Water and mosquitoes as key components of the infective cycle of Francisella tularensis in Europe: a review.

Francisella tularensis is the pathogen of tularemia, a zoonotic disease that have a broad range of hosts. Its epidemiology is related to aquatic environments, particularly in the subspecies holarctica. In this review, we explore the role of water and mosquitoes in the epidemiology of Francisella in Europe. F. tularensis epidemiology has been linked to natural waters, where its persistence has been associated with biofilm and amebas. In Sweden and Finland, the European countries where most human cases have been reported, mosquito bites are a main route of transmission. F. tularensis is present in other European countries, but to date positive mosquitoes have not been found. Biofilm and amebas are potential sources of Francisella for mosquito larvae, however, mosquito vector capacity has not been demonstrated experimentally, with the need to be studied using local species to uncover a potential transmission adaptation. Transstadial, for persistence through life stages, and mechanical transmission, suggesting contaminated media as a source for infection, have been studied experimentally for mosquitoes, but their natural occurrence needs to be evaluated. It is important to clear up the role of different local mosquito species in the epidemiology of F. tularensis and their importance in all areas where tularemia is present.

Diet and Spatial Ecology Influence Red-Legged Partridge Exposure to Pesticides Used as Seed Treatment.

Seed treatment with pesticides is an extended agricultural practice with a high risk to granivorous birds that consume those seeds. To characterize that risk, it is necessary to understand the ecological factors that determine the exposure chances of birds to treated seeds. We investigated how pesticide uptake by red-legged partridges was related to cultivated plant ingestion and to the use of recently sown fields. We analyzed pesticide residues in 144 fecal samples from 32 flocks and determined the plant diet composition using DNA metabarcoding. Habitat use was studied through the monitoring of 15 GPS-tagged partridges. We confirmed, through the analysis of seeds, that >80% of cereal fields from the area had seeds treated with triazole fungicides. Tebuconazole was detected in 16.6% of partridges' feces. During the sowing season, cultivated plants accounted for half of the plant diet, but no association was found between cultivated plant consumption and pesticide intake. GPS tracking revealed that tebuconazole was detected in feces when partridges had recently used sown fields, whereas nonexposed partridges showed no overlap with recently sown areas. Our results highlight the need to incorporate field ecology into the characterization of pesticide exposure to improve the efficacy of environmental risk assessment.

From pattern to process? Dual travelling waves, with contrasting propagation speeds, best describe a self-organised spatio-temporal pattern in population growth of a cyclic rodent.

The dynamics of cyclic populations distributed in space result from the relative strength of synchronising influences and the limited dispersal of destabilising factors (activators and inhibitors), known to cause multi-annual population cycles. However, while each of these have been well studied in isolation, there is limited empirical evidence of how the processes of synchronisation and activation-inhibition act together, largely owing to the scarcity of datasets with sufficient spatial and temporal scale and resolution. We assessed a variety of models that could be underlying the spatio-temporal pattern, designed to capture both theoretical and empirical understandings of travelling waves using large-scale (>35,000 km2 ), multi-year (2011-2017) field monitoring data on abundances of common vole (Microtus arvalis), a cyclic agricultural rodent pest. We found most support for a pattern formed from the summation of two radial travelling waves with contrasting speeds that together describe population growth rates across the region.

Viral Zoonoses in Small Wild Mammals and Detection of Hantavirus, Spain.

We screened 526 wild small mammals for zoonotic viruses in northwest Spain and found hantavirus in common voles (Microtus arvalis) (1.5%) and high prevalence (48%) of orthopoxvirus among western Mediterranean mice (Mus spretus). We also detected arenavirus among small mammals. These findings suggest novel risks for viral transmission in the region.

Do human infrastructures shape nest distribution in the landscape depending on individual personality in a farmland bird of prey?

