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Understanding animal movement behaviour is key to furthering our knowledge on intra- and inter-specific competition, group cohesion, energy expenditure, habitat use, the spread of zoonotic diseases or species management. We used a radial basis function surface approximation subject to minimum description length constraint to uncover the state-space dynamical systems from time series data. This approximation allowed us to infer structure from a mathematical model of the movement behaviour of sheep and red deer, and the effect of density, thermal stress and vegetation type. Animal movement was recorded using GPS collars deployed in sheep and deer grazing a large experimental plot in winter and summer. Information on the thermal stress to which animals were exposed was estimated using the power consumption of mechanical heated models and meteorological records of a network of stations in the plot. Thermal stress was higher in deer than in sheep, with less differences between species in summer. Deer travelled more distance than sheep, and both species travelled more in summer than in winter; deer travel distance showed less seasonal differences than sheep. Animal movement was better predicted in deer than in sheep and in winter than in summer; both species showed a swarming behaviour in group cohesion, stronger in deer. At shorter separation distances swarming repulsion was stronger between species than within species. At longer separation distances inter-specific attraction was weaker than intra-specific; there was a positive density-dependent effect on swarming, and stronger in deer than in sheep. There was not clear evidence which species attracted or repelled the other; attraction between deer at long separation distances was stronger when the model accounted for thermal stress, but in general the dynamic movement behaviour was hardly affected by the thermal stress. Vegetation type affected intra-species interactions but had little effect on inter-species interactions. Our modelling approach is useful in interpreting animal interactions, in order to unravel complex cooperative or competitive behaviours, and to the best of our knowledge is the first modelling attempt to make predictions of multi-species animal movement under different habitat mosaics and abiotic environmental conditions.

The definition of the most relevant parameters that describe the wild boar (WB) population dynamics is essential to guide African swine fever (ASF) control policies. These parameters should be framed considering different contexts, such as geographic, ecological and management contexts, and gaps of data useful for the parameter definition should be identified. This information would allow better harmonized monitoring of WB populations and higher impact of ASF management actions, as well as better parametrizing population dynamics and epidemiological models, which is key to develop more efficient cost‐benefit strategies. This report presents a comprehensive compilation and description of parameters of WB population dynamics, including general drivers, population demography, mortality, reproduction, and spatial behaviour. Beyond the collection of current available data, we provided an open data model to allow academics and wildlife professionals to continuously update new and otherwise hardly accessible data, e.g. those from grey literature which is often not publicly available or only in local languages. This data model, conceived as an open resource and collaborative approach, will be incorporated in the European Observatory of Wildlife (EOW) platform, and include all drivers and population parameters that should be specified in studies on wild boar, and wildlife in general, ecology and epidemiology at the most suitable spatio‐temporal resolution. This harmonized approach should be extended to other taxa in the future as an essential tool to improve European capacities to monitor, to produce risk assessment and to manage wildlife under an international perspective. [ABSTRACT FROM AUTHOR]

Abstract   Sarcoptes mite from collection to DNA extraction forms the cornerstone for studies on Sarcoptes scabiei. Whilst the new science era took a shy leap into the different facets of mite studies, the cornerstone was almost entirely neglected. Mite collection, cleaning, storage and DNA extraction were, basically, humble attempts to extrapolate, adapt, modify or ‘pirate’ those existing methods to the peculiarities of Sarcoptes research. These aspects usually constituted few lines, bashfully mentioned, in the materials and methods section of some papers, which arose in unique problems concerning cost-effectiveness, time profitability, safety and even worse, the credibility of the results, creating contradictory conclusions in some cases. This ‘noisy’ situation encouraged us to collect, classify and review, for the first time to our knowledge, some aspects relating to studies on Sarcoptes mite from collection to DNA extraction, which will be useful for further studies on Sarcoptes, and have implications for the effective control of the diseases Sarcoptes mite causes. Further studies are needed, especially to compare the profitability, safety, sensibility and specificity of the different methods of this neglected realm of the ubiquitous ectoparasite. [ABSTRACT FROM AUTHOR]

Abstract  The objective of the present study was to examine the extent of genetic diversity among Sarcoptes scabiei individuals belonging to different skin subunits of the body from individual mangy hosts. Ten microsatellite primers were applied on 44 individual S. scabiei mites from three mangy Iberian ibexes from Sierra Nevada Mountain in Spain. Dendrograms of the mites from the individual Iberian ibexes, showing the proportion of shared alleles between pairs of individual mites representing three skin subpopulations (head, back, and abdomen subunits), allowed the clustering of some mite samples up to their skin subunits. This genetic diversity of S. scabiei at skin-scale did not have the same pattern in all considered hosts: for the first Iberian ibex (Cp1), only mites from the head subunit were grouped together; in the second individual (Cp2), the clustering was detected only for mites from the abdomen subunit; and for the third one (Cp3), only mites from the back subunit were clustered together. Our results suggest that the local colonization dynamics of S. scabiei would have influenced the nonrandom distribution of this ectoparasite, after a single infestation. Another presumable explanation to this skin-scale genetic structure could be the repeated infestations. To our knowledge, this is the first documentation of genetic structuring among S. scabiei at individual host skin-scale. Further studies are warranted to highlight determining factors of such trend, but the pattern underlined in the present study should be taken into account in diagnosis and monitoring protocols for studying the population genetic structure and life cycle of this neglected but important ectoparasite. [ABSTRACT FROM AUTHOR]

Abstract  A total of 2,229 adults ticks (1,428 males and 801 females) belonging to the brown dog tick, Rhipicephalus sanguineus Latreille, 1806, collected from dogs in Seville province (Andalusia), distributed in 500 lots ranging from one to eight specimens per lot, were examined for the presence of rickettsiae by molecular techniques. Specific rickettsiae DNA were detected in 90 lots (18%) of ticks tested. Sequence analysis of amplicons revealed that R. sanguineus ticks were infected exclusively with Rickettsia massiliae (including the strain Bar-29). The results of this study extend the knowledge of the geographic distribution and prevalence of these spotted fever group (SFG) rickettsiae and indicate that at least two of them, with yet uncertain pathogenicity to humans, are present in brown dog ticks in south western Spain. Although Mediterranean spotted fever (MSF) is an endemic disease in Andalusia, Rickettsia conorii was not found, whereas R. massiliae, recently described as a pathogenic species, was highly prevalent in this area. Our data suggest that in Andalusia a number of MSF or MSF-like cases attributed to R. conorii could have been actually caused by other SFG rickettsia present in R. sanguineus, particularly, R. massiliae. [ABSTRACT FROM AUTHOR]

Abstract  The present study adapted the HotSHOT method, a technique which has been successfully applied on different kinds of tissues, to studies of Sarcoptes. Some modifications of this technique were made which allowed the quick preparation of PCR-quality Sarcoptes genomic DNA (gDNA), namely applying sodium hydroxide as a substrate for three cycles of thermal shock, followed by a short incubation and pH adjustment with a Tris solution (HotSHOT Plus ThermalSHOCK). The performance of this technique was tested by amplifying a ~450-bp rDNA fragment of the second internal transcribed spacer (ITS-2) and by multi-locus genotyping using ten microsatellites on 520 individual Sarcoptes samples. No difference in performance was observed between gDNA samples prepared using the HotSHOT Plus ThermalSHOCK technique and those prepared using a commercial kit utilizing proteinase K digestion. The results demonstrated that the HotSHOT Plus ThermalSHOCK technique is time-saving, economic, and easily automatable for the preparation of PCR-quality mite gDNA, which has implications for studying the molecular biology of mites with human and animal health significance. Although tested in the present study using Sarcoptes mites as a model, this technique may find broad applicability in extraction of gDNA from other parasites with small sizes and hard bodies. [ABSTRACT FROM AUTHOR]

