Fine-scale genetic structure in a high dispersal capacity raptor, the Montagu's harrier (Circus pygargus), revealed by a set of novel microsatellite loci.

The Montagu's harrier (Circus pygargus) is a semi-colonial raptor species widely but patchily distributed across the Palearctic region with recorded cases of philopatry and presence of extra-pair copulation. In order to assess Montagu's harrier spatial genetic structure and contemporary gene flow, we developed 16 new microsatellite markers using 454 pyrosequencing. Genotypes of 117 chicks sampled in a 200 × 300 km farmland area in Central Western France were analyzed to characterize genetic polymorphism at each locus and regional and fine-scale genetic structure. Fourteen markers were found polymorphic, with a number of alleles ranging from 3 to 11. The expected and observed heterozygosities ranged from 0.36 to 0.856 and from 0.35 to 0.868, respectively. A single genetic unit was found at the regional scale with higher genetic similarity observed at a small spatial scale (up to 10 km). Our results are consistent with overall large-scale juvenile and adult dispersal together with small-scale male philopatry. Cross-species amplification of this set of microsatellites makers has been successful in two closely related harrier species: the marsh harrier (Circus aeruginosus) and the Hen harrier (Circus cyaneus) for which 14 and 12 markers were polymorphic, respectively. These new microsatellite markers could be used to study the population genetic structure, contemporary gene flow and parentage analyses in these three species and to conduct microsatellite-based demographic inferences on the Montagu's harrier.

The consumption pattern of 28 species of carabid beetles (Carabidae) to a weed seed, Viola arvensis.

Seedbank control has been the cornerstone of agricultural management practices. Regulating weeds by using their predators, as a weed control strategy, may be a prerequisite of decreasing herbicide use, and has thus attracted much research investigating the possible contribution of both vertebrates and invertebrates as weed seed predators. Carabid beetles are considered as one of the most important invertebrate seed predators in agroecosystems. We aimed at investigating carabid beetle preferences to a single prey type, seeds of Viola arvensis. We measured the consumption of seeds in 28 species of carabid beetles, under controlled experimental conditions. Two main tribes are identified in tested species, Harpalini with 12 species and Pterostichini with ten species. We found no relationships between species body mass and Viola's seed consumption, nor with the ratio between mandible length and labrum width (ML/LW). However, trends became significant with the ratio ML/LW when restricting these analyses to species that ate at least five seeds. In addition, we detected a positive and significant relationship between consumption rate and occurrence in trapping sessions over the last 3 years. These results are in favor of weed seeds control by carabids. Clear limits of this study are the use of a single seed species and under control conditions. This experimentation calls for additional studies to check for consistencies in consumption against seed species.

The evolution of north-east Atlantic gadfly petrels using statistical phylogeography.

Macaronesia (north-east Atlantic archipelagos) has been host to complex patterns of colonization and differentiation in many groups of organisms including seabirds such as gadfly petrels (genus Pterodroma). Considering the subspecies of widely distributed soft-plumaged petrel for many years, the taxonomic status of the three gadfly petrel taxa breeding in Macaronesia is not yet settled, some authors advocating the presence of three, two or one species. These birds have already been the subject of genetic studies with only one mtDNA gene and relatively modest sample sizes. In this study, using a total of five genes (two mitochondrial genes and three nuclear introns), we investigated the population and phylogeographical histories of petrel populations breeding on Madeira and Cape Verde archipelagos. Despite confirming complete lineage sorting with mtDNA, analyses with nucDNA failed to reveal any population structuring and Isolation with Migration analysis revealed the absence of gene flow during the differentiation process of these populations. It appears that the three populations diverged in the late Pleistocene in the last 150 000 years, that is 10 times more recently than previous estimates based solely on one mtDNA gene. Finally, our results suggest that the Madeira petrel population is ancestral rather than that from Cape Verde. This study strongly advocates the use of nuclear loci in addition to mtDNA in demographical and phylogeographical history studies.

Glyphosate, AMPA and glufosinate in soils and earthworms in a French arable landscape.

