We further analysed the data in STRUCTURE v.2.3.2 (Pritchard et a/., 2000, Hubisz et al., 2009), which implements a model-based clustering method for inferring population structure. The programme classifies individuals into the set number of clusters so that the Hardy-Weinberg Equilibrium Figure 2.4: Genetic structure of Moehringia tommasinii from five sampled locations in Slovenia, Italy and Croatia. The graph is based on the program STRUCTURE. The colour segments in pie charts correspond to the membership in particular clusters (K=3). 33 % of variation, P = 0.011) mainly separated the Italian and Slovenian individuals. Table 4.2: Systematic list of recorded dung beetle species, number of specimens, chorotype and ecology. When < species was recorded exclusively in one study site, numbers are denoted in bold type. Figure 4.2: Position of Slovenia and two selected karst pastures [study sites] located near Zazid (Z] anc Hrastovlje (H}. Red stars represent the localities on which traps were set. D1 - open pasture; De - pasturt with overgrowth (both within the fenced pasture); D3 - meadow at the forest edge outside the fenced pasture Cartography: Peter Glasnovic. While developing time-efficient and cost-effective ecological research, taxonomic challenges in hyper-diverse invertebrate groups, such as dung beetles in our case, represent a major barrier (Samways, 2002]. The taxonomical relationship between European dung beetles has only been superficially studied, and mostly involved species The general conclusion is that for the majority of Natura 2000 species, this type of grazing was beneficial. Some species which have declined at a national level and are becoming increasingly rare, showed increases in population here. These include turtle dove Streptopelia turtur, hoopoe Upupa epops, wood lark Lullula arborea, red-backed shrike Lanius collurio, linnet Carduelis cannabina and corn bunting Emberiza calandra. Even though the area is relatively small, it still holds a substantial percentage of the national population for hoopoe, woodlark and corn bunting and an even more substantial part of the populations inside the SPA Kras {Table 5.3}. Tawny pipit Anthus campestris is also present here with three pairs. AS somewhat expected, scrubland species such as common whitethroat Sylvia communis, the population of melodious warbler Hippolais polyglotta and yellowhammer Emberiza citrinella declined. Grazing intensification negatively impacted ground nesters, most notably skylark Alauda arvensis and ortolan bunting Emberiza hortulana. Ortolan bunting Emberiza hortulana Figure 5.4: Northern (open) part of the research area 1 (Podgorski Kras). Photograph: Tomaz Mihelic. Table 5.2: Results of the surveys in 2002 and 2013 on the Golié plateau (research area 2); Index: indices of counted pairs 2013/2002 * 100 (100 = no change}; DOe, D13: breeding densities in 2002 and 2013, calculated where there were 20 or more counted birds; Natura 2000 species are shaded. The transect was wal and rain) permitting, fr beginning of July (13 ed weekly, weather (wind om the end of March to the visits], beginning not later than 90 minutes after dawn, stopping at the points 1-8 for 5 minutes. All ornithological contacts within 100 metres of the observation point (birds seen, heard and identified) were recorded, including fly-overs. Observations of interest on the walk between the points were also recorded but separately. The data obtained for 2013 was compared with that obtained using the same techniques in 2009 (when again 13 visits were made between March and July]. Two late evening visits were made in fine weather in the months of June and / or July in all the years of the survey (including 2012, when the post-dawn surveys were not carried out} to count Figure 6.1: Map of Karst edge with sampled ponds. Cartography: Peter Glasnovic and Sara Zupan. 6.2 AMPHIBIAN POPULATIONS IN PONDS OF THE KARST EDGE were visited many more times than others (see Table 6.2}. The presence of fish species in ponds was noted, 6.4 NEWTS ABUNDANCE Figure 6.2: Amphibians found in ponds of the Karst edge. A. Female smooth newt Lissotriton vulgaris and Italian crestec newt Triturus carnifex larva with prominent gills. B. Yellow bellied toad Bombina variegata. C. Common toad Bufo bufo D. Male tree frog Hyla arborea calling. Photographs: Peter Glasnovic (A and C) and Martina Luznik (B and D). & Arnason, 1996; White & Burnham, 1999]. For abundance estimation of the Italian crested newt a different approach was used, assuming a closed population within module CLOSED CAPTURES in programme MARK. Abundance in a further five ponds (SOCO2, KASO2, CRNO2, RAKO2 and MOVO1} with less intensive field sampling was evaluated using the Petersen Index modified after Chapman for small recapture samples. Other sites in the area were surveyed for newts within the broader study of amphibian diversity, with no evaluation of population size. Figure 6.3: Examples of Karst ponds. A - intensively studied pond near Kastelec (KASO1); B - visibly eutrophic pond near Crnotiée (CRNO2) with high water level; C - pond in Zazid (ZAZO1) with abundant surrounding vegetation; 0 - pond near Movraz (MOVO1) amidst cultivated