Mickaël Sage

Kinetics of bromadiolone in rodent populations and implications for predators after field control of the water vole, Arvicola terrestris

We document the kinetics of bromadiolone in two rodent populations after a field control of water voles, and their implications for predator exposure. Water voles and common voles were trapped aboveground and underground from 1 to 135 days after bromadiolone treatment in the field. Livers, digestive tracts, and rests of the body were analyzed separately. Our results indicate that 99.6% of the water voles trapped underground and 41% of the common voles trapped aboveground contain bromadiolone residues. Concentrations were maximal between 3.3 and 6.5 days after treatment, according to the tissues examined and the model applied for water voles, and after 1.3 to 3.7 days for common voles. Water voles appeared available almost exclusively for foraging predators. Common voles, found less likely to be poisoned and exhibiting weaker concentrations, were mainly sampled aboveground. The liver, primarily eaten by some predators and scavengers, contains a larger bromadiolone quantity (59% of the total amount found in water voles). The rejection of the digestive tract by those species may lead to a subsequent consumption of voles with higher bromadiolone concentrations (from +3.8 to +5.8% of concentration) and provide a moderate risk increase. After 135 days, eight of the ten water voles and one of the two common voles exhibited detectable residues. Additionally, one specimen presented higher concentrations than the others, and similar to those measured in Voles trapped between the first 15-20 days. This may have consequences on predator intoxications several months after treatment. These results integrate individual differences for the two main rodent species present in treated areas. Implications for predator exposure were investigated at the end of the study and suggest that, if the risk of secondary poisoning is maximal during the first 15-20 days when the rodent densities remain high, exposure conditions are maintained for at least 135 days.

Determination of bromadiolone residues in fox faeces by LC/ESI-MS in relationship with toxicological data and clinical signs after repeated exposure.

In many countries, the fox (Vulpes vulpes), predator of small mammals, is particularly affected by anticoagulant rodenticides such as bromadiolone due to secondary poisoning. Nevertheless, to date, no method of exposure monitoring is applicable in the field over large areas, and no toxicological data are available concerning sensitivity of foxes to bromadiolone. The aim of this work was to compare excretion kinetics of bromadiolone in fox faeces with clinical and haemostatic effects after repeated exposure to intoxicated voles. A sensitive method for the quantification of bromadiolone excretion in fox faeces and plasma was developed, using liquid chromatography combined with electrospray ionisation mass spectrometry (LC/ESI-MS). The LoD was 0.9microg/kg and 0.15microg/L, and the LoQ was 3.0microg/kg and 0.5microg/L, in faeces and in plasma, respectively. Four captive foxes were fed for 2 or 5 days with water voles (Arvicola terrestris Sherman) spiked with bromadiolone at concentrations close to those measured in the field. Faeces and blood were collected for bromadiolone titration, and blood-clotting tests were performed to monitor fox health daily during 10 days and then every 3-4 days until the end of the experiment (D28). Then, after euthanasia, a complete necropsy was performed, and levels of bromadiolone residues in the liver were determined. Bromadiolone residues were detected in faeces 15h after the first exposure. They increased dramatically during the exposure period and then gradually decreased, but they remained detectable at the end of the experiment, i.e., 26 days after the last exposure. Bromadiolone residues in plasma showed a similar pattern but were no longer detectable 7-24 days after the last exposure. Two foxes presented very severe external haemorrhages, requiring the administration of the antidote vitamin-K1. Bromadiolone residues in faeces and their relationships with exposure and other direct-markers that were measured are discussed. Liver residues and the toxicity data of our study will help to interpret data from fox carcasses collected by wildlife disease surveillance networks. These findings provide a basis for programs aiming to monitor the exposure of wild fox populations to bromadiolone using non-invasive methods based on standard sampling and analysis of residues in faeces.

Negative impact of urban habitat on immunity in the great tit Parus major.

Urban habitats are described as having an overall negative influence on many fitness-related traits in several bird species, but a vital function such as immunity remains poorly studied. The immune response is strongly linked to individual condition, which partly depends on resource availability and the parasitic context that often differ between urban and natural habitats. A difference between the immunity of populations dwelling in urban areas and populations from more natural habitats can, therefore, be hypothesized. We conducted a 2-year experimental study on great tits (Parus major) in urban and forest areas. We stimulated the constitutive immunity of nestlings and assessed both the inflammatory response by measuring the plasma levels of haptoglobin, an inflammatory marker, and its activation cost through the loss of body mass. In addition, we checked the nestlings for ectoparasites and assessed haemosporidian prevalence in adults. Nestlings from urban sites produced relatively less haptoglobin and lost more body mass than those from forest sites, which suggests that the activation of constitutive immunity is more costly for birds living in urban sites than for those living in the forest. We detected no ectoparasite in birds in both habitats. However, urban adults showed lower haemosporidian prevalence than forest ones, suggesting a reduced exposure to these parasites and their vectors in towns. Overall, our study provides evidence for an immune difference between urban and forest populations. Because immunity is crucial for organism fitness, it is of prime interest to identify causes and processes at the origin of this difference.

Multi-Element Analysis of Blood Samples in a Passerine Species: Excesses and Deficiencies of Trace Elements in an Urbanization Study

Urbanization is a growing phenomenon characterized by a complete restructuring of natural areas. In urban bird populations, a reduced offspring survival and body condition and an overall lower breeding success are often observed compared to populations inhabiting more natural habitats. Higher pollution levels and poorer quality of natural resources in cities are two environmental factors frequently mentioned in the literature to explain the differences between urban and non-urban populations. Pollution and poor quality of food may lead to an excess of non-essential elements such as Pb or Cd or to deficiencies of essential elements such as Cu or Zn, which may explain some of the impacts, notably on immunity, observed in urbanization studies. The present study compared the breeding parameters, brood body mass and condition, and haptoglobin levels, a marker of inflammatory immunity, in two urban and two forest populations of Great tits in Eastern France, together with a multi-element analysis (25 non-essential and essential trace elements) of blood samples from 13-day-old nestlings from the four populations. The concentration of NO2, a gaseous pollutant typical of urban pollution, was also measured. The NO2 concentrations were significantly higher in the urban areas, but no association with biological variables was detected. Non-essential metals were undetectable in the plasma of the birds from both habitats, except Pb, whose concentrations, however, did not differ between the urban and forest birds. A positive relationship was found between the plasmatic richness in essential elements (as assessed from the coordinates of the first axis of a PCA including 12 elements) and the average brood body mass and condition. We suggest that lower quality resources or/and a higher metabolic demand may be a causal mechanism for the reduced body condition often observed in urban bird nestlings. Finally, our exploratory study could promote more mechanistic experiments (e.g. supplementation) to explain the negative effect of urban conditions on bird populations.