Joxerra Aihartza

PCR-RFLP identification of mustelid species: European mink (Mustela lutreola), American mink (M. vison) and polecat (M. putorius) by analysis of excremental DNA

Recent advances in molecular scatology have allowed the development of reliable and non-invasive methods that can be applied in monitoring of small carnivores, without disturbance of the animals. Here a method is described that can be used to differentiate European mink Mustela lutreola , polecat M. putorius and American mink M. vison based on the analysis of DNA extracted from faeces. It consists of a nested PCR of a region of the mitochondrial D-loop followed by digestion of the resulting 240 bp amplicons with the restriction enzymes Rsa I and Msp I. The restriction patterns of both enzymes, when used together, are found to detect species-specific sequence variation. Two different haplotypes for European mink (AA, AB), another two for the polecat (AC, AD) and one for American mink (BC) can also be discriminated by this technique. Two new haplotypes for the mitocondrial D-loop of mustelids are described after DNA sequencing.

Detection of alpha and betacoronaviruses in multiple Iberian bat species

Bat coronaviruses (CoV) are putative precursors of the severe acute respiratory syndrome (SARS) CoV and other CoV that crossed the species barrier from zoonotic reservoirs into the human population. To determine the presence and distribution of CoV in Iberian bats, 576 individuals of 26 different bat species were captured in 13 locations in Spain. We report for the first time the presence of 14 coronaviruses in 9 Iberian bat species. Phylogenetic analysis of a conserved CoV genome region (RdRp gene) shows a wide diversity and distribution of alpha and betacoronavirus in Spain. Interestingly, although some of these viruses are related to other European BatCoV, or to Asian CoV, some of the viruses found in Spain cluster in new groups of α and β CoV.

The Foraging Ecology of the Mountain Long-Eared Bat Plecotus macrobullaris Revealed with DNA Mini-Barcodes

Molecular analysis of diet overcomes the considerable limitations of traditional techniques for identifying prey remains in bat faeces. We collected faeces from individual Mountain Long-eared Bats Plecotus macrobullaris trapped using mist nets during the summers of 2009 and 2010 in the Pyrenees. We analysed their diet using DNA mini-barcodes to identify prey species. In addition, we inferred some basic features of the bats foraging ecology that had not yet been addressed. P. macrobullaris fed almost exclusively on moths (97.8%). As prey we detected one dipteran genus (Tipulidae) and 29 moth taxa: 28 were identified at species level (23 Noctuidae, 1 Crambidae, 1 Geometridae, 1 Pyralidae, 1 Sphingidae, 1 Tortricidae), and one at genus level (Rhyacia sp., Noctuidae). Known ecological information about the prey species allowed us to determine that bats had foraged at elevations between 1,500 and 2,500 m amsl (above mean sea level), mostly in subalpine meadows, followed by other open habitats such as orophilous grasslands and alpine meadows. No forest prey species were identified in the diet. As 96.4% of identified prey species were tympanate moths and no evidence of gleaning behaviour was revealed, we suggest P. macrobullaris probably forages by aerial hawking using faint echolocation pulses to avoid detection by hearing moths. As we could identify 87.8% of the analysed sequences (64.1% of the MOTUs, Molecular Operational Taxonomic Units) at species level, we conclude that DNA mini-barcodes are a very useful tool to analyse the diet of moth-specialist bats.