land. Photographs: Martina Luznik (A and B) and Scot Mills (C and 0). Figure 6.6: Estimated abundance of the smooth newt with 95 % Cl in Kastelec pond (KASO1) during four years of stud (2006, 2007, 2008 and 2013}. Blue: males, purple: females. Results showed three divergent mtDNA lineages (H, Ll and M; names are consistent with Babik et Figure 6.10: Diapause hatching rates of 3 Cephalodella sp. rotifers over 21 days. Eggs were recovered from 60 grams of sediments (spread across 12 replicates} using sucrose flotation accelerated with a centrifuge to *980 m, s* for 5 minutes in a SOml Falcon™ tube. With zooplankton’s capacity for global vagility and stasis it may be hypothesised that species are likely to be widespread and even cosmopolitan, indeed this was the prevailing view until the rise of molecular genetic techniques. Instead, the monopolisation hypothesis of gene flow whereby founding populations adapting to the local conditions, form resident resting egg banks in the sediment and maintain large populations during favourable conditions is now favoured. Under these conditions the resident founding populations effectively buffer against migrant genotypes. It Table 6.5: Cladocerans (phylum Athropoda, class Brachiopoda, order Cladocera} sampled from each site, e indicate that the species was present. In the Alture di Polazzo area, the climate shows similar features. The analyses considered the lonc time series from Gradisca d’lsonzo [very close to the site] and have been complemented usinc data from the rain gauge station at Opatje Selo located nearby but in Slovenia. The average annua temperature is about 13.8°C; those of January anc July being about 4°C and 24°C, respectively. The annual temperature range is 20°C, therefore fallinc between the climatic zones of sub-littoral anc sub-continental. The number of days with frost {with T_,,<O°C) is about 45, with that of tropica days (with T_,.> 30°C) being 38 on average. The absolute minimum value is -15°C, while the maximum values recorded more than 38°C. The trend shows important rises in temperature o Figure 7.1: Digital elevation model of the study area and main geographic features of the area with weather station: and study sites. Cartography: Geotema srl. around 1.4°C in the last two decades. The reason for this can be identified in the rapid urbanization that has characterized the area. Nevertheless, the De Martonne aridity index is around 37 points, and therefore the climate is humid without obvious summer water deficits. Finally, according to the Koéppen-Geiger climate classification, the climate is defined as humid temperate sub-littoral with a very hot summer (the “Cfa” climate sub-group). Average annual rainfall is about 1350 mm, rather abundant in relation to the geographic location of the site, and distributed on about 98 days. Moving towards the “edge” of the Karst plateau, rainfall remains constant, 1374 mm at Opatje Selo, distributed on about 101 days. The rainfall regime in this area pertain also to a sub-littoral type. The rainiest season is autumn, while summer and spring receive quite the same cumulated total rainfall [301 mm vs. 300 mm). The trend for the last 25 years shows a decrease of about 3 mm/yr but looking at the data over the longer term and referring to the last SO years, it shows a significant increase of around 4 mm/yr. Finally, the recent intensity of daily and hourly maximums show a clear decline. Figure 7.2: Synthesis of the main informal stratigraphic schemes proposed in the last 30 years; formation names in the original languages;the scheme of Cucchi & Piano (2013) is explained in the main text (author: Stefano Furin Geotema srl]. Figure 7.4: Schematic illustration of a heterogeneous karst aquifer system (modified from Goldscheider, 2012; witt permission). and (2] water presence (i.e. Vipavska jama with permanent spring is most distant from the other caves set in limestone]. Although a similarity index (Jaccard’s index] yielded results that are well correlated to the geographic distances between the caves (compare Figures 7.9a and 7.9b, see also Figure 7.10), the bedrock seems to be most important factor influencing their diversity. Table 7.1: List of seven caves from SW Slovenia with basic data and numbers of taxa and troglobionts. Figure 7.11: Cave shrimps Troglocaris aggr. anophthalmus (Decapoda: Atyidae}. Photograph: Jure Jugovic. For the biodiversity assessments and _ its conservation, faunal lists and studies of species composition have fundamental aspects. Ecological analyses of the composition of cave biota from seven caves showed that different types of cave habitats host different faunal assemblages, regardless of the distance and rock type. As a result, the community assemblage of each cave is very unique (similarity rates being always below * TPC2e: total plate count at 22 °C; TPC37: total plate count at 37 °C; TCG: total coliform count; CP: Clostridium perfringens The subspecies A. m. mellifera (lineage MJ, going by the name of the German bee or the “black bee” for its characteristic dark colour, is widespread mainly in the north-west and is adapted to cold climates. It is not very productive and is prone to