Although Glyphosate-based herbicides are often marketed as environmentally friendly and easily biodegradable, its bioavailability and risks to wildlife raise significant concerns. Among non-target organisms, earthworms which live in close contact with the soil can be directly exposed to pesticides and harmed. We investigated soil contamination and the exposure of earthworms to glyphosate, its metabolite AMPA, and glufosinate in an arable landscape in France, both in treated (i.e. temporary grasslands and cereal fields under conventional farming), and nontreated habitats (i.e. hedgerows, permanent grasslands and cereal fields under organic farming) (n = 120 sampling sites in total). Glyphosate, AMPA and glufosinate were detected in 88%, 58% and 35% of the soil samples, and in 74%, 38% and 12% of the earthworm samples, respectively. For both glyphosate and AMPA, concentrations in soils were at least 10 times lower than predicted environmental concentrations. However, the maximum glyphosate soil concentration measured (i.e., 0.598 mg kg-1) was only 2 to 3 times lower than the concentrations revealed to affect earthworms (survival and avoidance) in the literature. These compounds were found both in conventional and organic farming fields, thus supporting a recent study, and for the first time they were detected in hedgerows and grasslands. However, glyphosate and AMPA were more frequently detected in soils from cereal fields and hedgerows than in grasslands, and median concentrations measured in soils from cereal fields were significantly higher than in the two other habitats. Bioaccumulation of glyphosate and AMPA in earthworms was higher than expected according to the properties of the molecules. Our findings raised issues about the high occurrence of glyphosate and AMPA in soils from cropped and more natural areas in arable landscapes. They also highlight the potential for transfer of these molecules in terrestrial food webs as earthworms are prey for numerous animals.

Maternal investment of female mallards is influenced by male carotenoid-based coloration.

The differential allocation hypothesis predicts that females modify their investment in a breeding attempt according to its reproductive value. One prediction of this hypothesis is that females will increase reproductive investment when mated to high-quality males. In birds, it was shown that females can modulate pre-hatch reproductive investment by manipulating egg and clutch sizes and/or the concentrations of egg internal compounds according to paternal attractiveness. However, the differential allocation of immune factors has seldom been considered, particularly with an experimental approach. The carotenoid-based ornaments can function as reliable signals of quality, indicating better immunity or ability to resist parasites. Thus, numerous studies show that females use the expression of carotenoid-based colour when choosing mates; but the influence of this paternal coloration on maternal investment decisions has seldom been considered and has only been experimentally studied with artificial manipulation of male coloration. Here, we used dietary carotenoid provisioning to manipulate male mallard (Anas platyrhynchos) bill coloration, a sexually selected trait, and followed female investment. We show that an increase of male bill coloration positively influenced egg mass and albumen lysozyme concentration. By contrast, yolk carotenoid concentration was not affected by paternal ornamentation. Maternal decisions highlighted in this study may influence chick survival and compel males to maintain carotenoid-based coloration from the mate-choice period until egg-laying has been finished.

Genetic consequences of interglacial isolation in a steppe bird.

In response to climate changes that have occurred during Pleistocene glacial cycles, taxa associated to steppe vegetation might have followed a pattern of historical evolution in which isolation and fragmentation of populations occurred during the short interglacials and expansion events occurred during the long glacial periods, in contrast to the pattern described for temperate species. Here, we use molecular genetic data to evaluate this idea in a steppe bird with Palaearctic distribution, the little bustard (Tetrax tetrax). Overall, extremely low genetic diversity and differentiation was observed among eight little bustard populations distributed in Spain and France. Mismatch distribution analyses showed that most little bustard populations expanded during cooling periods previous to, and just after, the last interglacial period (127,000-111,000 years before present), when steppe habitats were widespread across Europe. Coalescent-based methods suggested that glacial expansions have resulted in substantial admixture in Western Europe due to the existence of different interglacial refugia. Our results are consistent with a model of evolution and genetic consequences of Pleistocene cycles with low between-population genetic differentiation as a result of short-term isolation periods during interglacials and long-term exchange during glacial periods.

Sex-biased dispersal patterns depend on the spatial scale in a social rodent.

