Spatial distribution and temporal dynamics of invasive and native mosquitoes in a large Mediterranean city.

Mosquitoes, including invasive species like the Asian tiger mosquito Aedes albopictus, alongside native species Culex pipiens s.l., pose a significant nuisance to humans and serve as vectors for mosquito-borne diseases in urban areas. Understanding the impact of water infrastructure characteristics, climatic conditions, and management strategies on mosquito occurrence and effectiveness of control measures to assess their implications on mosquito occurrence is crucial for effective vector control. In this study, we examined data collected during the local vector control program in Barcelona, Spain, focusing on 234,225 visits to 31,334 different sewers, as well as 1817 visits to 152 fountains between 2015 and 2019. We investigated both the colonization and recolonization processes of mosquito larvae within these water infrastructures. Our findings revealed higher larval presence in sandbox-sewers compared to siphonic or direct sewers, and the presence of vegetation and the use of naturalized water positively influenced larval occurrence in fountains. The application of larvicidal treatment significantly reduced larvae presence; however, recolonization rates were negatively affected by the time elapsed since treatment. Climatic conditions played a critical role in the colonization and recolonization of sewers and urban fountains, with mosquito occurrence exhibiting non-linear patterns and, generally, increasing at intermediate temperatures and accumulated rainfall levels. This study emphasizes the importance of considering sewers and fountains characteristics and climatic conditions when implementing vector control programs to optimize resources and effectively reduce mosquito populations.

Determinants of the current and future distribution of the West Nile virus mosquito vector Culex pipiens in Spain.

Changes in environmental conditions, whether related or not to human activities, are continuously modifying the geographic distribution of vectors, which in turn affects the dynamics and distribution of vector-borne infectious diseases. Determining the main ecological drivers of vector distribution and how predicted changes in these drivers may alter their future distributions is therefore of major importance. However, the drivers of vector populations are largely specific to each vector species and region. Here, we identify the most important human-activity-related and bioclimatic predictors affecting the current distribution and habitat suitability of the mosquito Culex pipiens and potential future changes in its distribution in Spain. We determined the niche of occurrence (NOO) of the species, which considers only those areas lying within the range of suitable environmental conditions using presence data. Although almost ubiquitous, the distribution of Cx. pipiens is mostly explained by elevation and the degree of urbanization but also, to a lesser extent, by mean temperatures during the wettest season and temperature seasonality. The combination of these predictors highlights the existence of a heterogeneous pattern of habitat suitability, with most suitable areas located in the southern and northeastern coastal areas of Spain, and unsuitable areas located at higher altitude and in colder regions. Future climatic predictions indicate a net decrease in distribution of up to 29.55%, probably due to warming and greater temperature oscillations. Despite these predicted changes in vector distribution, their effects on the incidence of infectious diseases are, however, difficult to forecast since different processes such as local adaptation to temperature, vector-pathogen interactions, and human-derived changes in landscape may play important roles in shaping the future dynamics of pathogen transmission.

A spatial ecology study in a high-diversity host community to understand blood-feeding behaviour in Phlebotomus sandfly vectors of Leishmania.

Molecular studies indicate that Phlebotomine sandflies (Diptera: Psychodidae) blood feed on many vertebrate species, of which only a few are proven parasite reservoirs. Investigating sandfly vector feeding preferences is therefore important and requires taking into account the availability and accessibility of host species. In terms of the latter, it is necessary to consider the metabolic cost to the insect of reaching the host and moving on to a suitable breeding site. The present study used statistical modelling to compare the feeding patterns of Phlebotomus perniciosus (n = 150), Phlebotomus papatasi (n = 35) and Phlebotomus ariasi (n = 7) on each of an average of 30 host species in a wildlife park in Murcia, Spain. Sandfly feeding movement costs were estimated as a function of the distance and altitude gradients saved by the insect, assuming that they displayed 'site fidelity'. Most (87%) engorged females were caught <100 m from the host on which they had fed. Although the percentage of bloodmeals was highest on fallow deer (Dama dama) (30%) and red deer (Cervus elaphus) (26%), the predicted feeding probability after considering movement cost was highest for red deer and common eland (Taurotragus oryx), and positively associated with host census. These results suggest that, under similar circumstances, sandflies prefer to feed on some host species more than on others.

House sparrow uropygial gland secretions do not attract ornithophilic nor mammophilic mosquitoes.

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 CO2 -enriched airflow.

First molecular identification of the vertebrate hosts of Culicoides imicola in Europe and a review of its blood-feeding patterns worldwide: implications for the transmission of bluetongue disease and African horse sickness.

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.

Telomere dynamics in parasitic great spotted cuckoos and their magpie hosts.

