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Living organisms are exposed to a wide range of substances - internal and external - which act like reactive oxygen species (ROS). Oxidative damage accurs when the balance between ROS and antioxidant defenses is altered. Urbanization and parasite infection are both important sources of ROS with different harmful effects on wildlife health, but the potential synergies between both factors are poorly known. Here, we analyse the oxidative stress of wild juvenile male house sparrows (Passer domesticus) along an urbanization gradient in relation to the infection status by three common blood parasites (Plasmodium, Haemoproteus and Leucocytozoon) and bird body condition. We analysed samples from 688 birds captured at 45 localities from southern Spain grouped into triplets including an urban, a rural and a natural habitat, with 15 localities per habitat type. We measured i) thiobarbituric acid reactive substances (TBARS) levels as indicator of the oxidative damage to lipids, and the activity of three antioxidant enzymes ii) glutathione peroxidase (GPx), iii) superoxide dismutase (SOD) and iv) glutathione reductase (GR) as indicators of bird's antioxidant capacity. Birds infected with Haemoproteus and urban birds showed significantly and marginally higher levels of TBARS than uninfected and rural birds, respectively. The relationship between TBARS and body condition is different regarding the infection status (significative) and habitat (marginally significant) being negative for Haemoproteus infected and urban birds but positive for uninfected and non-urban birds. The antioxidant activity was significantly lower in Plasmodium infected birds but marginally higher in Leucocytozoon infected birds than in uninfected ones. Individuals with higher body condition had higher GPx and SOD activity in relation to a lower GR activity. Overall, these results suggest that blood parasites infections and urbanization affect the oxidative status of wild birds and highlight the role of bird's body condition on the regulation of the oxidative stress status., Competing Interests: Declaration of competing interest 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., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Urbanization is an important human-driven process that leads to biodiversity loss and alters the interactions between organisms, including disease transmission. Although urbanization affects both host and vector communities, the effects on vector-borne pathogens are still poorly understood. Here, we monitored variation in prevalence and richness of three common blood parasites in birds (Plasmodium, Haemoproteus and Leucocytozoon) from localities with different land uses (urban, rural and natural) during two consecutive years (2013 and 2014). Overall, 1400 juvenile house sparrows (Passer domesticus) from 15 localities in southern Spain were included in this study. Haemoproteus and Leucocytozoon prevalence was higher in 2013 than in 2014, particularly in urban and natural habitats for the case of Leucocytozoon. Prevalence was correlated between years for Haemoproteus and Leucocytozoon, independently of the habitat. Additionally, rural habitats harboured significantly higher Haemoproteus lineage richness compared to urban and natural habitats during 2014. Leucocytozoon lineage richness was negatively correlated between years in rural habitats but positively correlated in urban and natural habitats in comparison. Parasite lineages found in birds were homogeneously distributed along habitats and years and the common lineages prevalence were not influenced by them. Our results highlight different patterns of infection depending on the parasite genera probably related to the composition and density of vector communities. The specific reproductive environmental requirements of the different groups of vectors involved in the transmission may be affected by climatic conditions and landscape features., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Bagaza virus (BAGV; synonymous to Israel turkey meningoencephalomyelitis virus, ITV) is a relevant arthropod-borne epornitic flavivirus. In its first emergence in Europe (southern Spain, 2010) BAGV caused an outbreak, severely affecting red-legged partridges and common pheasants. The effects (pathogenicity, role as reservoir host) of BAGV in other European phasianids are unknown. To fill this gap, grey partridges were experimentally infected with BAGV. The clinical course of the disease was severe, with neurological signs, significant weight loss and 40% mortality. Low viral loads in the blood and the absence of contact transmission suggest a limited-if any-role on BAGV transmission for this European phasianid.

