Margarida Santos-Reis

Distinct combinations of Borrelia burgdorferi sensu lato genospecies found in individual questing ticks from Europe.

ABSTRACT The genetic diversity of Borrelia burgdorferi sensu lato was assessed in individual adult Ixodes ricinusticks from Europe by direct PCR amplification of spirochetal DNA followed by genospecies-specific hybridization. Analysis of mixed infections in the ticks showed that B.garinii and B. valaisianasegregate from B. afzelii. This and previous findings suggest that host complement interacts with spirochetes in the tick, thereby playing an important role in the ecology of Lyme borreliosis.

The hard-tick fauna of mainland Portugal (Acari: Ixodidae): an update on geographical distribution and known associations with hosts and pathogens

This work is an updated revision of the available information on Portuguese ixodid tick species. It includes data on tick biology, ecology, taxonomy and host/pathogen-associations. The current list of Portuguese ixodid ticks comprises twenty species: Dermacentor marginatus (Sulzer, 1776), Dermacentor reticulatus (Fabricius, 1794), Haemaphysalis hispanica Gil Collado, 1938, Haemaphysalis inermis Birula, 1895, Haemaphysalis punctata Canestrini & Fanzago, 1878, Hyalomma lusitanicum Koch, 1844, Hyalomma marginatum Koch, 1844, Ixodes acuminatus Neumann, 1901, Ixodes bivari Dias, 1990, Ixodes canisuga Johnston, 1849, Ixodes frontalis (Panzer, 1798), Ixodes hexagonus Leach, 1815, Ixodes ricinus (Linnaeus, 1758), Ixodes simplex Neumann, 1906, Ixodes ventalloi Gil Collado, 1936, Ixodes vespertilionis Koch, 1844, Rhipicephalus (Boophilus) annulatus (Say, 1821), Rhipicephalus bursa Canestrini & Fanzago, 1878, Rhipicephalus pusillus Gil Collado, 1938, and Rhipicephalus sanguineus (Latreille, 1806).

World Distribution and Status of the Genus Martes in 2000

The genus Martes is comprised of 7 species of martens, sables and fishers, most of them forest-dwelling animals with valuable fur, distributed throughout North America, Europe and Asia. The pine marten (Martes martes) is indigenous over most of Europe, from Mediterranean biotopes to Fennoscandian taiga, and to western Siberia and Iran. It is found in insular wooded areas, shrublands, and coniferous forests. The stone marten (M. foina) occurs from Mongolia and the northern Himalayas to most of Europe. It frequents forests, woodlands and pastures, and is expanding in suburban and urban areas. The sable (M. zibellina) occurs in Russia, Mongolia, China, North Korea, and Japan. Over most of its distribution, the sable inhabits coniferous taiga forests with late seral attributes. The yellow-throated marten (M. flavigula; including the Nilgiri marten, M. gwatkinsi) occurs in sub-tropical and tropical forests from the Himalayas to eastern Russia, south to the Malay Peninsula and Sunda Shelf to Taiwan. The Japanese marten (M. melampus) occurs in forests of the main Japanese archipelago and the Korean peninsula. The American marten (M. americana) occurs in large contiguous populations in forested habitats of North America north of 35° latitude. It is associated with mesic coniferous and mixed forests with overhead cover and structural complexity near the ground. The fisher (M. pennanti) occurs in large contiguous areas across Canada, and in disjunct areas within the United States, north of 35° latitude. Whereas the distribution of Martes significantly expanded in many parts of the world over the last 20 years, largely due to several reintroduction programs, many populations are threatened by habitat loss and alteration. There is a need to develop cost-effective survey methods, monitor populations and fur-harvest activities, and assess the effects of natural and anthropogenic disturbance agents on habitat use by Martes species.

Species-specific mitochondrial DNA markers for identification of non-invasive samples from sympatric carnivores in the Iberian Peninsula

Genetic species identification of non-invasively collected samples has become an important tool in ecological research, management and conservation and wildlife forensics. This is especially true for carnivores, due to their elusive nature, and is crucial when several ecologically and phylogenetically close species, with similar faeces, hairs, bones and/or pelts, occur in sympatry. This is the case of the Iberian Peninsula, a region with a carnivore community of 16 species—about two-thirds of the European carnivore fauna. Here we present a simple, efficient and reliable PCR-based protocol, using a novel set of species-specific primers, for the unambiguous identification to species of non-invasively collected samples or forensic materials from Iberian carnivores. For each species, from the consensus of all cytochrome b haplotypes, found here and previously reported, we designed species-specific primer pairs for short fragments, the most likely to persist in low-quantity and degraded DNA samples. The predicted specificity of each primer pair was assessed through PCR of positive DNA extracts from the carnivore species, from an exhaustive array of potential prey and from humans. The robustness of PCR amplification for non-invasively sampled DNA was tested with scat samples. The primers did not produce false positives and correctly identified all carnivore samples to the species level. In comparison with sequencing and PCR-RFLP assays, our method is, respectively, cost- and time-effective, and is especially suited for monitoring surveys targeting multiple populations/species. It also introduces an approach that works for a whole community of carnivores living sympatrically over a large geographic area.

