Modelling the spatial risk of malaria through probability distribution of Anopheles maculipennis s.l. and imported cases.

Malaria remains one of the most important infectious diseases globally due to its high incidence and mortality rates. The influx of infected cases from endemic to non-endemic malaria regions like Europe has resulted in a public health concern over sporadic local outbreaks. This is facilitated by the continued presence of competent Anopheles vectors in non-endemic countries.We modelled the potential distribution of the main malaria vector across Spain using the ensemble of eight modelling techniques based on environmental parameters and the Anopheles maculipennis s.l. presence/absence data collected from 2000 to 2020. We then combined this map with the number of imported malaria cases in each municipality to detect the geographic hot spots with a higher risk of local malaria transmission.The malaria vector occurred preferentially in irrigated lands characterized by warm climate conditions and moderate annual precipitation. Some areas surrounding irrigated lands in northern Spain (e.g. Zaragoza, Logroño), mainland areas (e.g. Madrid, Toledo) and in the South (e.g. Huelva), presented a significant likelihood of A. maculipennis s.l. occurrence, with a large overlap with the presence of imported cases of malaria.While the risk of malaria re-emergence in Spain is low, it is not evenly distributed throughout the country. The four recorded local cases of mosquito-borne transmission occurred in areas with a high overlap of imported cases and mosquito presence. Integrating mosquito distribution with human incidence cases provides an effective tool for the quantification of large-scale geographic variation in transmission risk and pinpointing priority areas for targeted surveillance and prevention.

Mosquitoes in urban green spaces and cemeteries in northern Spain.

BACKGROUND: Mosquitoes inhabiting urban green spaces and cemeteries in Europe represent a crucial facet of public health concern and contribute to the ecological balance. As urbanization intensifies, these areas increasingly serve as vital habitats for various mosquito species, fostering breeding grounds and increasing the risk of disease transmission. METHODS: A study was conducted in the three main cities (inland, coastal, and estuarine) of the Basque Country, northern Spain, to investigate the species composition, abundance, dynamic populations, larval habitats, and host preferences of mosquitoes in urban green spaces and cemeteries. CDC traps and dipping were used to collect mosquitoes for 2 years (2019-2020). RESULTS: A total of 21 mosquito species were identified, with Culex pipiens s.l. being the most abundant and widespread. The three ecological forms of Cx. pipiens were found, and Cx. pipiens pipiens was the most common in both green areas and cemeteries. Morphological identification together with molecular tools identified 65 COI sequences with high homology. The highest species richness was found in the inland city, followed by the coastal city and the estuarine city. Mosquito abundance was significantly higher in green areas compared to cemeteries and in the coastal and estuarine cities compared to the inland city. The investigation of larval breeding sites highlighted the dominance of Cx. pipiens s.l., particularly in semi-artificial ponds, diverse water-holding containers (tyres and buckets) and drainage systems in green areas; in cemeteries, most of the larvae were found in flowerpots and funerary urns. Seasonal activity exhibited variable peaks in mosquito abundance in the different cities, with a notable increase in July or August. Additionally, blood meal analysis revealed that Cx. pipiens s.l. fed on several common urban avian species. CONCLUSIONS: Studies on mosquitoes are essential to understand their role in disease transmission and to design targeted and sustainable management strategies to mitigate the associated risks.

Coxiella burnetii infection persistence in a goat herd during seven kidding seasons after an outbreak of abortions: the effect of vaccination.