FORAGING BY MEDITERRANEAN HORSESHOE BATS (RHINOLOPHUS EURYALE) IN RELATION TO PREY DISTRIBUTION AND EDGE HABITAT

Abstract We studied the effect of habitat type and prey availability on the foraging decisions of the Mediterranean horseshoe bat (Rhinolophus euryale), a species specialized for cluttered environments. We modeled seasonal habitat selection using radiotelemetry in relation to prey availability in a heterogeneous landscape, determined seasonal diet and prey selection, and used geographic information system data to characterize the landscape surrounding 10 breeding colonies in order to assess the radiotracking results at the population level. Although R. euryale typically has been associated with woodland, our results suggest that the existence of edge habitat, created by semicluttered structures such as hedgerows and woodland edges, was a significant factor in the choice of foraging areas by these bats. Edge habitat was associated with meadows and pastures, creating a landscape highly suited to moths, the preferred prey of R. euryale. In the study area, however, moths were evenly distributed among habitat types; therefore, distribution of moths cannot explain the preference of these bats for semicluttered habitats. The results of our study are consistent with the presumed origin of R. euryale in an edge-rich ecosystem (i.e., the savannahs of northern Africa) and establish a new paradigm for how this species uses habitat. This new paradigm, which might also apply to other members of the genus in Europe, should prompt reconsideration of the presumed habitat requirements for this species, and should be incorporated into the conservation policies for the Mediterranean horseshoe bat.

Landscape features in the habitat selection of European mink ( Mustela lutreola ) in south-western Europe

Habitat change is one of the main factors influencing the decline of the western population of European mink Mustela lutreola, but data on habitat selection are scarce. Landscape features influencing selection of habitat and resting sites of male European mink were studied using radio-tracking. None of the habitat descriptors accounted for the habitat selection of European mink during their activity periods. On the other hand, resting site selection was correlated with the presence of bramble patches. Intensive use of bramble patches is explained as a consequence of the need of mink for protection against predators. Moreover, the high availability of bramble patches provides the mink with easy resting sites.

Home ranges of European minkMustela lutreola in southwestern Europe

We radiotracked 7 European minkMustela lutreola Linnaeus, 1761 (5 males and 2 females), to determine their distribution, size and temporal changes of their home range in an area of southwestern Europe, where American minkM. vison was not established. Size of home ranges varied from 11 to 17 km along watercourses in males and were 0.6 and 3.6 km in females. Home ranges of males were larger than those found in previous studies. Most females captured (either radiotracked or not) were found within the home range of males. Males occupied adjoining river sections with minimal range overlap, suggesting an intrasexually exclusive spacing pattern for males. Each month males used new river stretches, mainly along tributaries; meanwhile they showed a steady use of their stem river section. Resting sites were mainly beneath dense brambles ofRubus patches located in the river bank.

Detection of rhabdovirus viral RNA in oropharyngeal swabs and ectoparasites of spanish bats

Rhabdoviruses infect a variety of hosts, including mammals, birds, reptiles, fish, insects and plants. As bats are the natural host for most members of the genus Lyssavirus, the specificity of the amplification methods used for active surveillance is usually restricted to lyssaviruses. However, the presence of other rhabdoviruses in bats has also been reported. In order to broaden the scope of such methods, a new RT-PCR, able to detect a diverse range of rhabdoviruses, was designed. The method detected 81 of 86 different rhabdoviruses. In total, 1488 oropharyngeal bat swabs and 38 nycteribiid samples were analysed, and 17 unique rhabdovirus-related sequences were detected. Phylogenetic analysis suggested that those sequences detected in bats did not constitute a monophyletic group, even when originating from the same bat species. However, all of the sequences detected in nycteribiids and one sequence obtained from a bat did constitute a monophyletic group with Drosophila melanogaster sigma rhabdovirus.

Fishing behaviour in the long-fingered bat Myotis capaccinii (Bonaparte, 1837): an experimental approach