Individuals' distribution across habitats may depend on their personality. Human activities and infrastructures are critical elements of the landscape that may impact the habitat selection process. However, depending on their personality, individuals may respond differently to these unnatural elements. In the present study, we first investigated whether some human infrastructures (buildings, roads and paths) shaped Montagu's harrier nest spatial distribution in the landscape according to female personality (boldness). Second, we tested if the reproductive success of females depended on their boldness and nest location regarding infrastructures. Using a long-term (19 years) dataset, we calculated, for each infrastructure type, the distance from each nest to the nearest infrastructure and the infrastructure density around the nest. We tested the effects of female boldness (bold vs. shy) and its interaction with egg-laying date on these six metrics. Nest location in the landscape depended on female personality and on some human infrastructures: the building density was smaller around nests from shy females than from bold ones. Nest distribution related to other infrastructure metrics did not depend on female boldness. The pattern related to building density is consistent with some habitat choice hypotheses, which are discussed. Path density around nests negatively affected reproductive success regardless of female boldness, and late breeders nested further away from paths than early breeders. Human activities on paths (more common later in the season) could lead to disturbance and a decrease in parental care, reducing reproductive success. Increasing human presence in farmlands implies a need to better understand its impact on population composition, in terms of personality. Our results suggest that individual behavioural differences should be taken into account in studies assessing the effects of human disturbance on animal populations, to propose more appropriate conservation measures.

La distribution des individus au sein de l'habitat peut dépendre de leur personnalité. Les activités et infrastructures humaines sont des éléments importants du paysage qui peuvent impacter le processus de sélection des habitats. Cependant, en fonction de leur personnalité, les individus peuvent répondre différemment à ces éléments non naturels. Dans cette étude, nous avons d'abord examiné si certaines infrastructures humaines (bâtiments, routes et chemins) influençaient la répartition spatiale des nids de busard cendré en fonction de la personnalité des femelles (témérité). Ensuite, nous avons testé si le succès reproducteur des femelles dépendait de leur témérité et de la localisation du nid par rapport aux infrastructures. En utilisant des données à long terme (19 ans), nous avons calculé, pour chaque type d'infrastructure, la distance de chaque nid à la plus proche infrastructure et la densité en infrastructures autour de chaque nid. Nous avons testé les effets de la témérité des femelles (téméraire vs. timide) et son interaction avec la date de ponte sur les six métriques. La répartition spatiale des nids dépendait de la personnalité des femelles et de certaines infrastructures humaines: la densité en bâtiments était plus faible autour des nids de femelles timides qu'autour de ceux des téméraires. La distribution des nids par rapport aux autres métriques d'infrastructure n'était pas influencée par la témérité des femelles. Le patron lié à la densité en bâtiments est en accord avec certaines hypothèses de choix d'habitat qui sont discutées. La densité en chemins autour des nids affectait négativement le succès reproducteur indépendamment de la témérité des femelles, et les femelles tardives nichaient plus loin des chemins que les précoces. Les activités humaines sur les chemins (plus fréquentes tard dans la saison de reproduction) pourraient entrainer une perturbation et une diminution des soins parentaux, réduisant ainsi le succès reproducteur. L'augmentation de la présence humaine dans les zones agricoles implique la nécessité de mieux comprendre son impact sur la composition de la population, en termes de personnalité. Nos résultats suggèrent que les différences comportementales interindividuelles devraient être prises en compte dans les études évaluant les effets des perturbations humaines sur les populations animales afin de proposer des mesures de conservations plus appropriées.

Egg Overspray with Herbicides and Fungicides Reduces Survival of Red-Legged Partridge Chicks.

