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molecular detection of leishmania (leishmania) infantum in phlebotomine sandflies from a visceral leishmaniasis endemic area in northwestern of são paulo state, brazil.this study identified the natural infection rate of leishmania (leishmania) infantum in lutzomyia longipalpis sandflies collected in a neighborhood around a kennel, in dracena, northwestern of são paulo state. this region is highly endemic for visceral leishmaniasis in brazil. insects were captured during 2 to 3 nights monthly for 11 months (january-november 2012) using 10 automatic light traps around a kennel in a transition between periurban and urban neighborhood. capture aimed the determinat ...201829373820
identification of secreted proteins involved in nonspecific dsrna-mediated lutzomyia longipalpis ll5 cell antiviral response.hematophagous insects transmit infectious diseases. sand flies are vectors of leishmaniasis, but can also transmit viruses. we have been studying immune responses of lutzomyia longipalpis, the main vector of visceral leishmaniasis in the americas. we identified a non-specific antiviral response in l. longipalpis ll5 embryonic cells when treated with non-specific double-stranded rnas (dsrnas). this response is reminiscent of interferon response in mammals. we are investigating putative effectors ...201829346269
masp-1 of the complement system enhances clot formation in a microvascular whole blood flow model.the complement and coagulation systems closely interact with each other. these interactions are believed to contribute to the proinflammatory and prothrombotic environment involved in the development of thrombotic complications in many diseases. complement masp-1 (mannan-binding lectin-associated serine protease-1) activates coagulation factors and promotes clot formation. however, this was mainly shown in purified or plasma-based static systems. here we describe the role of masp-1 and complemen ...201829324883
bionomic aspects of lutzomyia evansi and lutzomyia longipalpis, proven vectors of leishmania infantum in an endemic area of non-ulcerative cutaneous leishmaniasis in honduras.some lutzomyia species are the vectors of human leishmaniasis in the americas. visceral and cutaneous leishmaniasis are both endemic in the pacific region of honduras, but the non-ulcerative form is the more frequent clinical manifestation in this region, where lutzomyia longipalpis is the most abundant and the only incriminated vector. taxonomic identification and distribution studies of sand flies are important to understand the epidemiology and to control these neglected tropical diseases.201829304878
antibody response to sand fly saliva is a marker of transmission intensity but not disease progression in dogs naturally infected with leishmania infantum.antibody responses to sand fly saliva have been suggested to be a useful marker of exposure to sand fly bites and leishmania infection and a potential tool to monitor the effectiveness of entomological interventions. exposure to sand fly bites before infection has also been suggested to modulate the severity of the infection. here, we test these hypotheses by quantifying the anti-saliva igg response in a cohort study of dogs exposed to natural infection with leishmania infantum in brazil.201829301571
bacterial diversity of wild-caught lutzomyia longipalpis (a vector of zoonotic visceral leishmaniasis in brazil) under distinct physiological conditions by metagenomics analysis.the leishmaniases are a group of diseases caused by protozoans of the genus leishmania, which are transmitted by the bite of phlebotomine sand flies. in the new world, lutzomyia longipalpis is the most important vector of visceral leishmaniasis and is a proven vector for leishmania infantum chagasi in brazil. during development within the vector, leishmania can interact with a variety of microorganisms such as fungi and bacteria. the presence of bacteria in the midgut of sand flies can influence ...201729284535
detection of leishmania dna and blood meal identification in sand flies (diptera: psychodidae) from lençois maranhenses national park region, brazil.to elucidate portions of the transmission cycles of american tegumentary leishmaniasis (tl) and visceral leishmaniasis (vl) occurring in the region surrounding the lençóis maranhenses national park, an important tourist center in brazil, the present study objectives were to determine the rate of natural infection by leishmania spp. and the blood meal in caught sand flies species in the region. sand flies were captured over 36 mo in 62 locations of the municipality of barreirinhas, maranhão with ...201729281057
the effect of luminous intensity on the attraction of phlebotomine sand flies to light traps.to improve the efficiency of light traps in collecting phlebotomine sand flies, the potential effects of luminous intensity on the attraction of these insects to traps were evaluated. sand flies were collected with hooper pugedo (hp) light traps fitted with 5-mm light-emitting diodes (led) bulbs: green (520 nm wavelength-10,000, 15,000 and 20,000 millicandela (mcd) and blue (470 nm-4,000, 12,000 and 15,000 mcd). a total of 3,264 sand flies comprising 13 species were collected. the collected spec ...201729272438
a defined subunit vaccine that protects against vector-borne visceral leishmaniasis.vaccine development for vector-borne pathogens may be accelerated through the use of relevant challenge models, as has been the case for malaria. because of the demonstrated biological importance of vector-derived molecules in establishing natural infections, incorporating natural challenge models into vaccine development strategies may increase the accuracy of predicting efficacy under field conditions. until recently, however, there was no natural challenge model available for the evaluation o ...201729263878
lipophosphoglycan polymorphisms do not affect leishmania amazonensis development in the permissive vectors lutzomyia migonei and lutzomyia longipalpis.lipophosphoglycan (lpg) is a dominant surface molecule of leishmania promastigotes. its species-specific polymorphisms are found mainly in the sugars that branch off the conserved gal(β1,4)man(α1)-po4 backbone of repeat units. leishmania amazonensis is one of the most important species causing human cutaneous leishmaniasis in the new world. here, we describe lpg intraspecific polymorphisms in two le. amazonensis reference strains and their role during the development in three sand fly species.201729246180
alternative splicing originates different domain structure organization of lutzomyia longipalpis chitinases.background the insect chitinase gene family is composed by more than 10 paralogs, which can codify proteins with different domain structures. in lutzomyia longipalpis, the main vector of visceral leishmaniasis in brazil, a chitinase cdna from adult female insects was previously characterized. the predicted protein contains one catalytic domain and one chitin-binding domain (cbd). the expression of this gene coincided with the end of blood digestion indicating a putative role in peritrophic matri ...201829236932
a temporal comparison of sex-aggregation pheromone gland content and dynamics of release in three members of the lutzomyia longipalpis (diptera: psychodidae) species complex.lutzomyia longipalpis is the south american vector of leishmania infantum, the etiologic agent of visceral leishmaniasis (vl). male l. longipalpis produce a sex-aggregation pheromone that is critical in mating, yet very little is known about its accumulation over time or factors involved in release. this laboratory study aimed to compare accumulation of pheromone over time and determine factors that might influence release in three members of the l. longipalpis species complex.201729194438
