Sand fly transmitted Leishmania species are responsible for severe, wide ranging, visceral and cutaneous leishmaniases. Genetic exchange can occur among natural Leishmania populations and hybrids can now be produced experimentally, with limitations. Feeding Phlebotomus orientalis or Phlebotomus argentipes on two strains of Leishmania donovani yielded hybrid progeny, selected using double drug resistance and fluorescence markers. Fluorescence activated cell sorting of cultured clones derived from these hybrids indicated diploid progeny. Multilocus sequence typing of the clones showed hybridisation and nuclear heterozygosity, although with inheritance of single haplotypes in a kinetoplastid target. Comparative genomics showed diversity of clonal progeny between single chromosomes, and extraordinary heterozygosity across all 36 chromosomes. Diversity between progeny was seen for the HASPB antigen, which has been noted previously as having implications for design of a therapeutic vaccine. Genomic diversity seen among Leishmania strains and hybrid progeny is of great importance in understanding the epidemiology and control of leishmaniasis. As an outcome of this study we strongly recommend that wider biological archives of different Leishmania species from endemic regions should be established and made available for comparative genomics. However, in parallel, performance of genetic crosses and genomic comparisons should give fundamental insight into the specificity, diversity and limitations of candidate diagnostics, vaccines and drugs, for targeted control of leishmaniasis.
- MeSH
- genomika MeSH
- křížení genetické MeSH
- Leishmania donovani * genetika MeSH
- leishmanióza kožní * MeSH
- leishmanióza viscerální * diagnóza prevence a kontrola epidemiologie MeSH
- Phlebotomus * genetika MeSH
- Psychodidae * genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Visceral leishmaniasis (VL), the most severe form of the disease, is caused by Leishmania donovani in the Indian sub-continent (ISC). Whole genome sequencing studies revealed that two parasite populations exist in the ISC: a main population named the Core Group (CG) found mostly in the lowlands, and a new, genetically different subpopulation called ISC1. Parasites belonging to the CG were shown to be responsible for the recent epidemics, while the ISC1 variant was originally identified in hilly districts of Nepal and was later on increasingly found in the lowlands. Importantly, the ISC1 and CG isolates differ in their drug susceptibility and virulence signatures, suggesting that ISC1 constitutes an emerging and functionally different variant of L. donovani. In present study we aimed to address the potential of ISC1 transmission by the natural vector of L. donovani in the lowlands, Phlebotomus argentipes. By experimental infection of sand flies with parasites of the different genotypes, we demonstrate that ISC1 and CG strains are developing similarly in P. argentipes, suggesting that P. argentipes is a fully competent vector for ISC1 parasites. Integration of previous and current findings shows thus that ISC1 is a new and different variant of L. donovani, fully adapted to spread in the ISC through the main vector. This information is directly useful for managers of the elimination program. Furthermore, integration of our successive studies (genotyping, phenotyping and vector competence) demonstrates the relevance of molecular surveillance and should be of interest for scientists working on vector borne diseases and control managers.
- MeSH
- fylogeneze MeSH
- hmyz - vektory parazitologie MeSH
- Leishmania donovani klasifikace genetika izolace a purifikace MeSH
- leishmanióza viscerální přenos MeSH
- lidé MeSH
- Phlebotomus parazitologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Indie MeSH
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
- MeSH
- gastrointestinální trakt parazitologie MeSH
- hmyz - vektory parazitologie MeSH
- králíci MeSH
- Leishmania infantum genetika růst a vývoj izolace a purifikace MeSH
- Leishmania major genetika růst a vývoj izolace a purifikace MeSH
- leishmanióza kožní epidemiologie parazitologie MeSH
- lidé MeSH
- Phlebotomus parazitologie MeSH
- Psychodidae parazitologie MeSH
- tropismus MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Geografické názvy
- Turecko MeSH
BACKGROUND: Despite their importance in animal and human health, the epidemiology of species of the Leishmania enriettii complex remains poorly understood, including the identity of their biological vectors. Biting midges of the genus Forcipomyia (Lasiohelea) have been implicated in the transmission of a member of the L. enriettii complex in Australia, but the far larger and more widespread genus Culicoides has not been investigated for the potential to include vectors to date. METHODOLOGY/PRINCIPAL FINDINGS: Females from colonies of the midges Culicoides nubeculosus Meigen and C. sonorensis Wirth & Jones and the sand fly Lutzomyia longipalpis Lutz & Nevia (Diptera: Psychodidae) were experimentally infected with two different species of Leishmania, originating from Australia (Leishmania sp. AM-2004) and Brazil (Leishmania enriettii). In addition, the infectivity of L. enriettii infections generated in guinea pigs and golden hamsters for Lu. longipalpis and C. sonorensis was tested by xenodiagnosis. Development of L. enriettii in Lu. longipalpis was relatively poor compared to other Leishmania species in this permissive vector. Culicoides nubeculosus was not susceptible to infection by parasites from the L. enriettii complex. In contrast, C. sonorensis developed late stage infections with colonization of the thoracic midgut and the stomodeal valve. In hamsters, experimental infection with L. enriettii led only to mild symptoms, while in guinea pigs L. enriettii grew aggressively, producing large, ulcerated, tumour-like lesions. A high proportion of C. sonorensis (up to 80%) feeding on the ears and nose of these guinea pigs became infected. CONCLUSIONS/SIGNIFICANCE: We demonstrate that L. enriettii can develop late stage infections in the biting midge Culicoides sonorensis. This midge was found to be susceptible to L. enriettii to a similar degree as Lutzomyia longipalpis, the vector of Leishmania infantum in South America. Our results support the hypothesis that some biting midges could be natural vectors of the L. enriettii complex because of their vector competence, although not Culicoides sonorensis itself, which is not sympatric, and midges should be assessed in the field while searching for vectors of related Leishmania species including L. martiniquensis and "L. siamensis".