The common house mosquito Culex pipiens s.l., widely distributed in Europe, Africa, and North America has two recognized biotypes, Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus, which hybridize. Despite their morphological similarities, these biotypes may exhibit ecological differences. This complex ecological mosaic may affect the interaction of Cx. pipiens with pathogens like avian Plasmodium, which is transmitted to wildlife. Although the vector competence for Cx. pipiens biotype molestus has been well studied, there is a lack of studies comparing the vector competence of Cx. pipiens biotype pipiens and their hybrids for the transmission of avian Plasmodium. Here, we evaluated the vector competence of the Cx. pipiens biotypes pipiens, molestus and their hybrids for the transmission of two avian Plasmodium species. Mosquitoes were allowed to feed on blood of wild infected birds and the presence of DNA of Plasmodium in head-thorax and saliva of mosquitoes was molecularly evaluated at 13 day-post exposure. The transmission rates (i.e., the detection of parasite DNA in mosquito saliva) for Plasmodium cathemerium were similar for the two biotypes of Cx. pipiens and their hybrids while Plasmodium relictum DNA was only found in the saliva of Cx. pipiens biotype pipiens. In addition, P. cathemerium was significantly more prevalent than P. relictum in the saliva of Cx. pipiens biotype pipiens. Our results suggest that avian Plasmodium is transmitted by both Cx. pipiens biotypes and their hybrids although differences could be found depending of the parasite species studied. Differences in the abundance of each biotype and their hybrids within areas characterized by distinct environmental conditions, along with variations in their blood-feeding patterns and the parasites infecting birds, could ultimately determine differences in the relevance of each Cx. pipiens biotype in the transmission of avian Plasmodium., Competing Interests: The authors declare no competing interests., (Copyright: © 2024 Gutiérrez-López et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

West Nile virus (WNV) is the most widely distributed mosquito-borne flavivirus in the world. This flavivirus can infect humans causing in some cases a fatal neurological disease and birds are the main reservoir hosts. WNV is endemic in Spain, and human cases have been reported since 2004. Although different studies analyse how climatic conditions can affect the dynamics of WNV infection, very few use long-term datasets. Between 2003 and 2020 a total of 2,724 serum samples from 1,707 common coots ( Fulica atra ) were analysed for the presence of WNV-specific antibodies. Mean (SD) annual seroprevalence was 24.67% (0.28) but showed high year-to-year variations ranging from 5.06% (0.17) to 68.89% (0.29). Significant positive correlations ( p  < 0.01) were observed between seroprevalence and maximum winter temperature and mean spring temperature. The unprecedented WNV outbreak in humans in the south of Spain in 2020 was preceded by a prolonged period of escalating WNV local circulation. Given current global and local climatic trends, WNV circulation is expected to increase in the next decades. This underscores the necessity of implementing One Health approaches to reduce the risk of future WNV outbreaks in humans. Our results suggest that higher winter and spring temperatures may be used as an early warning signal of more intense WNV circulation among wildlife in Spain, and consequently highlight the need of more intense vector control and surveillance in human inhabited areas.

West Nile virus (WNV) is a flavivirus naturally circulating between mosquito vectors and birds, occasionally infecting horses and humans and causing epidemiologically relevant outbreaks. In Spain, the first big WNV outbreak was recorded in 2020, resulting in 77 people infected and 8 fatalities, most of them in southern Spain. Culex perexiguus was identified as the primary vector of WNV maintaining its enzootic circulation of the virus. Growing evidence highlights the role of mosquito microbiota as a key component determining the vectorial capacity of mosquitoes, largely contributing to disease epidemiology. Here, we develop, to our knowledge, the first identification of the microbiota composition of this mosquito vector under natural conditions and test for the potential relationship between mosquito microbiota composition and WNV infection. To do so, we collected mosquitoes in a natural area of southern Spain during the 2020 WNV outbreak and identified the microbiota composition of mosquitoes using a 16S rRNA gene metabarcoding approach. The microbiota of Cx. perexiguus was dominated by the phylum Proteobacteria. The most abundant families were Burkholderiaceae and Erwiniaceae, including the genera Burkholderia, Erwinia, and Pantoea. The genus Wolbachia, which use to dominate the microbiota of Cx. pipiens and negatively interact with WNV according to the literature, had a low prevalence and relative abundance in Cx. perexiguus and its abundance did not differ between WNV-positive and WNV-negative mosquito pools. The microbiota diversity and composition of Cx. perexiguus were not significantly related to the WNV infection status. These results provide the first identification of the mosquito microbiota in an endemic area of WNV circulation in Spain., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Garrigós et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

West Nile virus (WNV) is a re-emerging zoonotic pathogen with increasing incidence in Europe, producing a recent outbreak in 2020 in Spain with 77 human cases and eight fatalities. However, the factors explaining the observed changes in the incidence of WNV in Europe are not completely understood. Longitudinal monitoring of WNV in wild animals across Europe is a useful approach to understand the eco-epidemiology of WNV in the wild and the risk of spillover into humans. However, such studies are very scarce up to now. Here, we analysed the occurrence of WNV and Usutu virus (USUV) antibodies in 2102 samples collected between 2005 and 2020 from a population of feral horses in Doñana National Park. The prevalence of WNV antibodies varied between years, with a mean seroprevalence of 8.1% (range 0%-25%) and seasonally. Climate conditions including mean minimum annual temperatures and mean rainy days per year were positively correlated with WNV seroprevalence, while the annual rainfall was negatively. We also detected the highest incidence of seroconversions in 2020 coinciding with the human outbreak in southern Spain. Usutu virus-specific antibodies were detected in the horse population since 2011. The WNV outbreak in humans was preceded by a long period of increasing circulation of WNV among horses with a very high exposure in the year of the outbreak. These results highlight the utility of One Health approaches to better understand the transmission dynamics of zoonotics pathogens., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Understanding factors that shape tick population genetic structure is important as they may be exploited in crafting strategies for vector control. Amblyomma tholloni, or "elephant tick" is a three-host tick whose adults preferentially parasitize African elephants. The aim of this study was to determine the influence of fragmentation of the host populations on the genetic structure of this tick species from different ecosystems in Kenya, using the second internal transcribed spacer (ITS-2) and mitochondrial cytochrome oxidase 1 (CO1) loci. Population genetic analysis of ticks collected from four elephant populations using ITS-2 and CO1 loci revealed high gene diversity. Gene diversity at the ITS-2 locus was 0.91 and the nucleotide diversity was, 0.01. ITS-2 gene diversity was highest in Laikipia-Samburu ecosystem (0.947) and lowest in Tsavo (0.80). The CO1 locus also had high gene diversity, 0.790, and low nucleotide diversity, 0.006, and like ITS-2, gene diversity was higher in Laikipia-Samburu ecosystem (1.00) and lower in Tsavo (0.286). There was a modest statistically significant genetic differentiation among the four tick populations based on ITS-2 (F ST  = 0.104, P < 0.001; Φ ST  = 0.105, P < 0.001), and a 10% of molecular variance attributed to genetic variation between populations. There was also statistically significant differentiation among tick populations using haplotype frequencies for CO1 locus (F ST  = 0.167, P < 0.001) accounting for 17% of genetic variance among populations, but not modelled genetic distances (Φ ST  = 0.029, P = 0.095) suggesting very recent genetic differentiation. In addition, populations of A. tholloni in Kenya had a significantly negative Tajima D and Fu & Li's F* and D* at the CO1 locus suggesting recent positive selection. The extensive acaricide use in livestock, which host the larval stage, could be driving purifying selection and genetic hitchhiking of the CO1 locus. However, tests sensitive to demography such as Fu's F S , Ramos-Onsins & Rozas's R 2 and raggedness index r were statistically significant at the ITS-2 locus suggesting ancient demographic expansion. Elephant population fragmentation appears to shape the genetic structure of A. tholloni, while agro-ecological factors could influence the genetic diversity of ticks., (Copyright © 2022. Published by Elsevier GmbH.)