Coxiella burnetii infection was monitored during seven kidding seasons (2017-2023) in a dairy goat herd that after an outbreak of Q fever abortions was vaccinated with an inactivated phase I vaccine. Due to the high infection rate just after the outbreak, only the replacement stock was vaccinated during the first three kidding seasons, and when the average herd immunity had decreased (fourth kidding season onwards), the whole herd was vaccinated. Vaginal swabs, feces, and milk were analyzed by PCR to monitor infection, and dust and aerosols were analyzed to measure C. burnetii environmental contamination. One year after the onset of the outbreak, a significant reduction in C. burnetii shedding loads was observed, but the percentage of shedding animals remained high until the third kidding season. By the seventh kidding season, no shedders were detected. The bacterial load excreted was significantly lower in vaccinated compared with unvaccinated animals, and in yearlings compared with multiparous. C. burnetii was detected by PCR in aerosols collected inside the animal premises throughout the study period except in the last season; whereas, aerosols collected outdoors tested negative in the last three kidding seasons. Viable C. burnetii was detectable in environmental dust collected inside the barn until the third kidding season following the outbreak. These results indicate that after an outbreak of Q fever, the risk of infection for humans and susceptible animals can remain high for at least three kidding seasons when the number of C. burnetii animal shedders is still high, even when bacterial excretion is low. IMPORTANCE: Q fever is a zoonosis distributed worldwide. Ruminants are the main reservoir, and infection can cause high rates of abortion. After entering a farm, Coxiella burnetii infection can persist in the animal population over several lambing/kidding periods. Once infection is established in a herd, vaccination with the inactivated Phase I vaccine significantly reduces bacterial shedding, but although at low levels, excretion may continue to occur for several lambing/kidding seasons. The time that C. burnetii remains viable in the farm environment after an outbreak of Q fever determines the period when risk of infection is high for the people in close contact. This work showed that this period extends at least three kidding seasons after the outbreak. These results provided valuable information on the epidemiology of C. burnetii infection in goat herds and may help to develop guidelines for controlling the disease and reducing infection risk for susceptible people and animals.

Prevalence and Incidence of Parkinson's Disease in Latin America: A Meta-Analysis.

BACKGROUND: Parkinson's disease (PD) is a rapidly growing neurodegenerative disorder, but up-to-date epidemiological data are lacking in Latin America. We sought to estimate the prevalence and incidence of PD and parkinsonism in Latin America. METHODS: We searched Medline, Embase, Scopus, Web of Science, Scientific Electronic Library Online, and Literatura Latino-Americana e do Caribe em Ciências da Saúde or the Latin American and Caribbean Health Science Literature databases for epidemiological studies reporting the prevalence or incidence of PD or parkinsonism in Latin America from their inception to 2022. Quality of studies was assessed using the Joanna Briggs Institute (JBI) Critical Appraisal Checklist. Data were pooled via random-effects meta-analysis and analyzed by data source (cohort studies or administrative databases), sex, and age group. Significant differences between groups were determined by meta-regression. RESULTS: Eighteen studies from 13 Latin American countries were included in the review. Meta-analyses of 17 studies (nearly 4 million participants) found a prevalence of 472 (95% CI, 271-820) per 100,000 and three studies an incidence of 31 (95% CI, 23-40) per 100,000 person-years for PD; and seven studies found a prevalence of 4300 (95% CI, 1863-9613) per 100,000 for parkinsonism. The prevalence of PD differed by data source (cohort studies, 733 [95% CI, 427-1255] vs. administrative databases. 114 [95% CI, 63-209] per 100,000, P < 0.01), age group (P < 0.01), but not sex (P = 0.73). PD prevalence in ≥60 years also differed significantly by data source (cohort studies. 1229 [95% CI, 741-2032] vs. administrative databases, 593 [95% CI, 480-733] per 100,000, P < 0.01). Similar patterns were observed for parkinsonism. CONCLUSIONS: The overall prevalence and incidence of PD in Latin America were estimated. PD prevalence differed significantly by the data source and age, but not sex. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Synthesis, structure, and characterisation of a ferromagnetically coupled dinuclear complex containing Co(II) ions in a high spin configuration and thiodiacetate and phenanthroline as ligands and of a series of isomorphous heterodinuclear complexes containing different Co : Zn ratios.

