs) by Plasmodium falciparum sporozoites is an essential step in the malaria life cycle. How infection modulates gene expression, and affects hematophagy remains unclear. Principal Findings Using Affimetrix chip microarray, we found that at least 43 genes are differentially expressed in the glands of Plasmodium falciparum-infected Anopheles gambiae mosquitoes. Among the upregulated genes, one codes for Agaphelin, a 58-amino acid protein containing a single Kazal domain with a Leu in the P1 position. Agaphelin displays high homology to orthologs present in Aedes sp and Culex sp salivary glands, indicating an evolutionarily expanded family. Kinetics and surface plasmon resonance experiments determined that chemically synthesized Agaphelin behaves as a slow and tight inhibitor of neutrophil elastase (KD∼10 nM), but does not affect other enzymes, nor promotes vasodilation, or exhibit antimicrobial activity. TAXIscan chamber assay revealed that Agaphelin inhibits neutrophil chemotaxis toward fMLP, affecting several parameter associated with cell migration. In addition, Agaphelin reduces paw edema formation and accumulation of tissue myeloperoxidase triggered by injection of carrageenan in mice. Agaphelin also blocks elastase/cathepsin-mediated platelet aggregation, abrogates elastase-mediated cleavage of tissue factor pathway inhibitor, and attenuates neutrophil-induced coagulation. Notably, Agaphelin inhibits neutrophil extracellular traps (NETs) formation and prevents FeCl3-induced arterial thrombosis, without impairing hemostasis. Conclusions Blockade of neutrophil elastase emerges as a novel antihemostatic mechanism in hematophagy; it also supports the notion that neutrophils and the innate immune response are targets for antithrombotic therapy. In addition, Agaphelin is the first antihemostatic whose expression is induced by Plasmodium sp infection. These results suggest that an important interplay takes place in Parasite-Vector-host interactions.
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de novo ixodes ricinus salivary gland transcriptome analysis using two next generation sequencing methodologies
The FASEB Journal, 2013Co-Authors: Alexandra Schwarz, Andrezza C Chagas, Bjoern M. Von Reumont, Jan Erhart, Michalis KotsyfakisAbstract:Tick salivary gland (SG) proteins possess powerful pharmacologic properties that facilitate tick feeding and pathogen transmission. For the first time, SG transcriptomes of Ixodes ricinus, an important disease Vector for humans and animals, were analyzed using next-generation sequencing. SGs were collected from different tick life stages fed on various animal species, including cofeeding of nymphs and adults on the same host. Four cDNA samples were sequenced, discriminating tick SG transcriptomes of early- and late-feeding nymphs or adults. In total, 441,381,454 pyrosequencing reads and 67,703,183 Illumina reads were assembled into 272,220 contigs, of which 34,560 extensively annotated coding sequences are disclosed; 8686 coding sequences were submitted to GenBank. Overall, 13% of contigs were classified as secreted proteins that showed significant differences in the transcript representation among the 4 SG samples, including high numbers of sample-specific transcripts. Detailed phylogenetic reconstructions of two relatively abundant SG-secreted protein families demonstrated how this study improves our understanding of the molecular evolution of hematophagy in arthropods. Our data significantly increase the available genomic information for I. ricinus and form a solid basis for future tick genome/transcriptome assemblies and the functional analysis of effectors that mediate the feeding physiology and Parasite-Vector interaction of I. ricinus.—Schwarz, A., von Reumont, B.M., Erhart, J., Chagas, A.C., Ribeiro, J.M.C., Kotsyfakis, M. De novo Ixodes ricinus salivary gland transcriptome analysis using two next-generation sequencing methodologies.
