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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. [ABSTRACT FROM PUBLISHER]

The effect of insect vectors on avian exposure to infection by pathogens remains poorly studied. Here, we used an insect repellent treatment to reduce the number of blood-sucking flying insects in blue tit Cyanistes caeruleus nests and examined its effect on nestling health status measured as body mass, nestling phytohaemagglutinin (PHA) response and blood parasite prevalence. We found that (i) the insect repellent treatment significantly reduced the number of blood-sucking flying insects in nests and (ii) the number of blood-sucking flying insects had a significant effect on the prevalence of the blood parasite Trypanosoma independently of the treatment. In addition, we found support for an adverse effect of parasite infections on nestling PHA response. Nestlings infected by Trypanosoma mounted a weaker response against PHA than non-parasitized ones. In addition, the number of blowflies in the nest was negatively associated with nestling PHA response. Overall, we found support for the hypothesis that blood-sucking flying insects attacking nestlings increase their exposure to parasite infections. Our results further substantiate the adverse effect of parasites on nestling condition. [ABSTRACT FROM AUTHOR]

Mechanisms affecting patterns of vector distribution among host individuals may influence the population and evolutionary dynamics of vectors, hosts and the parasites transmitted. We studied the role of different factors affecting the species composition and abundance of Culicoides found in nests of the blue tit (Cyanistes caeruleus). We identified 1531 females and 2 males of 7 different Culicoides species in nests, with C. simulator being the most abundant species, followed by C. kibunensis, C. festivipennis, C. segnis, C. truncorum, C. pictipennis and C. circumscriptus. We conducted a medication × fumigation experiment randomly assigning bird's nests to different treatments, thereby generating groups of medicated and control pairs breeding in fumigated and control nests. Medicated pairs were injected with the anti-malarial drug Primaquine diluted in saline solution while control pairs were injected with saline solution. The fumigation treatment was carried out using insecticide solution or water for fumigated and control nests respectively. Brood size was the main factor associated with the abundance of biting midges probably because more nestlings may produce higher quantities of vector attractants. In addition, birds medicated against haemoparasites breeding in non-fumigated nests supported a higher abundance of C. festivipennis than the rest of the groups. Also, we found that the fumigation treatment reduced the abundance of engorged Culicoides in both medicated and control nests, thus indicating a reduction of feeding success produced by the insecticide. These results represent the first evidence for the role of different factors in affecting the Culicoides infracommunity in wild avian nests. [ABSTRACT FROM AUTHOR]

Multiple invasions (MIs) or infections, i.e. those by more than 1 parasite in the same erythrocyte, could be the result of parasite density or, alternatively, to parasite-related factors or host-related factors. According to the last possibility, to our knowledge, only 3 laboratory studies of malaria have found an increase in the occurrence of MIs when antibodies to parasite antigens were present. Therefore, we tested the possibility that MIs were influenced by the host immune status, using as model the malaria-like parasite Haemoproteus infecting blue tits (Cyanistes caeruleus). Avian hosts infected with Haemoproteus were medicated with primaquine or injected with saline solution and the density of infection and the presence of MIs counted. Medication treatment reduced significantly the density of infection by Haemoproteus in females but not in males. For females, the presence of MIs was positively associated with both the density of infection and the immunoglobulin levels on each capture, but no association was found between the treatment and the presence of MIs. For males, the density of infection but not the immunoglobulin levels was positively associated with the presence of MIs. In addition, medicated males supported more MIs than controls. Our results represent the first line of evidence in the wild for a possible role of the host immune system promoting MIs. [ABSTRACT FROM PUBLISHER]

Wild birds are hosts of Culicoides from as early on as the nesting stage when constrained to their nests. However, the environmental factors which determine the abundance and composition of Culicoides species within each bird nest are still understudied. We sampled Culicoides from Eurasian blue tit ( Cyanistes caeruleus ) nests found in 2 types of forests located in southern Spain. Firstly, we monitored the abundance of Culicoides species in bird nests from a dry Pyrenean oak deciduous forest and a humid mixed forest comprising Pyrenean and Holm oaks throughout 2 consecutive years. During the 3rd year, we performed a cross-fostering experiment between synchronous nests to differentiate the role of rearing environment conditions from that of the genetically determined or maternally transmitted cues released by nestlings from each forest. We found 147 female Culicoides from 5 different species in the birds' nests. The abundance of Culicoides was higher in the dry forest than in the humid forest. Culicoides abundance, species richness and prevalence were greater when the nestlings were hatched later in the season. The same pattern was observed in the cross-fostering experiment, but we did not find evidence that nestling's features determined by the forest of origin had any effect on the Culicoides collected. These results support the notion that habitat type has a strong influence on the Culicoides affecting birds in their nests, while some life history traits of birds, such as the timing of reproduction, also influence Culicoides abundance and species composition.

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.)

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.

Avian Plasmodium and Haemoproteus parasites are easily detected by DNA analyses of infected samples but only correctly assigned to each genus by sequencing and use of a phylogenetic approach. Here, we present a restriction site to differentiate between both parasite genera avoiding the use of those analyses. Alignments of 820 sequences currently listed in GenBank encoding a particular cytochrome B region of avian Plasmodium and Haemoproteus show a shared restriction site for both genera using the endonuclease Hpy CH4III. An additional restriction site is present in Plasmodium sequences that would initially allow differentiation of both genera by differential migration of digested products on gels. Overall 9 out of 326 sequences containing both potential restriction sites do not fit to the general rule. We used this differentiation of parasite genera based on Hpy CH4III restriction sites to evaluate the efficacy of 2 sets of general primers in detecting mixed infections. To do so, we used samples from hosts infected by parasites of both genera. The use of general primers was only able to detect 25% or less of the mixed infections. Therefore, parasite DNA amplification using general primers to determine the species composition of haemosporidian infections in individual hosts is not recommended. Specific primers for each species and study area should be designed until a new method can efficiently discriminate both parasites.

Multiple invasions (MIs) or infections, i.e. those by more than 1 parasite in the same erythrocyte, could be the result of parasite density or, alternatively, to parasite-related factors or host-related factors. According to the last possibility, to our knowledge, only 3 laboratory studies of malaria have found an increase in the occurrence of MIs when antibodies to parasite antigens were present. Therefore, we tested the possibility that MIs were influenced by the host immune status, using as model the malaria-like parasite Haemoproteus infecting blue tits (Cyanistes caeruleus). Avian hosts infected with Haemoproteus were medicated with primaquine or injected with saline solution and the density of infection and the presence of MIs counted. Medication treatment reduced significantly the density of infection by Haemoproteus in females but not in males. For females, the presence of MIs was positively associated with both the density of infection and the immunoglobulin levels on each capture, but no association was found between the treatment and the presence of MIs. For males, the density of infection but not the immunoglobulin levels was positively associated with the presence of MIs. In addition, medicated males supported more MIs than controls. Our results represent the first line of evidence in the wild for a possible role of the host immune system promoting MIs.

Multiple infections, those by more than one parasite in the same erythrocyte, may be adaptive for the malarial parasite as a means to ensure fertility. Alternatively they may simply be the consequence of a non-adaptive process forcing several parasites to compete for resources in one host cell. Avian hosts infected with Haemoproteus were medicated with primaquine or injected with saline solution and the density of infection and number, maturity and sex of mature multiple infections counted. Multiple infections depend on density of infection, and maturity is attained rarely and usually by gametocytes of the same sex. The role of multiple infections for fertility insurance is not supported by these results.