co-occurrence and seasonal and environmental distributions of the sandflies lutzomyia longipalpis and nyssomyia whitmani in the city of puerto iguazú, northeastern argentina.the aim of this work was to study the distribution of phlebotominae (diptera: psycodidade) abundance in time and space in an area in northeastern argentina with vector transmission of visceral and tegumentary leishmaniasis. for this, 51 households were selected using a 'worst scenario' criterion where one light trap was set during two consecutive nights in peridomiciles in the transitions between the four seasons, and the environment was surveyed simultaneously. the relationships of phlebotomine ...201729178206
pre-clinical antigenicity studies of an innovative multivalent vaccine for human visceral leishmaniasis.the notion that previous infection by leishmania spp. in endemic areas leads to robust anti-leishmania immunity, supports vaccination as a potentially effective approach to prevent disease development. nevertheless, to date there is no vaccine available for human leishmaniasis. we optimized and assessed in vivo the safety and immunogenicity of an innovative vaccine candidate against human visceral leishmaniasis (vl), consisting of virus-like particles (vlp) loaded with three different recombinan ...201729176865
clinical and immunopathological findings during long term follow-up in leishmania infantum experimentally infected dogs.canine visceral leishmaniasis (cvl) is caused by leishmania infantum, which in the new world is transmitted by lutzomyia longipalpis. while prospective clinical and immunological assessments of dogs experimentally challenged with l. infantum have been previously reported over a relatively short follow-up period, the long-term characterization of infected animals has not been performed to date. we evaluated dogs in a subclinical state for six years following experimental infection with l. infantu ...201729162847
human competence to transmit leishmania infantum to lutzomyia longipalpis and the influence of human immunodeficiency virus infection.visceral leishmaniasis (vl) caused by leishmania infantum is a lethal disease transmitted by sand flies. although, considered a zoonosis with dogs held as the main reservoirs, humans are also sources of infection. therefore, control policies currently focused on dog culling may need to consider that vl and human immunodeficiency virus (hiv)/vl patients may also be infectious, contributing to transmission. reservoir competence of patients with vl without and with hiv infection and of persons asym ...201829141704
immunization against full-length protein and peptides from the lutzomyia longipalpis sand fly salivary component maxadilan protects against leishmania major infection in a murine model.leishmaniasis is an arthropod vectored disease causing considerable human morbidity and mortality. vaccination remains the most realistic and practical means to interrupt the growing number and diversity of sand fly vectors and reservoirs of leishmania. since transmission of leishmania is achieved exclusively by sand fly vectors via immune-modulating salivary substances, conventional vaccination requiring an unmodified host immune response for success are potentially destined to fail unless immu ...201729079105
immunization with ljm11 salivary protein protects against infection with leishmania braziliensis in the presence of lutzomyia longipalpis saliva.leishmania is transmitted in the presence of sand fly saliva. protective immunity generated by saliva has encouraged identification of a vector salivary-based vaccine. previous studies have shown that immunization with ljm11, a salivary protein from lutzomyia longipalpis, is able to induce a th1 immune response and protect mice against bites of leishmania major-infected lutzomyia longipalpis. here, we further investigate if immunization with ljm11 recombinant protein is able to confer cross-prot ...201829037520
new records of phlebotomine sand flies (diptera: psychodidae) from the state of alagoas, northeast of brazil.phlebotomine sand flies are small insects of great medical importance. this study presents new records of phlebotomine sand flies, which augment the list of species, recorded in the state of alagoas, brazil. sand flies were collected using cdc miniature light traps. collections were made between november 2008 and april 2016, in 47 municipalities within the state of alagoas. twenty-seven thousand two-hundred forty-four sand flies were collected, representing 18 species in seven genera. lutzomyia ...201829029319
serological tests fail to discriminate dogs with visceral leishmaniasis that transmit leishmania infantum to the vector lutzomyia longipalpis.the control of reservoirs for leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. here, we assessed the performance of several assays to identify lutzomyia longipalpis infectious dogs.201728954069
the role of gallery forests in maintaining phlebotominae populations: potential leishmania spp. vectors in the brazilian savanna.knowledge on synanthropic phlebotomines and their natural infection by leishmania is necessary for the identification of potential areas for leishmaniasis occurrence.201728953996
environmental suitability for lutzomyia longipalpis in a subtropical city with a recently established visceral leishmaniasis transmission cycle, argentina.visceral leishmaniasis (vl) is an endemic disease in northeastern argentina including the corrientes province, where the presence of the vector and canine cases of vl were recently confirmed in december 2008.201728953995
ph control in the midgut of aedesaegypti under different nutritional conditions.aedes aegypti is one of the most important disease vectors in the world. because their gut is the first site of interaction with pathogens, it is important to understand a. aegypti gut physiology. in this study, we investigated the mechanisms of ph control in the midgut of a. aegypti females under different nutritional conditions. we found that unfed females have an acidic midgut (ph ∼6). the midgut of unfed insects is actively maintained at ph 6 regardless of the ingestion of either alkaline or ...201728931720
dog skin parasite load, tlr-2, il-10 and tnf-α expression and infectiousness.visceral leishmaniosis is a zoonotic disease that is transmitted by lutzomyia longipalpis sandflies. dogs are the main peri-urban reservoir of the disease, and progression of canine leishmaniosis is dependent on the type of immune response elaborated against the parasite. type 1 immunity is characterized by effective cellular response, with production of pro-inflammatory cytokines such as tumour necrosis factor alpha (tnf-α). in contrast, type 2 immunity is predominantly humoral, associated with ...201728929498
the sand fly salivary protein lufaxin inhibits the early steps of the alternative pathway of complement by direct binding to the proconvertase c3b-b.saliva of the blood feeding sand fly lutzomyia longipalpis was previously shown to inhibit the alternative pathway (ap) of the complement system. here, we have identified lufaxin, a protein component in saliva, as the inhibitor of the ap. lufaxin inhibited the deposition of c3b, bb, properdin, c5b, and c9b on agarose-coated plates in a dose-dependent manner. it also inhibited the activation of factor b in normal serum, but had no effect on the components of the membrane attack complex. surface p ...201728912782