Dispersal is a fundamental process in ecology because it influences the dynamics, genetic structure and persistence of populations. Furthermore, understanding the evolutionary causes of dispersal pattern, particularly when they differ between genders, is still a major question in evolutionary ecology. Using a panel of 10 microsatellite loci, we investigated at different spatial scales the genetic structure and the sex-specific dispersal patterns in the common vole Microtus arvalis, a small colonial mammal. This study was conducted in an intensive agricultural area of western France. Hierarchical F(ST) analyses, relatedness and assignment tests suggested (i) that females are strongly kin-clustered within colonies; (ii) that dispersal is strongly male-biased at a local scale; and (iii) long-distance dispersal is not rare and more balanced between genders. We conclude that males migrate continuously from colony to colony to reproduce, whereas females may disperse just once and would be mainly involved in new colony foundation.

Spatial genetic structure of a small rodent in a heterogeneous landscape.

Gene flow in natural populations may be strongly influenced by landscape features. The integration of landscape characteristics in population genetic studies may thus improve our understanding of population functioning. In this study, we investigated the population genetic structure and gene flow pattern for the common vole, Microtus arvalis, in a heterogeneous landscape characterised by strong spatial and temporal variation. The studied area is an intensive agricultural zone of approximately 500 km(2) crossed by a motorway. We used individual-based Bayesian methods to define the number of population units and their spatial borders without prior delimitation of such units. Unexpectedly, we determined a single genetic unit that covered the entire area studied. In particular, the motorway considered as a likely barrier to dispersal was not associated with any spatial genetic discontinuity. Using computer simulations, we demonstrated that recent anthropogenic barriers to effective dispersal are difficult to detect through analysis of genetic variation for species with large effective population sizes. We observed a slight, but significant, pattern of isolation by distance over the whole study site. Spatial autocorrelation analyses detected genetic structuring on a local scale, most probably due to the social organisation of the study species. Overall, our analysis suggests intense small-scale dispersal associated with a large effective population size. High dispersal rates may be imposed by the strong spatio-temporal heterogeneity of habitat quality, which characterises intensive agroecosystems.

Density dependence in a recovering osprey population: demographic and behavioural processes.

1. Understanding how density-dependent and independent processes influence demographic parameters, and hence regulate population size, is fundamental within population ecology. We investigated density dependence in growth rate and fecundity in a recovering population of a semicolonial raptor, the osprey Pandion haliaetus [Linnaeus, 1758], using 31 years of count and demographic data in Corsica. 2. The study population increased from three pairs in 1974 to an average of 22 pairs in the late 1990s, with two distinct phases during the recovery (increase followed by stability) and contrasted trends in breeding parameters in each phase. 3. We show density dependence in population growth rate in the second phase, indicating that the stabilized population was regulated. We also show density dependence in productivity (fledging success between years and hatching success within years). 4. Using long-term data on behavioural interactions at nest sites, and on diet and fish provisioning rate, we evaluated two possible mechanisms of density dependence in productivity, food depletion and behavioural interference. 5. As density increased, both provisioning rate and the size of prey increased, contrary to predictions of a food-depletion mechanism. In the time series, a reduction in fledging success coincided with an increase in the number of non-breeders. Hatching success decreased with increasing local density and frequency of interactions with conspecifics, suggesting that behavioural interference was influencing hatching success. 6. Our study shows that, taking into account the role of non-breeders, in particular in species or populations where there are many floaters and where competition for nest sites is intense, can improve our understanding of density-dependent processes and help conservation actions.

Estimating acridid densities in grassland habitats: a comparison between presence-absence and abundance sampling designs.

Sampling methods to estimate acridid density per surface area unit in grassland habitats were compared using presence-absence data and count data. Sampling plans based on 6 yr of surveys were devised to estimate the density of Chorthippus spp., Euchorthippus spp., and Calliptamus italicus L. These acridids represented >90% of species in the study area. Sampling plans based on count data provided a reasonable tool when densities were >1/m(2) and when the level of precision was 0.20-0.30. A binomial sampling plan can be used to estimate C. italicus density with a level of precision >or=0.28. Sampling characteristics, i.e., estimated mean, actual precision, and sample size, were established on validation data sets with bootstrapping analysis. Sampling costs were also calculated according to density-dependent functions. Comparison between binomial sampling and enumerative sampling of C. italicus showed that binomial sampling required less time than enumerative sampling when densities were 0.35. Plot area had no significant effect on sample variances of counts.