Mosquito host utilization is a key factor in the transmission of vector-borne pathogens given that it greatly influences host-vector contact rates. Blood-feeding patterns of mosquitoes are not random, as some mosquitoes feed on particular species and/or individuals more than expected by chance. Mosquitoes use a number of cues including visual, olfactory, acoustic, and thermal stimuli emitted by vertebrate hosts to locate and identify their blood meal sources. Thus, differences in the quality/intensity of the released cues may drive host selection by mosquitoes at both inter- and intra-specific levels. Such patterns of host selection by mosquitoes in space and time can be structured by factors related to mosquitoes (e.g. innate host preference, behavioural plasticity), to hosts (e.g. emission of host-seeking cues, host availability) or to both (e.g. pathogen infection). In this study, we review current evidence, from phenomena to mechanisms, of how these factors influence host utilization by mosquitoes. We also review the methodologies commonly used in this research field and identify the major challenges for future studies. To bridge the knowledge gaps, we propose improvements to strengthen traditional approaches and the use of a functional trait-based approach to infer mosquito host utilization in natural communities., (© 2021 Cambridge Philosophical Society.)

The Dilution Effect Hypothesis (DEH) argues that greater biodiversity lowers the risk of disease and reduces the rates of pathogen transmission since more diverse communities harbour fewer competent hosts for any given pathogen, thereby reducing host exposure to the pathogen. DEH is expected to operate most intensely in vector-borne pathogens and when species-rich communities are not associated with increased host density. Overall, dilution will occur if greater species diversity leads to a lower contact rate between infected vectors and susceptible hosts, and between infected hosts and susceptible vectors. Field-based tests simultaneously analysing the prevalence of several multi-host pathogens in relation to host and vector diversity are required to validate DEH. We tested the relationship between the prevalence in house sparrows (Passer domesticus) of four vector-borne pathogens-three avian haemosporidians (including the avian malaria parasite Plasmodium and the malaria-like parasites Haemoproteus and Leucocytozoon) and West Nile virus (WNV)-and vertebrate diversity. Birds were sampled at 45 localities in SW Spain for which extensive data on vector (mosquitoes) and vertebrate communities exist. Vertebrate censuses were conducted to quantify avian and mammal density, species richness and evenness. Contrary to the predictions of DEH, WNV seroprevalence and haemosporidian prevalence were not negatively associated with either vertebrate species richness or evenness. Indeed, the opposite pattern was found, with positive relationships between avian species richness and WNV seroprevalence, and Leucocytozoon prevalence being detected. When vector (mosquito) richness and evenness were incorporated into the models, all the previous associations between WNV prevalence and the vertebrate community variables remained unchanged. No significant association was found for Plasmodium prevalence and vertebrate community variables in any of the models tested. Despite the studied system having several characteristics that should favour the dilution effect (i.e., vector-borne pathogens, an area where vector and host densities are unrelated, and where host richness is not associated with an increase in host density), none of the relationships between host species diversity and species richness, and pathogen prevalence supported DEH and, in fact, amplification was found for three of the four pathogens tested. Consequently, the range of pathogens and communities studied needs to be broadened if we are to understand the ecological factors that favour dilution and how often these conditions occur in nature., Competing Interests: The authors have declared that no competing interests exist.

Vector-borne diseases, especially those transmitted by mosquitoes, have severe impacts on public health and economy. West Nile virus (WNV) and avian malaria parasites of the genus Plasmodium are mosquito-borne pathogens that may produce severe disease and illness in humans and birds, respectively, and circulate in an endemic form in southern Europe. Here, we used field-collected data to identify the impact of Culex pipiens, Cx. perexiguus and Cx. modestus, on the circulation of both WNV and Plasmodium in Andalusia (SW Spain) using mathematical modelling of the basic reproduction number (R 0 ). Models were calibrated with field-collected data on WNV seroprevalence and Plasmodium infection in wild house sparrows, presence of WNV and Plasmodium in mosquito pools, and mosquito blood-feeding patterns. This approach allowed us to determine the contribution of each vector species to pathogen amplification. Overall, 0.7% and 29.6% of house sparrows were positive to WNV antibodies and Plasmodium infection, respectively. In addition, the prevalence of Plasmodium was higher in Cx. pipiens (2.0%), followed by Cx. perexiguus (1.8%) and Cx. modestus (0.7%). Three pools of Cx. perexiguus were positive to WVN. Models identified Cx. perexiguus as the most important species contributing to the amplification of WNV in southern Spain. For Plasmodium models, R 0 values were higher when Cx. pipiens was present in the population, either alone or in combination with the other mosquito species. These results suggest that the transmission of these vector-borne pathogens depends on different Culex species, and consequently, their transmission niches will present different spatial and temporal patterns. For WNV, targeted surveillance and control of Cx. perexiguus populations appear as the most effective measure to reduce WNV amplification. Also, preventing Culex populations near human settlements, or reducing the abundance of these species, are potential strategies to reduce WNV spillover into human populations in southern Spain., (© 2020 Wiley-VCH GmbH.)

Zoological gardens are home to a large number of vertebrate species and as such are suitable sites for both mosquito breeding and maintenance. They are excellent places for entomological studies of mosquito phenology, diversity, and blood-feeding patterns, as well as for xenomonitoring. During 2016, we sampled mosquitoes in Barcelona Zoo and used molecular methods to determine their blood-feeding patterns and the prevalence and diversity of avian malaria parasites. We also estimated the flight distance of engorged mosquitoes in the area. Overall, 1,384 adult Culex pipiens s.l., Culiseta longiareolata , and Aedes albopictus were captured. Birds dominated the diet of Cx. pipiens s.l. ( n = 87) and Cs. longiareolata ( n = 6), while humans were the only blood-meal source of Ae. albopictus ( n = 3). Mosquitoes had a mean flight distance of 95.67 m after feeding on blood (range 38.71-168.51 m). Blood parasites were detected in the abdomen of 13 engorged Cx. pipiens s.l., eight of which had fed on magpies. Four Plasmodium lineages and a single lineage of the malaria-like parasite Haemoproteus were identified. These results suggest that Cx. pipiens s.l. is involved in the local transmission of avian Plasmodium , which potentially affects the circulation of parasites between and within wildlife and enclosed animals. Vigilance regarding possible mosquito breeding sites in this zoo is thus recommended., (Copyright © 2020 Martínez-de la Puente, Soriguer, Senar, Figuerola, Bueno-Mari and Montalvo.)

