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.

Department of Parasitology Faculty of Science Charles University Prague Czech Republic

Zobrazit více v PubMed

Elnaiem DEA (2011) Ecology and control of the sand fly vectors of Leishmania donovani in East Africa, with special emphasis on Phlebotomus orientalis . J Vector Ecol 36: S23–S31. 10.1111/j.1948-7134.2011.00109.x PubMed DOI

Maroli M, Feliciangeli MD, Bichaud L, Charrel RN, Gradoni L (2013) Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern. Med Vet Entomol 27: 123–147. 10.1111/j.1365-2915.2012.01034.x PubMed DOI

Bates PA, Rogers ME (2004) New insights into the developmental biology and transmission mechanisms of Leishmania . Curr Mol Med 4: 601–609. PubMed

Kamhawi S (2006) Phlebotomine sand flies and Leishmania parasites: friends or foes? Trends Parasitol 22: 439–445. PubMed

Dostalova A, Volf P (2012) Leishmania development in sand flies: parasite-vector interactions overview. Parasit Vectors 5: 276 10.1186/1756-3305-5-276 PubMed DOI PMC

Ramalho-Ortigao M, Saraiva EM, Traub-Cseko YM (2010) Sand fly-Leishmania interactions: long relationships are not necessarily easy. Open Parasitol J 4: 195–204. PubMed PMC

Dillon RJ, Lane RP (1993) Bloodmeal digestion in the midgut of Phlebotomus papatasi and Phlebotomus langeroni . Med Vet Entomol 7: 225–232. PubMed

Telleria EL, de Araujo APO, Secundino NF, d'Avila-Levy CM, Traub-Cseko YM (2010) Trypsin like serine proteases in Lutzomyia longipalpis—expression, activity and possible modulation by Leishmania infantum chagasi . PLoS One 5: e10697 10.1371/journal.pone.0010697 PubMed DOI PMC

Borovsky D, Schlein Y (1987) Trypsin and Chymotrypsin-like enzymes of the sandfly Phlebotomus papatasi infected with Leishmania and their possible role in vector competence. Med Vet Entomol 1: 235–242. PubMed

Schlein Y, Jacobson RL (1998) Resistance of Phlebotomus papatasi to infection with Leishmania donovani is modulated by components of the infective bloodmeal. Parasitology 117: 467–473. PubMed

Pimenta PFP, Modi GB, Pereira ST, Shahabuddin M, Sacks DL (1997) Novel role for the peritrophic matrix in protecting Leishmania from the hydrolytic activities of the sand fly midgut. Parasitology 115: 359–369. PubMed

Adler S (1938) Factors determining the behaviour of Leishmania sp. in sandflies. Harefuah 14: 1–6.

Schlein Y, Romano H (1986) Leishmania major and Leishmania donovani—effects on proteolytic enzymes of Phlebotomus papatasi (Diptera, Psychodidae). Exp Parasitol 62: 376–380. PubMed

Dostalova A, Votypka J, Favreau AJ, Barbian KD, Volf P, Valenzuela JG, et al. (2011) The midgut transcriptome of Phlebotomus (Larroussius) perniciosus, a vector of Leishmania infantum: comparison of sugar fed and blood fed sand flies. BMC Genomics 12: 223 10.1186/1471-2164-12-223 PubMed DOI PMC

Santos VC, Vale VF, Silva SM, Nascimento AA, Saab NA, Soares RPP, et al. (2014) Host modulation by a parasite: How Leishmania infantum modifies the intestinal environment of Lutzomyia longipalpis to favor its development. PLoS One 9: e111241 10.1371/journal.pone.0111241 PubMed DOI PMC

Lehane MJ (1997) Peritrophic matrix structure and function. Annu Rev Entomol 42: 525–550. PubMed

Pascoa V, Oliveira PL, Dansa-Petretski M, Silva JR, Alvarenga PH, Jacobs-Lorena M, et al. (2002) Aedes aegypti peritrophic matrix and its interaction with heme during blood digestion. Insect Biochem Mol Biol 32: 517–523. PubMed

de Araujo APO, Telleria EL, Dutra JDF, Julio RM, Traub-Cseko YM (2012) Disruption of the peritrophic matrix by exogenous chitinase feeding reduces fecundity in Lutzomyia longipalpis females. Mem Inst Oswaldo Cruz 107: 543–545. PubMed

Sadlova J, Volf P (2009) Peritrophic matrix of Phlebotomus duboscqi and its kinetics during Leishmania major development. Cell Tissue Res 337: 313–325. 10.1007/s00441-009-0802-1 PubMed DOI PMC

Bates PA (2008) Leishmania sand fly interaction: progress and challenges. Curr Opin Microbiol 11: 340–344. 10.1016/j.mib.2008.06.003 PubMed DOI PMC

Volf P, Myskova J (2007) Sand flies and Leishmania: specific versus permissive vectors. Trends Parasitol 23: 91–92. PubMed PMC

Myskova J, Svobodova M, Beverley SM, Volf P (2007) A lipophosphoglycan-independent development of Leishmania in permissive sand flies. Microbes Infect 9: 317–324. PubMed PMC

Hoogstraal H, Heyneman D (1969) Leishmaniasis in Sudan Republic. 30. Final Epidemiologic Report. Am J Trop Med Hyg 18: 1091–1210.

