OBJECTIVES: This study evaluated the implications of clinically acquired miltefosine resistance (MIL-R) by assessing virulence in mice and sand flies to reveal the potential of MIL-R strains to circulate. METHODS: Experimental infections with the MIL-R clinical Leishmania infantum isolate MHOM/FR/2005/LEM5159, having a defect in the LiROS3 subunit of the MIL-transporter, and its syngeneic experimentally reconstituted MIL-S counterpart (LEM5159LiROS3) were performed in BALB/c mice and Lutzomyia longipalpis and Phlebotomus perniciosus sand flies. In mice, the amastigote burdens in liver and spleen were compared microscopically using Giemsa smears and by bioluminescent imaging. During the sand fly infections, the percentage of infected flies, parasite load, colonization of the stomodeal valve and metacyclogenesis were evaluated. The stability of the MIL-R phenotype after sand fly and mouse passage was determined as well. RESULTS: The fitness of the MIL-R strain differed between the mouse and sand fly infection model. In mice, a clear fitness loss was observed compared to the LiROS3-reconstituted susceptible strain. This defect could be rescued by episomal reconstitution with a wildtype LiROS3 copy. However, this fitness loss was not apparent in the sand fly vector, resulting in metacyclogenesis and efficient colonization of the stomodeal valve. Resistance was stable after passage in both sand fly and mouse. CONCLUSION: The natural MIL-R strain is significantly hampered in its ability to multiply and cause a typical visceral infection pattern in BALB/c mice. However, this LiROS3-deficient strain efficiently produced mature infections and metacyclic promastigotes in the sand fly vector highlighting the transmission potential of this particular MIL-R clinical Leishmania strain.
- MeSH
- antiprotozoální látky farmakologie MeSH
- fosforylcholin analogy a deriváty farmakologie MeSH
- hmyz - vektory parazitologie MeSH
- Leishmania infantum * účinky léků genetika patogenita MeSH
- leishmanióza viscerální farmakoterapie patologie MeSH
- léková rezistence genetika MeSH
- membránové transportní proteiny genetika MeSH
- myši inbrední BALB C parazitologie MeSH
- myši MeSH
- Phlebotomus parazitologie MeSH
- protozoální geny MeSH
- Psychodidae parazitologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: The phlebotomine sand fly Phlebotomus perniciosus (Diptera: Psychodidae, Phlebotominae) is a major Old World vector of the protozoan Leishmania infantum, the etiological agent of visceral and cutaneous leishmaniases in humans and dogs, a worldwide re-emerging diseases of great public health concern, affecting 101 countries. Despite the growing interest in the study of this sand fly species in the last years, the development of genomic resources has been limited so far. To increase the available sequence data for P. perniciosus and to start studying the molecular basis of the sexual differentiation in sand flies, we performed whole transcriptome Illumina RNA sequencing (RNA-seq) of adult males and females and de novo transcriptome assembly. RESULTS: We assembled 55,393 high quality transcripts, of which 29,292 were unique, starting from adult whole body male and female pools. 11,736 transcripts had at least one functional annotation, including full-length low abundance salivary transcripts, 981 transcripts were classified as putative long non-coding RNAs and 244 transcripts encoded for putative novel proteins specific of the Phlebotominae sub-family. Differential expression analysis identified 8590 transcripts significantly biased between sexes. Among them, some show relaxation of selective constraints when compared to their orthologs of the New World sand fly species Lutzomyia longipalpis. CONCLUSIONS: In this paper, we present a comprehensive transcriptome resource for the sand fly species P. perniciosus built from short-read RNA-seq and we provide insights into sex-specific gene expression at adult stage. Our analysis represents a first step towards the identification of sex-specific genes and pathways and a foundation for forthcoming investigations into this important vector species, including the study of the evolution of sex-biased genes and of the sexual differentiation in phlebotomine sand flies.
- MeSH
- hmyz - vektory genetika MeSH
- Leishmania infantum genetika patogenita MeSH
- leishmanióza viscerální genetika parazitologie MeSH
- lidé MeSH
- Phlebotomus genetika parazitologie MeSH
- pohlavní dimorfismus MeSH
- psi MeSH
- sekvence aminokyselin MeSH
- transkriptom genetika MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- psi MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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.
We conducted a case-control study to evaluate risk factors for cutaneous leishmaniasis caused by Leishmania infantum outbreaks in villages in the Cukurova region, South Anatolia, Turkey. 282 respondents from eight villages were interviewed using structured questionnaires. Epidemiological and clinical characteristics, personal protection and knowledge of leishmania were analyzed. Young people, aged from 5-19 years, were found to be the most endangered group of villagers. The concurrent presence of both lesions and scars in nine persons may indicate repeated infections. Sleeping without bed nets, ownership of a dog and cattle ownership (living close to a barn and storage of dried dung according univariate analyses) were associated with a significantly increased risk of leishmania infection. Non-impregnated bed nets provided only partial protection, but their use decreased the risk approximately 1.6 times. Further research on the role of dogs in the transmission cycle and the effect of suitable interventions are needed to design the best strategy for disease control. Results suggest that personal protection should be increased, particularly among outdoor sleepers, with insecticide-treated bed nets suggested as the best choice.