Parasites can manipulate their hosts to increase their transmission success. Avian malaria parasites (Plasmodium) are thought to alter the cues such as host odour, used by host-seeking mosquitoes. Bird odour is affected by secretions from the uropygial gland and may play a role in modulating vector-host interactions. We tested the hypothesis that mosquitoes are more attracted to the uropygial secretions and/or whole-body odour (headspace) of Plasmodium-infected house sparrows (Passer domesticus) than to those of uninfected birds. We tested the attraction of nulliparous (e.g. uninfected mosquitoes without previous access to blood) Culex pipiens females towards these stimuli in a dual-choice olfactometer. We used Gas Chromatography-Mass Spectrometry (GC-MS) analyses to assess whether Plasmodium infection is associated with differences in the chemical composition of uropygial secretions. Mosquitoes were more attracted to the odours of infected than uninfected birds, regardless of sex. However, the significant interaction between infection status and the stimuli (uropygial secretion or headspace) showed that mosquitoes were more attracted to the headspace of infected birds; no differences were found in the case of uropygial secretions. The compounds in the volatile lipophilic fraction of the uropygial secretion did not differ between infected and uninfected birds. These results support the host manipulation hypothesis since avian Plasmodium parasites may be capable of altering their host's body odour, thereby making infected individuals more attractive to mosquitoes., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Mosquito feeding preferences determine host-vector contact rates and represent a key factor in the transmission of vector-borne pathogens. The semiochemical compounds of which vertebrate odours are composed probably play a role in mosquito host choice. Birds spread secretions from uropygial gland over their feathers to protect their plumage, comprising behaviour that may in turn affect odour profiles. Although uropygial secretions are expected to modify the attractiveness of birds to mosquitoes, contradictory findings have been reported. Mosquito species differ in their feeding preferences, with some species feeding mainly on birds (ornithophilic species) and others on mammals (mammophilic species). Consequently, it is possible that ornithophilic and mammophilic species differ in their response to uropygial gland secretions. Using a dual-choice olfactometer, the present study tests this hypothesis by comparing the behavioural response to uropygial gland secretions from juvenile male and female house sparrows (Passer domesticus) in the ornithophilic Culex pipiens and the mammophilic Aedes (Ochlerotatus) caspius mosquitoes. No differences were found in the response of either mosquito species to the uropygial gland secretions. Therefore, the preference of ornithophilic mosquitoes for avian hosts is apparently not explained by a greater attraction of mosquitoes to the uropygial gland secretion odour when presented in combination with a CO 2 -enriched airflow., (© 2019 The Royal Entomological Society.)

Factors such as the particular combination of parasite-mosquito species, their co-evolutionary history and the host's parasite load greatly affect parasite transmission. However, the importance of these factors in the epidemiology of mosquito-borne parasites, such as avian malaria parasites, is largely unknown. Here, we assessed the competence of two mosquito species [Culex pipiens and Aedes (Ochlerotatus) caspius], for the transmission of four avian Plasmodium lineages (Plasmodium relictum SGS1 and GRW11 and Plasmodium cathemerium-related lineages COLL1 and PADOM01) naturally infecting wild house sparrows. We assessed the effects of parasite identity and parasite load on Plasmodium transmission risk through its effects on the transmission rate and mosquito survival. We found that Cx. pipiens was able to transmit the four Plasmodium lineages, while Ae. caspius was unable to transmit any of them. However, Cx. pipiens mosquitoes fed on birds infected by P. relictum showed a lower survival and transmission rate than those fed on birds infected by parasites related to P. cathemerium. Non-significant associations were found with the host-parasite load. Our results confirm the existence of inter- and intra-specific differences in the ability of Plasmodium lineages to develop in mosquito species and their effects on the survival of mosquitoes that result in important differences in the transmission risk of the different avian malaria parasite lineages studied.

Environment determines the distribution and prevalence of vector-borne pathogens due to its direct and indirect effects on the hosts, vectors, and pathogens. To investigate the relationship between Usutu virus occurrence and host biodiversity and to characterize the nidus of infection, we used field-based measures of host diversity and density (all birds and only passerines), vector abundance, landscape and Usutu virus prevalence (mosquito infection rate), an emergent disease with a similar cycle to West Nile virus. We collected 908,237 female mosquitoes in an area of 54,984 ha in the Doñana National Park, southern Spain. We identified the mosquitoes and screened them for viruses, censused birds, characterized landscape and climatic variables, and then modeled the presence and infection rate of the virus in relation to host, vector, climatic, and landscape variables. Monthly Usutu presence, detected in Culex perexiguus, was positively related to Passeriformes richness and secondarily to NDVI in the previous month. Our results suggest that Usutu prevalence may be higher when and where host (passerine) richness was high, and thus challenging the conventional idea that host biodiversity reduces flavivirus amplification.

Background: Avian malaria parasites are a highly diverse group that commonly infect birds and have deleterious effects on their hosts. Some parasite lineages are geographically widespread and infect many host species in many regions. Bird migration, natural dispersal, invasive species and human-mediated introductions into areas where competent insect vectors are present, are probably the main drivers of the current distribution of avian malaria parasites., Methods: A total of 412 and 2588 wild house sparrows (Passer domesticus) were captured in 2012 and 2013 in two areas of the Iberian Peninsula (central and southern Spain, respectively). Genomic DNA was extracted from blood samples; parasite lineages were sequenced and identified by comparing with GenBank and/or MalAvi databases., Results: Thirteen Plasmodium lineages were identified in house sparrows corresponding to three major clades. Five individuals were infected by the African Plasmodium lineage PAGRI02, which has been proposed to actively circulate only in Africa., Conclusions: Despite the low prevalence of PAGRI02 in sparrows in Spain, our results suggest that the area of transmission of this parasite is more widespread than previously thought and covers both Africa and Europe. Further studies of the global distribution of Plasmodium lineages infecting wild birds are required to identify the current transmission areas of these parasites. This is vital given the current scenario of global change that is providing new opportunities for avian malaria transmission into areas where parasites were previously absent.

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., (© 2019 Blackwell Verlag GmbH.)

Mosquitoes are the main vectors of pathogens affecting wild animals, livestock and humans. Here, we used molecular tools to assess the local circulation of filarial parasites in mosquitoes collected during 2013 from natural, rural and urban habitats from southern Spain. We screened parasites in 22,791 female mosquitoes of the genera Aedes, Culex and Culiseta. Filarial worms were only detected in two mosquito pools. An Ae. caspius pool was positive for Setaria equina and an unidentified worm related to Onchocerca was detected in a Cx. pipiens pool. None of the mosquito pools were positive for Dirofilaria. These results underlay the role of Ae. caspius in the transmission of Setaria parasites among livestock and/or wildlife to humans in southern Spain., (© 2019 Blackwell Verlag GmbH.)