Within the environmental risk assessment conducted for pesticide registration in the European Union (EU), avian reproductive toxicity is characterized after exposing adults. However, eggs of ground-nesting species can be exposed when pesticide applications occur during laying or incubation. We simulated environmentally realistic exposure of red-legged partridge (Alectoris rufa) eggs to an herbicide (2,4-D) and a fungicide (tebuconazole) applied to winter cereal crops during the breeding season of most farmland birds. We analyzed the effects on hatching success, offspring survival, and physiology. Exposure by overspray led to greater pesticide accumulation in the eggshell or content than exposure through contact with treated soil (3.1-13.7 times higher, depending on the pesticide and target sample). Egg overspray with tebuconazole significantly increased chick mortality, which was 26% higher than that of controls. 2,4-D caused a similar but a close to significant increase (chick mortality 24% higher than controls). Exposure to either pesticide through contact with treated soils did not affect chick survival but altered some biochemical parameters posthatching. Our experiment shows that egg spraying with pesticides should be considered as a relevant exposure scenario in risk assessment procedures, given its potential to affect the reproductive success of ground-nesting farmland birds.

Multi-level analysis of exposure to triazole fungicides through treated seed ingestion in the red-legged partridge.

Triazole fungicides are the most widely used products to treat cereal seeds. Granivorous birds, such as red-legged partridges (Alectoris rufa), which consume seeds left on the surface of fields after sowing, have a high risk of exposure. As triazole fungicides can affect sterol synthesis, we tested the hypothesis that treated seed consumption could alter the synthesis of sex hormones and reduce the reproductive capacity of partridges. We exposed adult partridges to seeds treated with four different formulations containing triazoles as active ingredients (flutriafol, prothioconazole, tebuconazole, and a mixture of the latter two) simulating a field exposure during the late autumn sowing season. All treatments produced biochemical changes and an overexpression of genes encoding for enzymes involved in the biosynthesis of sterols and steroid hormones, such as PMVK, ABCA1, MVD, PSCK9, DHCR7 and HSD17B7. Plasma levels of oestradiol were reduced in partridges exposed to tebuconazole. We also monitored reproduction 3 months after exposure (laying date, egg fertilization and hatching rates). We observed a 14-day delay in the laying onset of partridges that had been exposed to flutriafol as compared to controls. These results show that the consumption of seeds treated with triazole fungicides has the potential to affect granivorous bird reproduction. We recommend the evaluation of lagged reproductive effects as part of the protocols of environmental risk assessment of pesticides in wild birds in light of the effects resulting from the exposure to triazole-treated seeds.

Zoonotic Bacteria in Fleas Parasitizing Common Voles, Northwestern Spain.

We detected Francisella tularensis and Bartonella spp. in fleas parasitizing common voles (Microtus arvalis) from northwestern Spain; mean prevalence was 6.1% for F. tularensis and 51% for Bartonella spp. Contrasted vector-host associations in the prevalence of these bacteria suggest that fleas have distinct roles in the transmission cycle of each pathogen in nature.

Zoonotic pathogens in fluctuating common vole (Microtus arvalis) populations: occurrence and dynamics.

Diseases and host dynamics are linked, but their associations may vary in strength, be time-lagged, and depend on environmental influences. Where a vector is involved in disease transmission, its dynamics are an additional influence, and we often lack a general understanding on how diseases, hosts and vectors interact. We report on the occurrence of six zoonotic arthropod-borne pathogens (Anaplasma, Bartonella, Borrelia, Coxiella, Francisella and Rickettsia) in common voles (Microtus arvalis) throughout a population fluctuation and how their prevalence varies according to host density, seasonality and vector prevalence. We detected Francisella tularensis and four species of Bartonella, but not Anaplasma, Borrelia, Coxiella or Rickettsia. Bartonella taylorii and B. grahamii prevalence increased and decreased with current host (vole and mice) density, respectively, and increased with flea prevalence. Bartonella doshiae prevalence decreased with mice density. These three Bartonella species were also more prevalent during winter. Bartonella rochalimae prevalence varied with current and previous vole density (delayed-density dependence), but not with season. Coinfection with F. tularensis and Bartonella occurred as expected from the respective prevalence of each disease in voles. Our results highlight that simultaneously considering pathogen, vector and host dynamics provide a better understanding of the epidemiological dynamics of zoonoses in farmland rodents.

Unintentional effects of environmentally-friendly farming practices: Arising conflicts between zero-tillage and a crop pest, the common vole (Microtus arvalis).