We report the synthesis, crystal structure, and characterisation of a dinuclear Co(II) compound with thiodiacetate (tda) and phenanthroline (phen) as ligands (1), and of a series of metal complexes isomorphous to 1 with different Co : Zn ratios (2, 4 : 1; 3, 1 : 1; 4, 1 : 4; 5, 1 : 10). General characterisation methodologies and X-ray data showed that all the synthesised complexes are isomorphous to Zn(II) and Cu(II) analogues (CSD codes: DUHXEL and BEBQII). 1 consists of centrosymmetric Co(II) ion dimers in which the ions are 3.214 Å apart, linked by two µ-O bridges. Each cobalt atom is in a distorted octahedral environment of the N2O3S type. UV-vis spectra of 1 and 5 are in line with high spin (S = 3/2) Co(II) ions in octahedral coordination and indicate that the electronic structure of both Co(II) ions in the dinuclear unit does not significantly change relative to that of the magnetically isolated Co(II) ion. EPR spectra of powder samples of 5 (Co : Zn ratio of 1 : 10) together with spectral simulation indicated high spin Co(II) ions with high rhombic distortion of the zfs [E/D = 0.31(1), D > 0]. DC magnetic susceptibility experiments on 1 and analysis of the data constraining the E/D value obtained by EPR yielded g = 2.595(7), |D| = 61(1) cm-1, and an intradimer ferromagnetic exchange coupling of J = 1.39(4) cm-1. EPR spectra as a function of Co : Zn ratio for both powder and single crystal samples confirmed that they result from two effective S' = 1/2 spins that interact through dipolar and isotropic exchange interactions to yield magnetically isolated S' = 1 centres and that interdimeric exchange interactions, putatively mediated by hydrophobic interactions between phen moieties, are negligible. The latter observation contrasts with that observed in the Cu(II) analogue, where a transition from S = 1 to S' = 1/2 was observed. Computational calculations indicated that the absence of the interdimeric exchange interaction in 1 is due to a lower Co(II) ion spin density delocalisation towards the metal ligands.

Efficacy of Protein Baits with Fipronil to Control Vespa velutina nigrithorax (Lepeletier, 1836) in Apiaries.

The yellow-legged hornet (Vespa velutina nigrithorax), outside its natural range, has become a major threat to domestic bees. Several control methods have been used to fight against V. velutina, but the results achieved are not satisfactory. The use of protein baits with biocides has shown to be an effective method to control invasive wasp populations, but they have not been used to control V. velutina. Thus, the efficacy of protein baits containing fipronil to reduce the presence of hornets in apiaries was evaluated in this study. After laboratory determination of the optimal efficacy of a protein bait at a 0.01% concentration of fipronil, field trials were conducted involving 222 beekeepers. The data reported by the 90 beekeepers who completed the requested questionnaire demonstrated that in the groups of apiaries with the highest pressure of hornets (groups with 10-30 and >30 hornets), there was a significant decrease in the presence of V. velutina, lasting at least two weeks. The reduction in the number of hornets was positively correlated with bait consumption, and bait consumption was positively correlated with the number of hornets present at the time of treatment. Although the method used has shown good efficacy and the concentration of fipronil used was very low; possible negative effects on the environment should also be evaluated.

A Q fever outbreak among visitors to a natural cave, Bizkaia, Spain, December 2020 to October 2021.

We describe a large Q fever outbreak reported in Spain, including 108 cases, 53 with pneumonia and 27 requiring hospitalisations. The first cases were detected in February 2021 among rock climbers visiting a cave in Bizkaia, and the last case was detected in October 2021. Most cases were notified after the Easter holiday (April-May 2021). More males (63.9%) than females (36.1%) were infected (median ages: 42 (1-68) and 39 years (6-61), respectively). We detected Coxiella burnetii by PCR in faecal, dust and/or aerosol samples taken inside the cave in March 2021, and in dust and aerosol samples collected between March 2021 and February 2023. Coxiella burnetii from dust samples were cultured on Vero cells, showing viability for 24 months. Based on serological and genotyping data, goats sheltering in the cave were the most likely source of infection. The cave was closed on 29 April 2021, movements of goats and sheep in the area were restricted (March-July 2021), and the animals were vaccinated in October 2021. Investigation of Q fever outbreaks requires a multidisciplinary One Health approach as these outbreaks can occur in unexpected places like natural sites where animals are present.

Invasive Aedes mosquitoes in an urban-peri-urban gradient in northern Spain: evidence of the wide distribution of Aedes japonicus.

