Mesocarnivore community structure under predator control: Unintended patterns in a conservation context.

Across the Mediterranean, conservation programmes often operate concomitantly with hunting interests within game-lands. Carnivore guilds lie at the interface between contrasting management goals, being simultaneously fundamental components of ecosystems and targets of predator control to reduce predation on game species. Here, we evaluate the composition and spatial structure of a mesocarnivore community in a protected area of Southeast Portugal, with high economic investment in conservation and significant hunting activity. Between June and August 2015, we deployed 77 camera-traps across a ~80 km2 area. We report on interspecific disparities in mesocarnivore occupancy and associated environmental determinants. Contrasting occupancy states suggest an unbalanced community, biased towards the widespread occurrence of the red fox Vulpes vulpes ([Formula: see text]) compared to other species (stone marten Martes foina, European badger Meles meles, Egyptian mongoose Herpestes ichneumon, common genet Genetta genetta, and Eurasian otter Lutra lutra) exhibiting spatially-restricted occupancy patterns ([Formula: see text]). The feral cat Felis silvestris catus was the exception ([Formula: see text]) and, together with the stone marten, exhibited a positive association with human settlements. These findings are consistent with theoretical predictions on how mesocarnivore communities are shaped by the effects of non-selective predator control, paradoxically favouring species with higher population growth rates and dispersal abilities, such as the red fox. Our results reinforce the need to understand the role of predator control as a community structuring agent with potential unintended effects, while exposing issues hindering such attempts, namely non-selective illegal killing or biased/concealed information on legal control measures.

Non-invasive hair sampling of Neotropical otters / Coleta não invasiva de pelos de lontra Neotropical

Abstract: Sampling wild animal populations using non-invasive techniques is advised when dealing with threatened species. Hair samples provide ecological information like species and individual identification. However, hair trapping is scarcely used in otters, due to their aquatic habits. Most studies are with captive individuals, so there is the need to test non-invasive hair trapping methods in otters in the wild. The aim of this study was to develop a simple and cost-effective method to collect hair from otter species in a non-invasive way. The study was carried out in the Paranapanema River, São Paulo State, Brazil, with the Neotropical otter (Lontra longicaudis Olfers, 1818), a protected species. Hair traps (wooden sticks and tree roots with adhesive tape or wax bands) were set during six nights on river banks, otter trails and scent-marking sites. Traps were baited with otter fresh spraints from other river locations. From the 23 traps, 10 (43.7%) were successful in collecting otter hairs, mostly guard-hair. The sticks were much more efficient than the roots at capturing otter hair (70.6.% vs. 0%) as well as adhesive tape when compared to wax (71.4% vs. 0%). Method simplicity and efficiency suggest that it can be a cost-effective way for collecting otter hairs without the need for capturing individuals. This method can be used for: assessment of local otter distribution; collecting otter hair samples for sex and individual identification (by molecular analysis), trophic ecology (by isotopic analyses), ecotoxicology (by contamination analysis) or behaviour ecology (by hormonal and stress levels analysis). More trapping campaigns should be implemented to further test the method's efficiency.

Resumo: O uso de técnicas de amostragem não-invasivas é aconselhado quando se trabalha com espécies ameaçadas de animais selvagens. Amostras de pelo fornecem informações ecológicas, como a identificação ao nível da espécie e do indivíduo. No entanto, a coleta de pelo é pouco usada em lontras, devido aos seus hábitos aquáticos. A maioria dos estudos é feita com indivíduos em cativeiro, existindo por isso a necessidade de testar métodos não invasivos de coleta de pelos de lontras na natureza. O objetivo deste estudo foi desenvolver um método simples e com uma boa relação custo-benefício para coletar pelos de espécies de lontra de maneira não invasiva. O estudo foi realizado no rio Paranapanema, Estado de São Paulo, Brasil, com a lontra Neotropical (Lontra longicaudis Olfers, 1818), uma espécie protegida. Armadilhas de pelo (estacas de madeira e raízes de árvores com fita adesiva ou bandas de cera depilatória) foram colocadas durante seis noites nas margens do rio, em trilhas e locais de marcação de lontra. As armadilhas foram iscadas com dejetos frescos de lontra de outros locais do rio. Das 23 armadilhas, 10 (43.7%) foram eficazes na coleta de pelos de lontra, maioritariamente pelos-guarda. As estacas foram muito mais eficientes que as raízes na captura de pelos de lontra (70.6.% vs. 0%) tal como a fita adesiva quando comparada com a cera (71,4% vs. 0%). A simplicidade e a eficiência do método sugerem que esta pode ser uma maneira econômica de coletar pelo de lontra sem a necessidade de capturar indivíduos. Este método pode ser usado para: levantamento da distribuição local da lontra; coleta de amostras de pelo de lontra para identificação sexual e individual (por meio de análise molecular); ecologia trófica (por meio de análise isotópica); ecotoxicologia (por meio de análise de contaminantes); e ecologia comportamental (por meio da determinação de níveis hormonais reprodutivos e ligados ao estresse). Mais campanhas de armadilhagem devem ser implementadas para melhor avaliar a eficiência do método.

