BACKGROUND: Phlebotomine sand flies are blood-sucking insects that can transmit Leishmania parasites. Hosts bitten by sand flies develop an immune response against sand fly salivary antigens. Specific anti-saliva IgG indicate the exposure to the vector and may also help to estimate the risk of Leishmania spp. transmission. In this study, we examined the canine antibody response against the saliva of Phlebotomus perniciosus, the main vector of Leishmania infantum in the Mediterranean Basin, and characterized salivary antigens of this sand fly species. METHODOLOGY/PRINCIPAL FINDINGS: Sera of dogs bitten by P. perniciosus under experimental conditions and dogs naturally exposed to sand flies in a L. infantum focus were tested by ELISA for the presence of anti-P. perniciosus antibodies. Antibody levels positively correlated with the number of blood-fed P. perniciosus females. In naturally exposed dogs the increase of specific IgG, IgG1 and IgG2 was observed during sand fly season. Importantly, Leishmania-positive dogs revealed significantly lower anti-P. perniciosus IgG2 compared to Leishmania-negative ones. Major P. perniciosus antigens were identified by western blot and mass spectrometry as yellow proteins, apyrases and antigen 5-related proteins. CONCLUSIONS: Results suggest that monitoring canine antibody response to sand fly saliva in endemic foci could estimate the risk of L. infantum transmission. It may also help to control canine leishmaniasis by evaluating the effectiveness of anti-vector campaigns. Data from the field study where dogs from the Italian focus of L. infantum were naturally exposed to P. perniciosus bites indicates that the levels of anti-P. perniciosus saliva IgG2 negatively correlate with the risk of Leishmania transmission. Thus, specific IgG2 response is suggested as a risk marker of L. infantum transmission for dogs.

Department of Parasitology Faculty of Science Charles University Prague Prague Czech Republic

Zobrazit více v PubMed

Gramiccia M, Gradoni L. The current status of zoonotic leishmaniases and approaches to disease control. Int J Parasitol. 2005;35:1169–1180. PubMed

Miró G, Cardoso L, Pennisi MG, Oliva G, Baneth G. Canine leishmaniosis – new concepts and insights on an expanding zoonosis: part two. Trends Parasitol. 2008;24:371–377. PubMed

Molina R, Amela C, Nieto J, Sanandres M, Gonzalez F, et al. Infectivity of dogs naturally infected with Leishmania infantum to colonized Phlebotomus perniciosus. Trans R Soc Trop Med Hyg. 1994;88:491–493. PubMed

Otranto D, Paradies P, de Caprariis D, Stanneck D, Testini G, et al. Toward diagnosing Leishmania infantum infection in asymptomatic dogs in an area where leishmaniasis is endemic. Clin Vaccine Immunol. 2009;16:337–343. PubMed PMC

Maroli M, Bigliocchi F, Khoury C. Sandflies in Italy: observations on their distribution and methods for control. Parassitologia. 1994;36:251–264. PubMed

Kilick-Kendrick R. The biology and control of Phlebotomine sand flies. Clin Dermatol. 1999;17:279–289. PubMed

Hostomska J, Rohousova I, Volfova V, Stanneck D, Mencke N, et al. Kinetics of canine antibody response to saliva of the sand fly Lutzomyia longipalpis. Vector Borne Zoonotic Dis. 2008;8:443–450. PubMed

Gomes RB, Mendonça IL, Silva VC, Ruas J, Silva MB, et al. Antibodies against Lutzomyia longipalpis saliva in the fox Cerdocyon thous and the sylvatic cycle of Leishmania chagasi. Trans R Soc Trop Med Hyg. 2007;101:127–133. PubMed

Oliveira F, Jochim RC, Valenzuela JG, Kamhawi S. Sand flies, Leishmania, and transcriptome-borne solutions. Parasitol Int. 2009;58:1–5. PubMed PMC

Baneth G, Koutinas AF, Solano-Gallego L, Bourdeau P, Ferrer L. Canine leishmaniosis-new concepts and insights on an expanding zoonosis:part one. Trends Parasitol. 2008;24:324–330. PubMed

Collin N, Gomes R, Teixeira C, Cheng L, Laughinghouse A, et al. Sand fly salivary proteins induce strong cellular immunity in a natural reservoir of visceral leishmaniasis with adverse consequences for Leishmania. PLoS Pathog. 2009;5:e1000441. PubMed PMC

Volf P, Volfova V. Establishment and maintenance of sand fly colonies. J Vector Ecol. 2011;36:S1–S9. PubMed

Mencke N, Volf P, Volfova V, Stanneck D. Repellent efficacy of a combination containing imidacloprid and permethrin against sand flies (Phlebotomus papatasi) on dogs. Parasitol Res. 2003;90:S108–S111. PubMed

Tarallo VD, Dantas-Torres F, Lia RP, Otranto D. Phlebotomine sand fly population dynamics in a leishmaniasis endemic peri-urban area in southern Italy. Acta Trop. 2010;116:227–234. PubMed

Otranto D, de Caprariis D, Lia RP, Tarallo V, Lorusso V, et al. Prevention of endemic canine vector-borne diseases using imidacloprid 10% and permethrin 50% in young dogs: a longitudinal field study. Vet Parasitol. 2010;172:323–332. PubMed