Sadlova J, Dvorak V, Seblova V, Warburg A, Votypka J, Volf P (2013) Sergentomyia schwetzi is not a competent vector for Leishmania donovani and other Leishmania species pathogenic to humans. Parasit Vectors 6: 186 10.1186/1756-3305-6-186 PubMed DOI PMC

Volf P, Volfova V (2011) Establishment and maintenance of sand fly colonies. J Vector Ecol 36: S1–S9. 10.1111/j.1948-7134.2011.00106.x PubMed DOI

Sadlova J, Reishig J, Volf P (1998) Prediuresis in female Phlebotomus sandflies (Diptera: Psychodidae). Eur J Entomol 95: 643–647.

Briegel H, Lea AO, Klowden MJ (1979) Hemoglobinometry as a method for measuring blood meal sizes of mosquitos (Diptera, Culicidae). J Med Entomol 15: 235–238. PubMed

Benkova I, Volf P (2007) Effect of temperature on metabolism of Phlebotomus papatasi (Diptera: Psychodidae). J Med Entomol 44: 150–154. PubMed

Gemetchu T (1974) Morphology and fine structure of midgut and peritrophic membrane of adult female, Phlebotomus longipes Parrot and Martin (Diptera-Psychodidae). Ann Trop Med Parasitol 68: 111–124. PubMed

Seblova V, Volfova V, Dvorak V, Pruzinova K, Votypka J, Kassahun A, et al. (2013) Phlebotomus orientalis sand flies from two geographically distant ethiopian localities: biology, genetic analyses and susceptibility to Leishmania donovani . PLoS Negl Trop Dis 7: e2187 10.1371/journal.pntd.0002187 PubMed DOI PMC

Myskova J, Votypka J, Volf P (2008) Leishmania in sand flies: Comparison of quantitative polymerase chain reaction with other techniques to determine the intensity of infection. J Med Entomol 45: 133–138. PubMed

Mary C, Faraut F, Lascombe L, Dumon H (2004) Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 42: 5249–5255. PubMed PMC

Wilson R, Bates MD, Dostalova A, Jecna L, Dillon RJ, Volf P, et al. (2010) Stage-specific adhesion of Leishmania promastigotes to sand fly midguts assessed using an Iimproved comparative binding assay. PLoS Negl Trop Dis 4: e816 10.1371/journal.pntd.0000816 PubMed DOI PMC

Sant'Anna MRV, Diaz-Albiter H, Mubaraki M, Dillon RJ, Bates PA (2009) Inhibition of trypsin expression in Lutzomyia longipalpis using RNAi enhances the survival of Leishmania . Parasit Vectors 2: 62 10.1186/1756-3305-2-62 PubMed DOI PMC

Sand fly blood meal volumes and their relation to female body weight under experimental conditions

RNAi-mediated gene silencing of Phlebotomus papatasi defensins favors Leishmania major infection

Experimental feeding of Sergentomyia minuta on reptiles and mammals: comparison with Phlebotomus papatasi

Genomic analysis of two phlebotomine sand fly vectors of Leishmania from the New and Old World

Molecular Rationale of Insect-Microbes Symbiosis-From Insect Behaviour to Mechanism

Phlebotomus papatasi Antimicrobial Peptides in Larvae and Females and a Gut-Specific Defensin Upregulated by Leishmania major Infection

Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

The Phlebotomus papatasi systemic transcriptional response to trypanosomatid-contaminated blood does not differ from the non-infected blood meal

Role for the flagellum attachment zone in Leishmania anterior cell tip morphogenesis

A novel MALDI-TOF MS-based method for blood meal identification in insect vectors: A proof of concept study on phlebotomine sand flies

Experimental Infection of Sand Flies by Massilia Virus and Viral Transmission by Co-Feeding on Sugar Meal

Refractoriness of Sergentomyia schwetzi to Leishmania spp. is mediated by the peritrophic matrix

Leishmania mortality in sand fly blood meal is not species-specific and does not result from direct effect of proteinases

Leishmania donovani development in Phlebotomus argentipes: comparison of promastigote- and amastigote-initiated infections

The Development of Leishmania tropica in Sand Flies (Diptera: Psychodidae): A Comparison of Colonies Differing in Geographical Origin and a Gregarine Coinfection