Human-Related Factors Regulate the Spatial Ecology of Domestic Cats in Sensitive Areas for Conservation

Background Domestic cats ranging freely in natural areas are a conservation concern due to competition, predation, disease transmission or hybridization with wildcats. In order to improve our ability to design effective control policies, we investigate the factors affecting their numbers and space use in natural areas of continental Europe. Methodology/Principal Findings We describe the patterns of cat presence, abundance and space use and analyse the associated environmental and human constraints in a well-preserved Mediterranean natural area with small scattered local farms. We failed in detecting cats in areas away from human settlements (trapping effort above 4000 trap-nights), while we captured 30 individuals near inhabited farms. We identified 130 cats, all of them in farms still in use by people (30% of 128 farms). All cats were free-ranging and very wary of people. The main factor explaining the presence of cats was the presence of people, while the number of cats per farm was mostly affected by the occasional food provisioning with human refuse and the presence of people. The home ranges of eight radio tagged cats were centred at inhabited farms. Males went furthest away from the farms during the mating season (3.8 km on average, maximum 6.3 km), using inhabited farms as stepping-stones in their mating displacements (2.2 km of maximum inter-farm distance moved). In their daily movements, cats notably avoided entering in areas with high fox density. Conclusions The presence, abundance and space use of cats were heavily dependent on human settlements. Any strategy aiming at reducing their impact in areas of conservation concern should aim at the presence of settlements and their spatial spread and avoid any access to human refuse. The movements of domestic cats would be limited in areas with large patches of natural vegetation providing good conditions for other carnivore mammals such as red foxes.

Does riparian habitat condition influence mammalian carnivore abundance in Mediterranean ecosystems

The severe loss or degradation of riparian habitats has led to their impoverishment and impaired function, which may have severe consequences on both the riparian habitats themselves and their associated biota, including mammalian carnivores. We selected 70 riparian habitat reaches to evaluate the condition of the riparian habitats in southern Portugal and their use by carnivores. These sites were assessed for riparian condition using the stream visual assessment protocol (SVAP) and surveyed for carnivore presence along the riparian zones and across the surrounding matrix landscape, both in the wet (winter) and the dry (summer) season. Results show that carnivore surveys adjacent to riparian habitats consistently had significantly higher species richness than the matrix habitats, in both sampling seasons. Carnivore relative abundance and relative abundance of stone marten, common genet and Egyptian mongoose also showed higher values in riparian habitats, with significant differences in at least one season. The Eurasian badger, on the other hand, showed higher relative abundance values in the landscape matrix, though differences were not significant. The SVAP index ranked about 83% riparian reaches as poor or fair condition, and species richness was significantly higher in fair condition reaches during the wet season. These results reflect the importance of riparian habitats in Mediterranean ecosystems for mammalian carnivores. However, the generalized poor condition of these habitats suggests that direct measures for riparian restoration could be appropriate. The preservation or improvement of riparian habitats would certainly benefit the mammalian carnivore populations and consequently their conservation.

Individual Spatial Responses towards Roads: Implications for Mortality Risk

Background Understanding the ecological consequences of roads and developing ways to mitigate their negative effects has become an important goal for many conservation biologists. Most mitigation measures are based on road mortality and barrier effects data. However, studying fine-scale individual spatial responses in roaded landscapes may help develop more cohesive road planning strategies for wildlife conservation. Methodology/Principal Findings We investigated how individuals respond in their spatial behavior toward a highway and its traffic intensity by radio-tracking two common species particularly vulnerable to road mortality (barn owl Tyto alba and stone marten Martes foina). We addressed the following questions: 1) how highways affected home-range location and size in the immediate vicinity of these structures, 2) which road-related features influenced habitat selection, 3) what was the role of different road-related features on movement properties, and 4) which characteristics were associated with crossing events and road-kills. The main findings were: 1) if there was available habitat, barn owls and stone martens may not avoid highways and may even include highways within their home-ranges; 2) both species avoided using areas near the highway when traffic was high, but tended to move toward the highway when streams were in close proximity and where verges offered suitable habitat; and 3) barn owls tended to cross above-grade highway sections while stone martens tended to avoid crossing at leveled highway sections. Conclusions Mortality may be the main road-mediated mechanism that affects barn owl and stone marten populations. Fine-scale movements strongly indicated that a decrease in road mortality risk can be realized by reducing sources of attraction, and by increasing road permeability through measures that promote safe crossings.