sand fly (diptera, psychodidae, phlebotominae) abundance and diversity in areas affected by the são francisco river transposition project in ceará state, brazil.entomological surveillance of sand fly vectors was carried out to support leishmaniasis prevention and control measures in areas affected by the são francisco river transposition project.201728851424
leishmania infection and blood food sources of phlebotomines in an area of brazil endemic for visceral and tegumentary leishmaniasis.the aims of the study were to determine the blood feeding preferences of sandflies and to identify species of leishmania that infected phlebotomines in caxias, maranhão, brazil, an area that is highly endemic for leishmaniasis. sandflies were captured in light traps located in the peridomiciliary environments of randomly selected houses in urban and rural settings between 1800 and 0600 hours on new moon days between march 2013 and february 2015. dna extracts from 982 engorged female sandflies we ...201728837565
molecular detection of wolbachia pipientis in natural populations of sandfly vectors of leishmania infantum in endemic areas: first detection in lutzomyia longipalpis.a polymerase chain reaction-based method was used to screen sandflies for infection with wolbachia (rickettsiales: rickettsiaceae), an intracellular bacterial endosymbiont found in many arthropods and filarial hosts. positive results were obtained in five of 200 field-collected sandflies and were confirmed by sequencing. all sandflies were lutzomyia longipalpis (diptera: psychodidae) captured in a region endemic for visceral leishmaniasis in brazil. this is the first study to identify wolbachia ...201728799248
evaluation of chemical spraying and environmental management efficacy in areas with minor previous application of integrated control actions for visceral leishmaniasis in brazil.leishmaniases are vector-borne diseases that are transmitted to humans through the bite of leishmania-infected phlebotomine sand flies (diptera:psychodidae). the main proved vector of visceral leishmaniais (vl) in the new world - lutzomyia longipalpis - is well-adapted to urban areas and has extensive distribution within the five geographical regions of brazil. integrated public health actions directed for the vector, domestic reservoir and humans for the control of vl are preferentially applied ...201728760482
spatial population dynamics and temporal analysis of the distribution of lutzomyia longipalpis (lutz & neiva, 1912) (diptera: psychodidae: phlebotominae) in the city of clorinda, formosa, argentina.lutzomyia longipalpis, the vector for the causal agent of visceral leishmaniasis (vl), has extended its distribution in the southern cone in the americas. the first urban record of lu. longipalpis in argentina was from the city of clorinda in 2004. the aim of this study was to analyse the monthly distribution and abundance of lu. longipalpis and to evaluate its association with environmental and climatic variables in clorinda city, province of formosa.201728743283
after infection with leishmania infantum, golden hamsters (mesocricetus auratus) become more attractive to female sand flies (lutzomyia longipalpis).in brazil, human and canine visceral leishmaniasis is caused by infection with leishmania infantum, a protist parasite transmitted by blood-feeding female lutzomyia longipalpis sand flies. the objective of this study was to determine if the odour of hamsters, infected with le. infantum, was more attractive than the odour of the same hamsters, before they were infected. the attractiveness of odour collected from individual hamsters (n = 13), before they were infected, was compared in a longitudin ...201728733676
occurrence of lutzomyia longipalpis lutz & neiva 1912 and cerdocyon thous linnaeus 1977, in a visceral leishmaniasis endemic area in brazil.cerdocyon thous presents a wide geographic distribution in brazil and its role as a possible leishmania infantum reservoir in a visceral leishmaniasis (vl) transmission cycle regardless of dogs (canis familiaris) has been discussed. from this perspective, this work describes the occurrence and use of the habitat by cerdocyon thous in a lutzomyia longipalpis occurrence area teresina (piaui - brazil), vl endemic region. three specimens of c. thous were monitored with the use of radio telemetry and ...201728712973
the role of wing geometric morphometrics in the identification of sandflies within the subgenus lutzomyia.the lutzomyia subgenus (diptera: psychodidae) includes sibling species with morphologically indistinguishable females. the aims of this study were to analyse variations in the size and shape of wings of species within the lutzomyia subgenus and to assess whether these analyses might be useful in their identification. wings (n = 733) of 18 species deposited in brazilian collections were analysed by geometric morphometrics, using other genera and subgenera as outgroups. shape variation was summari ...201728707301
visceral leishmaniasis in an environmentally protected area in southeastern brazil: epidemiological and laboratory cross-sectional investigation of phlebotomine fauna, wild hosts and canine cases.leishmaniasis is a rapidly expanding zoonosis that shows increasing urbanization. concern exists regarding the role of wildlife in visceral leishmaniasis (vl) transmission, due to frequent natural or anthropogenic environmental changes that facilitate contact between wildlife, humans and their pets. the municipality of campinas, in southeastern brazil, initially recorded vl in 2009, when the first autochthonous case was confirmed in a dog living in an upscale residential condominium, located ins ...201728704391
influences of climate change on the potential distribution of lutzomyia longipalpis sensu lato (psychodidae: phlebotominae).this study explores the present day distribution of lutzomyia longipalpis in relation to climate, and transfers the knowledge gained to likely future climatic conditions to predict changes in the species' potential distribution. we used ecological niche models calibrated based on occurrences of the species complex from across its known geographic range. anticipated distributional changes varied by region, from stability to expansion or decline. overall, models indicated no significant north-sout ...201728668326
ecological parameters of the (s)-9-methylgermacrene-b population of the lutzomyia longipalpis complex in a visceral leishmaniasis area in são paulo state, brazil.visceral leishmaniasis (vl) is an important public health challenge in brazil because of the high number of human and canine cases reported annually. leishmania infantum is the etiological agent of vl and lutzomyia longipalpis is its main vector. however, evidence suggests that this taxon constitutes a species complex. in sao paulo state, there are two populations of lu. longipalpis, each secreting distinct pheromones, (s)-9-methylgermacrene-b and cembrene 1; both have been associated with diffe ...201728558760
standardization of a continuous assay for glycosidases and its use for screening insect gut samples at individual and populational levels.glycoside hydrolases (ghs) are enzymes able to recognize and cleave glycosidic bonds. insect ghs play decisive roles in digestion, in plant-herbivore, and host-pathogen interactions. gh activity is normally measured by the detection of a release from the substrate of products as sugars units, colored, or fluorescent groups. in most cases, the conditions for product release and detection differ, resulting in discontinuous assays. the current protocols result in using large amounts of reaction mix ...028553236