Virulence traits and antibiotic resistance among enterococci isolated from Eurasian otter (Lutra lutra).

Enterococci are ubiquitous microorganisms found as part of the normal intestinal microbiota of many animals such as the free-ranging Eurasian otter (Lutra lutra Linnaeus, 1758). In this work, twenty-nine enterococci isolated from fecal samples of Eurasian otters free-living in reservoirs and associated river stretches in South Portugal were identified and typed by conventional/molecular methods and screened for virulence factors and antibiotic resistance. Identification allocated the isolates to the species Enterococcus faecalis (19), E. faecium (9) and E. durans (1) and PCR-fingerprinting revealed their high genomic diversity. Regarding virulence factors, three isolates produced cytolysin and six were gelatinase-positive. Genes ace and acm were detected in five enterococci each, ebpABC in seventeen, gelE in fourteen and cylA in three. All isolates showed resistance patterns and antibiotic resistance genes tet(M) and pbp5 were detected in seventeen isolates each, whereas vanB and vanD were identified in thirteen and five, respectively, being most van-harboring isolates members of E. faecium. The aac(6')-Ie-aph (2″) gene, encoding for gentamicin resistance, was observed in all gentamicin-resistant enterococci. Since all isolates harbor virulence and/or antibiotic resistance traits, the role of free-living Eurasian otters in the dissemination of virulent/resistant enterococci among other animals sharing the same ecological niche cannot be disregarded, as well as the health risk they may represent for humans directly interacting with them or their habitat.

The plasma membrane-enriched fraction proteome response during adaptation to hydrogen peroxide in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, adaptation to hydrogen peroxide (H2O2) decreases plasma membrane permeability to H2O2, changes its lipid composition and reorganizes ergosterol-rich microdomains by a still unknown mechanism. Here we show, by a quantitative analysis of the H2O2-induced adaptation effect on the S. cerevisiae plasma membrane-enriched fraction proteome, using two-dimensional gel electrophoresis, that 44 proteins are differentially expressed. Most of these proteins were regulated at a post-transcriptional level. Fourteen of these proteins contain redox-sensitive cysteine residues and nine proteins are associated with lipid and vesicle traffic. In particular, three proteins found in eisosomes and in the eisosome-associated membrane compartment occupied by Can1p were up-regulated (Pil1p, Rfs1p and Pst2p) during adaptation to H2O2. Survival studies after exposure to lethal H2O2 doses using yeast strains bearing a gene deletion corresponding to proteins associated to lipid and vesicle traffic demonstrated for the first time that down-regulation of Kes1p, Vps4p and Ynl010wp and up-regulation of Atp1 and Atp2 increases resistance to H2O2. Moreover, for the pil1Δ strain, H2O2 at low levels produces a hormetic effect by increasing proliferation. In conclusion, these data further confirms the plasma membrane as an active cellular site during adaptation to H2O2 and shows that proteins involved in lipid and vesicle traffic are important mediators of H2O2 adaptation.

Antimicrobial-resistant Salmonella isolated from Eurasian otters (Lutra lutra Linnaeus, 1758) in Portugal.

Eurasian otters (Lutra lutra Linnaeus, 1758) are present in a wide range of aquatic environments. Salmonella isolates have been obtained from otters at rehabilitation centers and in the wild and are sometimes associated with serious illnesses. Antimicrobial-resistant Salmonella were isolated from otter fecal samples collected in March 2006, September 2007, and March 2008 in two river basins in southern Portugal. From 67 samples tested, five were positive for Salmonella (7.58%). None of the isolates were susceptible to all antimicrobials tested, and all were resistant to multiple antimicrobials. Our results confirm the role of otters as potential carriers of Salmonella and the importance of environmental exposure to antimicrobial agents in selection for resistance in bacteria.