Features of DNA fragments obtained by random amplified polymorphic DNA (RAPD) assays

Random amplified polymorphic DNA (RAPD) fragments were prepared from samples of Calonectris diomedea (Cory's shearwater, Aves) and Haemonchus contortus (Nematoda) DNA by polymerase chain reaction (PCR) using decamers containing two restriction enzyme sites as primers. Six of 19 studied RAPD fragments probably originated from traces of commensal microorganisms. Many rearranged fragments, absent in the original genomic DNA, were synthesized and amplified during the processing of all the DNA samples, indicating that interactions occur within and between strands during the annealing step of PCR. The model of interactions between molecular species during DNA amplification with a single arbitrary oligonucleotide primer was modified to include nested primer annealing and interactions within and between strands. The presence of these artefacts in the final RAPD have a major effect on the interpretation of polymorphism studies.

Effect of preen oil on plumage bacteria: an experimental test with the mallard.

Feathers are essential for avian thermoregulation, communication or flight and a reduction of plumage condition may alter these functions and reduce individual fitness. Recently, descriptive studies provided evidence that birds carry feather-degrading bacteria on their plumage that have the ability to degrade feathers rapidly under laboratory conditions. If such bacteria reduce avian fitness, natural selection should favour the evolution of anti-bacterial defences to limit the effects of these detrimental microorganisms. Preening behaviour and associated preen gland secretions have been proposed as the main factor used by birds to limit feather-degrading bacterial growth and some recent in vitro studies provided evidence that uropygial oil inhibited the growth of some keratinolytic strains in passerines. However, preen oil antimicrobial properties remained to be experimentally tested in vivo. We conducted an experiment with mallards (Anas platyrhynchos) onto which we fixed a removable mechanism that blocked access to the uropygial gland in a first group of mallards, whilst birds in a second group had the same removable mechanism but access to their gland and a third group of birds had normal access to their gland. We found no significant effect of our treatment on total and feather-degrading bacterial loads. Three hypotheses may explain the discrepancy between our results and previous in vitro studies. First, in vitro studies may have over-estimated the bactericidal properties of the preen oil. Second, preen gland deprivation may have affected only a small portion of the feather-degrading bacterial community. Third, ducks and passerine oils might have different bactericidal properties.

Spatial autocorrelation in farmland grasshopper assemblages (Orthoptera: Acrididae) in western France.

Agricultural intensification in western Europe has caused a dramatic loss of grassland surfaces in farmlands, which have resulted in strong declines in grassland invertebrates, leading to cascade effects at higher trophic levels among consumers of invertebrates. Grasshoppers are important components of grassland invertebrate assemblages in European agricultural ecosystems, particularly as prey for bird species. Understanding how grasshopper populations are distributed in fragmented landscapes with low grassland availability is critical for both studies in biodiversity conservation and insect management. We assessed the range and strength of spatial autocorrelation for two grasshopper taxa (Gomphocerinae subfamily and Calliptamus italicus L.) across an intensive farmland in western France. Data from surveys carried out over 8 yr in 1,715 grassland fields were analyzed using geostatistics. Weak spatial patterns were observed at small spatial scales, suggesting important local effects of management practices on grasshopper densities. Spatial autocorrelation patterns for both grasshopper taxa were only detected at intermediate scales. For Gomphocerinae, the range of spatial autocorrelation varied from 802 to 2,613 m according to the year, depending both on grasshopper density and on grassland surfaces in the study site, whereas spatial patterns for the Italian locust were more variable and not related to grasshopper density or grassland surfaces. Spatial patterns in the distribution of Gomphocerinae supported our hypothesis that habitat availability was a major driver of grasshopper distribution in the landscape, and suggested it was related to density-dependent processes such as dispersal.

Adaptive significance of seabird coloration: the case of procellariiforms.

The function of bird coloration has been debated over recent decades, but no general theory has yet emerged. Most studies have only analyzed the influence of a single factor, but multifactorial analyses may be required to understand the biological function of coloration. On procellariiforms (albatrosses and petrels), which comprise 105 seabird species, all with simple coloration (mainly black and/or white), correspondence analysis was used to find correlations between coloration and species body size, diet, feeding techniques, foraging group sizes, and climate at breeding localities. Correlations with taxonomic classification were also examined. No single factor played an overwhelming part in determining color patterns, but whereas diet, size, and climate had little effect, feeding techniques and foraging group sizes had important influences. Classification, at the genus or family level, and color pattern were also correlated. Clusters of species, whether derived from their color patterns or their feeding ecologies, were similar. Coloration may therefore be related to procellariiform life histories, possibly acting through competition and predation risk. Relative uniformity of procellariiform coloration, compared with other seabirds, may relate to their peculiar foraging techniques.