BACKGROUND: The expansion of invasive mosquitoes throughout Europe has increased in recent decades. In northern Spain, Aedes albopictus was detected for the first time in 2014, and Aedes japonicus was detected in the three Basque provinces in 2020. This study aimed to evaluate the distribution of these mosquito species and their association with factors related to urbanization. METHODS: In 2021, a total of 568 ovitraps were deployed in 113 sampling sites from 45 municipalities with > 10,000 inhabitants. Oviposition substrate sticks were replaced each fortnight and examined for Aedes eggs from June to November. Aedes eggs were counted, and the eggs from a selection of positive oviposition sticks, encompassing at least one stick from each positive ovitrap, were hatched following their life cycle until the adult stage. When egg hatching was not successful, PCR targeting the COI gene and sequencing of amplicons were carried out. RESULTS: Eggs were detected in 66.4% of the sampling sites and in 32.4% of the ovitraps distributed in the three provinces of the Basque Country. Aedes albopictus and Ae. japonicus were widespread in the studied area, confirming their presence in 23 and 26 municipalities, respectively. Co-occurrence of both species was observed in 11 municipalities. The analysis of the presence of Aedes invasive mosquitoes and the degree of urbanization (urban, suburban, peri-urban) revealed that Ae. albopictus showed a 4.39 times higher probability of being found in suburban areas than in peri-urban areas, whereas Ae. japonicus had a higher probability of being found in peri-urban areas. Moreover, the presence of Ae. albopictus was significantly associated with municipalities with a higher population density (mean = 2983 inh/km2), whereas Ae. japonicus was associated with lower population density (mean = 1590 inh/km2). CONCLUSIONS: The wide distribution of Ae. albopictus and Ae. japonicus observed confirmed the spread and establishment of these species in northern Spain. A new colonization area of Ae. japonicus in Europe was confirmed. Due to the potential impact of Aedes invasive mosquitoes on public health and according to our results, surveillance programs and control plans should be designed considering different urbanization gradients, types of environments, and population density.

Mosquito community composition in two major stopover aquatic ecosystems used by migratory birds in northern Spain.

Mosquitoes (Diptera: Culicidae) are common bloodsucking Diptera frequently found in aquatic environments, which are valuable ecosystems for many animal species, particularly migrating birds. Therefore, interactions between these animal species and mosquitoes may play a critical role in pathogen transmission. During 2018-2019, mosquitoes were collected from two aquatic ecosystems in northern Spain using different methodologies and identified using classical morphology and molecular tools. A total of 1529 males and females of 22 native mosquito species (including eight new records for the region) were trapped using CO2 -baited Centers for Disease Control and Prevention (CDC) traps and sweep netting. Among the blood-fed female mosquitoes, 11 vertebrate host species-six mammals and five birds-were identified using DNA barcoding. The developmental sites of eight mosquito species were determined across nine microhabitats, and 11 mosquito species were caught landing on humans. The flight period varied among mosquito species, with some peaking in the spring and others in the summer. Our study highlights the advantages of mosquito sampling using various techniques to comprehensively characterise species composition and abundance. Information on the trophic preferences, biting behaviour and influence of climatic variables on the ecology of mosquitoes is also provided.

Geographic distribution of the V1016G knockdown resistance mutation in Aedes albopictus: a warning bell for Europe.

BACKGROUND: Colonization of large part of Europe by the Asian tiger mosquito Aedes albopictus is causing autochthonous transmission of chikungunya and dengue exotic arboviruses. While pyrethroids are recommended only to reduce/limit transmission, they are widely implemented to reduce biting nuisance and to control agricultural pests, increasing the risk of insurgence of resistance mechanisms. Worryingly, pyrethroid resistance (with mortality < 70%) was recently reported in Ae. albopictus populations from Italy and Spain and associated with the V1016G point mutation in the voltage-sensitive sodium channel gene conferring knockdown resistance (kdr). Genotyping pyrethroid resistance-associated kdr mutations in field mosquito samples represents a powerful approach to detect early signs of resistance without the need for carrying out phenotypic bioassays which require availability of live mosquitoes, dedicated facilities and appropriate expertise. METHODS: Here we report results on the PCR-genotyping of the V1016G mutation in 2530 Ae. albopictus specimens from 69 sampling sites in 19 European countries. RESULTS: The mutation was identified in 12 sites from nine countries (with allele frequencies ranging from 1 to 8%), mostly distributed in two geographical clusters. The western cluster includes Mediterranean coastal sites from Italy, France and Malta as well as single sites from both Spain and Switzerland. The eastern cluster includes sites on both sides of the Black Sea in Bulgaria, Turkey and Georgia as well as one site from Romania. These results are consistent with genomic data showing high connectivity and close genetic relationship among West European populations and a major barrier to gene flow between West European and Balkan populations. CONCLUSIONS: The results of this first effort to map kdr mutations in Ae. albopictus on a continental scale show a widespread presence of the V1016G allele in Europe, although at lower frequencies than those previously reported from Italy. This represents a wake-up call for mosquito surveillance programs in Europe to include PCR-genotyping of pyrethroid resistance alleles, as well as phenotypic resistance assessments, in their routine activities.

