Nature protection areas of Europe are insufficient to preserve the threatened beetle Rosalia alpina (Coleoptera: Cerambycidae): evidence from species distribution models and conservation gap analysis

1. Natura 2000 network (N2000) and national protected areas (NPAs) are recognised as the most important core ‘units’ for biological conservation in Europe. 2. Species distribution models (SDMs) were developed to detect the potential distribution of the rare and threatened cerambycid beetle Rosalia alpina L. in Europe, and the amount of suitable habitat within the N2000 network [special areas of conservation (SACs) and special protection areas (SPAs)], NPAs (e.g. national parks, regional parks, state reserves, natural monuments and protected landscapes) and the overall European protected area network (EPAN) (N2000 + NPAs) was quantified. 3. According to this analysis, the suitable habitat for R. alpina in Europe amounts to c. 754 171 km² and stretches across substantially uninterrupted areas from Portugal to Romania (west to east) and from Greece to Germany (south to north). The overlay between the existing system of conservation areas in Europe (N2000 and NPAs) and the binary map for R. alpina showed that only c. 42% of potential habitat is protected. SACs and SPAs protect c. 25% and 21% of potential habitat, respectively. However, because the two site types often spatially overlap, when taken together the entire N2000 network protects c. 31% of potential habitat. Instead, NPAs offer a degree of protection of c. 29%. Overall, almost 60% of the area potentially suitable for the species is unprotected by the EPAN, an aspect that should be considered carefully when planning the conservation of this beetle at a large scale. 4. These results may also help to focus field surveys in selected areas where greater chances of success are encountered to save resources and increase survey effectiveness.     

Genetic structure of expanding wolf (Canis lupus) populations in Italy and Croatia,and the early steps of the recolonization of the Eastern Alps

After centuries of range contraction and demographic declines wolves are now expanding in Europe, colonizing regions from where they have been absent for centuries. Wolf colonizing the western Alps originate by the expansion of the Italian population. Vagrant wolves of Italian and Dinaric-Balkan origins have been recently observed in the Eastern Alps. In this study we compared the genetic structure of wolf populations in Italy and Croatia, aiming to identify the sources of the ongoing recolonization of the Eastern Alps. DNA samples, extracted from 282 Italian and 152 Croatian wolves, were genotyped at 12 autosomal microsatellites (STR), four Y-linked STR and at the hypervariable part of the mitochondrial DNA control-region (mtDNA CR1). Wolves in Croatia and Italy underwent recent demographic bottlenecks, but they differ in genetic diversity and population structure. Wolves in Croatia were more variable at STR loci (N_A = 7.4, H_O = 0.66, H_E = 0.72; n = 152) than wolves in Italy (N_A = 5.3, H_O = 0.57, H_E = 0.58; n = 282). We found four mitochondrial DNA (mtDNA CR1) and 11 Y-STR haplotypes in Croatian wolves, but only one mtDNA CR1 and three Y-STR haplotypes in Italy. Wolves in Croatia were subdivided into three genetically distinct subpopulations (in Dalmatia, Gorski kotar and Lika regions), while Italian wolves were not sub-structured. Assignment testing shows that the eastern and central Alps are recolonized by wolves dispersing from both the Italian and Dinaric populations. The recolonization of the Alps will predictably continue in the future and the new population will be genetically admixed and very variable with greater opportunities for local adaptations and survival.     

Tree selection by the endangered beetle <Emphasis Type="Italic">Rosalia alpina</Emphasis> in a lapsed pollard beech forest

Abandoned pollard beech forests are particular habitats that may require special conservation efforts to preserve the endangered beetle Rosalia alpina, a model species whose protection may perpetuate the habitat of many other saproxylic species. Forest use can determine the tree selection and population size of R. alpina, variables previously not researched in pollard forests. Selected tree traits and population size, indicated by presence of adult specimens and recent exit holes, were determined in a pollard beech forest before the implementation of habitat manipulations targeted to preserve the species. Multivariate analysis showed tree condition (living, snag, fallen) and clearing size and aspect as influential variables, with the former explaining more variance than the latter. R. alpina positively selected pollard snags and trees located in big and dry clearings, avoiding those in small clearings or shaded areas. Snags showed most adults (90 %) and recent emergence holes (84 %). Distance to the nearest occupied tree, trunk diameter, bark thickness and presence of sap leaks had no effect on the occurrence of R. alpina. Population size was estimated in 0.1 individuals day^?1 ha^?1, and 38 % of available habitat trees showed exit holes, values lower than those observed in other European countries. These results show the need to start re-pollardings in the forest to extend pollard standing life and enhance exposure to sunlight. Pinpointing pollard snags and clearing the surrounding vegetation are recommended as the first measures to be taken in order to favor R. alpina populations on pollard stands with closed canopies.     