leishmania infantum dna detected in phlebotomine species from puerto iguazú city, misiones province, argentina.in puerto iguazú city, argentina, human and canine visceral leishmaniasis cases have been recorded since the year 2010, with leishmania infantum as the etiological agent and lutzomyia longipalpis as its main vector. in the present study, polymerase chain reaction and sequencing were used to detect l. infantum dna in 3.9% of the female sandflies captured in puerto iguazú city. this is the first report of l. infantum dna detection in micropygomyia quinquefer, and the second one in lu. longipalpis ...201728476601
in vitro inhibition of leishmania attachment to sandfly midguts and ll-5 cells by divalent metal chelators, anti-gp63 and phosphoglycans.leishmania braziliensis and leishmania infantum are the causative agents of cutaneous and visceral leishmaniasis, respectively. several aspects of the vector-parasite interaction involving gp63 and phosphoglycans have been individually assayed in different studies. however, their role under the same experimental conditions was not studied yet. here, the roles of divalent metal chelators, anti-gp63 antibodies and purified type i phosphoglycans (pgs) were evaluated during in vitro parasite attachm ...201728472733
occurrence and probability maps of lutzomyia longipalpis and lutzomyia cruzi (diptera: psychodidae: phlebotominae) in brazil.leishmaniases are serious diseases caused by trypanosomatid protozoans of the genus leishmania transmitted by the bite of phlebotomine sand flies. we analyzed records pertaining to lutzomyia longipalpis (lutz and neiva, 1912) and lutzomyia cruzi (mangabeira, 1938) in brazil from the following sources: the collection of phlebotomine sand flies of the centro de pesquisas rené rachou/fiocruz (fiocruz-colfleb), the "specieslink" (cria) database, from systematic surveys of scientific articles and gra ...201728472338
first record of lutzomyia (lutzomyia) longipalpis (diptera: psychodidae: phlebotominae) on the trinational frontier (brazil-peru-bolivia) of south-western amazonia.in south america, the main sand fly species involved in the transmission of leishmania infantum chagasi (cunha & chagas, 1937), etiological agent of the visceral leishmaniasis (vl), is lutzomyia longipalpis (lutz & neiva, 1912). the species has been recorded in colombia, venezuela, bolivia, argentina, paraguay, uruguay, and brazil, where it is recorded in 24 of the 27 brazilian states, except acre, amazonas, and santa catarina. collections were carried out for one year (april 2013 to march 2014) ...201728453779
bloodmeal identification in field-collected sand flies from casa branca, brazil, using the cytochrome b pcr method.pcr-based identification of vertebrate host bloodmeals has been performed on several vectors species with success. in the present study, we used a previously published pcr protocol followed by dna sequencing based on primers designed from multiple alignments of the mitochondrial cytochrome b gene used to identify avian and mammalian hosts of various hematophagous vectors. the amplification of a fragment encoding a 359 bp sequence of the cyt b gene yielded recognized amplification products in 192 ...201728399200
increased transmissibility of leishmania donovani from the mammalian host to vector sand flies after multiple exposures to sand fly bites.patients with active visceral leishmaniasis are important reservoirs in the anthroponotic transmission cycle of leishmania donovani. the role of the blood or skin as a source of infection to sand flies remains unclear, and the possible effect of multiple exposures to fly bites on transmissibility has not been addressed.201728329329
seasonality of sand flies (diptera: psychodidae) and leishmania dna detection in vector species in an area with endemic visceral leishmaniasis.leishmaniases are a serious health problem in southeast brazil, including the city of belo horizonte (bh), minas gerais state (mg), where there are high rates of incidence and mortality due to visceral leishmaniases. bh is divided into nine sanitary districts (sd) of which one, the venda nova sd, was selected for this study because it has high rates of positivity for canine leishmaniasis and high incidence of human leishmaniasis.201728327794
structure of salo, a leishmaniasis vaccine candidate from the sand fly lutzomyia longipalpis.immunity to the sand fly salivary protein salo (salivary anticomplement of lutzomyia longipalpis) protected hamsters against leishmania infantum and l. braziliensis infection and, more recently, a vaccine combination of a genetically modified leishmania with salo conferred strong protection against l. donovani infection. because of the importance of salo as a potential component of a leishmaniasis vaccine, a plan to produce this recombinant protein for future scale manufacturing as well as knowl ...201728278244
the current status of the lutzomyia longipalpis (diptera: psychodidae: phlebotominae) species complex.lutzomyia longipalpis s.l. is a complex of sibling species and is the principal vector of american visceral leishmaniasis. the present review summarises the diversity of efforts that have been undertaken to elucidate the number of unnamed species in this species complex and the phylogenetic relationships among them. a wide variety of evidence, including chemical, behavioral and molecular traits, suggests very recent speciation events and complex population structure in this group. although signi ...201728225906
autochthonous outbreak and expansion of canine visceral leishmaniasis, uruguay.we report an outbreak of canine visceral leishmaniasis in uruguay. blood specimens from 11/45 dogs tested positive for leishmania spp. specimens of lutzomyia longipalpis sand flies were captured; typing revealed leishmania infantum. our findings document an expansion of visceral leishmaniasis to southern south america and risk for vectorborne transmission to humans.201728221113
maxadilan-simile expression in nyssomyia neivai, a sandfly vector in an endemic region of brazil, and its immunogenicity in patients with american tegumentary leishmaniasis.maxadilan (max) is a salivary component in the sandfly lutzomyia longipalpis (lutz & neiva 1912), a vector of visceral leishmaniasis. max has a powerful vasodilatory effect and is a candidate vaccine that has been tested in experimental leishmaniasis. nyssomyia neivai (pinto 1926) is a vector of the pathogen responsible for american tegumentary leishmaniasis (atl) in brazil.201728177045
risk analysis and prediction of visceral leishmaniasis dispersion in são paulo state, brazil.visceral leishmaniasis (vl) is an important neglected disease caused by a protozoan parasite, and represents a serious public health problem in many parts of the world. it is zoonotic in europe and latin america, where infected dogs constitute the main domestic reservoir for the parasite and play a key role in vl transmission to humans. in brazil this disease is caused by the protozoan leishmania infantum chagasi, and is transmitted by the sand fly lutzomyia longipalpis. despite programs aimed a ...201728166251
