Background: This report describes L. infantum infection seroprevalence in dogs in Spain through data obtained from peer-reviewed literature and a cross-sectional serological survey assessing epidemiological and habitat variables as risk factors for infection. The study also provides preliminary sand fly species distribution data and indicates factors affecting their distribution and density. Methods: Three different studies were conducted in Spain: (i) a peer-reviewed literature seroprevalence survey (1985–2019); (ii) a cross-sectional serological survey (2011–2016); and (iii) a preliminary entomological survey (2013–2014). In the cross-sectional serological survey, 1739 dogs from 74 different locations including 25 Spanish provinces were tested for L. infantum by indirect immunofluorescence antibody test (IFAT) (antibody titre ≥ 1:100). Seroprevalence of L. infantum infection was analysed by province and bioclimatic zone. Statistics were used to analyse relationships between several dog- and environment-related variables and L. infantum seroprevalence. In parallel, during 2013–2014, sand flies were collected across the Iberian Peninsula and the Balearic Islands using CDC light traps to examine relationships between habitat-related factors and sand fly species densities (number of sand flies per trap per hour). Results: The literature review revealed that the provinces showing the highest seroprevalence were Balearic Islands (57.1%), Ourense (35.6%), Málaga (34.6%) and Cáceres (34.2%), and those showing the lowest seroprevalence were Vizcaya (0%), Cantabria (2.0%) and Álava (3.3%). In our survey, anti-Leishmania IgG antibodies were detected in 176 of the 1739 dogs rendering a seroprevalence of 10.12%. Percentage seroprevalence distributions significantly varied among bioclimatic belts. Seropositivity for L. infantum was related to size (large breed dogs versus small) and were significantly higher in younger dogs (≤ 1 years-old). In the entomological survey, 676 sand flies of five species were captured: 562 (83.13%) Phlebotomus perniciosus; 64 (9.47%) Sergentomyia minuta; 38 (5.62%) P. ariasi: 6 (0.89%) P. sergenti; and 6 (0.89%) P. papatasi. Phlebotomus perniciosus showed a greater density in the thermo-Mediterranean than in the meso-Mediterranean zone. Densities of S. minuta and P. ariasi were significantly higher in rural habitats. Conclusions: This updated seroprevalence map of L. infantum infection in dogs in Spain defines non-endemic, hypoendemic, endemic and hyperendemic areas, and confirms P. perniciosus as the most abundant sand fly vector in Spain.

The cat flea, Ctenocephalides felis, is the most prevalent flea species detected on dogs and cats in Europe and other world regions. The status of flea infestation today is an evident public health concern because of their cosmopolitan distribution and the flea-borne diseases transmission. This study determines the spatial distribution of the cat flea C. felis infesting dogs in Spain. Using geospatial tools, models were constructed based on entomological data collected from dogs during the period 2013–2015. Bioclimatic zones, covering broad climate and vegetation ranges, were surveyed in relation to their size. The models builded were obtained by negative binomial regression of several environmental variables to show impacts on C. felis infestation prevalence: land cover, bioclimatic zone, mean summer and autumn temperature, mean summer rainfall, distance to urban settlement and normalized difference vegetation index. In the face of climate change, we also simulated the future distributions of C. felis for the global climate model (GCM) “GFDL-CM3” and for the representative concentration pathway RCP45, which predicts their spread in the country. Predictive models for current climate conditions indicated the widespread distribution of C. felis throughout Spain, mainly across the central northernmost zone of the mainland. Under predicted conditions of climate change, the risk of spread was slightly greater, especially in the north and central peninsula, than for the current situation. The data provided will be useful for local veterinarians to design effective strategies against flea infestation and the pathogens transmitted by these arthropods.

Background: Chronic schistosomiasis can lead to significant morbidity. Serology is highly sensitive; however, its role in assessing treatment response is controversial. This study aimed to analyze serological values following treatment of chronic imported schistosomiasis. Methods: A retrospective observational study was performed including patients treated for chronic imported schistosomiasis from 2018 to 2022 who had at least one serological result at baseline and during follow-up. Demographic, clinical, and laboratory data were evaluated. Generalized estimating equation (GEE) models and Kaplan–Meier curves were used to analyze the evolution of serological values. Results: Of the 83 patients included, 72 (86.7%) were male, and the median age was 26 years (IQR 22–83). Most patients, 76 (91.6%), were migrants from sub-Saharan Africa. While 24 cases (28.9%) presented with urinary symptoms, the majority (59; 71.1%) were asymptomatic. Schistosoma haematobium eggs were observed in five cases (6.2%). Eosinophilia was present in 34 participants (40.9%). All patients had an initial positive Schistosoma ELISA serology, median ODI 2.3 (IQR 1.5–4.4); the indirect hemagglutination (IHA) test was positive/indeterminate in 34 cases (43.1%). Following treatment with praziquantel, serology values significantly decreased: −0.04 (IC95% −0.073, −0.0021) and −5.73 (IC95% −9.92, −1.53) units per month for ELISA and IHA, respectively. A quarter of patients (25%) had negative ELISA results 63 weeks after treatment. All symptomatic cases were clinically cured. Conclusions: Serial serological determinations could be helpful for monitoring chronic schistosomiasis in non-endemic regions. The ideal timing for these follow-up tests is yet to be determined. Further research is needed to determine the factors that influence a negative result during follow-up.