Common voles are a main European facultative, fossorial, farmland rodent pest that can greatly reduce crop yields during population outbreaks. Crop protection against common voles is a complex task that requires the consideration of a set of preventive and control measures within an integrated pest management strategy. A possible option could be to modify farming practices to reduce the availability of refuges for rodents and the damage to crops that they subsequently cause. Farming, however, must simultaneously meet multiple goals including the reduction of the carbon (C) emissions, soil erosion and water use, and the improvement of soil quality. Crop establishment through conservation agriculture strategies, like zero-tillage, would reduce crop management investment, but is also promoted in many regions to reduce C emissions and increase soil organic matter. It could, however, create favourable refuge habitats for fossorial rodent crop pests, like common voles, benefitting from reduced soil disturbance between crop rotations and thus increasing burrow persistence. Assessing the impact that tillage practices, their interaction with different crops and the influence of proximity to potential common vole sources, have on common vole occupancy could provide a valuable tool within an integrated management strategy. Using a 2-ha experimental field with 62 plots 180 m2 (each roughly matching common vole home range size) located experimental plots in north-western Spain, we tested how tillage practices, crop type (wheat, barley, vetch, Narbonne vetch, pea and fallow) and distances from possible colonization sources affect field use by common vole during low population density conditions. Our results show that tillage practices have more influence on common vole occurrence (zero tillage > reduced and conventional tillage) than other aspects such as crop type thus supporting the hypothesis that tillage practices play a key role in common vole habitat use.

"Got rats?" Global environmental costs of thirst for milk include acute biodiversity impacts linked to dairy feed production.

Rodents damaging alfalfa crops typically destined for export to booming Eastern markets often cause economical losses to farmers, but management interventions attempting to control rodents (i.e., use of rodenticides) are themselves damaging to biodiversity. These damages resonate beyond dairy feed producing regions through animal migration and are an overlooked part of the transferred environmental burden caused by a growing thirst for milk in China and elsewhere.

The fractal dimension of a conspicuous ornament varies with mating status and shows assortative mating in wild red-legged partridges (Alectoris rufa).

Complex body designs, such as plumage ornaments in birds, can be described by fractal geometry. These complex patterns could have a role as visual signals during courtship and social interactions, but an empirical validation in the wild is currently lacking. Here, we investigated whether the fractal dimension (FD) of a complex plumage pattern displayed by red-legged partridges Alectoris rufa could function as a potential sexual signal. We captured wild birds early in the breeding season and tested if mated and unmated birds differed in the FD of their conspicuous melanin-based black bib. We also tested if the FD of the black bib was correlated within the pair, looking for evidence of assortative mating based on the expression of this trait. We simultaneously assessed similar effects in other ornamental traits (black bib size, white throat patch and black flank band surface, redness of the eye rings and bill). Mated birds showed higher black bib FD values than unmated ones. Mated males, but not females, also displayed a larger black bib. Moreover, the black bib FD (but not the trait size) and the white throat patch surface showed assortative mating. Finally, females with higher black bib FD showed smaller black flank band surface, suggesting a trade-off in the expression of the two melanin-pigmented plumage traits. This provides unique and novel indication for the shape complexity of a pigmented trait, here described by its fractal dimension, to be potentially under sexual selection in a wild animal.

Individual variation in behavioural responsiveness to humans leads to differences in breeding success and long-term population phenotypic changes.

Whether human disturbance can lead to directional selection and phenotypic change in behaviour in species with limited behavioural plasticity is poorly understood in wild animal populations. Using a 19-year study on Montagu's harrier, we report a long-term increase in boldness towards humans during nest visits. The probability of females fleeing or being passive during nest visits decreased, while defence intensity steadily increased over the study period. These behavioural responses towards humans were significantly repeatable. The phenotypic composition of the breeding population changed throughout the study period (4-5 harrier generations), with a gradual disappearance of shy individuals, leading to a greater proportion of bolder ones and a more behaviourally homogeneous population. We further show that nest visit frequency increased nest failure probability and reduced productivity of shy females, but not of bold ones. Long-term research or conservation programmes needing nest visits can therefore lead to subtle but relevant population compositional changes that require further attention.