canine visceral leishmaniasis in the metropolitan area of são paulo: pintomyia fischeri as potential vector of leishmania infantum.american visceral leishmaniasis is a zoonosis caused by leishmania infantum and transmitted mainly by lutzomyia longipalpis. however, canine cases have been reported in the absence of this species in the greater são paulo region, where pintomyia fischeri and migonemyia migonei are the predominant species. this raises the suspicion that they could be acting as vectors. therefore, this study sought to investigate specific vector capacity parameters of these species and to compare them with those o ...201728134092
the gut microbiome of the vector lutzomyia longipalpis is essential for survival of leishmania infantum.the vector-borne disease leishmaniasis, caused by leishmania species protozoa, is transmitted to humans by phlebotomine sand flies. development of leishmania to infective metacyclic promastigotes in the insect gut, a process termed metacyclogenesis, is an essential prerequisite for transmission. based on the hypothesis that vector gut microbiota influence the development of virulent parasites, we sequenced midgut microbiomes in the sand fly lutzomyia longipalpis with or without leishmania infant ...201728096483
evidence of neofunctionalization after the duplication of the highly conserved polycomb group gene caf1-55 in the obscura group of drosophila.drosophila caf1-55 protein is a subunit of the polycomb repressive complex prc2 and other protein complexes. it is a multifunctional and evolutionarily conserved protein that participates in nucleosome assembly and remodelling, as well as in the epigenetic regulation of a large set of target genes. here, we describe and analyze the duplication of caf1-55 in the obscura group of drosophila. paralogs exhibited a strong asymmetry in evolutionary rates, which suggests that they have evolved accordin ...201728094282
comparative genomics of canine-isolated leishmania (leishmania) amazonensis from an endemic focus of visceral leishmaniasis in governador valadares, southeastern brazil.leishmaniasis is a highly diverse group of diseases caused by kinetoplastid of the genus leishmania. these parasites are taxonomically diverse, with human pathogenic species separated into two subgenera according to their development site inside the alimentary tract of the sand fly insect vector. the disease encompasses a variable spectrum of clinical manifestations with tegumentary or visceral symptoms. among the causative species in brazil, leishmania (leishmania) amazonensis is an important e ...201728091623
nycthemeral rhythm of phlebotominae (diptera: psychodidae) in a craggy region, transitioning between the wetland and the plateau, brazil.recording the nycthemeral rhythm of sand flies allows the evaluation of the daily activity in different ecotypes, the period of greatest activity, and their degree of anthropophily. we investigated the fauna and the rhythm of sand fly activity in an ecotourism region in mato grosso do sul (ms) state, brazil. sand flies were captured monthly, using a shannon trap for 24 h periods between july 2012 and june 2014. we collected 1,815 sand flies, in which lutzomyia whitmani (=nyssomyia whitmani, sens ...201728082638
photoperiod differences in sand fly (diptera: psychodidae) species richness and abundance in caves in minas gerais state, brazil.caves are unique habitats that are inhabited by a diverse and singular biota. among these inhabitants are sand flies, which are of great epidemiological interest in the neotropical region because they are vectors of leishmania the period of activity of these insects is usually crepuscular and nocturnal, but there are reports of diurnal activity of sand flies in caves. thus, the aim of this study was to evaluate the periodicity of daily activity of sand flies in cave environments in the municipal ...201728082636
first description of leishmania (viannia) infection in evandromyia saulensis, pressatia sp. and trichophoromyia auraensis (psychodidae: phlebotominae) in a transmission area of cutaneous leishmaniasis in acre state, amazon basin, brazil.studies on the sandfly fauna to evaluate natural infection indexes are still limited in the brazilian amazon, a region with an increasing incidence of cutaneous leishmaniasis. here, by using a multiplex polymerase chain reaction directed to leishmania kdna and hybridisation, we were able to identify l. (viannia) subgenus in 12 out of 173 sandflies captured in the municipality of rio branco, acre state, revealing a positivity of 6.94%. by sequencing the leishmania 234 bp-hsp70 amplified products ...201728076470
the search for novel insecticide targets in the post-genomics era, with a specific focus on g-protein coupled receptors.insects are considered pests globally, implicated in the destruction of agricultural fields and transmission of pathogens that cause deadly human diseases, such as dengue, zika and malaria. the diversity of the insecticide arsenal has remained stagnant for decades, but the recent rise of insecticide resistance fueled the discovery of novel modes of action, and the power of genomics has reinvigorated this search. this review discusses the importance of comparative and functional insect genomics i ...201728076467
insect vectors of disease: untapped reservoirs for new antimicrobials?with the increase in antibiotic resistance among infectious diseases, the need for new strategies for identifying compounds with inhibitory effects is dire. traditional methods of genome sequencing and systematic characterization of potential antimicrobial gene clusters, although effective, are unfortunately not yielding results at a speed consistent with the rise in antimicrobial resistance. one approach could be to use a more targeted approach to antimicrobial compound discovery. insect vector ...201628066398
leg loss in lutzomyia longipalpis (diptera: psychodidae) due to pyrethroid exposure: toxic effect or defense by autotomy?phlebotomine sandflies lose their legs after exposure to pyrethroids. in some insects leg loss helps to defend them from intoxication and predation, a phenomenon known as autotomy. a field observation has shown that sandflies that have lost some legs are still able to blood-feed. the aims of the study were to determine whether leg loss in sandflies, after exposure to deltamethrin, is due to autotomy and to establish the effect of the leg loss on blood-feeding.201728035108
increasing potential risk for american visceral leishmaniasis in amapá, brazil.the present note discusses some evidence on the increasing potential risk for american visceral leishmaniasis (avl) transmission in the northern brazilian state of amapá, the guianan-amazon biome.201728001227
sandflies (diptera: psychodidae) in an urban area of northeastern brazil.the sandfly fauna is well studied globally. in brazil, sandfly fauna is very diverse in the northeast region, especially in states such as maranhão, ceará, and bahia. however, in the state of pernambuco, the distribution of these insects is still not well known. therefore, the objective of this study was to identify the different species that constitute the sandfly fauna in an urban area in the northeast region of brazil, where an outbreak of visceral leishmaniasis (vl) was recently reported.201728001216
sand flies (diptera: psychodidae) in an endemic area of leishmaniasis in aquidauana municipality, pantanal of mato grosso do sul , brazil.the aquidauana municipality is considered an endemic area of leishmaniasis and an important tourist site in mato grosso do sul state. the aim of this study was to investigate the sand fly fauna in the city of aquidauana. captures were carried out twice a month, from april 2012 to march 2014 with automatic light traps and active aspiration, in the peridomicile and domicile of six residences. a total of 9,338 specimens were collected, 3,179 and 6,159 using light traps and active aspiration, respec ...201627982353