- MeSH
- bydlení MeSH
- dítě MeSH
- dobytek MeSH
- domácí zvířata MeSH
- dospělí MeSH
- kojenec MeSH
- Leishmania infantum patogenita MeSH
- leishmanióza kožní epidemiologie prevence a kontrola MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- předškolní dítě MeSH
- průzkumy a dotazníky MeSH
- psi MeSH
- rizikové faktory MeSH
- senioři MeSH
- sítě nad lůžka napuštěné insekticidy využití MeSH
- skot MeSH
- studie případů a kontrol MeSH
- zdraví - znalosti, postoje, praxe MeSH
- zvířata MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- psi MeSH
- senioři MeSH
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Turecko MeSH
We quantified Leishmania infantum parasites transmitted by natural vectors for the first time. Both L. infantum strains studied, dermotropic CUK3 and viscerotropic IMT373, developed well in Phlebotomus perniciosus and Lutzomyia longipalpis. They produced heavy late-stage infection and colonized the stomodeal valve, which is a prerequisite for successful transmission. Infected sand fly females, and especially those that transmit parasites, feed significantly longer on the host (1.5-1.8 times) than non-transmitting females. Quantitative PCR revealed that P. perniciosus harboured more CUK3 strain parasites, while in L. longipalpis the intensity of infection was higher for the IMT373 strain. However, in both sand fly species the parasite load transmitted was higher for the strain with dermal tropism (CUK3). All but one sand fly female infected by the IMT373 strain transmitted less than 600 promastigotes; in contrast, 29% of L. longipalpis and 14% of P. perniciosus infected with the CUK3 strain transmitted more than 1000 parasites. The parasite number transmitted by individual sand flies ranged from 4 up to 4.19×10(4) promastigotes; thus, the maximal natural dose found was still about 250 times lower than the experimental challenge dose used in previous studies. This finding emphasizes the importance of determining the natural infective dose for the development of an accurate experimental model useful for the evaluation of new drugs and vaccines.
- MeSH
- infekce přenášené vektorem MeSH
- Leishmania infantum izolace a purifikace patogenita MeSH
- leishmanióza viscerální přenos MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- Phlebotomus parazitologie MeSH
- Psychodidae parazitologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We have developed a PCR assay that in a single reaction distinguishes between Leishmania infantum and Leishmania donovani strains on the basis of different size of the amplicon. The targeted intergenic region between putative biopterin transporter and nucleotide binding protein on chromosome 35 is highly variable, species-specific and can be amplified from clinical samples. Based on the assay, five tested Leishmania archibaldi and L. infantum strains from the Sudan and Ethiopia clearly belong to L. donovani, which is in accordance with a recent multifactorial analysis of these strains. The nucleotide sequence reported in this paper has been submitted to the GenBankTM with accession number EU068004.
Leishmanióza, parazitární onemocnění přenášené sáním krve některých druhů hmyzu rodu Phlebotomus, postihuje různé věkové skupiny v závislosti na druhu Leishmania spp., geografické lokalitě, rezervoáru nemoci a imunokompetenci hostitele. Viscerální leishmanióza je nejtěžší forma nemoci, která, není-li léčena, ohrožuje život. Rozsah a projev onemocnění závisí na několika faktorech, mezi které zahrnujeme protilátkovou a na buňky vázanou imunitní odpověď hostitele, virulenci infikujících druhů a velikost parazitární nálože. Intermitentní horečka, splenomegalie, hepatomegalie, zvětšení lymfatických uzlin, trombocytopenie, rychlý úbytek tělesné hmotnosti, anémie, leukopenie a hypergamaglobulinemie jsou nejčastější příznaky viscerální leishmaniózy. Autoři popisují dvě děti s viscerální leishmaniózou diagnostikovanou v České republice. Obě děti byly infikovány v oblasti Středozemního moře několik měsíců před prvními klinickými projevy onemocnění.
Leishmaniasis, a parasitic disease transmitted by the suction of blood of some species of sandflies, affects various age groups depending on the infecting Leishmania species, geographic location, disease reservoir, and host immunocompetence. Visceral leishmaniasis is the most severe form of the disease which, if left untreated, could have a life-threatening course. The extent and presentation of the disease depend on several factors, including the humoral and cell-mediated immune response of the host, the virulence of the infecting species, and the parasite burden. Intermittent fever, splenomegaly, hepatomegaly, lymph node enlargement, thrombocytopaenia, rapid weight loss, anaemia, leukopaenia and hypergammaglobulinaemia are the most common findings of visceral leishmaniasis. The authors describe two cases of paediatric visceral leishmaniasis diagnosed in the Czech Republic. Both children were infected in Mediterranean countries several months before clinical presentation.
- Klíčová slova
- Pentostam, Glucantime, Pentacarinat, Am Bisome, Amphocit, Abetcet,
- MeSH
- alopurinol aplikace a dávkování MeSH
- amfotericin B aplikace a dávkování farmakologie MeSH
- antimon aplikace a dávkování MeSH
- dějiny MeSH
- dítě MeSH
- horečka neznámého původu MeSH
- klinické laboratorní techniky MeSH
- Leishmania donovani patogenita růst a vývoj MeSH
- Leishmania infantum patogenita růst a vývoj MeSH
- leishmanióza viscerální diagnóza epidemiologie genetika MeSH
- lidé MeSH
- paromomycin aplikace a dávkování MeSH
- Phlebotomus patogenita MeSH
- splenomegalie terapie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- přehledy MeSH
- Geografické názvy
- Středomoří MeSH
- MeSH
- amfotericin B aplikace a dávkování MeSH
- dítě MeSH
- hepatomegalie etiologie farmakoterapie ultrasonografie MeSH
- klinické laboratorní techniky MeSH
- kojenec MeSH
- Leishmania infantum patogenita MeSH
- lidé MeSH
- splenomegalie etiologie farmakoterapie ultrasonografie MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- Publikační typ
- kazuistiky MeSH
- kongresy MeSH