- MeSH
- Ceratopogonidae parazitologie MeSH
- gastrointestinální trakt parazitologie MeSH
- hmyz - vektory * MeSH
- křeček rodu Mesocricetus MeSH
- Leishmania enriettii izolace a purifikace MeSH
- leishmanióza parazitologie patologie přenos MeSH
- morčata MeSH
- Psychodidae parazitologie MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The early stage of Leishmania development in sand flies is closely connected with bloodmeal digestion. Here we compared various parameters of bloodmeal digestion in sand flies that are either susceptible (Phlebotomus argentipes and P. orientalis) or refractory (P. papatasi and Sergentomyia schwetzi) to Leishmania donovani, to study the effects on vector competence. The volume of the bloodmeal ingested, time of defecation of bloodmeal remnants, timing of formation and degradation of the peritrophic matrix (PM) and dynamics of proteolytic activities were compared in four sand fly species. Both proven vectors of L. donovani showed lower trypsin activity and slower PM formation than refractory species. Interestingly, the two natural L. donovani vectors strikingly differed from each other in secretion of the PM and midgut proteases, with P. argentipes possessing fast bloodmeal digestion with a very high peak of chymotrypsin activity and rapid degradation of the PM. Experimental infections of P. argentipes did not reveal any differences in vector competence in comparison with previously studied P. orientalis; even the very low initial dose (2×103 promastigotes/ml) led to fully developed late-stage infections with colonization of the stomodeal valve in about 40% of females. We hypothesise that the period between the breakdown of the PM and defecation of the bloodmeal remnants, i.e. the time frame when Leishmania attach to the midgut in order to prevent defecation, could be one of crucial parameters responsible for the establishment of Leishmania in the sand fly midgut. In both natural L. donovani vectors this period was significantly longer than in S. schwetzi. Both vectors are equally susceptible to L. donovani; as average bloodmeal volumes taken by females of P. argentipes and P. orientalis were 0.63 μl and 0.59 μl, respectively, an infective dose corresponding to 1-2 parasites was enough to initiate mature infections.
- MeSH
- druhová specificita MeSH
- feces parazitologie MeSH
- hmyz - vektory imunologie metabolismus parazitologie fyziologie MeSH
- krev metabolismus MeSH
- Leishmania donovani růst a vývoj fyziologie MeSH
- leishmanióza viscerální imunologie MeSH
- membrány parazitologie MeSH
- náchylnost k nemoci MeSH
- přijímání potravy * MeSH
- proteolýza MeSH
- Psychodidae imunologie metabolismus parazitologie fyziologie MeSH
- trávení * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
BACKGROUND: In areas endemic for visceral leishmaniasis (VL), the majority of infected hosts remain asymptomatic but potentially infectious to biting sand flies. Their infectiousness for sand fly vectors is crucial for the transmission of the disease and can be quantified only by xenodiagnosis. However, in the case of human hosts, xenodiagnosis can be problematic for ethical and logistic reasons. The BALB/c mouse model described in this paper was designed to enable xenodiagnostic studies on VL hosts circumventing the need for human volunteers, it permits xenodiagnosis using the same individual host repeatedly, over several months. METHODS: BALB/c mice were intradermally inoculated in the ear pinnae with Leishmania donovani, primarily metacyclic stages isolated from the thoracic midguts of experimentally-infected Phlebotomus orientalis females. Naïve sand flies were allowed to feed on anaesthetized mice in 1-3-weeks- interval, firstly on the site of inoculation of L. donovani (weeks 2-8 post infection, p.i.), later on the whole body of mice (weeks 9 - 15 p.i.). Infections of sand flies were evaluated microscopically or by PCR analysis. RESULTS: Although infected mice did not show any signs of disease, 19% (N = 876) of the P. orientalis females that fed at the site of inoculation, became infected. The majority of L. donovani-positive females (76%) had heavy infections with their stomodeal valves colonized by attached parasites. Inoculated mouse ears remained infective for sand flies until week 15 p.i. Females feeding on other parts of the body remained negative with exception of two groups feeding on contralateral ears by week 12 p.i. On week 15, however, these two mice returned negative at xenodiagnosis of the contralateral ears. In sacrificed mice, the highest parasite numbers were found in inoculated ears and their draining lymph nodes. Infections were detected also in the spleen, liver, blood and rarely in the contralateral ear. CONCLUSIONS: The study showed that BALB/c mice harbored parasites in sufficient numbers to promote heavy infections in P. orientalis and thus comprised a suitable laboratory model for xenodiagnoses of L. donovani. Parasites persisted in the inoculation site and were found transmissible for months to sand flies biting on the same site.
