Detection and Molecular Characterization of Giardia and Cryptosporidium spp. Circulating in Wild Small Mammals from Portugal.

Cryptosporidium spp. and Giardia spp. are important diarrhea-causing protozoan parasites worldwide that exhibit broad host ranges. Wild small mammals can harbor host-adapted and potentially zoonotic species of both parasites. The aim of this study was to investigate Cryptosporidium spp. and Giardia spp. in wild rodents and shrews in Portugal, focusing on the protist's occurrence and genetic diversity. Molecular screening by PCR at the small subunit (SSU) rRNA gene locus of 290 fecal samples from wood mice (Apodemus sylvaticus), southwestern water voles (Arvicola sapidus), Cabrera's voles (Microtus cabrerae), Lusitanian pine voles (Microtus lusitanicus), Algerian mice (Mus spretus) and greater white-toothed shrews (Crocidura russula) in Northeast Portugal revealed the low occurrence of Cryptosporidium spp. (1%) and high occurrence of Giardia spp. (32.8%). The analysis revealed that "species" was the only significant factor associated with the increasing probability of Giardia spp. infection, with the highest prevalence reported in southwestern water voles and Lusitanian pine voles. Cryptosporidium and Giardia species determination at the SSU rRNA gene locus revealed C. muris and G. microti as the only circulating species, respectively. Subtyping of the glutamate dehydrogenase (gdh) and beta-giardin (bg) genes provided evidence of the high genetic diversity within the G. microti clade. This study suggests that rodent-adapted G. microti occurs to a large extent in cricetid hosts and supports the limited role of wild rodents and shrews as natural sources of human infections in Northeast Portugal regarding the investigated parasites. Moreover, this is the first record of G. microti in southwestern water voles, Lusitanian pine voles, Algerian mice, wood mice and Cabrera's voles and C. muris in Cabrera's voles. Finally, this study improves the database of sequences relevant for the sequence typing of G. microti strains and provides new insights about the epidemiology of Giardia spp. and Cryptosporidium spp. in wild rodents and shrews, two parasite genera of high importance for public and animal health.

Crowding after sudden habitat loss affects demography and social structure in a bat population.

The sudden loss of habitats due to natural or anthropogenic disturbances causes displacement of mobile animals from affected areas to refuge habitats, where large but often transitory concentrations of individuals may occur. While these local density increases have been previously described, the hypothesis that crowding disrupts demographic processes remains largely untested. Here we used the sudden flooding of a river valley by a hydroelectric reservoir as a quasi-experiment to investigate the consequences of crowding on demography, fecundity and social structure in the European free-tailed bat Tadarida teniotis. We monitored bat populations at roosts near and far from the flooded area, before (2013-2014), during (2015) and after (2016) habitat flooding. We assessed population demographic parameters using Capture-Mark-Recapture (CMR) models (3,821 PIT-tagged individuals), and used genetic relatedness among individuals (1,407 individuals genotyped for 14 microsatellite markers) to infer changes in social structure. Habitat loss through flooding was associated with significant but transitory increases in the number of bats using nearby roosts. This may be related to the higher probability of individuals arriving at those roosts during flooding, together with increases in individual local residency through time, particularly among males. Individual apparent survival was highest during flooding and lowest in the following year, while the probability of leaving a roost safe from flooding was higher near the impact area than farther away. Crowding did not negatively affect fecundity, but the arrival of new individuals led to changes in social structure as revealed by lower genetic relatedness between individuals after disturbance at roosts near the flooding area, but not in those farther afield. Our study documents a clear example of crowding effects, suggesting that bats losing roosts due to a hydroelectric reservoir moved to alternative roosts, where local increases in population size and the arrival of new individuals reduced genetic relatedness and apparent survival, but not fecundity. These results support the hypothesis that crowding after habitat loss can disrupt population processes, even though effects may be subtle and short-lived. Also, they point out the need to duly consider crowding effects when assessing and mitigating anthropogenic impacts on animal populations.

Adenovirus emergence in a red squirrel (Sciurus vulgaris) in Iberian Peninsula.

Adenoviruses (AdV) are pathogens capable of infecting animals and humans leading to a wide spectrum of diseases. They have a widespread geographical dissemination and infect several species, including red squirrels (Sciurus vulgaris). In the Iberian Peninsula, there are no reports of infection of red squirrels by AdV. In this study, we report for the first time the detection of squirrel AdV in a red squirrel trapped in central Portugal. The phylogenetic analysis showed that the sequences obtained for the hexon and the penton base genes were similar to the Korean squirrel AdV strain and strongly divergent from the strains present in red squirrels in Europe. These findings indicate the emergence of a new strain circulating in Europe and raise concerns regarding the conservation of the red squirrel.