Density-Dependent Prevalence of Francisella tularensis in Fluctuating Vole Populations, Northwestern Spain.

Tularemia in humans in northwestern Spain is associated with increases in vole populations. Prevalence of infection with Francisella tularensis in common voles increased to 33% during a vole population fluctuation. This finding confirms that voles are spillover agents for zoonotic outbreaks. Ecologic interactions associated with tularemia prevention should be considered.

Changes in behaviour and faecal glucocorticoid levels in response to increased human activities during weekends in the pin-tailed sandgrouse.

Human recreational activities are becoming increasingly widespread and frequent, a fact that may potentially exacerbate their effects on wildlife. These human-related disturbances on animals may induce behavioural and physiological changes that can ultimately affect their fitness, showing a similar anti-predator response that against natural predator or other threats. Here, we combine the use of behavioural and physiological approaches to assess the potential effect of winter human activities on a threatened farmland bird in Europe, the pin-tailed sandgrouse (Pterocles alchata). We compared before, during and after weekend variations in human activity rates, pin-tailed sandgrouse behaviour (flocking and flying behaviour, interspecific association in mixed flocks and habitat use) and faecal glucocorticoid metabolite concentrations. Human disturbances, in particular those associated with hunting activities, peaked during weekends. Sandgrouse showed significant behavioural changes (increased sandgrouse-only flock sizes, increased proportion of birds flying and changes in habitat use) during weekends and higher faecal glucocorticoid metabolite concentrations after the weekends compared with during or before weekends. Therefore, physiological stress levels could be modulated by behavioural adjustments such as increased flock sizes and changes in habitat use that may allow sandgrouse to cope with increased human disturbance rates during weekends. Nevertheless, temporal and spatial organization of hunting days among groups of estates might be good strategies to buffer these potential adverse effects on wintering pin-tailed sandgrouse and other steppe species of conservation concern, while preserving a socio-economically important activity such as hunting.

Effects of Lead Exposure on Sperm Quality and Reproductive Success in an Avian Model.

Lead (Pb) poisoning via ingestion of shot pellets is a frequent cause of death in wild birds and also has a wide range of subclinical effects. Here we report on the sublethal effects Pb exposure has on the breeding performance of red-legged partridges (Alectoris rufa). We studied the effects of Pb exposure on sperm quality, reproductive success, egg properties, laying performance, antioxidant levels, and carotenoid-based coloration. Birds were exposed by oral gavage to one or three No. 6 Pb shot pellets (2.8 mm in diameter, mean mass ± SD: 109 ± 7.97 mg). We show that exposure to three pellets (330 mg) reduced the hatching rate of females and decreased the acrosome integrity and sperm motility of males. In addition, females exposed to 1 pellet (110 mg) produced heavier eggs and chicks, whereas males exposed to 1 pellet presented an increase in sperm vigor. Sperm viability, concentration, progressiveness or fecundation rate were not affected by Pb treatment. Pb exposure increased circulating antioxidant levels in males, whereas the percentage of carotenoid-pigmented eye-ring area decreased in exposed females. Several sperm parameters showed positive relationships with coloration and antioxidant levels, suggesting that males displaying redder ornaments may be more capable of protecting sperm from oxidative stress in the event of sublethal Pb exposure.

Sublethal Pb exposure produces season-dependent effects on immune response, oxidative balance and investment in carotenoid-based coloration in red-legged partridges.