Efficacy of Iranian diatomaceous earth deposits against Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae)

Laboratory bioassays were carried out to evaluate the insecticidal efficacy of two Iranian deposits of diatomaceous earths (DEs) and a commercial formulation, SilicoSec®, against adult confused flour beetle (Tribolium confusum Jacquelin du Val). The Iranian DEs were dried and sieved to get particle sizes of 0–149, 74–149, 0–74 μm, and 0–37 μm. First, DE samples were applied at the four concentration levels of 500, 1000, 1500 and 2000 ppm and each concentration was replicated four times. Tests were carried out at 27 ± 1 °C and 55 ± 5% r.h. in continuous darkness. The beetle mortality was counted at 2, 7, and 14 days after exposure. Moreover, another experiment was conducted to estimate the LC₅₀ values of the DEs. For the first experiment, adult beetle mortality exceeded 51% when exposed for 2 d to 2000 ppm of SilicoSec®. Complete mortality was recorded at each concentration of SilicoSec® at an exposure longer than 7 days except for 500 ppm; while mortalities of T. confusum at 2000 ppm of Maragheh and Mamaghan samples with the 0–149 μm particle size were 40.62% and 85.41%, respectively. Mortality of T. confusum was influenced by concentration and time of exposure to the DEs. SilicoSec® was the most effective DE followed by Mamaghan samples. The Maragheh samples were the least effective. In addition, in most cases fractions with smaller particles were more effective than larger ones. More experiments are necessary to process natural DEs and make them commercially exploitable.     

How far a protected area contributes to conserve habitat species composition and population structure of endangered African tree species (Benin,West Africa)

The Pendjari Biosphere Reserve located in the Sudanian zone of Bénin, is a protected area well managed, but mainly aimed at wild animal conservation. This study assessed its effectiveness to conserve habitat species composition and population structure of three endangered African tree species: Afzelia africana Sm., Pterocarpus erinaceus Poir. and Khaya senegalensis (Desv.) A. Juss. We randomly sampled 120 plots in the protected and surrounding unprotected habitats by inventorying plant species. For the three target species, we estimated adult and juvenile densities and recorded size classes. According to floristic composition four habitats groups were recognized in relation to human disturbance, vegetation type, and moisture. These were protected savannas, unprotected savannas, old fallows and gallery forests. The estimated adult densities of A. africana were similar between protected (14 ± 1.2 tree/ha) and unprotected savannas (17 ± 0.9 tree/ha) while for P. erinaceus the adult density was significantly higher in protected (12 ± 3.7 tree/ha) than in unprotected savannas (5 ± 1.9 tree/ha). Estimated adult density of K. senegalensis was also significantly higher in protected gallery forest (40 ± 5.8 tree/ha) than in unprotected one (29 ± 4.8 tree/ha). Juvenile densities of A. africana, K. senegalensis and P. erinaceus were higher in protected habitats than in unprotected ones but the difference was not significant. Skewness coefficient indicated that populations of investigated trees were declining in their protected habitats. However, in the case of A. africana and K. senegalensis populations seemed to be mostly threatened in the protected area. We concluded that although the studied protected area is effective to conserve some habitats species compositions, protection is not sufficient to guarantee future conservation of some threatened tree species.     