Modulation of plasma membrane lipid profile and microdomains by H2O2 in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, the rate of hydrogen peroxide (H(2)O(2)) diffusion through the plasma membrane decreases during adaptation to H(2)O(2) by a still unknown mechanism. Here, adaptation to H(2)O(2) was observed to modulate rapidly the expression of genes coding for enzymes involved in ergosterol and lipid metabolism. Adaptation to H(2)O(2) also alters plasma membrane lipid composition. The main changes were the following: (a) there was a decrease in oleic acid (30%) and in the ratio between unsaturated and saturated long-chain fatty acids; (b) the phosphatidylcholine:phosphatidylethanolamine ratio increased threefold; (c) sterol levels were unaltered but there was an increased heterogeneity of sterol-rich microdomains and increased ordered domains; (d) the levels of the sterol precursor squalene increased twofold, in agreement with ERG1 gene down-regulation; and (e) C26:0 became the major very long chain fatty acid owing to an 80% decrease in 2-hydroxy-C26:0 levels and a 50% decrease in C20:0 levels, probably related to the down-regulation of fatty acid elongation (FAS1, FEN1, SUR4) and ceramide synthase (LIP1, LAC1) genes. Therefore, H(2)O(2) leads to a reorganization of the plasma membrane microdomains, which may explain the lower permeability to H(2)O(2), and emerges as an important regulator of lipid metabolism and plasma membrane lipid composition.

H2O2 induces rapid biophysical and permeability changes in the plasma membrane of Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, the diffusion rate of hydrogen peroxide (H2O2) through the plasma membrane decreases during adaptation to H2O2 by means of a mechanism that is still unknown. Here, evidence is presented that during adaptation to H2O2 the anisotropy of the plasma membrane increases. Adaptation to H2O2 was studied at several times (15min up to 90min) by applying the steady-state H2O2 delivery model. For wild-type cells, the steady-state fluorescence anisotropy increased after 30min, or 60min, when using 2-(9-anthroyloxy) stearic acid (2-AS), or diphenylhexatriene (DPH) membrane probe, respectively. Moreover, a 40% decrease in plasma membrane permeability to H2O2 was observed at 15min with a concomitant two-fold increase in catalase activity. Disruption of the ergosterol pathway, by knocking out either ERG3 or ERG6, prevents the changes in anisotropy during H2O2 adaptation. H2O2 diffusion through the plasma membrane in S. cerevisiae cells is not mediated by aquaporins since the H2O2 permeability constant is not altered in the presence of the aquaporin inhibitor mercuric chloride. Altogether, these results indicate that the regulation of the plasma membrane permeability towards H2O2 is mediated by modulation of the biophysical properties of the plasma membrane.

Assessing dam implementation impact on threatened carnivores: the case of Alqueva in SE Portugal.

Large dam construction in water deficient areas is a management decision often controversial. Besides providing water storage, economical benefits, and a source of renewable energy, the construction and flooding caused by large dams cause disruptions in natural systems. We monitored the pre- and post-Alqueva dam impacts on the threatened carnivore species (polecat, otter, wildcat and Iberian lynx) populations in SE Portugal, and assessed which factors mostly contribute to post-dam distribution. Major short term impacts of large dams are: (1) increase in accessibility and human presence; (2) movement of heavy machinery and dam-workers; (3) deforestation with habitat loss and fragmentation; (4) change from lotic to lentic system; (5) lower prey availability and harsher capture; and (6) changes in land use adjacent to the reservoir. Thus, the response to those impacts can be predicted as a decline of polecat, wildcat and lynx distribution ranges, and a recovery of the otter from the severe short term impacts. Our results corroborate this hypothesis for all the species, especially during deforestation/early flooding. Otter's distribution range increased in the phase of greater impact, with a subsequent decrease with flooding. Our results suggest carnivores used "escape" areas with favourable habitat and prey conditions, however, the areas with higher probability of species presence decreased by two fold showing a drastic range reduction. To ensure populations' survival of these charismatic threatened carnivore populations of Mediterranean landscapes of south-east Portugal, we propose continuing the monitoring program and the development of a conservation program for the subsisting areas of optimal and suboptimal habitats.

Down-regulation of fatty acid synthase increases the resistance of Saccharomyces cerevisiae cells to H2O2.

Changes in plasma membrane permeability caused by H2O2 were recently found to be involved in the adaptation to H2O2, but the mechanism responsible for this change remains largely unknown. Here this mechanism was addressed and two lines of evidence showed for the first time that fatty acid synthase (Fas) plays a key role during the cellular response of Saccharomyces cerevisiae to H2O2: (1) adaptation was associated with a decrease in both Fas expression and activity; (2) more importantly, decreasing Fas activity by 50% through deletion of one of the FAS alleles increased the resistance to lethal doses of H2O2. The mechanism by which a decrease of Fas expression causes a higher resistance to H2O2 was not fully elucidated. However, the fas1Delta strain plasma membrane had large increases in the levels of lignoceric acid (C24:0) (40%) and cerotic acid (C26:0) (50%), suggesting that alterations in the plasma membrane composition are involved. Very-long-chain fatty acids (VLCFA) through interdigitation or by modulating formation of lipid rafts may decrease the overall or localized plasma membrane permeability to H2O2, respectively, thus conferring a higher resistance to H2O2.