Plasmodium transmission success depends upon the trade-off between the use of host resources to favour parasite reproduction and the negative effects on host health, which can be mediated by infection intensity. Despite its potential influence on parasite dynamics, the effects of infection intensity on both, birds and vectors, and on Plasmodium transmission success are still poorly understood. Here, we experimentally reduced the Plasmodium load in naturally infected wild house sparrows with the antimalarial primaquine to assess the effects of intensity of infection in the vertebrate hosts on Plasmodium transmission to and by mosquitoes. We monitored the survival of Culex pipiens mosquitoes throughout the development of the parasite and the infection status of the mosquitoes by analysing the head-thorax and saliva at 13 days post-exposure to birds. The proportion of mosquitoes infected by Plasmodium and the presence of Plasmodium in saliva were not associated with the medication treatment of birds. However, the experimental treatment affected vector survival with mosquitoes fed on medicated birds showing a higher survival rate than those fed on control individuals. These results provide strong experimental evidence of the impact of parasite load of vertebrate hosts on the survival probability of malaria vectors.

Background: The transmission of mosquito-borne pathogens is strongly influenced by the contact rates between mosquitoes and susceptible hosts. The biting rates of mosquitoes depend on different factors including the mosquito species and host-related traits (i.e. odour, heat and behaviour). However, host characteristics potentially affecting intraspecific differences in the biting rate of mosquitoes are poorly known. Here, we assessed the impact of three host-related traits on the biting rate of two mosquito species with different feeding preferences: the ornithophilic Culex pipiens and the mammophilic Ochlerotatus (Aedes) caspius. Seventy-two jackdaws Corvus monedula and 101 house sparrows Passer domesticus were individually exposed to mosquito bites to test the effect of host sex, body mass and infection status by the avian malaria parasite Plasmodium on biting rates., Results: Ochlerotatus caspius showed significantly higher biting rates than Cx. pipiens on jackdaws, but non-significant differences were found on house sparrows. In addition, more Oc. caspius fed on female than on male jackdaws, while no differences were found for Cx. pipiens. The biting rate of mosquitoes on house sparrows increased through the year. The bird infection status and body mass of both avian hosts were not related to the biting rate of both mosquito species., Conclusions: Host sex was the only host-related trait potentially affecting the biting rate of mosquitoes, although its effect may differ between mosquito and host species.

Human landscape transformation, especially urbanization, strongly affects ecosystems worldwide. Both urban stressors and parasites have negative effects on organism health, however the potential synergy between those factors has been poorly investigated. We analysed the body condition (i.e. body mass after controlling for wing chord) of 2043 house sparrows (adults and yearlings) captured in 45 localities along an urbanization gradient in relation to Plasmodium, Haemoproteus and Leucocytozoon infection status. Body condition was negatively related to urbanization level and to urbanized land coverage but only in yearling birds from urban habitats. In addition, bird body condition tended to increase in rural habitats, significantly in the case of yearlings. Infected individuals by Plasmodium or Haemoproteus had higher body condition than un-infected birds, but this pattern could be due to a selective disappearance of infected individuals with lower body condition as suggested by the reduced variance in body condition in infected birds in urban habitats. These results provide support for a negative impact of urbanization on bird body condition, while Plasmodium and Haemoproteus may exert selection against individuals with lower body condition living in urban habitats, especially during earlier life stages, underlining the synergistic effects that urbanization and parasites may have on wild birds., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Aedes vittatus is currently found in Africa, Asia and Europe, where it acts as a vector of pathogens causing animal and human diseases (e.g. chikungunya, Zika and dengue). Like other Aedes species, Ae. vittatus is able to breed in artificial containers. The ECDC has recently highlighted the need for molecular tools (i.e. barcoding characterization) that enable Aedes species to be identified in entomological surveys., Results: We sampled mosquito larvae and adults in southern Spain and used a molecular approach to amplify and sequence a fragment of the cytochrome c oxidase subunit 1 gene (barcoding region) of the mosquitoes. The blast comparison of the mosquito sequences isolated from Spain with those deposited in public databases provided a ≥ 99% similarity with sequences for two Aedes mosquitoes, Ae. vittatus and Ae. cogilli, while similarities with other Aedes species were ≤ 94%. Aedes cogilli is only present in India and there are no records of this species from Europe., Conclusions: Due to the low genetic differences between Ae. vittatus and Ae. cogilli, the barcoding region should not be used as the only method for identifying Ae. vittatus, especially in areas where both of these Aedes species are present. This type of analysis should thus be combined with morphological identification using available keys and/or the characterization of other molecular markers. In addition, further entomological surveys should be conducted in order to identify the fine-scale distribution of this mosquito species in Europe.

Pathogen-induced host phenotypic changes are widespread phenomena that can dramatically influence host-vector interactions. Enhanced vector attraction to infected hosts has been reported in a variety of host-pathogen systems, and has given rise to the parasite manipulation hypothesis whereby pathogens may adaptively modify host phenotypes to increase transmission from host to host. However, host phenotypic changes do not always favour the transmission of pathogens, as random host choice, reduced host attractiveness and even host avoidance after infection have also been reported. Thus, the effects of hosts' parasitic infections on vector feeding behaviour and on the likelihood of parasite transmission remain unclear. Here, we experimentally tested how host infection status and infection intensity with avian Plasmodium affect mosquito feeding patterns in house sparrows (Passer domesticus). In separate experiments, mosquitoes were allowed to bite pairs containing (i) one infected and one uninfected bird and (ii) two infected birds, one of which treated with the antimalarial drug, primaquine. We found that mosquitoes fed randomly when exposed to both infected and uninfected birds. However, when mosquitoes were exposed only to infected individuals, they preferred to bite the non-treated birds. These results suggest that the malarial parasite load rather than the infection itself plays a key role in mosquito attraction. Our findings partially support the parasite manipulation hypothesis, which probably operates via a reduction in defensive behaviour, and highlights the importance of considering parasite load in studies on host-vector-pathogen interactions., (Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Background: Host selection by mosquitoes plays a central role in the transmission of vector-borne infectious diseases. Although interspecific variation in mosquito attraction has often been reported, the mechanisms underlying intraspecific differences in hosts' attractiveness to mosquitoes are still poorly known. Metabolic rate is related to several physiological parameters used as location cues by mosquitoes, and so potentially affect host-vector contact rates. Therefore, individual hosts with higher metabolic rates should be more attractive to host-seeking mosquitoes. Here, we experimentally investigated the role of bird metabolic rate in the feeding preferences of Culex pipiens (Linnaeus), a widespread mosquito vector of many pathogens affecting human and wildlife health., Results: Passer domesticus (Linnaeus) pairs containing one bird treated with 2,4-dinitrophenol (DNP) and the other injected with phosphate-buffered saline solution (PBS) (i.e. control) were simultaneously exposed overnight to mosquitoes. The treatment did not affect the proportion of mosquitoes biting on each individual. However, mosquito feeding preference was negatively associated with bird resting metabolic rate but positively with bird body mass. These two variables explained up to 62.76% of the variations in mosquito feeding preference., Conclusions: The relationships between mosquito feeding preferences and individual host characteristics could be explained by enhanced anti-mosquito behaviour associated with higher metabolic rates. The potential role of cues emitted by hosts is also discussed. Thus, individuals with high metabolism may actively avoid being bitten by mosquitoes, despite releasing more attractant cues. Since metabolic rates can be related to individual differences in personality and life history traits, differences in mosquitoes' feeding preferences may be related to intraspecific differences in exposure to vector-borne pathogens.