Malcolmia littorea: The isolated Italian population in the European context

We compared the coenological information of the only Italian population of Malcolmia littorea (L.) R. Br. with published phytosociological relevés, including ones of this species, throughout its European range. With the aim of highlighting the main climatic features influencing the distribution patterns of M. littorea, we integrated coenological data with some climatic variables and considered major drivers for plant distribution at the European scale. Finally, we analysed the population extent of M. littorea in Italy, in order to assess its conservation status at regional level. The DCA analysis, performed on a matrix 139 relevés × 183 species, separated the relevés according to their floristic composition, showing a geographic gradient from Portugal to Mediterranean coasts, until Italy; with Mediterranean relevés clearly separated from Atlantic ones as well. Along the beach-inland gradient, the analysis highlights that the species is typical of fixed dunes habitats (more inland, mainly stabilised dunes), although in the Atlantic it can also be found in mobile dunes. The analysis of climatic variables in relation to M. littorea distribution, suggested that the species is sensitive to low winter temperature and to summer drought. The only Italian population of M. littorea is subjected to many threats, due to its small dimensions (<1 ha), isolation from the rest of its distribution area, that ranges from Portugal to France (until the Camargue region) and intensive human disturbances. Using both field and remote sensing information, we showed a considerable decrease of the occupied surface in Italy, leading us to suggest that the IUCN threat category of M. littorea in Italy should be reassessed from endangered to critically endangered.     

The importance of relative humidity and trophic resources in governing ecological niche of the invasive carabid beetle Merizodus soledadinus in the Kerguelen archipelago

Comprehensive studies to identify species-specific drivers of survival to environmental stress, reproduction, growth, and recruitment are vital to gaining a better understanding of the main ecological factors shaping species habitat distribution and dispersal routes. The present study performed a field-based assessment of habitat distribution in the invasive carabid beetle Merizodus soledadinus for the Kerguelen archipelago. The results emphasised humid habitats as a key element of the insect’s realised niche. In addition, insects faced food and water stress during dispersal events. We evaluated quantitatively how water availability and trophic resources governed the spatial distribution of this invasive predatory insect at Îles Kerguelen. Food and water stress survival durations [in 100%, 70%, and 30% relative humidity (RH) conditions] and changes in a set of primary metabolic compounds (metabolomics) were determined. Adult M. soledadinus supplied with water ad libitum were highly tolerant to prolonged starvation (LT₅₀ = 51.7 ± 6.2 d). However, food-deprived insect survival decreased rapidly in moderate (70% RH, LT₅₀ = 30.37 ± 1.39 h) and low (30% RH, LT₅₀ = 13.03 ± 0.48 h) RH conditions. Consistently, body water content decreased rapidly in insects exposed to 70% and 30% RH. Metabolic variation evidenced the effects of food deprivation in control insects (exposed to 100% RH), which exhibited a progressive decline of most glycolytic sugars and tricarboxylic acid cycle intermediates. Most metabolite levels were elevated levels during the first few hours of exposure to 30% and 70% RH. Augmented alanine and lactate levels suggested a shift to anaerobic metabolism. Simultaneously, peaks in threonine and glycolytic sugars pointed to metabolic disruption and a progressive physiological breakdown in dehydrating individuals. Overall, the results of our study indicate that the geographic distribution of M. soledadinus populations is highly dependent on habitat RH and water accessibility.     

Natural parasitism of Romanomermis iyengari (Welch) (Nematoda: Mermithidae) on various species of mosquitoes breeding in rice fields in Pondicherry,India

Natural habitat of Romanomermis iyengari, an entomophilic mermithid nematode, is rice field. Parasitism of this nematode on mosquito larvae breeding in rice fields in Pondicherry, India, has been studied in 1979. The present study was to find out its persistence in the same habitat, after many years of agricultural practices. Evaluation of parasitism on mosquito larvae was carried out during two rice cultivation seasons at a schedule of 1 week prior, and 1 and 2 weeks after transplantation of paddy. Larval stages of mosquito species such as Culex tritaeniorhynchus, Culex vishnui, Anopheles vagus, and Anopheles subpictus showed parasitism of R. iyengari. Parasitism was the highest on C. tritaeniorhynchus with a maximum of 15.38% during season I and 17.85% during season II. Followed by this, it was on A. vagus (10.82% and 9.43%), C. vishnui (7.37% and 8.69%), and A. subpictus (3.70% and 6.36%). The overall density of mosquito larvae and level of parasitism of R. iyengari on them during season II was significantly higher than that of season I. The parasitism was maximum at 1 week before transplantation and showed a decreasing trend towards 2 weeks after transplantation, during both the seasons. The level of natural parasitism of R. iyengari during the present investigation was higher than that observed in the same ecosystem almost 35 years ago. This is clearly an indication towards the capability of the nematode to withstand several years of agricultural practices, including pesticide and agrichemical application. Hence, if applied at sufficient level and frequency, this nematode could act as a successful biocontrol agent against mosquitoes breeding in rice fields.     