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De novo Ixodes ricinus salivary gland transcriptome analysis using two next-generation sequencing methodologies
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2013Co-Authors: Alexandra Schwarz, Andrezza C Chagas, Jan Erhart, Björn M. Von Reumont, Michalis KotsyfakisAbstract:Tick salivary gland (SG) proteins possess powerful pharmacologic properties that facilitate tick feeding and pathogen transmission. For the first time, SG transcriptomes of Ixodes ricinus, an important disease Vector for humans and animals, were analyzed using next-generation sequencing. SGs were collected from different tick life stages fed on various animal species, including cofeeding of nymphs and adults on the same host. Four cDNA samples were sequenced, discriminating tick SG transcriptomes of early- and late-feeding nymphs or adults. In total, 441,381,454 pyrosequencing reads and 67,703,183 Illumina reads were assembled into 272,220 contigs, of which 34,560 extensively annotated coding sequences are disclosed; 8686 coding sequences were submitted to GenBank. Overall, 13% of contigs were classified as secreted proteins that showed significant differences in the transcript representation among the 4 SG samples, including high numbers of sample-specific transcripts. Detailed phylogenetic reconstructions of two relatively abundant SG-secreted protein families demonstrated how this study improves our understanding of the molecular evolution of hematophagy in arthropods. Our data significantly increase the available genomic information for I. ricinus and form a solid basis for future tick genome/transcriptome assemblies and the functional analysis of effectors that mediate the feeding physiology and Parasite-Vector interaction of I. ricinus.
Eunice Aparecida Bianchi Galati - One of the best experts on this subject based on the ideXlab platform.
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experimental infection and transmission of leishmania by lutzomyia cruzi diptera psychodidae aspects of the ecology of Parasite Vector interactions
PLOS Neglected Tropical Diseases, 2017Co-Authors: Alessandra Gutierrez De Oliveira, Everton Falcao De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Paula Guerra Murat, Marcio Jose De Medeiros, Alda Izabel De Souza, Eunice Aparecida Bianchi GalatiAbstract:Several parameters should be addressed before incriminating a Vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain Parasite infection under laboratory conditions. This study evaluated the Vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both Parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis–infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both Parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis–infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of Vectorial capacity for L. infantum transmission. Moreover, it was also permissive to L. amazonensis.
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alternative method for the mass rearing of lutzomyia lutzomyia cruzi diptera psychodidae in a laboratory setting
Journal of Medical Entomology, 2015Co-Authors: Everton Falcao De Oliveira, Alessandra Gutierrez De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Eunice Aparecida Bianchi GalatiAbstract:ABSTRACT The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to Parasite—Vector—host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of Vector competence and parameters of the Vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumba, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the...
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alternative method for the mass rearing of lutzomyia lutzomyia cruzi diptera psychodidae in a laboratory setting
Journal of Medical Entomology, 2015Co-Authors: Everton Falcao De Oliveira, Alessandra Gutierrez De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Eunice Aparecida Bianchi GalatiAbstract:The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to Parasite-Vector-host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of Vector competence and parameters of the Vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumba, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the larvae to light was reduced in comparison with Method 1. With Method 2, a significantly greater number of specimens of Lu. cruzi was obtained. The ratio between the number of emerged adults and the females followed for oviposition was 0.42 with Method 1 and 2.75 with Method 2. The optimization of the rearing conditions for Lu. cruzi will enable the establishment of a colony providing a sufficient number of specimens to develop experimental infection by Leishmania as well as Vectorial competence and some parameters of the Vectorial capacity of this sand fly.
Everton Falcao De Oliveira - One of the best experts on this subject based on the ideXlab platform.
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experimental infection and transmission of leishmania by lutzomyia cruzi diptera psychodidae aspects of the ecology of Parasite Vector interactions
PLOS Neglected Tropical Diseases, 2017Co-Authors: Alessandra Gutierrez De Oliveira, Everton Falcao De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Paula Guerra Murat, Marcio Jose De Medeiros, Alda Izabel De Souza, Eunice Aparecida Bianchi GalatiAbstract:Several parameters should be addressed before incriminating a Vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain Parasite infection under laboratory conditions. This study evaluated the Vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both Parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis–infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both Parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis–infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of Vectorial capacity for L. infantum transmission. Moreover, it was also permissive to L. amazonensis.
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alternative method for the mass rearing of lutzomyia lutzomyia cruzi diptera psychodidae in a laboratory setting
Journal of Medical Entomology, 2015Co-Authors: Everton Falcao De Oliveira, Alessandra Gutierrez De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Eunice Aparecida Bianchi GalatiAbstract:ABSTRACT The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to Parasite—Vector—host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of Vector competence and parameters of the Vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumba, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the...