Molecular methods are increasingly used for both species identification of sandflies and assessment of their population structure. In general, they are based on DNA sequence analysis of targets previously amplified by PCR. However, this approach requires access to DNA sequence facilities, and in some circumstances, it is time-consuming. Though DNA sequencing provides the most reliable information, other downstream PCR applications are explored to assist in species identification. Thus, it has been recently proposed that the amplification of a DNA region encompassing partially both the cytochrome-B (cytb) and the NADH dehydrogenase 1 (nd1) genes followed by RFLP analysis with the restriction enzyme Ase I allows the rapid identification of the most prevalent species of phlebotomine sandflies in the Mediterranean region. In order to confirm the suitability of this method, we collected, processed, and molecularly analyzed a total of 155 sandflies belonging to four species including Phlebotomus ariasi, P. papatasi, P. perniciosus, and Sergentomyia minuta from different regions in Spain. This data set was completed with DNA sequences available at the GenBank for species prevalent in the Mediterranean basin and the Middle East. Additionally, DNA sequences from 13 different phlebotomine species (P. ariasi, P. balcanicus, P. caucasicus, P. chabaudi, P. chadlii, P. longicuspis, P. neglectus, P. papatasi, P. perfiliewi, P. perniciosus, P. riouxi, P. sergenti, and S. minuta), from 19 countries, were added to the data set. Overall, our molecular data revealed that this PCR-RFLP method does not provide a unique and specific profile for each phlebotomine species tested. Intraspecific variability and similar RFLP patterns were frequently observed among the species tested. Our data suggest that this method may not be applicable throughout the Mediterranean region as previously proposed. Other molecular approaches like DNA barcoding or phylogenetic analyses would allow a more precise molecular species identification.

Highly active antiretroviral therapy (HAART) has reduced the incidence of VL/HIV dramatically. However, HAART only partially prevents relapses, with one-year relapse rates ranging from 30 to 60%. Consequently, secondary prophylaxis is recommended for patients with <200 CD4+ cells/μL. In clinical practice, characterizing cellular immune response could help estimate the risk of relapse in VL/HIV coinfected patients. In this study, the lymphoproliferative response after stimulation with soluble Leishmania antigen was assessed in 2022 and 2023 in three cases of VL/HIV coinfection with long-term follow-up (17, 8 and 19 years). PCR and rK-39 results for Leishmania, HIV viral load, CD4 cell count, proliferation index, IFN-γ, IL-2, IP-10, IL-10 and TNF-α were determined. Heterogeneous results were obtained, with only one patient having developed specific cellular immunity against Leishmania. No cases of relapse were observed. The heterogeneity of lymphoproliferative test results in the three cases described highlights the need to identify surrogate markers of cure to guide maintenance or withdrawal of prophylaxis.

Background: Schistosomiasis remains a significant neglected tropical disease in Kenya, but its presence in the western/northern Lake Turkana region is poorly characterised. We conducted an exploratory serosurvey to assess evidence of Schistosoma spp. exposure and a diagnostic accuracy study to evaluate dried blood spots (DBSs) for field serology. Methods: We performed a cross-sectional survey in adults (≥18 years) from six communities in the western/northern and shoreline area of Turkana Lake, excluding individuals with >6 months of residence in other Kenyan endemic areas. Capillary blood was collected on DBSs and tested centrally using ELISA for Schistosoma spp. IgG. In parallel, DBS cards performance was compared with paired routine serum ELISA in 23 patients assessed for suspected schistosomiasis at our centre. Results: We enrolled 155 participants (60% men; median age 30 years), with nearly universal reported freshwater contact (154/155, 99.4%). In the validation study, DBS values were lower than serum values (mean bias 0.27), with moderate correlation (r = 0.54) and modest discrimination (AUC 0.65; sensitivity 80% and specificity 50% at OD index >0.8). The median DBS ELISA OD index for the 155 participants was 0.55 (IQR, 0.34–0.79). Six samples exceeded 0.8, but these values were low, and all had negative IHA (<1/80), yielding no confirmed seropositive cases. Conclusions: These findings suggest low or absent sustained transmission in the sampled communities during the study period and indicate that DBS-based serology is operationally feasible but requires careful calibration and confirmatory testing for robust field inference.