effect of the saliva from different triatomine species on the biology and immunity of tlr-4 ligand and trypanosoma cruzi-stimulated dendritic cells.triatomines are blood-sucking vectors of trypanosoma cruzi, the causative agent of chagas disease. during feeding, triatomines surpass the skin host response through biomolecules present in their saliva. dendritic cells (dcs) play a crucial role in the induction of the protection to aggressive agents, including blood-sucking arthropods. here, we evaluated if salivary components of triatomines from different genera evade the host immunity by modulating the biology and the function of lps- or t. c ...201627938380
prevalence of cutaneous leishmaniasis in districts of high and low endemicity in mali.historically the western sahelian dry regions of mali are known to be highly endemic for cutaneous leishmaniasis (cl) caused by leishmania major, while cases are rarely reported from the southern savanna forest of the country. here, we report baseline prevalence of cl infection in 3 ecologically distinct districts of mali (dry sahelian, north savanna and southern savanna forest areas). we screened 195 to 250 subjects from 50 to 60 randomly selected households in each of the 6 villages (four from ...201627898671
association between leishmania infantum dna in the hair of dogs and their infectiousness to lutzomyia longipalpis.diagnosis of infection with leishmania infantum by dna detection in the hair has been recently demonstrated in dogs and wild animals. our objective was to investigate if polymerase chain reaction (pcr) in hair might be used to identify infectious dogs. thus, we assessed the infectiousness to lutzomyia longipalpis by xenodiagnosis in comparison with the detection of l. infantum dna by pcr in the hair, and with serology for anti-leishmania igg by elisa in 15 positive dogs for l. infantum infection ...201627890081
proteomic profile of the bradysia odoriphaga in response to the microbial secondary metabolite benzothiazole.benzothiazole, a microbial secondary metabolite, has been demonstrated to possess fumigant activity against sclerotinia sclerotiorum, ditylenchus destructor and bradysia odoriphaga. however, to facilitate the development of novel microbial pesticides, the mode of action of benzothiazole needs to be elucidated. here, we employed itraq-based quantitative proteomics analysis to investigate the effects of benzothiazole on the proteomic expression of b. odoriphaga. in response to benzothiazole, 92 of ...201627883048
essential oil composition, antimicrobial and pharmacological activities of lippia sidoides cham. (verbenaceae) from são gonçalo do abaeté, minas gerais, brazil.lippia sidoides (verbenaceae) is used in brazilian folk medicine as an antiseptic, and it is usually applied topically on skin, mucous membranes, mouth, and throat, or used for vaginal washings.027867267
screening of fungi for biological control of a triatomine vector of chagas disease: temperature and trypanosome infection as factors.entomopathogenic fungi have been investigated as an alternative tool for controlling various insects, including triatomine vectors of the protozoan trypanosoma cruzi, the etiological agent of chagas disease. here we tested the pathogenicity and virulence of ten isolates of the fungi metarhizium spp. and beauveria bassiana against rhodnius prolixus and found all of the isolates to be virulent. we used two isolates (urpe-11 metarhizium anisopliae and ent-1 beauveria bassiana) for further screening ...201627855217
evolutionarily conserved odorant receptor function questions ecological context of octenol role in mosquitoes.olfaction is a key insect adaptation to a wide range of habitats. in the last thirty years, the detection of octenol by blood-feeding insects has been primarily understood in the context of animal host-seeking. the recent discovery of a conserved octenol receptor gene in the strictly nectar-feeding elephant mosquito toxorhynchites amboinensis (taor8) suggests a different biological role. here, we show that taor8 is a functional ortholog of its counterparts in blood-feeding mosquitoes displaying ...201627849027
the lutzomyia longipalpis complex: a brief natural history of aggregation-sex pheromone communication.in this paper we review the natural history of pheromone communication and the current diversity of aggregation-sex pheromones in the sand fly lutzomyia longipalpis. this species complex is the main vector of leishmania infantum, the agent of visceral leishmaniasis in the americas. the identification of variation in pheromone chemotypes combined with molecular and sound analyses have all contributed to our understanding of the extent of divergence among cryptic members of this complex. the impor ...201627842601
prevalence of american trypanosomiasis and leishmaniases in domestic dogs in a rural area of the municipality of são joão do piauí, piauí state, brazil.chagas disease and the leishmaniases are endemic zoonoses of great importance to public health in the state of piauí, brazil. the domestic dog (canis familiaris) is a major reservoir, host of trypanosoma cruzi and leishmania spp. in both urban and rural areas, playing an important role in the transmission of these parasites. the present study evaluated the prevalence of both infectious diseases in dogs of a rural area in the municipality of são joão do piauí, piauí state. one hundred twenty-nine ...201627828620
leishmania infantum infection in dogs from the southern region of minas gerais state, brazil.visceral leishmaniasis is a systemic and chronic disease and dogs are the main reservoir of the etiologic agent, leishmania infantum (syn l. chagasi). a serological and molecular investigation of canine visceral leishmaniasis (cvl) was performed in the municipality of alfenas, located in the southern region of minas gerais, where the disease is not endemic. samples from 87 dogs were submitted to serological tests including the dual path platform (dpp (r) ) cvl bio-manguinhos rapid test, an in-ho ...201627828616
diversity of bacteriome associated with phlebotomus chinensis (diptera: psychodidae) sand flies in two wild populations from china.sand fly phlebotomus chinensis is a primary vector of transmission of visceral leishmaniasis in china. the sand flies have adapted to various ecological niches in distinct ecosystems. characterization of the microbial structure and function will greatly facilitate the understanding of the sand fly ecology, which would provide critical information for developing intervention strategy for sand fly control. in this study we compared the bacterial composition between two populations of ph. chinensis ...201627819272
insecticide resistance in phlebotomine sandflies in southeast asia with emphasis on the indian subcontinent.visceral leishmaniasis, commonly known as kala-azar in india, is a global public health problem. in southeast asia, bangladesh, bhutan, india, nepal, sri lanka and thailand are endemic for visceral leishmaniasis. the role of sandflies as the vector of kala-azar was first confirmed in 1942 in india. insecticide resistance in phlebotomus argentipes annandale and brunetti, the vector of kala-azar in the indian subcontinent, was first reported in 1987 in bihar, india. this article provides a scoping ...201627817749