Courtship behaviour in a lekking species: individual variations and settlement tactics in male little bustard.

We analysed the display behaviour of male little bustard Tetrax tetrax to identify displays that are used in the context of male-male competition and those that are used for attracting females. Courtship was the main activity of males during the breeding season. Calling activity occurred throughout the day, and leks were attended for more than 4 months. Male sexual displays included snort call, wing-flash, and jump display. Snort call was performed throughout the day and mainly involved male-male interactions. In contrast, the wing-flash display was given only at twilight, and was performed most commonly when a female was present, supporting an inter-sexual function for this display. The jump display was performed only in the presence of female at anytime of the day. Analysis of individual variations in display behaviour revealed that intra-individual variation was low compared to inter-individual variation, especially for the jump display. It is, therefore, possible that display rates provide information on male quality. Four male settlement patterns could be defined, singles, paired, lekking and satellite lekking, but only wing-flash display and stamped snort call differed among those categories. We suggest that satellite males are attempting to benefit from proximity to higher status males, in accordance with the hotshot hypothesis of lek evolution.

Predation as a cost of sexual communication in nocturnal seabirds: an experimental approach using acoustic signals.

Conspicuous behaviour, such as sexual advertisement, exposes animals to predation; mate attraction thus often conflicts with antipredator behaviour. We investigated whether an avian predator, the brown skua, Catharacta antarctica lönnbergi, uses the mate attraction calls of colonial seabirds, the petrels. The majority of petrels attract mates at night and vocalizations are their main way of communicating. At our study sites, skua predation on nocturnal petrels was heavy, and concentrated particularly on a single species, the blue petrel, Halobaena caerulea. Using playback experiments, we showed that skuas can use male petrel calls as a cue for prey location and selection. This listening behaviour of skuas probably imposes a major constraint on advertising petrels, and especially on single males which face a trade-off between attracting females (which respond by calling in flight) and avoiding predation. We also investigated the consequences of this predation risk on the behaviour of petrels: a second set of playback experiments showed that the most heavily preyed on petrel species could use skua territorial calls to infer predation risk and stop calling thereafter, which may reduce conspicuousness and predation risk. Copyright 2000 The Association for the Study of Animal Behaviour.

Lek mating systems: a case study in the Little Bustard Tetrax tetrax.

Leks have recently been defined as male display aggregations that females attend primarily for the purpose of mating. This is an extended version of previous definitions, as a clear-cut definition of leks is difficult to obtain. Four criteria should be verified to identify a lekking species: (i) there is no male parental investment beyond the sperm; (ii) males aggregate at specific sites for display; (iii) the only resource females find on the lek is the male, i.e. the male genes; (iv) females can select her mate(s), although the necessity of this latter condition for lekking species has been highly debated. We applied these criteria to the endangered little bustard Tetrax tetrax, a species that is claimed to show an exploded lek mating system, but for which this has never been fully investigated. We monitored a population of little bustards in western France during 2 years to investigate the two central criteria in the assessment of their mating system: male aggregation in arenas and lack of consistent resources in male territories. We analysed the spatial distribution of little bustard male territories, the individual variation in size, and the land use characteristics of male territories, with particular attention to the habitats that may be considered as defensible resources. Displaying males showed an aggregated spatial distribution over the study area during the 2 years of survey. Male territories were rather large (19+/-16 ha), but a large among-male variability in territory size was observed. Land use within the territories included mainly permanent and semi-permanent crops. The variability in land use among territories suggests also that resources found within male territories were selected according to male needs (food and display) rather than to female needs (permanent crops that are more appropriate for reproduction). The mating system of the little bustard seems to match the general (and extended) definition of leks, at least in some populations. However, limits between resource defence polygyny and extreme exploded or resource-based leks are thin and unclear, and the little bustard is a good example of how lek definitions may be difficult to apply in non clear-cut empirical situations.