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alternative method for the mass rearing of lutzomyia lutzomyia cruzi diptera psychodidae in a laboratory setting
Journal of Medical Entomology, 2015Co-Authors: Everton Falcao De Oliveira, Alessandra Gutierrez De Oliveira, Wagner De Souza Fernandes, Elisa Teruya Oshiro, Eunice Aparecida Bianchi GalatiAbstract:The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to Parasite-Vector-host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of Vector competence and parameters of the Vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumba, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the larvae to light was reduced in comparison with Method 1. With Method 2, a significantly greater number of specimens of Lu. cruzi was obtained. The ratio between the number of emerged adults and the females followed for oviposition was 0.42 with Method 1 and 2.75 with Method 2. The optimization of the rearing conditions for Lu. cruzi will enable the establishment of a colony providing a sufficient number of specimens to develop experimental infection by Leishmania as well as Vectorial competence and some parameters of the Vectorial capacity of this sand fly.
Petr Volf - One of the best experts on this subject based on the ideXlab platform.
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lutzomyia migonei is a permissive Vector competent for leishmania infantum
Parasites & Vectors, 2016Co-Authors: Vanessa Cristina Fitipaldi Veloso Guimaraes, Katerina Pruzinova, Jovana Sadlova, Vera Volfova, Jitka Myskova, Sinval Pinto Brandao Filho, Petr VolfAbstract:Leishmania infantum is the most widespread etiological agent of visceral leishmaniasis (VL) in the world, with significant mortality rates in human cases. In Latin America, this Parasite is primarily transmitted by Lutzomyia longipalpis, but the role of Lutzomyia migonei as a potential Vector for this protozoan has been discussed. Laboratory and field investigations have contributed to this hypothesis; however, proof of the Vector competence of L. migonei has not yet been provided. In this study, we evaluate for the first time the susceptibility of L. migonei to L. infantum. Females of laboratory-reared L. migonei were fed through a chick-skin membrane on rabbit blood containing L. infantum promastigotes, dissected at 1, 5 and 8 days post-infection (PI) and checked microscopically for the presence, intensity and localisation of Leishmania infections. In addition, morphometric analysis of L. infantum promastigotes was performed. High infection rates of both L. infantum strains tested were observed in L. migonei, with colonisation of the stomodeal valve already on day 5 PI. At the late-stage infection, most L. migonei females had their cardia and stomodeal valve colonised by high numbers of Parasites, and no significant differences were found compared to the development in L. longipalpis. Metacyclic forms were found in all Parasite-Vector combinations since day 5 PI. We propose that Lutzomyia migonei belongs to sand fly species permissive to various Leishmania spp. Here we demonstrate that L. migonei is highly susceptible to the development of L. infantum. This, together with its known anthropophily, abundance in VL foci and natural infection by L. infantum, constitute important evidence that L. migonei is another Vector of this Parasite in Latin America.
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natural hybrid of leishmania infantum l donovani development in phlebotomus tobbi p perniciosus and lutzomyia longipalpis and comparison with non hybrid strains differing in tissue tropism
Parasites & Vectors, 2015Co-Authors: Veronika Seblova, Jitka Myskova, Jan Votýpka, Jana Hlavacova, Maria Antoniou, Petr VolfAbstract:Infection caused by Parasites from L. donovani complex can manifest as a serious visceral disease or a self-healing milder cutaneous form. The different tropism and pathology in humans is caused by the interaction between Parasites, host and Vector determinants but the mechanisms are not well understood. In Cukurova region in Turkey we previously identified a major focus of cutaneous leishmaniasis caused by L. donovani/infantum hybrids (CUK strain) and isolated this Parasite from the locally abundant sand fly, Phlebotomus tobbi. Here, we present the first experimental study with P. tobbi. We tested the susceptibility of this species to various Leishmania under laboratory conditions, characterized glycoproteins in the P. tobbi midgut putatively involved in Parasite-Vector interaction and compared the development of the CUK strain in the sand fly with one other dermotropic and three viscerotropic strains belonging to the L. donovani complex. Females of laboratory reared P. tobbi, P. perniciosus and Lutzomyia longipalpis were infected using membrane feeding on rabbit blood containing promastigotes of various Leishmania species with different tropisms. The individual guts were checked microscopically for presence and localization of Leishmania Parasites; the number of Parasites was assessed more precisely by qPCR. In addition, glycosylation of midgut proteins of P. tobbi was studied by lectin blotting of midgut lysate with lectins specific for terminal sugars of N-type and O-type glycans. High infection rates, heavy Parasite loads and late-stage infection with colonization of the stomodeal valve were observed in P. tobbi infected by Leishmania major or L. infantum CUK hybrid. In parallel, lectin blotting revealed the presence of O-glycosylated proteins in the P. tobbi midgut. In P. perniciosus and L. longipalpis all five Leishmania strains tested developed well. In both Vectors, significantly higher Parasite numbers were detected by qPCR for dermotropic L. donovani from Cyprus, however, in all other parameters studied, including localization of infection and colonization of stomodeal valve, dermotropic and viscerotropic strains were not significantly different. We showed high susceptibility of P. tobbi to various Leishmania spp. This, together with the presence of O-glycosylated midgut proteins in their midguts demonstrate that P. tobbi is a permissive Vector. Two dermotropic and three viscerotropic strains from the L. donovani complex developed late-stage infections in natural L. infantum Vectors, P. perniciosus and L. longipalpis and none of the parameters studied seem to be linked with different tropism of Parasites in the vertebrate host.