presence of lutzomyia longipalpis (diptera: psychodidae) in the parque estadual da serra da tiririca, state of rio de janeiro, southeastern brazil.the sand fly, lutzomyia longipalpis, is the main vector of leishmania infantum in the americas, primarily occurring in areas of apparent anthropomorphic modifications in several regions of brazil.201627812657
the diversity of yellow-related proteins in sand flies (diptera: psychodidae).yellow-related proteins (yrps) present in sand fly saliva act as affinity binders of bioamines, and help the fly to complete a bloodmeal by scavenging the physiological signals of damaged cells. they are also the main antigens in sand fly saliva and their recombinant form is used as a marker of host exposure to sand flies. moreover, several salivary proteins and plasmids coding these proteins induce strong immune response in hosts bitten by sand flies and are being used to design protecting vacc ...201627812196
parasite load in the blood and skin of dogs naturally infected by leishmania infantum is correlated with their capacity to infect sand fly vectors.the sand fly lutzomyia longipalpis is primarily responsible for the transmission of visceral leishmaniasis (vl) in the new world, and dogs are considered to be the main urban reservoir of this disease. in order to improve the efficacy of control measures, it is essential to assess the transmission capacity of leishmania infantum to the sand fly vector by naturally infected dogs. the present study investigated the existence of correlations between canine clinical presentation and the intensity of ...201627809965
field evaluation of a new strategy to control lutzomyia longipalpis, based on simultaneous application of an adulticide-larvicide mixture.leishmania infantum (syn. chagasi) is the etiologic agent of visceral leishmaniasis in argentina, and the phlebotomine fly lutzomyia longipalpis is its main vector. the objective of this study was to evaluate the effectiveness of dragon max(®), an emulsifiable concentrate formulation containing the pyrethroid permethrin and the larvicide pyriproxyfen, for lu. longipalpis control under field conditions. the work was conducted in the city of posadas (misiones province, argentina). comparisons were ...201627802395
monthly distribution of phlebotomine sand flies, and biotic and abiotic factors related to their abundance, in an urban area to which visceral leishmaniasis is endemic in corumbá, brazil.the monthly distribution and abundance of sand flies are influenced by both biotic and abiotic factors. the present study aimed to evaluate the seasonal distribution of sand flies and the relation between their abundance and environmental parameters, including vegetation and climate. this study was conducted over a 2-year period (april 2012 to march 2014). monthly distribution was evaluated through the weekly deployment of cdc light traps in the peridomicile area of 5 residences in an urban area ...201627783667
environmental niche modelling of phlebotomine sand flies and cutaneous leishmaniasis identifies lutzomyia intermedia as the main vector species in southeastern brazil.cutaneous leishmaniasis (cl) is caused by a protozoan of the genus leishmania and is transmitted by sand flies. the state of espírito santo (es), an endemic area in southeast brazil, has shown a considerably high prevalence in recent decades. environmental niche modelling (enm) is a useful tool for predicting potential disease risk. in this study, enm was applied to sand fly species and cl cases in es to identify the principal vector and risk areas of the disease. sand flies were collected in 46 ...201627783641
molecular phylogeny of the megadiverse insect infraorder bibionomorpha sensu lato (diptera).the phylogeny of the insect infraorder bibionomorpha (diptera) is reconstructed based on the combined analysis of three nuclear (18s, 28s, cad) and three mitochondrial (12s, 16s, coi) gene markers. all the analyses strongly support the monophyly of bibionomorpha in both the narrow (sensu stricto) and the broader (sensu lato) concepts. the major lineages of bibionomorpha sensu lato (sciaroidea, bibionoidea, anisopodoidea, and scatopsoidea) and most of the included families are supported as monoph ...201627781163
an increase of larval rearing temperature does not affect the susceptibility of phlebotomus sergenti to leishmania tropica but effectively eliminates the gregarine psychodiella sergenti.in mosquitoes, it has previously been shown that rearing conditions of immature stages have an effect on the vector competence of adults. here, we studied the impact of different larval rearing temperatures (27 °c versus 32 °c) on the sand fly phlebotomus sergenti parrot, 1917 and its susceptibility to two parasites: leishmania tropica wright, 1903, a dixenous trypanosomatid transmissible from sand flies to humans, and psychodiella sergenti lantova, volf & votypka, 2010, a monoxenous sand fly gr ...201627756429
nycthemeral rhythm of phlebotominae (diptera: psychodidae) in a craggy region, transitioning between the wetland and the plateau, brazil.recording the nycthemeral rhythm of sand flies allows the evaluation of the daily activity in different ecotypes, the period of greatest activity, and their degree of anthropophily. we investigated the fauna and the rhythm of sand fly activity in an ecotourism region in mato grosso do sul (ms) state, brazil. sand flies were captured monthly, using a shannon trap for 24 h periods between july 2012 and june 2014. we collected 1,815 sand flies, in which lutzomyia whitmani (=nyssomyia whitmani, sens ...201627688269
proceedings of the ix international symposium on phlebotomine sandflies (isops ix), reims, france, june 28th-july 1st, 2016. 201627677701
two in one: cryptic species discovered in biological control agent populations using molecular data and crossbreeding experiments.there are many examples of cryptic species that have been identified through dna-barcoding or other genetic techniques. there are, however, very few confirmations of cryptic species being reproductively isolated. this study presents one of the few cases of cryptic species that has been confirmed to be reproductively isolated and therefore true species according to the biological species concept. the cryptic species are of special interest because they were discovered within biological control ag ...201627648231
dispersal of lutzomyia longipalpis and expansion of canine and human visceral leishmaniasis in são paulo state, brazil.visceral leishmaniasis (vl), a neglected disease, is a serious public health problem that affects millions of people worldwide. the objectives of the study were to evaluate the sensitivity of lutzomyia longipalpis and canine vl (cvl) autochthony early detection and describe the spatial and temporal dispersal of vector and expansion of vl in a brazilian state. we obtained data on the leishmaniasis vector and vl cases in são paulo state (sp), brazil, from the division of endemic disease control an ...201627640322
structural differences in gut bacteria communities in developmental stages of natural populations of lutzomyia evansi from colombia's caribbean coast.lutzomyia evansi, a phlebotomine insect endemic to colombia's caribbean coast, is considered to be the main vector of visceral and cutaneous leishmaniasis in the region. although insects of this species can harbor pathogenic and non-pathogenic microorganisms in their intestinal microbiota, there is little information available about the diversity of gut bacteria present in lutzomyia evansi. in this study, conventional microbiological methods and molecular tools were used to assess the compositio ...201627618991