Species traits, patch turnover and successional dynamics: when does intermediate disturbance favour metapopulation occupancy?

BACKGROUND: In fragmented landscapes, natural and anthropogenic disturbances coupled with successional processes result in the destruction and creation of habitat patches. Disturbances are expected to reduce metapopulation occupancy for species associated with stable habitats, but they may benefit species adapted to transitory habitats by maintaining a dynamic mosaic of successional stages. However, while early-successional species may be favoured by very frequent disturbances resetting successional dynamics, metapopulation occupancy may be highest at intermediate disturbance levels for species with mid-successional habitat preferences, though this may be conditional on species traits and patch network characteristics. Here we test this 'intermediate disturbance hypothesis' applied to metapopulations (MIDH), using stochastic patch occupancy simulation modelling to assess when does intermediate disturbance favour metapopulation occupancy. We focused on 54 virtual species varying in their habitat preferences, dispersal abilities and local extinction and colonization rates. Long-term metapopulation dynamics was estimated in landscapes with different habitat amounts and patch turnover rates (i.e. disturbance frequency). RESULTS: Equilibrium metapopulation occupancy by late-successional species strongly declined with increasing disturbance frequency, while occupancy by early-successional species increased with disturbance frequency at low disturbance levels and tended to level-off thereafter. Occupancy by mid-successional species tended to increase along with disturbance frequency at low disturbance levels and declining thereafter. Irrespective of habitat preferences, occupancy increased with the amount of habitat, and with species dispersal ability and colonisation efficiency. CONCLUSIONS: Our study suggests that MIDH is verified only for species associated with mid-successional habitats. These species may be particularly sensitive to land use changes causing either increases or decreases in disturbance frequency. This may be the case, for instance, of species associated with traditional agricultural and pastoral mosaic landscapes, where many species disappear either through intensification or abandonment processes that change disturbance frequency.

Hierarchical spatial segregation of two Mediterranean vole species: the role of patch-network structure and matrix composition.

According to ecological theory, the coexistence of competitors in patchy environments may be facilitated by hierarchical spatial segregation along axes of environmental variation, but empirical evidence is limited. Cabrera and water voles show a metapopulation-like structure in Mediterranean farmland, where they are known to segregate along space, habitat, and time axes within habitat patches. Here, we assess whether segregation also occurs among and within landscapes, and how this is influenced by patch-network and matrix composition. We surveyed 75 landscapes, each covering 78 ha, where we mapped all habitat patches potentially suitable for Cabrera and water voles, and the area effectively occupied by each species (extent of occupancy). The relatively large water vole tended to be the sole occupant of landscapes with high habitat amount but relatively low patch density (i.e., with a few large patches), and with a predominantly agricultural matrix, whereas landscapes with high patch density (i.e., many small patches) and low agricultural cover, tended to be occupied exclusively by the small Cabrera vole. The two species tended to co-occur in landscapes with intermediate patch-network and matrix characteristics, though their extents of occurrence were negatively correlated after controlling for environmental effects. In combination with our previous studies on the Cabrera-water vole system, these findings illustrated empirically the occurrence of hierarchical spatial segregation, ranging from within-patches to among-landscapes. Overall, our study suggests that recognizing the hierarchical nature of spatial segregation patterns and their major environmental drivers should enhance our understanding of species coexistence in patchy environments.

Influence of land mosaic composition and structure on patchy populations: the case of the water vole (Arvicola sapidus) in Mediterranean farmland.

The ability of patchy populations to persist in human-dominated landscapes is often assessed using focal patch approaches, in which the local occurrence or abundance of a species is related to the properties of individual patches and the surrounding landscape context. However, useful additional insights could probably be gained through broader, mosaic-level approaches, whereby whole land mosaics with contrasting patch-network and matrix characteristics are the units of investigation. In this study we addressed this issue, analysing how the southern water vole (Arvicola sapidus) responds to variables describing patch-network and matrix properties within replicated Mediterranean farmland mosaics, across a gradient of agricultural intensification. Patch-network characteristics had a dominant effect, with the total amount of habitat positively influencing both the occurrence of water voles and the proportion of area occupied in land mosaics. The proportions of patches and area occupied by the species were positively influenced by mean patch size, and negatively so by patch isolation. Matrix effects were weak, although there was a tendency for a higher proportion of occupied patches in more intensive, irrigated agricultural landscapes, particularly during the dry season. In terms of conservation, results suggest that water voles may be able to cope well with, or even be favoured by, the on-going expansion of irrigated agriculture in Mediterranean dry-lands, provided that a number of patches of wet herbaceous vegetation are maintained within the farmland mosaic. Overall, our study suggests that the mosaic-level approach may provide a useful framework to understand the responses of patchy populations to land use change.