Mosquito community composition plays a central role in the transmission of zoonotic vector-borne pathogens. We evaluated how the mosquito community affects the seroprevalence of West Nile virus (WNV) in house sparrows along an urbanisation gradient in an area with the endemic circulation of this virus. We sampled 2544 birds and 340829 mosquitoes in 45 localities, analysed in 15 groups, each containing one urban, one rural and one natural area. WNV seroprevalence was evaluated using an epitope-blocking ELISA kit and a micro virus-neutralization test (VNT). The presence of WNV antibodies was confirmed in 1.96% and 0.67% of birds by ELISA and VNT, respectively. The VNT-seropositive birds were captured in rural and natural areas, but not in urban areas. Human population density was zero in all the localities where VNT-positive birds were captured, which potentially explains the low incidence of human WNV cases in the area. The prevalence of neutralizing antibodies against WNV was positively correlated with the abundance of the ornithophilic Culex perexiguus but negatively associated with the abundance of the mammophilic Ochlerotatus caspius and Anopheles atroparvus. These results suggest that the enzootic circulation of WNV in Spain occurs in areas with larger populations of Cx. perexiguus and low human population densities.

Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens that affect wildlife, livestock and, occasionally, humans. Culicoides imicola (Kieffer, 1913) is considered to be the main vector of the pathogens that cause bluetongue disease (BT) and African horse sickness (AHS) in southern Europe. The study of blood-feeding patterns in Culicoides is an essential step towards understanding the epidemiology of these pathogens. Molecular tools that increase the accuracy and sensitivity of traditional methods have been developed to identify the hosts of potential insect vectors. However, to the present group's knowledge, molecular studies that identify the hosts of C. imicola in Europe are lacking. The present study genetically characterizes the barcoding region of C. imicola trapped on farms in southern Spain and identifies its vertebrate hosts in the area. The report also reviews available information on the blood-feeding patterns of C. imicola worldwide. Culicoides imicola from Spain feed on blood of six mammals that include species known to be hosts of the BT and AHS viruses. This study provides evidence of the importance of livestock as sources of bloodmeals for C. imicola and the relevance of this species in the transmission of BT and AHS viruses in Europe., (© 2017 The Royal Entomological Society.)

Host choice by mosquitoes affects the transmission dynamics of vector-borne infectious diseases. Although asymmetries in mosquito attraction to vertebrate species have been reported, the relative importance of host characteristics in mosquito blood-feeding behavior is still poorly studied. Here, we investigate the relationship between avian phenotypic traits-in particular, morphometry, plumage coloration, and nesting and roosting behavior-and the blood-feeding patterns in two common Culex mosquito species on a North American avian community. Forage ratios of the mosquito species were unrelated to the phylogenetic relationships among bird species. Culex pipiens fed preferably on birds with lighter-colored plumage and longer tarsi; furthermore, solitary roosting avian species were both bitten by Cx. pipiens and Cx. restuans more often than expected. These associations may be explained by greater mosquito attraction towards larger birds with a greater color contrast against the background. Although communally roosting birds may release more cues and attract more mosquitoes, individuals may in fact receive fewer bites due to the encounter-dilution effect. Mosquito feeding behavior is a highly complex phenomenon, and our results may improve understanding of the non-random interaction between birds and mosquitoes in natural communities.

The current magnitude of big-game hunting has outpaced the natural growth of populations, making artificial breeding necessary to rapidly boost hunted populations. In this study, we evaluated if the rapid increase of red deer (Cervus elaphus) abundance, caused by the growing popularity of big-game hunting, has impacted the natural genetic diversity of the species. We compared several genetic diversity metrics between 37 fenced populations subject to intensive management and 21 wild free-ranging populations. We also included a historically protected population from a national park as a baseline for comparisons. Contrary to expectations, our results showed no significant differences in genetic diversity between wild and fenced populations. Relatively lower genetic diversity was observed in the protected population, although differences were not significant in most cases. Bottlenecks were detected in both wild and fenced populations, as well as in the protected population. Assignment tests identified individuals that did not belong to their population of origin, indicating anthropogenic movement. We discuss the most likely processes, which could have led to the observed high levels of genetic variability and lack of differentiation between wild and fenced populations and suggest cautionary points for future conservation. We illustrate our comparative approach in red deer. However, our results and interpretations can be largely applicable to most ungulates subject to big-game hunting as most of them share a common exploitation-recovery history as well as many ecological traits., (© The American Genetic Association 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Avipoxvirus (APV) is a fairly common virus affecting birds that causes morbidity and mortality in wild and captive birds. We studied the prevalence of pox-like lesions and genetic diversity of APV in house sparrows (Passer domesticus) in natural, agricultural and urban areas in southern Spain in 2013 and 2014 and in central Spain for 8 months (2012-2013). Overall, 3.2% of 2,341 house sparrows visually examined in southern Spain had cutaneous lesions consistent with avian pox. A similar prevalence (3%) was found in 338 birds from central Spain. Prevalence was higher in hatch-year birds than in adults. We did not detect any clear spatial or temporal patterns of APV distribution. Molecular analyses of poxvirus-like lesions revealed that 63% of the samples were positive. Molecular and phylogenetic analyses of 29 DNA sequences from the fpv167 gene, detected two strains belonging to the canarypox clade (subclades B1 and B2) previously found in Spain. One of them appears predominant in Iberia and North Africa and shares 70% similarity to fowlpox and canarypox virus. This APV strain has been identified in a limited number of species in the Iberian Peninsula, Morocco and Hungary. The second one has a global distribution and has been found in numerous wild bird species around the world. To our knowledge, this represents the largest study of avian poxvirus disease in the broadly distributed house sparrow and strongly supports the findings that Avipox prevalence in this species in South and central Spain is moderate and the genetic diversity low., Competing Interests: The authors have declared that no competing interests exist.

Very little is known about the influence of massive and long distance migration on parasite epidemiology. Migration can simultaneously minimize exposure to common parasites in their habitats and increase exposure to novel pathogens from new environments and habitats encountered during migration, while physiological stress during long distance movement can lead to immune suppression, which makes migrants vulnerable to parasites. In this paper, we investigated the diversity, prevalence, parasite load, co-infection patterns and predilection sites of adult gastrointestinal helminths in 130 migrating wildebeests and tested for their relation with animal age, sex and migration time (which also could indicate different migration routes), and compared them with the non-migratory wildebeest. Surprisingly, only four parasite species were found, Oesophagostomum columbianum, Haemonchus placei, Calicophoron raja and Moniezia expansa, which were lower than in non-migratory wildebeest reported in the literature. These parasites were generalists, infecting livestock, and suggests that wildebeest and livestock, because of their interaction during migration, have a cross-infection risk. There was a negative relation between parasites diversity, prevalence and intensity of infection, and host age, which suggests that wildebeests acquire protective immunity against these parasites as they get older. Prevalence and intensity of infection were higher among wildebeest crossing the Mara Bridge (early migrants) compared to those crossing the Serena (late migrants), which suggests that early migrants (or migrants originating from different areas) have varying infection intensities. The prevalence and intensity of infection were higher in males compared to females and may be due to ecological, behavioural, or physiological differences between males and females. Our findings compared to those of previous studies suggest that migration may provide a mechanism to minimize exposure of hosts to common parasites through migratory escape, but this result awaits examination of helminths epidemiology of non-migratory wildebeests from areas of migrant origins. The potential parasitic cross-infection between wildebeests and livestock is a real risk to be taken into account in the management of wildebeest migration corridors.