The red-legged partridge (Alectoris rufa) is a competent host for West Nile virus (WNV) replication and highly susceptible to WNV disease. With the aim to assess in this species whether the inoculation of non-structural protein NS1 from WNV elicits a protective immune response against WNV infection, groups of partridges were inoculated with recombinant NS1 (NS1 group) or an unrelated recombinant protein (mock group), and challenged with infectious WNV. A third group received no inoculation prior to challenge (challenge group). The NS1 group failed to elicit detectable antibodies to NS1 while in the mock group a specific antibody response was observed. Moreover, no protection against WNV disease was observed in the NS1 group, but rather, it showed significantly higher viral RNA load and delayed neutralizing antibody response, and suffered a more severe clinical disease, which resulted in higher mortality. This adverse effect has not been observed before and warrants further investigations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The wide spread mosquito Culex pipiens pipiens have two forms molestus and pipiens which frequently hybridize. The two forms have behavioural and physiological differences affecting habitat requirements and host selection, which may affect the transmission dynamic of Cx. p. pipiens-borne diseases., Methods: During 2013, blood engorged Cx. p. pipiens mosquitoes were captured in urban, rural and natural areas from Southern Spain. In 120 mosquitoes, we identified the blood meal origin at vertebrate species/genus level and the mosquito form. The presence and molecular lineage identity of avian malaria parasites in the head-thorax of each mosquito was also analysed., Results: Mosquitoes of the form pipiens were more frequently found in natural than in urban areas. The proportion of Cx. pipiens form molestus and hybrids of the two forms did not differ between habitat categories. Any significant difference in the proportion of blood meals on birds between forms was found. Birds were the most common feeding source for the two forms and their hybrids. Among mammals, dogs and humans were the most common hosts. Two Plasmodium and one Haemoproteus lineages were found in mosquitoes, with non-significant differences between forms., Conclusion: This study supports a differential distribution of Cx. p. pipiens form pipiens between urban and natural areas. Probably due to the similar feeding sources of both mosquito forms and their hybrids here, all of them may frequently interact with avian malaria parasites playing a role in the transmission of Plasmodium.

Background: The life-cycle of many vector-borne pathogens includes an asexual replication phase in the vertebrate host and sexual reproduction in the insect vector. However, as only a small array of parasites can successfully develop infective phases inside an insect, few insect species are competent vectors for these pathogens. Molecular approaches have identified the potential insect vectors of blood parasites under natural conditions. However, the effectiveness of this methodology for verifying mosquito competence in the transmission of avian malaria parasites and related haemosporidians is still under debate. This is mainly because positive amplifications of parasite DNA in mosquitoes can be obtained not only from sporozoites, the infective phase of the malaria parasites that migrate to salivary glands, but also from different non-infective parasite forms in the body of the vector. Here, we assessed the vectorial capacity of the common mosquito Culex pipiens in the transmission of two parasite genera., Methods: A total of 1,560 mosquitoes were allowed to feed on five house sparrows Passer domesticus naturally infected by Haemoproteus or co-infected by Haemoproteus/Plasmodium. A saliva sample of the mosquitoes that survived after 13 days post-exposure was taken to determine the presence of parasite DNA by PCR., Results: Overall, 31.2% mosquito's head-thorax and 5.8% saliva samples analysed showed positive amplifications for avian malaria parasites. In contrast to Haemoproteus DNA, which was not found in either the body parts or the saliva, Plasmodium DNA was detected in both the head-thorax and the saliva of mosquitoes. Parasites isolated from mosquitoes feeding on the same bird corresponded to the same Plasmodium lineage., Conclusions: Our experiment provides good evidence for the competence of Cx. pipiens in the transmission of Plasmodium but not of Haemoproteus. Molecular analyses of saliva are an effective method for testing the vector competence of mosquitoes and other insects in the transmission of vector-borne pathogens.