Otranto D, Paradies P, de Caprariis D, Stanneck D, Testini G, et al. Toward diagnosing Leishmania infantum infection in asymptomatic dogs in an area where leishmaniasis is endemic. Clin Vaccine Immunol. 2009;16:337–343. PubMed PMC

Anderson JM, Oliveira F, Kamhawi S, Mans BJ, Reynoso D, et al. Comparative salivary gland transcriptomics of sandfly vectors of visceral leishmaniasis. BMC Genomics. 2006;7:a.n. 52. PubMed PMC

Gerstman BB. Epidemiology kept simple: an introduction to traditional and modern epidemiology. Willey-Liss, Hoboken, New Yersey; 2003. 417

Katz J, Baptista J, Azen SP, Pike MC. Obtaining confidence intervals for the risk ratio in cohort studies. Biometrics. 1978;34:469–474.

Mazza G, Whiting AH, Day MJ, Duffus WPH. Development of an enzyme-linked immunosorbent assay for the detection of IgG subclasses in the serum of normal and diseased dogs. Res Vet Sci. 1994;57:133–139. PubMed

Rohousova I, Volf P. Sand fly saliva: effects on host immune response and Leishmania transmission. Folia Parasitol. 2006;53:161–171. PubMed

Rohousova I, Ozensoy S, Ozbel Y, Volf P. Detection of species-specific antibody response of humans and mice bitten by sand flies. Parasitology. 2005;130:493–499. PubMed

de Moura TR, Oliveira F, Novais FO, Miranda JC, Clarencio J, et al. Enhanced Leishmania braziliensis Infection Following Pre-Exposure to Sandfly Saliva. PloS Negl Trop Dis. 2007;1:e84. PubMed PMC

Barral A, Honda E, Caldas A, Costa J, Vinhas V, et al. Human immune response to sand fly salivary gland antigens: a useful epidemiological marker? Am J Trop Med Hyg. 2000;62:740–745. PubMed

Gomes RB, Brodskyn C, de Oliveira CI, Costa J, Miranda JC, et al. Seroconversion against Lutzomyia longipalpis saliva concurrent with the development of anti-Leishmania chagasi delayed-type hypersensitivity. J Infect Dis. 2002;186:1530–1534. PubMed

Bahia D, Gontijo NF, Leon IR, Perales J, Pereira MH, et al. Antibodies from dogs with canine visceral leishmaniosis recognise two proteins from the saliva of Lutzomyia longipalpis. Parasitol Res. 2007;100:449–454. PubMed

Teixeira C, Gomes R, Collin N, Reynoso D, Jochim R, et al. Discovery of markers of exposure specific to bites of Lutzomyia longipalpis, the vector of Leishmania infantum chagasi in Latin America. PLoS Negl Trop Dis. 2010;4:e638. PubMed PMC

Exposure to Phlebotomus perniciosus sandfly vectors is positively associated with Toscana virus and Leishmania infantum infection in human blood donors in Murcia Region, southeast Spain

Phlebotomus perniciosus Recombinant Salivary Proteins Polarize Murine Macrophages Toward the Anti-Inflammatory Phenotype

Interactions between host biogenic amines and sand fly salivary yellow-related proteins

Monitoring Leishmania infection and exposure to Phlebotomus perniciosus using minimal and non-invasive canine samples

Exploring the relationship between susceptibility to canine leishmaniosis and anti-Phlebotomus perniciosus saliva antibodies in Ibizan hounds and dogs of other breeds in Mallorca, Spain

Field study of the improved rapid sand fly exposure test in areas endemic for canine leishmaniasis

Antibody response to Phlebotomus perniciosus saliva in cats naturally exposed to phlebotomine sand flies is positively associated with Leishmania infection

Human antibody reaction against recombinant salivary proteins of Phlebotomus orientalis in Eastern Africa

Seasonal dynamics of canine antibody response to Phlebotomus perniciosus saliva in an endemic area of Leishmania infantum

Evaluation of the rSP03B sero-strip, a newly proposed rapid test for canine exposure to Phlebotomus perniciosus, vector of Leishmania infantum

A fine scale eco-epidemiological study on endemic visceral leishmaniasis in north ethiopian villages

Insights into the sand fly saliva: Blood-feeding and immune interactions between sand flies, hosts, and Leishmania

The Diversity of Yellow-Related Proteins in Sand Flies (Diptera: Psychodidae)

Recombinant Salivary Proteins of Phlebotomus orientalis are Suitable Antigens to Measure Exposure of Domestic Animals to Sand Fly Bites

Exposure to Leishmania spp. and sand flies in domestic animals in northwestern Ethiopia

Canine Antibodies against Salivary Recombinant Proteins of Phlebotomus perniciosus: A Longitudinal Study in an Endemic Focus of Canine Leishmaniasis

Comparative analysis of salivary gland transcriptomes of Phlebotomus orientalis sand flies from endemic and non-endemic foci of visceral leishmaniasis

Recombinant antigens from Phlebotomus perniciosus saliva as markers of canine exposure to visceral leishmaniases vector

Kinetics of antibody response in BALB/c and C57BL/6 mice bitten by Phlebotomus papatasi

Salivary gland transcriptomes and proteomes of Phlebotomus tobbi and Phlebotomus sergenti, vectors of leishmaniasis