Cancer surveillance in Italian army peacekeeping troops deployed in Bosnia and Kosovo, 1996–2007: Preliminary results

Objective: An excess of cancer cases was reported in 2000 among Italian troops involved in peacekeeping operations in Bosnia and Kosovo. A preliminary assessment of cancer risk in this cohort was done for the period 1996–2007, based on cancer surveillance activity, which was started in 2001. Methods: A retrospective cohort study was done among 27 361 Italian Army soldiers deployed in Bosnia, 31 052 deployed in Kosovo, and a control group of non-deployed soldiers, whose number varies from 130 275 in 1996 to 40 967 in 2007. Standardized incidence ratio and 95% confidence intervals (SIR [95% CI]) of the main types of tumours were computed as the ratio of the observed and expected cases. Results: 98 incident cancers were reported among soldiers deployed in Bosnia, 66 in those stationed in Kosovo and 388 in non-deployed personnel. Overall, cancer incidence was lower than expected in all groups. Annual SIR was significantly higher than expected in 2000 for Hodgkin's lymphoma only among troops deployed in Bosnia (4.34 [1.18–11.12]) and non-deployed personnel (3.48 [1.67–6.39]); and in 2001 for thyroid cancer only among troops deployed in Bosnia (5.28 [1.44–13.51]). Conclusion: Although further investigation is needed, these preliminary results give no indication of an increased risk of cancer for Italian soldiers who were stationed in Bosnia and Kosovo. The cluster of Hodgkin's lymphoma in 2000 and of thyroid cancer in 2001 were sporadic events, they did not specifically affect deployed personnel and are unlikely to be related to environmental exposures in the Balkans.     

Knowledge-based framework for delineation and classification of ephemeral plant communities in riverine landscapes to support EC Habitat Directive assessment

Riverine landscapes are shaped by the spatio-temporal dynamics of the water regime. Water level transitions induce a shift in plant species composition from aquatic to ephemeral vegetation communities in riparian habitats. Hence, the occurrence of these ephemerals is strongly related to the hydrological connectivity and therefore used as indicator for the assessment of riparian habitat types. The delineation and assessment of such habitat types is time-consuming due to the indifferent occurrence of the plants. Therefore, in this study a knowledge-based framework is presented to provide readily usable polygons to support subsequent field surveys on species level. Different hierarchical scales range from hydrological connectivity classes to watercourses and to the micro-morphological classification of riparian habitats. The object-based image analysis approach was used to extract information from terrain and groundwater models, aerial images, and thematic data. The study site is located in the Danube floodplains east of Vienna Natura 2000 site. The micro-morphological classification of the watercourses resulted in the delineation of the classes Waterbodies, Riparian Habitats and the remaining Transition Zones. Watercourses with high flow velocity or with low hydrological connectivity show a small portion of potentially suitable riparian habitats for ephemeral vegetation communities. The framework with focus on terrain models delineating the shape of the riparian habitats performed well with an overall accuracy of 90% (kappa = 0.74). The thresholds in the framework were set fixed or calculated automatically to facilitate an application by spatial ecologist due to the combination of remote sensing techniques and GIS functionalities. The knowledge-based framework can be adapted to provide a harmonised and standardised dataset for any riverine study area.     

Genome size variation and polyploidy in the resurrection plant genus Ramonda: Cytogeography of living fossils

The genus Ramonda includes three preglacial paleoendemic species surviving as the rare resurrection angiosperms of the Northern hemisphere in refugia habitats in the Balkan (Ramonda nathaliae and Ramonda serbica) and Iberian Peninsulas (Ramonda myconi). This study focuses on: assessing genome size and base composition, determining chromosome number and ploidy level in several populations, evaluating inter- and intra-specific variations in DNA content and chromosome number as well as looking for the possible hybridization in the sympatric zones of Balkan species. R. nathaliae and R. myconi are diploid species (2n = 2x = 48) while R. serbica is hexaploid (2n = 6x = 144). The mean 2C DNA values ranged from 2.30 pg for R. nathaliae to 2.59 pg for R. myconi compared to 7.91 pg for R. serbica. The base composition for R. nathaliae was 42.1% GC, for R. myconi 39.9% and for R. serbica 41.2%. In one population of R. serbica the DNA content ranged from 2C = 7.65 to 11.82 pg, revealing different ploidy levels among its individuals. In sympatric populations genome size was intermediary (～5 pg) between the diploid and hexaploid classes which indicates the hybridization ability between R. serbica and R. nathaliae. It appears that polyploidization is the major evolutionary mechanism in the genus Ramonda.     