- MeSH
- Leishmania donovani izolace a purifikace MeSH
- leishmanióza viscerální diagnóza parazitologie přenos MeSH
- lidé MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- Phlebotomus parazitologie MeSH
- xenodiagnóza metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: An important factor influencing the transmission dynamics of vector-borne diseases is the contribution of hosts with different parasitemia (no. of parasites per ml of blood) to the infected vector population. Today, estimation of this contribution is often impractical since it relies exclusively on limited-scale xenodiagnostic or artificial feeding experiments (i.e., measuring the proportion of vectors that become infected after feeding on infected blood/host). METHODOLOGY: We developed a novel mechanistic model that facilitates the quantification of the contribution of hosts with different parasitemias to the infection of the vectors from data on the distribution of these parasitemias within the host population. We applied the model to an ample data set of Leishmania donovani carriers, the causative agent of visceral leishmaniasis in Ethiopia. RESULTS: Calculations facilitated by the model quantified the host parasitemias that are mostly responsible for the infection of vector, the sand fly Phlebotomus orientalis. Our findings indicate that a 3.2% of the most infected people were responsible for the infection of between 53% and 79% (mean - 62%) of the infected sand fly vector population. SIGNIFICANCE: Our modeling framework can easily be extended to facilitate the calculation of the contribution of other host groups (such as different host species, hosts with different ages) to the infected vector population. Identifying the hosts that contribute most towards infection of the vectors is crucial for understanding the transmission dynamics, and planning targeted intervention policy of visceral leishmaniasis as well as other vector borne infectious diseases (e.g., West Nile Fever).
- MeSH
- hmyz - vektory parazitologie MeSH
- kohortové studie MeSH
- Leishmania donovani izolace a purifikace MeSH
- leishmanióza viscerální parazitologie přenos MeSH
- lidé MeSH
- logistické modely MeSH
- parazitemie přenos MeSH
- Phlebotomus parazitologie MeSH
- Psychodidae parazitologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Etiopie MeSH
We studied the development of antimony-resistant Leishmania infantum in natural vectors Lutzomyia longipalpis and Phlebotomus perniciosus to ascertain the risk of parasite transmission by sand flies. All three resistant strains produced fully mature late-stage infections in sand flies; moreover, the resistant phenotype was maintained after the passage through the vector. These results highlight the risk of circulation of resistant Leishmania strains and question the use of human drugs for treatment of dogs as Leishmania reservoirs.
Sand flies remain the only proven vectors of Leishmania spp. but recent implementation of PCR techniques has led to increasing speculation about "alternative vectors", including biting midges. Here, we summarize that PCR has considerable limits for studing the role of bloodsucking arthropods in the epidemiology of leishmaniasis. The Leishmania life cycle in the sand fly includes a complex series of interactions which are in many cases species-specific, the early phase of the infection is, however, non-specific to sand flies. These facts should be considered in detection of Leishmania in ,"alternative" or "new" vectors to avoid mistaken speculation about their vector competence.
BACKGROUND: Sand fly species of the genus Sergentomyia are proven vectors of reptilian Leishmania that are non-pathogenic to humans. However, a consideration of the role of Sergentomyia spp. in the circulation of mammalian leishmaniasis appears repeatedly in the literature and the possibility of Leishmania transmission to humans remains unclear. Here we studied the susceptibility of colonized Sergentomyia schwetzi to Leishmania donovani and two other Leishmania species pathogenic to humans: L. infantum and L. major. METHODS: Females of laboratory-reared S. schwetzi were infected by cultured Leishmania spp. by feeding through a chicken membrane, dissected at different time intervals post bloodmeal and examined by light microscopy for the abundance and location of infections. RESULTS: All three Leishmania species produced heavy late stage infections in Lutzomyia longipalpis or Phlebotomus duboscqi sand flies used as positive controls. In contrast, none of them completed their developmental cycle in Sergentomyia females; Leishmania promastigotes developed within the bloodmeal enclosed by the peritrophic matrix (PM) but were defecated together with the blood remnants, failing to establish a midgut infection. In S. schwetzi, the PM persisted significantly longer than in L. longipalpis and it was degraded almost simultaneously with defecation. Therefore, Leishmania transformation from procyclic to long nectomonad forms was delayed and parasites did not attach to the midgut epithelium. CONCLUSIONS: Sergentomyia schwetzi is refractory to human Leishmania species and the data indicate that the crucial aspect of the refractoriness is the relative timing of defecation versus PM degradation.
- MeSH
- hmyz - vektory parazitologie MeSH
- Leishmania donovani růst a vývoj patogenita fyziologie MeSH
- leishmanióza viscerální parazitologie přenos MeSH
- lidé MeSH
- Psychodidae parazitologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