Although little is known on the impact of environment on telomere length dynamics, it has been suggested to be affected by stress, lifestyle and/or life-history strategies of animals. We here compared telomere dynamics in erythrocytes of hatchlings and fledglings of the brood parasite great spotted cuckoos (Clamator glandarius) and of magpies (Pica pica), their main host in Europe. In magpie chicks, telomere length decreased from hatching to fledging, whereas no significant change in telomere length of great spotted cuckoo chicks was found. Moreover, we found interspecific differences in the association between laying date and telomere shortening. Interspecific differences in telomere shortening were interpreted as a consequence of differences in lifestyle and life-history characteristics of magpies and great spotted cuckoos. In comparison with magpies, cuckoos experience reduced sibling competition and higher access to resources and, consequently, lower stressful environmental conditions during the nestling phase. These characteristics also explain the associations between telomere attrition and environmental conditions (i.e. laying date) for magpies and the absence of association for great spotted cuckoos. These results therefore fit expectations on telomere dynamics derived from interspecific differences in lifestyle and life history of brood parasites and their bird hosts.

Vector abundance determines Trypanosoma prevalence in nestling blue tits.

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.

Interacting effects of aromatic plants and female age on nest-dwelling ectoparasites and blood-sucking flies in avian nests.

Some avian species incorporate aromatic plants to their nests. The "nest protection hypothesis", which posits that volatile secondary compounds contained in these plants may have antiparasite properties, has not received full support. All previous tests of this hypothesis have only considered effects on nest-dwelling ectoparasites, but not on blood-sucking flies. The "drug hypothesis" posits that aromatic plants may stimulate nestling immune system, development, or condition. We tested these hypotheses experimentally in wild blue tits, Cyanistes caeruleus, a species that adds aromatic plants to their nests. We supplemented aromatic plants to half of a sample of nests, while adding grass to the other half of nests. We quantified abundance of two groups of blood-sucking flies (blackflies and biting midges) at two different stages of the reproductive period, and abundance of three nest-dwelling ectoparasites (fleas, mites, and blowflies). Experimental supplementation of aromatic plants reduced abundance of fleas only in nests of yearling females and not in nests of older females. Blackflies and biting midges were both more numerous in nests of yearling females than in nests of older females. Mass of aromatic plants added by females was negatively related with abundance of fleas in control nests but not in experimental nests supplied with aromatic plants. Mass of plants added by females was also positively related with abundance of blackflies during the nestling stage. Finally, aromatic plants did not affect nestling growth or immune responses. We conclude that several factors such as female experience and their ability to add plants to the nest interact to explain effects of aromatic plants on different parasites.

Effects of feeding management and time of day on the occurrence of self-suckling in dairy goats.

The occurrence of self-suckling was recorded in 21 dairy goats during periods of 20 minutes at three different times per day (immediately after milking and the first feed, immediately after the second feed and in the afternoon) for 27 days (divided into three experimental periods of nine days). As expected, negative associations between milk yield and the frequency of self-suckling were observed (P<0.05). Goats suckled on their own right teat more frequently than on the left teat. The width of the right teat (measured at the middle of the teat) was positively associated with the frequency of self-suckling after controlling for the width of the left teat. A higher self-suckling frequency was observed immediately after milking than in the other two periods of the day. The frequency of self-suckling by each goat was reduced when animals were supplemented ad libitum with wheat straw in addition to their ordinary feed.

On the heritability of blue-green eggshell coloration.

Avian blue-green eggshell coloration has been proposed as a female signal of genetic or phenotypic quality to males. However, little is known about the relative importance of additive genetic and environmental effects as sources of eggshell colour variation in natural populations. Using 5 years of data and animal models, we explored these effects in a free-living population of pied flycatchers. Permanent environmental and year effects were negligible, although year environmental variance (V(Year)) was significant for all but one of the traits. However, we found high-moderate narrow-sense heritabilities for some colour parameters. Within-clutch colour variability showed the highest coefficient of additive genetic variation (i.e. evolvability). Previous evidence suggests that eggshell colour is sexually selected in this species, males enhancing parental effort in clutches with higher colour variability and peak values. Eggshell colour could be driven by good-genes selection in pied flycatchers although further genetic studies should confirm this possibility.

Factors affecting Culicoides species composition and abundance in avian nests.

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 medicationxfumigation 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.

A restriction site to differentiate Plasmodium and Haemoproteus infections in birds: on the inefficiency of general primers for detection of mixed infections.

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.

Can the host immune system promote multiple invasions of erythrocytes in vivo? Differential effects of medication and host sex in a wild malaria-like model.

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.

Are multiple gametocyte infections in malarial parasites an adaptation to ensure fertility?

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.