Bioactivity of essential oil of Litsea cubeba from China and its main compounds against two stored product insects

During our screening program for agrochemicals from Chinese medicinal herbs and wild plants, the essential oil of Litsea cubeba fruits was found to possess strong contact toxicity against the cigarette beetle Lasioderma serricorne adults and the booklouse Liposcelis bostrychophila, with LD₅₀ values of 27.33 μg/adult and 71.56 μg/cm², respectively, and also showed strong fumigant toxicity against the two stored product insects with LC₅₀ values of 22.97 and 0.73 mg/L, respectively. The essential oil obtained by hydrodistillation was investigated by GC MS. The main components of the essential oil were identified to be E-citral (geranial) (27.49%), Z-citral (neral) (23.57%) and d-limonene (18.82%) followed by β-thujene (3.34%), β-pinene (2.85%), α-pinene (2.57%), 6-methyl-5-hepten-2-one (2.40%) and linalool (2.36%). Citral (Z/E-citral), d-limonene, β-pinene, α-pinene and linalool were separated and purified by silica gel column chromatography and preparative thin layer chromatography, and further identified by means of physicochemical and spectrometric analysis. Citral and linalool showed strong contact toxicity against L. serricorne and L. bostrychophila (LD₅₀ = 11.76, 12.74 μg/adult and 20.15, 99.97 μg/cm², respectively) and fumigant toxicity against L. serricorne and L. bostrychophila (16.54, 18.04 mg/L air and 0.14, 0.71 mg/L air, respectively). Otherwise, citral, d-limonene and linalool were strongly repellent against the cigarette beetle L. serricorne as the essential oil whereas β-pinene and α-pinene exhibited weaker repellency against the cigarette beetle compared with the positive control, DEET. Moreover, except α-pinene and linalool, the other three compounds as well as the essential oil exhibited comparable repellency against the booklouse relative to DEET.     

Familial malignant mesothelioma: A population-based study in Central Italy (1980–2012)

Malignant mesothelioma is a sporadic cancer linked to asbestos exposure. Its occurrence among blood relatives (familial mesothelioma) may point to genetic susceptibility or shared exposures. The burden of the familial disease is unknown. The aims of the study were to assess at population level the proportion of familial mesotheliomas among all mesotheliomas and to investigate the family history of cancer among relatives of mesothelioma cases. We actively searched familial clusters based on a mesothelioma registry from central Italy (5.5 million people, 10% of the Italian population) of the National Mesothelioma Register network (ReNaM) as well as a pathology-based archive. Among 997 incident mesotheliomas recorded in a 32-year-period (1980–2012), we detected 13 clusters and 34 familial cases, accounting for 3.4% of all mesotheliomas. The most common clusters where those with affected siblings and unaffected parents. Asbestos exposure was occupational (n = 7 clusters), household (n = 2), environmental (n = 1), or not attributable for insufficient information (n = 3). There were 25 additional cancers in nine families. Some were cancer sites for which there is sufficient evidence (lung and larynx) or limited evidence (stomach and colon) of causal association with asbestos. The results suggest potential genetic recessive effects in mesothelioma that interact with asbestos exposure, but it is not possible to estimate the specific proportion attributable to each of these components.     

Crucial factors affecting the nitrile hydratase production of Mesorhizobium sp. F28

Mesorhizobium sp. F28 contains cobalt-NHase, which effectively converts acrylonitrile into acrylamide. When urea was added to the culture medium, the NHase activity was 62.3 U ml^?1 (R2A–R2A/urea) after 22.5 h of cultivation, which was similar to that in the medium without addition (R2A–R2A, 70.0 U ml^?1). The relative activity of the purified NHase was 100%, 92%, 94%, and 92% in the medium containing, respectively, 0 mM, 2 mM, 5 mM, and 10 mM of urea. Urea had no significant effect on the purified NHase activity of Mesorhizobium sp. F28. This research did not observe the NHase production by Mesorhizobium sp. F28 when acrylonitrile was supplemented in the culture medium except that cobalt ions existed. The highest enzyme activity was 328.5 U ml^?1 as cobalt ions were added in the pre-culture and culture medium after 22.5 h of cultivation (R2A/Co-R2A/Co); compared to media without cobalt ions (R2A–R2A, 22.5 h, 70.5 U ml^?1) this is an almost five-fold enhancement. It can be concluded that culture media containing cobalt ions was beneficial for the formation of active NHase of Mesorhizobium sp. F28.     

