Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.

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BackgroundLeishmaniosis is associated with Phlebotomus sand fly vector density, but our knowledge of the environmental framework that regulates highly overdispersed vector abundance distributions is limited. We used a standardized sampling procedure in the bioclimatically diverse Murcia Region in Spain and multilevel regression models for count data to estimate P. perniciosus abundance in relation to environmental and anthropic factors.MethodsTwenty-five dog and sheep premises were sampled for sand flies using adhesive and light-attraction traps, from late May to early October 2015. Temperature, relative humidity and other animal- and premise-related data recorded on site and other environmental data were extracted from digital databases using a geographical information system. The relationship between sand fly abundance and explanatory variables was analysed using binomial regression models.ResultsThe total number of sand flies captured, mostly with light-attraction traps, was 3,644 specimens, including 80% P. perniciosus, the main L. infantum vector in Spain. Abundance varied between and within zones and was positively associated with increasing altitude from 0 to 900 m above sea level, except from 500 to 700 m where it was low. Populations peaked in July and especially during a 3-day heat wave when relative humidity and wind speed plummeted. Regression models indicated that climate and not land use or soil characteristics have the greatest impact on this species density on a large geographical scale. In contrast, micro-environmental factors such as animal building characteristics and husbandry practices affect sand fly population size on a smaller scale.ConclusionsA standardised sampling procedure and statistical analysis for highly overdispersed distributions allow reliable estimation of P. perniciosus abundance and identification of environmental drivers. While climatic variables have the greatest impact at macro-environmental scale, anthropic factors may be determinant at a micro-geographical scale. These finding may be used to elaborate predictive distribution maps useful for vector and pathogen control programs.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2135-3) contains supplementary material, which is available to authorized users.

The epidemiological cycle of zoonotic phlebotomine‐borne Leishmania infantum is a complex system in which domestic animals and wildlife interact and participate in its maintenance and transmission. In this study, we combined entomological surveillance, xenomonitoring of L. infantum and identification of host feeding sources of engorged females to investigate the potential contribution of a periurban wildlife park to leishmaniosis in neighbouring residential areas. Overall, 7,309 sand flies were collected in 111 trap‐days during the summers of 2016–2018 in an endemic area in south‐east Spain. Five different sand fly species were captured, with Phlebotomus perniciosus, the main L. infantum vector in this region, representing the most common species. Sand fly distribution was spatially heterogeneous in terms of species, sexes and female physiological stage (unfed, gravid and engorged females) and related to host distribution and management, and environmental features. None of the 602 sand flies analysed for L. infantum infection by kinetoplast real‐time PCR were positive. We used molecular tools to identify the vertebrate hosts of sand flies and identified 17 host species, mainly mammals. Human DNA was not identified in engorged sand flies. This study provides evidence that wildlife parks in south‐east Spain are ideal grounds for sand fly vectors but do not necessarily increase L. infantum infection risk to humans and dogs living in surrounding residential areas. This is probably because vectors feed mostly on non‐L. infantum competent hosts and this should be investigated for a better understanding of the contribution of wildlife parks to the local epidemiology of L. infantum.