Solid-State Characterization of Acetylpyridine Copper Complexes for the Activation of H2 O2 in Advanced Oxidation Processes.

This work describes the synthesis of 4-(4-AcPy) and 3-acetylpyridine (3-AcPy) copper soluble complexes for the activation of hydrogen peroxide and the concomitant generation of reactive oxygen species (ROS). Given the paramagnetic effects of copper ions in the Nuclear Magnetic Resonance (NMR) lines, we aimed at demonstrating that the combination of high-resolution 2D solid-state NMR experiments, Electron Paramagnetic Resonance (EPR), single-crystal X-ray crystallography and Density Functional Theory (DFT) calculations allows a detailed study of the chemical structure of the ligands and the surrounding metal ions. The copper complexes synthesized with CuCl2 were useful for the activation of H2 O2 during which the only ROS was the hydroxyl one, as demonstrated by EPR experiments. A removal of methyl orange (MO) azo-dye higher than 85 % was achieved in 200 minutes, combining 1.7 mM of copper complexes with 60 mM of H2 O2 and 40 µM of MO.

Culicoides species community composition and feeding preferences in two aquatic ecosystems in northern Spain.

BACKGROUND: Aquatic ecosystems provide breeding sites for blood-sucking insects such as Culicoides biting midges (Diptera: Ceratopogonidae), but factors affecting their distribution and host choice are poorly understood. A study was undertaken at two nature reserves in northern Spain to examine the abundance, species composition, population dynamics and feeding patterns of biting midges between 2018 and 2019. METHODS: Culicoides were captured by light suction traps baited with CO2 and by sweep netting vegetation. Blood meals and species identification of blood-fed specimens were determined using cytochrome c oxidase I subunit (COI) DNA barcoding. Multivariate generalized linear models were used to evaluate the associations between the abundance of Culicoides, the species richness and other parameters. RESULTS: The 4973 identified specimens comprised 28 species of Culicoides. These included two species reported for the first time in northern Spain, thus raising to 54 the number of Culicoides species described in the region. Specimens of all 28 species and 99.6% of the total specimens collected were caught in suction traps, while sweep netting vegetation revealed just 11 species and 0.4% of the total specimens. Midge abundance peaked in June/early July, with five species comprising > 80% of the captures: Culicoides alazanicus (24.9%), Culicoides griseidorsum (20.3%), Culicoides poperinghensis (16.2%), Culicoides kibunensis (10.7%) and Culicoides clastrieri (9.6%). DNA barcode analysis of blood meals from eight Culicoides species revealed that they fed on 17 vertebrate species (3 mammals and 14 birds). Species in the subgenus Avaritia were primarily ornithophilic, except for C. griseidorsum and C. poperinghensis. Host DNA from blood meals was successfully amplified from 75% of blood-fed females. A pictorial blood meal digestion scale is provided to accurately assess the blood-fed status of female Culicoides. CONCLUSIONS: The large number of different blood meal sources identified in the midges captured in this study signals the likely importance of wild birds and mammals (e.g. red deer and wild boar) as reservoir/amplifying hosts for pathogens. Available hosts are more exposed to being bitten by biting midge populations in aquatic ecosystems in late spring and early summer.

Molecular identification of tick-borne pathogens (Rickettsia spp., Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Coxiella burnetii and piroplasms) in questing and feeding hard ticks from North-Western Spain.