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leishmania development in sand flies Parasite Vector interactions overview
Parasites & Vectors, 2012Co-Authors: Anna Dostalova, Petr VolfAbstract:Leishmaniases are Vector-borne parasitic diseases with 0.9 – 1.4 million new human cases each year worldwide. In the Vectorial part of the life-cycle, Leishmania development is confined to the digestive tract. During the first few days after blood feeding, natural barriers to Leishmania development include secreted proteolytic enzymes, the peritrophic matrix surrounding the ingested blood meal and sand fly immune reactions. As the blood digestion proceeds, Parasites need to bind to the midgut epithelium to avoid being excreted with the blood remnant. This binding is strictly stage-dependent as it is a property of nectomonad and leptomonad forms only. While the attachment in specific Vectors (P. papatasi, P. duboscqi and P. sergenti) involves lipophosphoglycan (LPG), this Leishmania molecule is not required for Parasite attachment in other sand fly species experimentally permissive for various Leishmania. During late-stage infections, large numbers of Parasites accumulate in the anterior midgut and produce filamentous proteophosphoglycan creating a gel-like plug physically obstructing the gut. The Parasites attached to the stomodeal valve cause damage to the chitin lining and epithelial cells of the valve, interfering with its function and facilitating reflux of Parasites from the midgut. Transformation to metacyclic stages highly infective for the vertebrate host is the other prerequisite for effective transmission. Here, we review the current state of knowledge of molecular interactions occurring in all these distinct phases of Parasite colonization of the sand fly gut, highlighting recent discoveries in the field.
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the midgut transcriptome of phlebotomus larroussius perniciosus a Vector of leishmania infantum comparison of sugar fed and blood fed sand flies
BMC Genomics, 2011Co-Authors: Anna Dostalova, Petr Volf, Jan Votýpka, Amanda J Favreau, Kent D Barbian, Jesus G Valenzuela, Ryan C JochimAbstract:Parasite-Vector interactions are fundamental in the transmission of Vector-borne diseases such as leishmaniasis. Leishmania development in the Vector sand fly is confined to the digestive tract, where sand fly midgut molecules interact with the Parasites. In this work we sequenced and analyzed two midgut-specific cDNA libraries from sugar fed and blood fed female Phlebotomus perniciosus and compared the transcript expression profiles. A total of 4111 high quality sequences were obtained from the two libraries and assembled into 370 contigs and 1085 singletons. Molecules with putative roles in blood meal digestion, peritrophic matrix formation, immunity and response to oxidative stress were identified, including proteins that were not previously reported in sand flies. These molecules were evaluated relative to other published sand fly transcripts. Comparative analysis of the two libraries revealed transcripts differentially expressed in response to blood feeding. Molecules up regulated by blood feeding include a putative peritrophin (PperPer1), two chymotrypsin-like proteins (PperChym1 and PperChym2), a putative trypsin (PperTryp3) and four putative microvillar proteins (PperMVP1, 2, 4 and 5). Additionally, several transcripts were more abundant in the sugar fed midgut, such as two putative trypsins (PperTryp1 and PperTryp2), a chymotrypsin (PperChym3) and a microvillar protein (PperMVP3). We performed a detailed temporal expression profile analysis of the putative trypsin transcripts using qPCR and confirmed the expression of blood-induced and blood-repressed trypsins. Trypsin expression was measured in Leishmania infantum-infected and uninfected sand flies, which identified the L. infantum-induced down regulation of PperTryp3 at 24 hours post-blood meal. This midgut tissue-specific transcriptome provides insight into the molecules expressed in the midgut of P. perniciosus, an important Vector of visceral leishmaniasis in the Old World. Through the comparative analysis of the libraries we identified molecules differentially expressed during blood meal digestion. Additionally, this study provides a detailed comparison to transcripts of other sand flies. Moreover, our analysis of putative trypsins demonstrated that L. infantum infection can reduce the transcript abundance of trypsin PperTryp3 in the midgut of P. perniciosus.