Diurnal patterns of abundance of aphidophagous lady beetle species on chili (Capsicum annuum L.)

The influence of temperature on the diurnal activity of five species of aphidophagous lady beetles (Coleoptera: Coccinellidae) was investigated between 0700 and 1900 h in chili (Capsicum annuum L.) agroecosystems and neighboring vegetation (goose grass, Eleusine indica L.). The lady beetle species observed were Menochilus sexmaculatus Fabricius, Coelophora inaequalis F., Coccinella transversalis F., Harmonia octomaculata F. and Coelophora bissellata Mulsant. More lady beetles (of all species) were found during cooler periods (at 0700, 0900, 1100, and 1900 h). The diurnal pattern of lady beetle adult was temperature dependent. On chili plants, numbers were higher at temperatures between 22 to 30 °C (at 0700, 0900, 1100 and 1900 h) and numbers decreased when temperatures were above 30 °C. When temperature was above 30 °C under the chili plant canopy, numbers were higher in neighboring goose grass, where temperatures were cooler (< 30 °C). Numbers of all species were negative correlated between chili plant and goose grass.     

A simulation study of subterranean termites' territory formation

The process through which the Formosan subterranean termite, Coptotermes formosanus Shiraki, establishes territory was simulated using a lattice model in order to understand how such territories are formed from their seeds-founding pairs that fall to the ground every year. The model incorporated, summer–winter cycles, and fourteen years were simulated. Simulated pairs fell to random sites within the lattice space at the beginning of every summer, and their territories grew during the summer season, and shrunk during the winter season. Fourteen years were sufficient for territory size and shape to become stable over time. The simulation revealed that only pairs introduced at t = 0 had established large territories by the end of the simulation (t = 14). Pairs that were introduced later expanded their territory a little at first, but ultimately shrunk back into a single-cell-sized or small-sized territory. This means that stable-state territory size is mostly determined by when that territory was initially established.     

Pond and landscape determinants of Rana dalmatina population sizes in a Romanian rural landscape

Amphibians are good indicators of human impact, declining steadily worldwide. We explored the relationships between the ponds and nearby landscape parameters and population size of the Agile Frog (Rana dalmatina), estimated from the number of egg masses, in a cultural landscape within the central section of Târnava Mare Basin, Romania. Forty-three permanent ponds were surveyed in a 2600 km² area. The average number of egg masses per pond was 211.13 (SD = 426.41). The egg mass number was significantly and positively related to the emergent aquatic macrophyte cover (its effect peaks at around 50%) and the green connecting corridors between the ponds and forests, and negatively related to the extent of nearby urban areas. The proximity of the forest (positive effect) and the presence of high traffic roads (negative effect) were highly correlated with green corridors and further eliminated from the model due to multicollinearity. Both these variables had significant effects when incorporated in univariate models and multivariate models without green corridors. Since a large part of our study area was currently declared as Natura 2000 site, there is an increased need for management proposals and conservation applications for biodiversity, including amphibians. Rana dalmatina is an important species for monitoring because it is common in the studied area and is suited for short surveys.     

Future land use effects on the connectivity of protected area networks in southeastern Spain

Land-use change is a major driver of the global biodiversity crisis, mainly via the fragmentation and loss of natural habitat. Although land-use changes will accelerate to meet humankind's growing demand for agricultural products, conservation planning rarely considers future land uses and how they may affect the connectivity of ecological networks. Here, we integrate land-use models with landscape fragmentation and connectivity analyses, to assess the effects of past and future land-use changes on the connectivity of protected area networks for a highly dynamic region in southeast Spain. Our results show a continued geographical polarisation of land use, with agricultural intensification and urban development in the coastal areas, and the abandonment of traditional land use in the mountains (e.g., 1100 km² of natural vegetation are projected to be lost in coastal areas whereas 32 km² of natural vegetation would recover in interior areas from 1991 to 2015). As a result, coastal protected areas will experience increasing isolation. The connectivity analyses reveal that the two protected area networks in place in the study area, the European “Natura 2000” and the Andalusian “RENPA” networks, include many landscape connectors. However, we identify two areas that currently lack protection but contain several important patches for maintaining the region's habitat connectivity: the northwestern and the southwestern slopes of the Sierra Cabrera and Bédar protected area. Our results highlight the importance of considering future land-use trajectories in conservation planning to maintain connectivity at the regional scale, and to improve the resilience of conservation networks.     