robbing but it is a big producer of propolis (Figure 9.2a). strongly attributed to the hybrid carnica-ligustica group (K3], supporting the hypothesis that the presence of this hybrid can be considered typical of an area that represents a natural boundary between the two subspecies. To extract DNA from museum specimens, we developed a non-destructive method to preserve the integrity of the specimens, so that they can be placed back in the collections (Figure 9.4). Samples from Trieste (1934) and Friuli (1966) are and K4, having all been assigned to the variety termed the “local hybrid” due to the presence of characteristics intermediate between the Italian and carnica bee subspecies. it constitutes from a taxonomic perspective, a geographical race of the carnica bee or a sub- species in itself? Figure 10.3: Linear models of the enzymatic activity for PO and proPO of the bees from the site of Domio and Sa Giovanni. The activity is indicated as absorbance per ul of haemolymph for 30 min at 5 min intervals. Bees from the polluted site also exhibited significantly lower enzymatic activity of both PQ and proPO. It should be mentioned, however, that 52 % of the bees collected from hives at Domio hac one or two individuals of Varroa destructor on the thoracic tergites. The higher activities of PO anc Figure 11.1: Methodological scheme (author: Katia Zanatta). Figure 11.3: Principal Component Analyses (PCA) of of associations of the landa carsica and proportion of chorotypes Legend: C1: Salvio officinalis-Euphorbietum fragiferae; Ce: Lactuco vimineae - Bothryochloetum ischaemum C3: Seseli gouanii-Artemisietum albae; C4: Genisto sericeae - Seslerietum tenuifoliae; C5: Genisto-Caricetun mucronatae; C6: Centaureo rupestris-Caricetum humilis; C7: Centaureo cristate - Chrysopogonetum grylli; CE Danthonio alpinae - Scorzoneretum villosae. Endem (Endemic]: northern Adriatic Karst; Orof (Orophyte}: stricth mountainous and rocky environments; Eur-Art-Alp: Alps; Eurosib (Eurosiberian) cold-temperate and cold regions o Eurasia; Stenom (Stenomediterranean): the Mediterranean coasts; Eurim (Mediterranean): Eurasia influenced by tht Mediterranean; Europ (Europe): Europe; Paleot (Paleotemperate): Eurasian and North Africa; Circum (Circumboreal] Eurasia and N America; P-SE-Eur (Pontic/SE-European): Ukraine, Pannonia, north of the Black Sea/south-eastert Europe; Med-Mon (Mediterranean-Montane]: mountain ranges of south-east Europe, lllir [Illyria]: Former Yugoslavia SE-Alps; Subatl (Subatlantic): Europe and western Siberia; Eur-Cauc (European-Caucasian): Europe with significan presences to the CaucasusCosmop (Cosmopolitan): a large part of the globe. Aerial photogrammetry is a technique that allows the acquisition of important metric data for an object {shape and position} through the analysis of pairs of frames using the principle of stereoscopy. On the basis of the photographs acquired by the drone and as a result of the partial overlap of the frames it was possible to create a three-dimensional model of the ground surface Figure 12.2: Digital Surface Model (DSM) obtained from the analysis of the photos taken by the drone. distinguished by the classifier due to their very similar chromaticity. Taking advantage of this parameter, it was possible to thematise the two objects well. Table 12.4: Distribution of OGU in five classes of density on the basis of the percentage of scrubbing-over In formerly wooded areas subjected to grazing for seven years, monitoring was also carried out using continuous belt transects to assess the two main groundcover species of the karstic scrub and woodland, the grasses Sesleria autumnalis, typical of the well-lit woodlands of downy oak, and Brachypodium rupestre, a herbaceous species that dominates in the herbaceous fringes. These two large herbs represent poor forage for grazing animals and reproduce vegetatively, forming a compact herbaceous layer, which is competitive in relation to other species. The two transects, one dominant in Ses/leria and the other featuring large quantities of Brachypodium, are 7 m long, divided into 7 subplots of 1 x 1 m, and were positioned within formerly wooded areas [respectively in downy oak woodland and a black pine plantation dominated by broadleaf trees} and grazed. With this sampling, the different behaviour over time of Brachypodium and Sesleria following the variation of two limiting factors, light (increased at ground Figure 13.2: Transect dominated by Brachypodium rupestre detected from 2007 to 2013. Figures 13.2 and 13.3 show the projections of the selected analyzed species within the two transect over time: it can be seen that both the grasses undergo a considerable reduction. Looking at the measurements on these surfaces from 2007 to 2013, Brachypodium rupestre and Sesleria autumnalis suffer a significant reduction in favour of some potential forage species such as. Potentilla acaulis subsp. tommasiniana, Thymus longicaulis, Dorycnium germanicum and Bromopsis erecta (aggr.