The occurrence of tick-borne pathogens (TBPs) of human and veterinary interest was studied in questing and feeding ticks collected from wild animals in a region in North-Western Spain. A total of 529 ticks (489 questing, 40 feeding) of seven different species (386 Ixodes ricinus, 53 Haemaphysalis concinna, 27 Haemaphysalis punctata, 25 Dermacentor marginatus, 21 Haemaphysalis inermis, 15 Dermacentor reticulatus, and two Rhipicephalus bursa) were analyzed. Molecular analysis of the 16S rRNA gene in I. ricinus ticks, revealed the presence of two phylogenetic groups in the region. Most of the sequenced ticks (96%) were assigned to I. ricinus haplogroup and 4% of the ticks were phylogenetically related to I. inopinatus haplogroup. Feeding ticks were removed from 17 animals from seven wild species (seven roe deer -Capreolus capreolus-, three wolves -Canis lupus-, two Iberian red deer -Cervus elaphus hispanicus-, two European wild boar -Sus scrofa-, one Cantabrian brown bear -Ursus arctos-, one Eurasian badger -Meles meles-, and one red fox -Vulpes vulpes-). Presence of Rickettsia spp., Anaplasma phagocytophilum, piroplasms, Borrelia burgdorferi sensu lato (s.l.) and Coxiella burnetii were tested in ticks by specific PCR. A total of 92 (17.4%) of the 529 ticks analyzed were positive for at least one of the TBPs tested. Sequencing revealed the presence of the genospecies "Candidatus Rickettsia rioja", Rickettsia raoultii, and Anaplasma phagocytophilum in both questing and feeding ticks. Rickettsia slovaca, Borrelia lusitaniae, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto and Babesia bigemina were only detected in questing ticks, while Babesia sp. badger type A, Theileria OT3 and Hepatozoon canis occurred only in engorged ticks. None of the ticks were positive for C. burnetii. The analysis of the 16S rRNA gene sequences of A. phagocytophilum revealed the presence of three variants (I, X and W) circulating in the region. New host-tick-pathogen interactions have been revealed, finding for the first time the human pathogen R. raoultii in D. reticulatus removed from a Cantabrian brown bear. Co-occurrence between different TBPs were detected in 4.3% of the ticks. The association B. burgdorferi s.l./Rickettsia spp. was detected in questing ticks; and Rickettsia spp./piroplasms and A. phagocytophilum/Theileria OT3 in feeding ticks. The presence of pathogenic agents constitutes a threat to human and animal health, and should be considered in the diagnosis and treatment after a tick bite. This study increases the knowledge on TBPs diversity of medical and veterinary interest circulating between ticks and their hosts in North-Western Spain.

Poly-L-Lysine-Based αGal-Glycoconjugates for Treating Anti-αGal IgE-Mediated Diseases.

Anti-αGal IgE antibodies mediate a spreading allergic condition known as αGal-syndrome (AGS). People exposed to hard tick bites are sensitized to αGal, producing elevated levels of anti-αGal IgE, which are responsible for AGS. This work presents an immunotherapy based on polymeric αGal-glycoconjugates for potentially treating allergic disorders by selectively inhibiting anti-αGal IgE antibodies. We synthesized a set of αGal-glycoconjugates, based on poly-L-lysine of different degrees of polymerization (DP1000, DP600, and DP100), to specifically inhibit in vitro the anti-αGal IgE antibodies in the serum of αGal-sensitized patients (n=13). Moreover, an animal model for αGal sensitization in GalT-KO mice was developed by intradermal administration of hard tick' salivary gland extract, mimicking the sensitization mechanism postulated in humans. The in vitro exposure to all polymeric glycoconjugates (5-10-20-50-100 µg/mL) mainly inhibited anti-αGal IgE and IgM isotypes, with a lower inhibition effect on the IgA and IgG, respectively. We demonstrated a differential anti-αGal isotype inhibition as a function of the length of the poly-L-lysine and the number of αGal residues exposed in the glycoconjugates. These results defined a minimum of 27 αGal residues to inhibit most of the induced anti-αGal IgE in vitro. Furthermore, the αGal-glycoconjugate DP1000-RA0118 (10 mg/kg sc.) showed a high capacity to remove the anti-αGal IgE antibodies (≥75% on average) induced in GalT-KO mice, together with similar inhibition for circulating anti-αGal IgG and IgM. Our study suggests the potential clinical use of poly-L-lysine-based αGal-glycoconjugates for treating allergic disorders mediated by anti-αGal IgE antibodies.