Bagaza virus (BAGV) is a mosquito-borne flavivirus belonging to the Ntaya serocomplex. In 2010, a disease outbreak was reported in Cádiz (Southern Spain) affecting game birds (red-legged partridges and common pheasants). In this work, red-legged partridges were inoculated experimentally with infectious BAGV isolated from this outbreak in order to make a complete clinical and analytical assessment of the disease caused by the pathogen in this species. Viral load (by real-time RT-PCR) in blood, oral and cloacal swabs, and feathers, and neutralizing antibody titres (by VNT) were measured. In order to determine direct contact transmission, non-inoculated partridges were caged together with the inoculated ones. To assess infectiousness in other species, house sparrows and mice were also inoculated with the virus. All the inoculated partridges were clinically affected, and 30% of them died. All the infected individuals lost weight, with larger losses being recorded in females. Conversely, no mortality or disease symptoms were observed in the sparrows or mice. Remarkably, all the contact partridges acquired the infection by direct (non-vectored) transmission. This study confirms that the red-legged partridge is a susceptible host for BAGV infection, and that this pathogen is transmitted by direct contact. Long-lasting viral loads detected in calami of immature feathers demonstrate that feather sampling could be a useful strategy in active surveillance programs for early detection of BAGV.

West Nile virus (WNV) is a zoonotic pathogen which is maintained in an enzootic cycle between mosquitoes and birds; humans, equines, other mammals and some bird species are dead-end hosts. Lineage 1 WNV strains have predominated in Europe since the 1960s. However, in 2004 lineage 2 strains emerged in Hungary and Russia, respectively, spreading since then to a number of neighbouring countries (e.g., Austria, Greece, Italy, Serbia and Romania). Wild bird mortality is a hallmark of North American WNV outbreaks, a feature uncommon in Europe. This study aimed to compare the course of infection of lineage 1 (NY99) and lineage 2 (Austria/2008) WNV strains in the house sparrow, a bird species common in Europe and North America. House sparrows were inoculated with either NY99 or Austria/2008 WNV strains, or sham-inoculated, and clinical and analytic parameters (viraemia, viral load, antibodies) were examined until 14 days after inoculation. Although all inoculated sparrows became infected, no mortality or clinical signs were observed due to the infection. However, the magnitude and duration of viraemia were higher for NY99 - than for Austria/2008 - infected birds. The house sparrow proved to be a competent host for both strains, although the competence index calculated for NY99 was higher than for Austria/2008. Viral load in tissues and swabs was also higher in NY99-inoculated sparrows. In conclusion, the house sparrow is a convenient avian model for studying host competence of WNV strains. The observed differences between NY99 and Austria/2008 strains might have important epidemiological consequences for disease incidence and dispersal capacity., (Copyright © 2014 Elsevier B.V. All rights reserved.)

West Nile virus (WNV) is a zoonotic arboviral pathogen transmitted by mosquitoes in a cycle involving wild birds as reservoir hosts. The virus has recently emerged in North America and re-emerged in Europe. North American WNV outbreaks are often accompanied by high mortality in wild birds, a feature that is uncommon in Europe. The reason for this difference is unknown, but the intrinsic virulence of the viruses circulating in each continent and/or the susceptibility to the disease of Palearctic as opposed to Nearctic wild bird species could play a role. To assess this question, experimental inoculations with four lineage 1 WNV strains, three from southern Europe (Italy/2008, Italy/2009 and Spain/2007) and one from North America (NY99) were performed on house sparrows (Passer domesticus), a wild passerine common in both continents. Non-significant differences which ranged from 0% to 25% were observed in mortality for the different WNV strains. Viremias lasted from 1 to 5-6 days post-inoculation (dpi) in all cases; individuals inoculated with NY99 had significantly higher titres than those inoculated with any of the Euro-Mediterranean strains. Remarkably, host competence was found to be higher for NY99 than for the other strains. Consequently, albeit being pathogenic for house sparrows, some Euro-Mediterranean strains had reduced capacity for replication in -and transmission from- this host, as compared to the NY99 strain. If applicable also to other wild bird host species, this relatively reduced transmission capacity of the Euro-Mediterranean strains could explain the lower incidence of this disease in wild birds in the Euro-Mediterranean area.