Reactive oxygen and nitrogen species (ROS-RNS) are important defence substances involved in the immune response against pathogens. An excessive increase in ROS-RNS, however, can damage the organism causing oxidative stress (OS). The organism is able to neutralise OS by the production of antioxidant enzymes (AE); hence, tissue damage is the result of an imbalance between oxidant and antioxidant status. Though some work has been carried out in humans, there is a lack of information about the oxidant/antioxidant status in the presence of tuberculosis (TB) in wild reservoirs. In the Mediterranean Basin, wild boar (Sus scrofa) is the main reservoir of TB. Wild boar showing severe TB have an increased risk to Mycobacterium spp. shedding, leading to pathogen spreading and persistence. If OS is greater in these individuals, oxidant/antioxidant balance in TB-affected boars could be used as a biomarker of disease severity. The present work had a two-fold objective: i) to study the effects of bovine TB on different OS biomarkers (namely superoxide dismutase (SOD), catalasa (CAT), glutathione peroxidase (GPX), glutathione reductase (GR) and thiobarbituric acid reactive substances (TBARS)) in wild boar experimentally challenged with Mycobacterium bovis, and ii) to explore the role of body weight, sex, population and season in explaining the observed variability of OS indicators in two populations of free-ranging wild boar where TB is common. For the first objective, a partial least squares regression (PLSR) approach was used whereas, recursive partitioning with regression tree models (RTM) were applied for the second. A negative relationship between antioxidant enzymes and bovine TB (the more severe lesions, the lower the concentration of antioxidant biomarkers) was observed in experimentally infected animals. The final PLSR model retained the GPX, SOD and GR biomarkers and showed that 17.6% of the observed variability of antioxidant capacity was significantly correlated with the PLSR X's component represented by both disease status and the age of boars. In the samples from free-ranging wild boar, however, the environmental factors were more relevant to the observed variability of the OS biomarkers than the TB itself. For each OS biomarker, each RTM was defined as a maximum by one node due to the population effect. Along the same lines, the ad hoc tree regression on boars from the population with a higher prevalence of severe TB confirmed that disease status was not the main factor explaining the observed variability in OS biomarkers. It was concluded that oxidative damage caused by TB is significant, but can only be detected in the absence of environmental variation in wild boar., Competing Interests: The authors have declared that no competing interests exist.

The aims of the present study were to investigate Pb and Cd levels in tissues of wild boar (Sus scrofa) from the Sierra Nevada Natural Space (SNNS) (southern Spain). Heavy metal concentrations in livers, kidneys and bones from 111 animals were determined by inductively coupled plasma optical emission spectrometry (ICP-OES). Bones and kidneys were the most Pb- and Cd-contaminated tissues, respectively; Cd concentrations were 5.6 times higher in kidneys than in livers. This is the first biomonitoring study of these pollutants in wild boar tissues in the SNNS, and findings indicate that this population is chronically exposed to these heavy metals. The detected Pb and Cd concentrations were lower than those found in many studies performed in Europe on the same species.

Anthropogenic landscape transformation has an important effect on vector-borne pathogen transmission. However, the effects of urbanization on mosquito communities are still only poorly known. Here, we evaluate how land-use characteristics are related to the abundance and community composition of mosquitoes in an area with endemic circulation of numerous mosquito-borne pathogens. We collected 340 829 female mosquitoes belonging to 13 species at 45 localities spatially grouped in 15 trios formed by 1 urban, 1 rural and 1 natural area. Mosquito abundance and species richness were greater in natural and rural areas than in urban areas. Environmental factors including land use, vegetation and hydrological characteristics were related to mosquito abundance and community composition. Given the differing competences of each species in pathogen transmission, these results provide valuable information on the transmission potential of mosquito-borne pathogens that will be of great use in public and animal health management by allowing, for instance, the identification of the priority areas for pathogen surveillance and vector control.

West Nile virus (WNV) is an emerging vector-borne arbovirus with a zoonotic life-cycle whose main reservoir hosts are birds. In humans and horses, WNV infections rarely result in clinical disease but on occasions - depending on factors such as climatic conditions, insect communities and background immunity levels in local populations - they can lead to outbreaks that threaten public and animal health. We tested for the presence of WNV antibodies in 149 birds belonging to 32 different species. Samples were first tested using a bird-specific ELISA kit and then both positive and doubtful results were confirmed by neutralization tests using WNV and Usutu virus. WNV antibodies were confirmed in a resident Sylvia melanocephala juvenile, supporting the idea of local transmission of WNV in southern Spain in 2013. In addition, the serum from an adult blackbird (Turdus merula) showed neutralization of both WNV and Usutu virus. We discuss our results in light of the occurrence of WNV on horse farms in southern Spain in 2013.

Understanding animal movement behaviour is key to furthering our knowledge on intra- and inter-specific competition, group cohesion, energy expenditure, habitat use, the spread of zoonotic diseases or species management. We used a radial basis function surface approximation subject to minimum description length constraint to uncover the state-space dynamical systems from time series data. This approximation allowed us to infer structure from a mathematical model of the movement behaviour of sheep and red deer, and the effect of density, thermal stress and vegetation type. Animal movement was recorded using GPS collars deployed in sheep and deer grazing a large experimental plot in winter and summer. Information on the thermal stress to which animals were exposed was estimated using the power consumption of mechanical heated models and meteorological records of a network of stations in the plot. Thermal stress was higher in deer than in sheep, with less differences between species in summer. Deer travelled more distance than sheep, and both species travelled more in summer than in winter; deer travel distance showed less seasonal differences than sheep. Animal movement was better predicted in deer than in sheep and in winter than in summer; both species showed a swarming behaviour in group cohesion, stronger in deer. At shorter separation distances swarming repulsion was stronger between species than within species. At longer separation distances inter-specific attraction was weaker than intra-specific; there was a positive density-dependent effect on swarming, and stronger in deer than in sheep. There was not clear evidence which species attracted or repelled the other; attraction between deer at long separation distances was stronger when the model accounted for thermal stress, but in general the dynamic movement behaviour was hardly affected by the thermal stress. Vegetation type affected intra-species interactions but had little effect on inter-species interactions. Our modelling approach is useful in interpreting animal interactions, in order to unravel complex cooperative or competitive behaviours, and to the best of our knowledge is the first modelling attempt to make predictions of multi-species animal movement under different habitat mosaics and abiotic environmental conditions.