Stable prevalence of Coxiella burnetii in wildlife after a decade of surveillance in northern Spain.

Coxiella burnetii is an obligate intracellular zoonotic bacterium widespread in nature that causes Q fever in animals and humans. The most common sources of human infection are domestic ruminants, but wildlife can also act as reservoir. Here, spleen samples from 652 wild ungulates and 218 wild birds collected in 2011-2019 in the Basque Country (northern Spain) were analysed by real-time PCR (IS1111 gene) and the results compared with data from a past study in 2001-2006. Among wild ungulates, C. burnetii DNA was detected in 7.0% (6/86) of roe deer (Capreolus capreolus), 1.9% (9/484) of wild boar (Sus scrofa) and 2.4% (2/82) of red deer (Cervus elaphus). The prevalence in roe deer was significantly higher compared to wild boar (p = 0.006). Among wild birds, only one white stork (Ciconia ciconia) tested positive. SNP-typing of C. burnetii-positive samples showed that wild ungulates shared SNP 2, SNP 6 and SNP 8 genotypes with domestic ruminants of the region. However, the white stork harboured a C. burnetii genotype (SNP 3) never identified in the studied area before. Comparing these results with those obtained in the same area a decade before (2001-2006), no significant differences were observed in the prevalence of C. burnetii in any of the wildlife species, indicating stability in C. burnetii prevalence. Nevertheless, continuous surveillance is needed to monitor any future changes in the reservoir role of roe deer and wild boar considering the increase in density of both species observed in Europe in the last decades.

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases.

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named "AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108". AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species.

Analysis of environmental dust in goat and sheep farms to assess Coxiella burnetii infection in a Q fever endemic area: Geographical distribution, relationship with human cases and genotypes.

Real-time PCR analysis of environmental samples (dust and aerosols) is an easy tool to investigate the presence of Coxiella burnetii in the farm environment. The aim of this study was to assess the distribution of C. burnetii DNA in dust collected inside animal premises from 272 small ruminant farms in Bizkaia (northern Spain), a region with recent reports of human Q fever cases and outbreaks. Within each farm, 5 samples of dust were collected from difference surfaces, and data on animal census, management procedures, characteristics of the premises and geographic location were collected. Real-time PCR analysis of the dust samples detected presence of C. burnetii DNA in 98 farms (36.0%), flock-prevalence being higher in sheep (38.9%) or mixed ovine-caprine production systems (36.8%), compared to goats (25.0%). Larger bacterial burdens were observed in mixed farms, compared to sheep (p < .05). Single nucleotide polymorphism (SNP) analysis identified 5 different genotypes, with SNP8 being the predominant genotype (73%), followed by SNP6 (11%), SNP2 (9%), SNP4 (5%) and SNP1 (2%). Proportion of farms where C. burnetii DNA was detected differed among the different agricultural counties, and a higher proportion of C. burnetii DNA positive farms was associated with the occurrence of recent human Q fever outbreaks at several geographical locations. Dust sampling in domestic ruminant farms coupled with real-time PCR to screen for the presence of C. burnetii and estimate bacterial load can be a useful tool to identify herds and regions with high prevalence, define priority actions and monitor the effect of control measures. If combined with molecular genotyping and spatial distribution maps, it can help to identify farm contamination sources and trace the origin of human outbreaks.

At the tip of an iceberg: citizen science and active surveillance collaborating to broaden the known distribution of Aedes japonicus in Spain.