differences in immune responses against leishmania induced by infection and by immunization with killed parasite antigen: implications for vaccine discovery.the leishmaniases are a group of diseases caused by different species of the protozoan genus leishmania and transmitted by sand fly vectors. they are a major public health problem in almost all continents. there is no effective control of leishmaniasis and its geographical distribution is expanding in many countries. great effort has been made by many scientists to develop a vaccine against leishmaniasis, but, so far, there is still no effective vaccine against the disease. the only way to gener ...201627600664
circulating biomarkers of immune activation, oxidative stress and inflammation characterize severe canine visceral leishmaniasis.clinical manifestations in canine visceral leishmaniasis (cvl) have not been clearly associated with immunological status or disease progression. we simultaneously assessed biomarkers of inflammation, immune activation, oxidative stress, and anti-sand fly saliva igg concentrations in dog sera with different clinical manifestations to characterize a biosignature associated with cvl severity. in a cross-sectional exploratory study, a random population of 70 dogs from an endemic area in brazil was ...201627595802
crosstalk between purinergic receptors and lipid mediators in leishmaniasis.leishmaniasis is a neglected tropical disease affecting millions of people around the world caused by organisms of the genus leishmania. parasite escape mechanisms of the immune system confer the possibility of resistance and dissemination of the disease. a group of molecules that has become a target for leishmania survival strategies are lipid mediators. among them, leukotriene b4 (ltb4) has been described as a pro-inflammatory molecule capable of activating cells of the immune system to combat ...201627595742
sandflies in an urban area of transmission of visceral leishmaniasis in midwest brazil.the phlebotomine fauna of campo grande city, capital of mato grosso do sul state in brazil, an endemic area for visceral leishmaniasis, has been thoroughly investigated, but all the insect collections were undertaken with automatic light traps. the present study sought to investigate the fauna in this city using shannon and disney traps, having human beings and hamsters, respectively, as bait. both types of traps were installed in forest fragment and peridomiciliary areas in the period from 2007 ...201627593433
photoperiod differences in sand fly (diptera: psychodidae) species richness and abundance in caves in minas gerais state, brazil.caves are unique habitats that are inhabited by a diverse and singular biota. among these inhabitants are sand flies, which are of great epidemiological interest in the neotropical region because they are vectors of leishmania the period of activity of these insects is usually crepuscular and nocturnal, but there are reports of diurnal activity of sand flies in caves. thus, the aim of this study was to evaluate the periodicity of daily activity of sand flies in cave environments in the municipal ...201627559123
impact of lbsapsal vaccine in canine immunological and parasitological features before and after leishmania chagasi-challenge.dogs represent the most important domestic reservoir of l. chagasi (syn. l. infantum). a vaccine against canine visceral leishmaniasis (cvl) would be an important tool for decreasing the anxiety related to possible l. chagasi infection and for controlling human visceral leishmaniasis (vl). because the sand fly salivary proteins are potent immunogens obligatorily co-deposited during transmission of leishmania parasites, their inclusion in an anti-leishmania vaccine has been investigated in past d ...201627556586
could phlebotomus mascittii play a role as a natural vector for leishmania infantum? new data.the occurrence of phlebotomine sand flies in central europe was questioned until they were recorded for the first time in germany in 1999, and ten years later also in austria. the aim of this study was to investigate sand flies collected in austria for their carrier status of leishmania spp.201627542911
visceral leishmaniasis in the state of sao paulo, brazil: spatial and space-time analysis.to perform both space and space-time evaluations of visceral leishmaniasis in humans in the state of sao paulo, brazil.201627533364
saliva of rhipicephalus (boophilus) microplus (acari: ixodidae) inhibits classical and alternative complement pathways.rhipicephalus (boophilus) microplus is the main ectoparasite affecting livestock worldwide. for a successful parasitism, ticks need to evade several immune responses of their hosts, including the activation of the complement system. in spite of the importance of r. microplus, previous work only identified one salivary molecule that blocks the complement system. the current study describes complement inhibitory activities induced by r. microplus salivary components and mechanisms elicited by puta ...201627515662
lipophosphoglycans from leishmania amazonensis strains display immunomodulatory properties via tlr4 and do not affect sand fly infection.the immunomodulatory properties of lipophosphoglycans (lpg) from new world species of leishmania have been assessed in leishmania infantum and leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. this glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved gal(β1,4)man(α1)-po4 backbone of repeat units. here, the immunomodulatory activity of lpgs from leishmania amazonensis, the causative agent of di ...201627508930
canine-based strategies for prevention and control of visceral leishmaniasis in brazil.visceral leishmaniasis (vl) is a zoonosis found worldwide. its incidence has increased in brazil in recent years, representing a serious public and animal health problem. the strategies applied in brazil are questionable and are not sufficient to control the disease. thus, we have compared the efficacy of some of the currently available strategies focused on dogs to prevent and control zoonotic vl in endemic areas by optimizing a mathematical model. the simulations showed that the elimination of ...201627471852
argentinian phlebotomine fauna, new records of phlebotominae (diptera: psychodidae) for the country and the province of chaco.sand flies are insects of medical and veterinary importance, because some species are able to transmit several pathogens such as bartonella spp., phlebovirus spp., and protozoan parasites of the genus leishmania (ross). they are widely distributed in the americas, with recordings ranging from canada to argentina. approximately 500 phlebotominae species are known in the americas, of which it is considered that at least 56 are involved in the transmission of leishmaniasis (maroli et al. 2012). pre ...201627470814
characterization of a midgut mucin-like glycoconjugate of lutzomyia longipalpis with a potential role in leishmania attachment.leishmania parasites are transmitted by phlebotomine sand flies and a crucial step in their life-cycle is the binding to the sand fly midgut. laboratory studies on sand fly competence to leishmania parasites suggest that the sand flies fall into two groups: several species are termed "specific/restricted" vectors that support the development of one leishmania species only, while the others belong to so-called "permissive" vectors susceptible to a wide range of leishmania species. in a previous s ...201627457627
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