Abstract To study the fishing behaviour of Myotis capaccinii, we performed an experiment in a flight tent containing an artificial pond. We recorded the behaviour of two groups of bats — eight individuals from two different roosts — using IR video camcorders and ultrasound detectors, and evaluated diet by analyzing faeces. Nightly, increasing amounts of fish were released in the pond. Our data show that M. capaccinii is able to exhibit fishing behaviour when fish occur in high densities in shallow waters, gaffing live fish from the water using their hind feet. They were attracted neither by dead fish floating, nor by ripples made by fishes feeding on the water surface. Bats showed a specific fishing behaviour with two main foraging patterns: A) long series of circular flights, skimming along the water and dipping in softly twice or three times in each roundabout; B) long figure-eight loops with bats flying faster and higher, swooping down on the centre of the pond, where they snapped their hind feet hard into the water. Compared with the echolocation calls used to catch insects from the waters surface in the wild, terminal buzzes were incomplete during the dips made to fish. Buzz II were always lacking, and buzz I had much longer inter-pulse intervals. This suggests that they were not pursuing specific targets but dipping randomly. We propose a scenario in which fishing behaviour occurs in the wild, linked to the seasonal drought of small ponds, marshes, or channels where large numbers of small fish become readily available and thus a profitable resource.

What mechanism of niche segregation allows the coexistence of sympatric sibling rhinolophid bats

IntroductionOur purpose was to assess how pairs of sibling horseshoe bats coexists when their morphology and echolocation are almost identical. We collected data on echolocation, wing morphology, diet, and habitat use of sympatric Rhinolophus mehelyi and R. euryale. We compared our results with literature data collected in allopatry with similar protocols and at the same time of the year (breeding season).ResultsEcholocation frequencies recorded in sympatry for R. mehelyi (mean = 106.8 kHz) and R. euryale (105.1 kHz) were similar to those reported in allopatry (R. mehelyi 105–111 kHz; R. euryale 101–109 kHz). Wing parameters were larger in R. mehelyi than R. euryale for both sympatric and allopatric conditions. Moths constitute the bulk of the diet of both species in sympatry and allopatry, with minor variation in the amounts of other prey. There were no inter-specific differences in the use of foraging habitats in allopatry in terms of structural complexity, however we found inter-specific differences between sympatric populations: R. mehelyi foraged in less complex habitats. The subtle inter-specific differences in echolocation frequency seems to be unlikely to facilitate dietary niche partitioning; overall divergences observed in diet may be explained as a consequence of differential prey availability among foraging habitats. Inter-specific differences in the use of foraging habitats in sympatry seems to be the main dimension for niche partitioning between R. mehelyi and R. euryale, probably due to letter differences in wing morphology.ConclusionsCoexistence between sympatric sibling horseshoe bats is likely allowed by a displacement in spatial niche dimension, presumably due to the wing morphology of each species, and shifts the niche domains that minimise competition. Effective measures for conservation of sibling/similar horseshoe bats should guarantee structural diversity of foraging habitats.

Foraging Ecology in Mehely's Horseshoe Bats: Influence of Habitat Structure and Water Availability

The climate in Mediterranean ecosystems is characterised by aestival hot temperatures and water shortages which may affect the behaviour of bats living in these environments. We evaluated the influence of habitat type, habitat structure, and water availability on the foraging behaviour of the Mehelys horseshoe bats (Rhinolophus mehelyi) using classification and regression tree (CART) analysis. Twenty-five individuals were successively radio-tracked during May and June in two colonies (Marias and Aurora) of south-western Spain. Twelve females were tracked in Marias and 13 males in Aurora. Both localities differed in the availability of habitat types: in Marias savannah-like oak woodland dominated the landscape (81% of the study area), whereas in Aurora pasture were dominant (64% of the area). Rhinolophus mehelyi always foraged in woodland; preferentially in cluttered spaces, but also in less-cluttered/more-open ones, while open spaces were completely avoided. Bats foraged close to water bodies, preferentially at distances below 500 m, where they may readily have access to drinking water or may encounter higher insect abundances. The similarity between sexes on the preferences of habitat structure and distance to water suggest that the disparity in the habitat types used by males in Aurora and females in Marias may be due to local availability. Riparian forest is an important habitat for foraging and commuting, and should be scrupulously protected. Conservation strategies should also include the protection of woodland of diverse structure and linear landscape elements. We recommend the creation of water bodies in woodlands to enhance the suitability of foraging sites that might be underused.