BACKGROUND: Active surveillance aimed at the early detection of invasive mosquito species is usually focused on seaports and airports as points of entry, and along road networks as dispersion paths. In a number of cases, however, the first detections of colonizing populations are made by citizens, either because the species has already moved beyond the implemented active surveillance sites or because there is no surveillance in place. This was the case of the first detection in 2018 of the Asian bush mosquito, Aedes japonicus, in Asturias (northern Spain) by the citizen science platform Mosquito Alert. METHODS: The collaboration between Mosquito Alert, the Ministry of Health, local authorities and academic researchers resulted in a multi-source surveillance combining active field sampling with broader temporal and spatial citizen-sourced data, resulting in a more flexible and efficient surveillance strategy. RESULTS: Between 2018 and 2020, the joint efforts of administrative bodies, academic teams and citizen-sourced data led to the discovery of this species in northern regions of Spain such as Cantabria and the Basque Country. This raised the estimated area of occurrence of Ae. japonicus from < 900 km2 in 2018 to > 7000 km2 in 2020. CONCLUSIONS: This population cluster is geographically isolated from any other population in Europe, which raises questions about its origin, path of introduction and dispersal means, while also highlighting the need to enhance surveillance systems by closely combining crowd-sourced surveillance with public health and mosquito control agencies' efforts, from local to continental scales. This multi-actor approach for surveillance (either passive and active) shows high potential efficiency in the surveillance of other invasive mosquito species, and specifically the major vector Aedes aegypti which is already present in some parts of Europe.

One Health Approach: An Overview of Q Fever in Livestock, Wildlife and Humans in Asturias (Northwestern Spain).

This study aimed to investigate the seroprevalence of C. burnetii in domestic ruminants, wild ungulates, as well as the current situation of Q fever in humans in a small region in northwestern Spain where a close contact at the wildlife-livestock-human interface exists, and information on C. burnetii infection is scarce. Seroprevalence of C. burnetii was 8.4% in sheep, 18.4% in cattle, and 24.4% in goats. Real-time PCR analysis of environmental samples collected in 25 livestock farms detected Coxiella DNA in dust and/or aerosols collected in 20 of them. Analysis of sera from 327 wild ungulates revealed lower seroprevalence than that found in domestic ruminants, with 8.4% of Iberian red deer, 7.3% chamois, 6.9% fallow deer, 5.5% European wild boar and 3.5% of roe deer harboring antibodies to C. burnetii. Exposure to the pathogen in humans was determined by IFAT analysis of 1312 blood samples collected from patients admitted at healthcare centers with Q fever compatible symptoms, such as fever and/or pneumonia. Results showed that 15.9% of the patients had IFAT titers ≥ 1/128 suggestive of probable acute infection. This study is an example of a One Health approach with medical and veterinary institutions involved in investigating zoonotic diseases.

Pathological Study of Facial Eczema (Pithomycotoxicosis) in Sheep.

Facial eczema (FE) is a secondary photosensitization disease of farm ruminants caused by the sporidesmin A, produced in the spores of the saprophytic fungus Pithomyces chartarum. This study communicates an outbreak of ovine FE in Asturias (Spain) and characterizes the serum biochemical pattern and the immune response that may contribute to liver damage, favoring cholestasis and the progression to fibrosis and cirrhosis. Animals showed clinical signs of photosensitivity, with decrease of daily weight gain and loss of wool and crusting for at least 6 months after the FE outbreak. Serum activity of γ-glutamyltransferase and alkaline phosphatase were significantly increased in sheep with skin lesions. In the acute phase, edematous skin lesions in the head, hepatocytic and canalicular cholestasis in centrilobular regions, presence of neutrophils in small clumps surrounding deposits of bile pigment, ductular proliferation, as well as cholemic nephrosis, were observed. Macrophages, stained positively for MAC387, were found in areas of canalicular cholestasis. In the chronic phase, areas of alopecia and crusting were seen in the head, and the liver was atrophic with large regeneration nodules and gallstones. Fibrosis around dilated bile ducts, "typical" and "atypical" ductular reaction and an inflammatory infiltrate composed of lymphocytes and pigmented macrophages, with iron deposits and lipofuscin, were found. The surviving parenchyma persisted with a jigsaw pattern characteristic of biliary cirrhosis. Concentric and eccentric myointimal proliferation was found in arteries near damaged bile ducts. In cirrhotic livers, stellated cells, ductular reaction, ectatic bile ducts and presence of M2 macrophages and lymphocytes, were observed in areas of bile ductular reaction.