Relationships between Stone Martens, Genets and Cork Oak Woodlands in Portugal

Although the stone marten (Martes foina) is widely distributed in Europe, little is known about its ecological and spatial requirements. Using radiotelemetry and food habits, we investigated the use of cork oak (Quercus suber) woodlands by an unharvested population of stone martens in SW Portugal. We also evaluated spatial ecology and food habits of the common genet, (Genetta genetta), and its niche overlap with stone martens. Home range size (minimum convex polygon) of martens (n=5) and genets (n=7) averaged 2.6 and 3.3 km 2 , respectively. Variation in home range area during the year was associated with breeding activities. Tolerance among stone martens and genets was greater than between conspecifics of the same sex. Both stone martens and genets were nocturnal and used riparian vegetation and cultivated fields more than expected. However, genets also used oak woodland more than expected for foraging and resting. Stone martens and genets used similar rest sites. The seasonal variation in resource exploitation by stone martens (58 scats) and genets (75 scats) followed the same pattern. However, in summer and autumn, genets ate more crayfish (Procambarus clarkii) and mammals, while stone martens ate more fruits. Despite similarities in habitat utilization and food habits, core areas of both species were mutually exclusive, thus suggesting the existence of some present or past interspecific interaction. This study suggests that, except for resting, stone martens do not depend on cork oak trees and its products for existence. However, intensive destruction of understory cover and riparian corridors might alter use of cork oak trees by stone martens and their interspecific interactions with genets.

Otters and fish farms in the Sado estuary: ecological and socio-economic basis of a conflict

The degree of conflict between otter conservation and fish farming was assessed at Sado estuary (SW Portugal), using ecological (otter visiting rates to fish farms and consumption of commercial fish) and socio-economic (past and current instruments and policies addressing the conflict and a social impact assessment, including a discourse analysis of relevant stakeholders) parameters. The study concerned 14 fish farms producing Sparus aurata, Dicentrarchus labrax, Solea senegalensis and Solea solea. Results indicate high visiting rates in most fish farms (average: 76%), although in only 29%, species stocked were the most consumed prey. Other marine species and freshwater prey were the basis of otter diet in the remaining fish farms. The conflict is quite consensual among most fish farmers, and the results indicate that the perceived conflict by fish farmers has an ecological basis, although there are large gaps between effective and perceived predation. No specific instruments exist in Portugal to address the conflict, but some not specifically targeted can have an effect (e.g. species protection legislation and aquaculture licensing), although with limitations to effectively contribute to its mitigation (e.g. lack of enforcement and supervision). Formulating and assessing solutions is the following step, using a participatory approach to the development and evaluation of mitigation/compensation strategies, capable of providing an effective reconciliation of the conflict.

Stone marten (Martes foina) habitat in a Mediterranean ecosystem: effects of scale, sex, and interspecific interactions

Mediterranean ecosystems are inherently patchy, challenging habitat-use behavior. Certain mammalian carnivores take advantage of this patchiness by a strategy of habitat complementation/supplementation, which is invariant to the scale of analysis. To test if the same behavior is adopted by the stone marten, we used a combined data set of capture and radio-tracking data at three scales of analysis (1-m, 25-m, and 452-m radius plots). We used compositional analysis to test if there were sex-specific differences in foraging and resting habitat use of stone martens and if these patterns were affected by the presence of other mesocarnivores. Our results showed that stone martens are found both in rural and forested landscapes. Foraging and resting activities occurred far from roads in large and complex patches of cork oak woodlands, riparian vegetation, orchards, and pastureland. Use varied with the scale of analysis and the sex. At smaller scales, females use pastures for foraging and orchards for resting, whereas riparian vegetation and sparse cork oak forests influenced this use at larger scales. Males, on the other hand, were more consistent across scales, using riparian areas and dense cork oak woodlands for foraging and pastures for resting. Stone martens shared the same areas with other coexisting mesocarnivores. Stone martens use cork oak woodlands and complement/supplement this use with other land cover types. The consistent use of cork oak woodlands across scales emphasizes the importance of this land cover to the preservation of functional Mediterranean ecosystems in southern Portugal.