Environment determines the distribution of mosquito-borne diseases in that it influences the vector-host-pathogen transmission cycle, including vector distribution, abundance and diversity. In this study, we analyse the relationship between environmental variables estimated by remote sensing and the spatial distribution (presence, abundance and diversity) of seven mosquito species vectors of West Nile and other pathogens (Usutu, avian malaria and dirofilariasis) in the Doñana Natural Park, Spain. Traps were distributed over an area of 54,984 ha divided into six ecological units: marshland, sand dunes, scrubland, ricefields, crops and fishponds. We collected mosquitoes once a month from up to 112 locations using BG-Sentinel traps baited with BG-lure and CO2 during March-November 2010. Hydroperiod, NDVI and Inundation surface were estimated at several resolution scales (100, 250, 500, 1000 and 2000 metres) from corrected and normalized Landsat Images. We sampled 972,346 female mosquitoes, the most abundant species being Culex theileri, Ochlerotatus caspius, Culex modestus, Culex perexiguus, Culex pipiens, Anopheles atroparvus and Ochlerotatus detritus. Our results suggest that: (1) hydroperiod, inundation surface and NDVI are strongly related to the spatial distribution of mosquitoes; (2) the spatial scales used to measure these variables affected quantification of these relationships, the larger scale being more informative; (3) these relationships are species-specific; (4) hydroperiod is negatively related to mosquito presence and richness; (5) Culex abundance is positively related to hydroperiod; (6) NDVI is positively related to mosquito diversity, presence and abundance, except in the case of the two salt marsh species (Oc. caspius and Oc. detritus); and (7) inundation surfaces positively condition the abundance and richness of most species except the salt marsh mosquitoes. Remote sensing data provided reliable information for monitoring mosquito populations. Landscape significantly affected mosquito distribution and abundance, and as a result may alter disease risk. These results suggest that while environmental conditions affect the distribution and abundance of mosquitoes, other factors such as human modification of landscapes may give rise to significant changes in mosquito populations and consequently disease risk.

Background: Understanding wildlife disease ecology is becoming an urgent need due to the continuous emergence and spread of several wildlife zoonotic diseases. West Nile Virus (WNV) is the most widespread arthropod-borne virus in the world, and in recent decades there has been an increase both in geographic range, and in the frequency of symptomatic infections in humans and wildlife. The principal vector for WNV in Europe is the common house Culex pipiens mosquito, which feeds on a wide variety of vertebrate host species. Variation in mosquito feeding preference has been described as one of the most influential parameters driving intensity and timing of WNV infection in the United States, but feeding preferences for this species have been little studied in Europe., Methods: Here, we estimated feeding preference for wild Cx. pipiens in northern Italy, using molecular analysis to identify the origin of blood meals, and avian census to control host abundance variations. Additionally, we used host bird odour extracts to test experimentally mosquito preferences in the absence of environmental variations., Results: For the first time, we demonstrate a clear feeding preference for the common blackbird (Turdus merula), both for wild collected specimens and in the lab, suggesting a potential important role for this species in the WNV epidemiology in Europe. A seasonal decrease in abundance of blackbirds is associated with increased feeding on Eurasian magpies (Pica pica), and this may be linked to seasonal emergence of WNV in humans. Feeding preferences on blackbirds are more marked in rural areas, while preference for magpies is higher in peridomestic areas. Other species, such as the house sparrow (Passer domesticus) appear to be selected by mosquitoes opportunistically in relation to its abundance., Conclusions: Our findings provide new insights into the ecology of Cx. pipiens in Europe and may give useful indications in terms of implementing targeted WNV surveillance plans. However, a clearer understanding of spatio-temporal variations of Cx. pipiens feeding preferences, and targeted studies on reservoir competence for WNV for these species are therefore now urgently needed as this is essential to describe disease dynamics and quantify virus transmission risk.

Background: The invasive Asian tiger mosquito Aedes albopictus has dramatically expanded its distribution range, being catalogued as one of the world's 100 worst invasive alien species. As vectors of pathogens, Ae. albopictus may create novel epidemiological scenarios in the invaded areas., Methods: Here, the frequency of encounters of Ae. albopictus with the avian malaria parasite Plasmodium and the related Haemoproteus was studied in an area with established populations in northeastern Italy and compared with those from four native mosquito species, Anopheles maculipennis s.l., Culex hortensis, Culex pipiens, and Ochlerotatus caspius. The abdomens of mosquitoes with a recent blood meal were used to identify both the blood meal source and the parasites harboured., Results: Aedes albopictus had a clear antropophilic behaviour while An. maculipennis and Oc. caspius fed mainly on non-human mammals. Birds were the most common hosts of Cx. pipiens and reptiles of Cx. hortensis. Parasites were isolated from three mosquito species, with Cx. pipiens (30%) showing the highest parasite prevalence followed by Cx. hortensis (9%) and Ae. albopictus (5%)., Conclusions: These results are the first identifying the avian malaria parasites harboured by mosquitoes in Italy and represent the first evidence supporting that, although Ae. albopictus could be involved in the transmission of avian malaria parasites, the risk of avian malaria parasite spread by this invasive mosquito in Europe would be minimal.

Understanding the feeding preferences of haematophagous insects is critical to depicting the amplification and transmission networks of pathogens and identifying key vector species for surveillance programs. In the case of species from genus Culicoides, many of which are important vectors of pathogens causing animal diseases, information from molecular studies on the feeding habits of females is expanding but still limited for a significant fraction of competent vectors of Culicoides-borne pathogens. In spite of these limitations, recent studies highlight that most Culicoides species are able to feed on several vertebrate species, but present clear preferences for mammals or birds., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: The impact of climate change on vector-borne diseases is highly controversial. One of the principal points of debate is whether or not climate influences mosquito abundance, a key factor in disease transmission., Methods: To test this hypothesis, we analysed ten years of data (2003-2012) from biweekly surveys to assess inter-annual and seasonal relationships between the abundance of seven mosquito species known to be pathogen vectors (West Nile virus, Usutu virus, dirofilariasis and Plasmodium sp.) and several climatic variables in two wetlands in SW Spain., Results: Within-season abundance patterns were related to climatic variables (i.e. temperature, rainfall, tide heights, relative humidity and photoperiod) that varied according to the mosquito species in question. Rainfall during winter months was positively related to Culex pipiens and Ochlerotatus detritus annual abundances. Annual maximum temperatures were non-linearly related to annual Cx. pipiens abundance, while annual mean temperatures were positively related to annual Ochlerotatus caspius abundance. Finally, we modelled shifts in mosquito abundances using the A2 and B2 temperature and rainfall climate change scenarios for the period 2011-2100. While Oc. caspius, an important anthropophilic species, may increase in abundance, no changes are expected for Cx. pipiens or the salt-marsh mosquito Oc. detritus., Conclusions: Our results highlight that the effects of climate are species-specific, place-specific and non-linear and that linear approaches will therefore overestimate the effect of climate change on mosquito abundances at high temperatures. Climate warming does not necessarily lead to an increase in mosquito abundance in natural Mediterranean wetlands and will affect, above all, species such as Oc. caspius whose numbers are not closely linked to rainfall and are influenced, rather, by local tidal patterns and temperatures. The final impact of changes in vector abundance on disease frequency will depend on the direct and indirect effects of climate and other parameters related to pathogen amplification and spillover on humans and other vertebrates.