Pharmacological evaluation of R(+)-pulegone on cardiac excitability: Role of potassium current blockage and control of action potential waveform

IntroductionR(+)-pulegone is a ketone monoterpene and it is the main constituent of essential oils in several plants. Previous studies provided some evidence that R(+)-pulegone may act on isolated cardiac myocytes. In this study, we evaluated in extended detail, the pharmacological effects of R(+)-pulegone on cardiac tissue.MethodsUsing in vivo measurements of rat cardiac electrocardiogram (ECG) and patch-clamp technique in isolated myocytes we determinate the influence of R(+)-pulegone on cardiac excitability.ResultsR(+)-pulegone delayed action potential repolarization (APR) in a concentration-dependent manner (EC₅₀ = 775.7 ± 1.48, 325.0 ± 1.30, 469.3 ± 1.91 μM at 10, 50 and 90% of APR respectively). In line with prolongation of APR R(+)-pulegone, in a concentration-dependent manner, blocked distinct potassium current components (transient outward potassium current (I_to), rapid delayed rectifier potassium current (I_Kr), inactivating steady state potassium current (I_ss) and inward rectifier potassium current (I_K1)) (EC₅₀ = 1441 ± 1.04; 605.0 ± 1.22, 818.7 ± 1.22; 1753 ± 1.09 μM for I_to, I_Kr, I_ss and I_K1, respectively). The inhibition occurred in a fast and reversible way, without changing the steady-state activation curve, but instead shifting to the left the steady-state inactivation curve (V_1/2 from −56.92 ± 0.35 to −67.52 ± 0.19 mV). In vivo infusion of 100 mg/kg R(+)-pulegone prolonged the QTc (∼40%) and PR (∼62%) interval along with reducing the heart rate by ∼26%.ConclusionTaken together, R(+)-pulegone prolongs the APR by inhibiting several cardiomyocyte K⁺ current components in a concentration-dependent manner. This occurs through a direct block by R(+)-pulegone of the channel pore, followed by a left shift on the steady state inactivation curve. Finally, R(+)-pulegone induced changes in some aspects of the ECG profile, which are in agreement with its effects on potassium channels of isolated cardiomyocytes.     

Actividad antifúngica de los enjuagues bucales frente a Candida albicans y Rhodotorula mucilaginosa: un estudio in vitro

BackgroundCandida albicans and Rhodotorula mucilaginosa are yeasts of clinical importance in the oral cavity. In immunocompromised patients they can cause some pathologies that must be controlled with antimicrobials.AimsTo evaluate and compare the antimicrobial efficacy of commercially available mouthrinses against strains of C. albicans and R. mucilaginosa.MethodsThe six mouthwashes studied in vitro were formulated (alone or in combination) with chlorhexidine (CHX) 0.12%, CHX 0.1%, CHX 0.05%, cetylpyridinium chloride (CPC) 0.075%, CPC 0.05%, and essential oils. Ten C. albicans and R. mucilaginosa isolates each were studied. The agar diffusion method (Mueller Hinton II), with incubation at 32 °C was used to evaluate the antifungal activity.ResultsThe results of this study indicate that mouthwashes with CHX 0.1%, CHX 0.12%, CHX 0.05% + CPC 0.05%, CHX 0.12% + CPC 0.05% and CPC 0.075% have an antifungal effect against C. albicans and R. mucilaginosa. CHX 0.1% led to the broadest inhibition zone for C. albicans and R. mucilaginosa (25.65 ± 2.39 mm and 40.05 ± 3.31 mm). Essential oils did not show any antifungal activity. Statistical analysis showed no statistical difference between mouth rinses CHX 0.1%, CHX 0.12% and CHX 0.12% + CPC 0.05% (p = 0.0001) against C. albicans and R. mucilaginosa.ConclusionsMouthwashes with CHX showed higher antifungal activity against C. albicans and R. mucilaginosa than other mouthwashes studied.