Ingestion of lead (Pb) shot pellets constitutes the main cause of Pb poisoning in avifauna. We studied the effects of sublethal Pb exposure on immunity, carotenoid-based coloration, oxidative stress and trade-offs among these types of responses during spring and autumn in red-legged partridges (Alectoris rufa). We evaluated constitutive immunity testing lysozyme and natural antibody levels, and blood bactericidal and phagocytic activities. We studied induced immunity by testing PHA and humoral responses. We analyzed fecal parasite and bacterial abundance and oxidative stress biomarkers. Pb exposure in spring reduced natural antibody levels, whereas in autumn, it reduced lysozyme levels and increased phagocytic activity. Pb exposure increased PHA response in both seasons, and decreased T-independent humoral response in autumn. Pb exposure also increased noncoliform and decreased coliform Gram-negative gut bacteria. In spring, Pb exposure decreased antioxidant levels and increased coloration in males, whereas in autumn, it increased retinol levels but reduced coloration in both genders. Our results suggest that in spring, Pb-exposed females used antioxidants to cope with oxidative stress at the expense of coloration, whereas Pb-exposed males increased coloration, which may reflect an increased breeding investment. In autumn, both genders prioritized oxidative balance maintenance at the expense of coloration.

Assessing the Risk of Fipronil-Treated Seed Ingestion and Associated Adverse Effects in the Red-Legged Partridge.

Fipronil is an insecticide commonly used in agriculture, but there are growing concerns over its environmental impacts (e.g., harmful effects on pollinators). Fipronil-treated seed ingestion might threaten granivorous farmland birds, in particular, Gallinaceous birds that are particularly sensitive to this insecticide. We report here on exposure risk and effects in a game bird of high socioeconomic importance, the red-legged partridge (Alectoris rufa). We fed captive birds with untreated maize (controls) or with a mixture of untreated-treated maize (ratio 80:20; exposed birds) during 10 days at the beginning of the breeding period (n = 12 pairs in each group). We first show that exposed partridges did not reject treated seeds but reduced food intake and lost body condition. We further studied the effects of treated seed ingestion on adult survival, oxidative balance, plasma biochemistry, carotenoid-based coloration, cellular immune response, steroid hormone levels, and reproduction. Fipronil exposure altered blood biochemistry and sexual hormone levels and reduced cellular immune response, antioxidant levels, and carotenoid-based coloration. Exposed pairs also had reduced egg fecundation rate and produced eggs with fewer antioxidants and offspring that had reduced cellular immune response. These negative effects on adult partridges, their reproductive performance, and offspring quality highlight that fipronil-treated seed ingestion is a significant threat to wild birds.

Imidacloprid-treated seed ingestion has lethal effect on adult partridges and reduces both breeding investment and offspring immunity.

The ingestion of imidacloprid treated seeds by farmland birds may result in exposure to toxic amounts of this insecticide. Here we report on the effects that the exposure to the recommended application rate and to 20% of that rate may produce on birds feeding on treated seeds. Experimental exposure to imidacloprid treated seeds was performed on red-legged partridges (Alectoris rufa) (n=15 pairs per treatment group: control, 20% or 100% of the recommended application rate) during two periods that corresponded to the autumn (duration of exposure: 25 days) and late winter (10 days) cereal sowing times in Spanish farmlands. We studied effects on the survival, body condition, oxidative stress biomarkers, plasma biochemistry, carotenoid-based coloration, T-cell mediated immune response and reproduction of exposed adult partridges, and on the survival and T-cell immune response of their chicks. The high dose (recommended application rate) killed all partridges, with mortality occurring faster in females than in males. The low dose (20% the recommended application rate) had no effect on mortality, but reduced levels of plasma biochemistry parameters (glucose, magnesium and lactate dehydrogenase), increased blood superoxide dismutase activity, produced changes in carotenoid-based integument coloration, reduced the clutch size, delayed the first egg lay date, increased egg yolk vitamins and carotenoids and depressed T-cell immune response of chicks. Moreover, the analysis of the livers of dead partridges revealed an accumulation of imidacloprid during exposure time. Despite the moratorium on the use of neonicotinoids in the European Union, birds may still be at high risk of poisoning by these pesticides through direct sources of exposure to coated seeds in autumn and winter.