Alexandra Schwarz - One of the best experts on this subject based on the ideXlab platform.
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de novo ixodes ricinus salivary gland transcriptome analysis using two next generation sequencing methodologies
The FASEB Journal, 2013Co-Authors: Alexandra Schwarz, Andrezza C Chagas, Bjoern M. Von Reumont, Jan Erhart, Michalis KotsyfakisAbstract:Tick salivary gland (SG) proteins possess powerful pharmacologic properties that facilitate tick feeding and pathogen transmission. For the first time, SG transcriptomes of Ixodes ricinus, an important disease Vector for humans and animals, were analyzed using next-generation sequencing. SGs were collected from different tick life stages fed on various animal species, including cofeeding of nymphs and adults on the same host. Four cDNA samples were sequenced, discriminating tick SG transcriptomes of early- and late-feeding nymphs or adults. In total, 441,381,454 pyrosequencing reads and 67,703,183 Illumina reads were assembled into 272,220 contigs, of which 34,560 extensively annotated coding sequences are disclosed; 8686 coding sequences were submitted to GenBank. Overall, 13% of contigs were classified as secreted proteins that showed significant differences in the transcript representation among the 4 SG samples, including high numbers of sample-specific transcripts. Detailed phylogenetic reconstructions of two relatively abundant SG-secreted protein families demonstrated how this study improves our understanding of the molecular evolution of hematophagy in arthropods. Our data significantly increase the available genomic information for I. ricinus and form a solid basis for future tick genome/transcriptome assemblies and the functional analysis of effectors that mediate the feeding physiology and Parasite-Vector interaction of I. ricinus.—Schwarz, A., von Reumont, B.M., Erhart, J., Chagas, A.C., Ribeiro, J.M.C., Kotsyfakis, M. De novo Ixodes ricinus salivary gland transcriptome analysis using two next-generation sequencing methodologies.
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De novo Ixodes ricinus salivary gland transcriptome analysis using two next-generation sequencing methodologies
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2013Co-Authors: Alexandra Schwarz, Andrezza C Chagas, Jan Erhart, Björn M. Von Reumont, Michalis KotsyfakisAbstract:Tick salivary gland (SG) proteins possess powerful pharmacologic properties that facilitate tick feeding and pathogen transmission. For the first time, SG transcriptomes of Ixodes ricinus, an important disease Vector for humans and animals, were analyzed using next-generation sequencing. SGs were collected from different tick life stages fed on various animal species, including cofeeding of nymphs and adults on the same host. Four cDNA samples were sequenced, discriminating tick SG transcriptomes of early- and late-feeding nymphs or adults. In total, 441,381,454 pyrosequencing reads and 67,703,183 Illumina reads were assembled into 272,220 contigs, of which 34,560 extensively annotated coding sequences are disclosed; 8686 coding sequences were submitted to GenBank. Overall, 13% of contigs were classified as secreted proteins that showed significant differences in the transcript representation among the 4 SG samples, including high numbers of sample-specific transcripts. Detailed phylogenetic reconstructions of two relatively abundant SG-secreted protein families demonstrated how this study improves our understanding of the molecular evolution of hematophagy in arthropods. Our data significantly increase the available genomic information for I. ricinus and form a solid basis for future tick genome/transcriptome assemblies and the functional analysis of effectors that mediate the feeding physiology and Parasite-Vector interaction of I. ricinus.