}. On the other hand there is an increase of spiny species, favoured by negative grazing selection such as Carduus nutans and, to a lesser extent, Amethyst Eryngo (Eryngium amethvstinum). Figure 13.4: Target species: Centaurea rupestris L. (a), Pulsatilla montana (Hoppe) Rchb. (b), Orchis morio L. (c) anc Orchis tridentata Scop. (d}. Photograph: Cristiano Francescato. Data on populations of these species were collected using GPS to identify the location of the plants. For each species, the individuals within a radius of about 1.5 m were counted, which corresponds to the error of the instrument, in order to evaluate any fluctuations in populations {increase and number of individuals} resulting from the activity of deforestation and grazing. Figures 13.5 and 13.6 show the distributions of target species detected across the study area before the intervention (2006) and after seven years of grazing (2013). Figure 13.6: Distribution of Orchis morio L. {up} and Orchis tridentata Scop. [down] throughout the study area befor the tree and scrub clearance and the reintroduction of grazing (2006), and after seven years of grazing (2013). These are two invasive species with high environmental impact, the presence of which must be controlled and their distribution limited as far as possible (see also the LR 17/2010 of the Figure 13.8: Population of Senecio inaequidens DC. which developed out of material resulting from the tree and scr clearance (wood chips]. Photograph: Cristiano Francescato. The covers in terms of area [m°} and percentage area occupied (%] of the two alien species Ailanthus altissima and Senecio inaequidens were monitored from 2005 to 2013 as reported in two separate diagrams to highlight the trend towards expansion over time (Figures 13.9 and 13.10). Figure 13.11 shows the distribution of Ailanthus altissima from 2005 [before the intervention of deforestation and grazing} through to 2013. One can note the increase in this species distributed mainly along the roadside leading to the border- crossing where one can encounter both single individuals and large blocks completely occupied by this species. A total of 40 punctiform stations were measured, for a total of 165 trunks, and 3 stations with coverage of between 60 and 75 Figure 13.11: Monitoring of Ailanthus altissima Mill. from 2005 to 2013 in the study area Senecio inaequidens, with very few individuals present in 2005, has spread throughout the study area (Figure 13.12]. It is most abundant at the points where materials were accumulated during the tree clearance phase and which constitute a very fertile substrate. 86-point stations were sampled for a total of 624 individuals and two areas health, being highly allergenic. The individuals of this species can easily be eliminated by annua eradication and must be removed and disposed of as waste. Figure 14.1: Meadow at Podpeé, Municipality of Koper, Slovenia. Meadows are forage crops maintained for two or more years, which provide every year one or more harvests. They are managed through phytomass cutting, its collection and removal as fodder for herbivores, not used in situ and usually after it has undergone transformation [e.g.: haymaking, drying, etc.). Since meadows are cut at least once a year, their vegetation is made up exclusively of herbaceous species. Figure 14.7: The graph on the left shows the dry matter (DM) yield at first harvest of 10 meadows in Podpeé (muni- cipality of Koper, Slovenia} in 2012 and 2013. The graphs on the right show the daily rainfall recorded between March and June for the same locality in 2012 (top) and 2013 (bottom). Figure 14.8: Values of the nitrogen-free extract (NFE], crude fibre (CF], crude protein (CP], ether extract (EE], ash neutral detergent fibre [NDF], acid detergent fibre [ADF] and acid detergent lignin [ADL] for two sites in Podpet (municipality of Koper, Slovenia) for the first and second harvest, respectively (June and September]. The picture: represent a meadow with mesic species [above] and a meadow with xeric species [below]. igure 14.9. Dry meadow on terracing at Podpeé, Municipality of Koper, Slovenia Table 15.2: Total annual yield (t ha? DM) and stocking rates (LU) of some karst pastures in Bazovizza (Trieste, NE Italy) from 2005-2009. The aetermination of pasture productivity Is the basis for the stocking rate calculation and the choice of grazing techniques. One of the aims of the BioDiNet project was to evaluate the biomass produced by some pastures in the Italian and “igure 15.5: Data collection phases for the study of pasture productivity: a} subdivision of the area into plots, b} measuring the vegetation height, c) cutting the plot, d) biomass collection. Slovenian karst. The Corrall-Fenlon method (Corall & Fenlon, 1978) was used for the determination of the daily dry matter production, involving the weekly biomass removal fram randomly selected plots set out on the grazing surface (Figure 15.5]. This method permits reconstruction of the seasonal pattern of daily productivity and quantification of the total annual production. Figure 15.6: Total pasture production (t ha? DM) of eight sites in the Italian and Slovenian karst studied in 2012 anc 2013. For the abbreviations in the figure and site characteristics see the table below the figure. 19.9 PRODUCTION AND SEASONALITY OF KARST PASTURES It is clear that the studied sites are very different, not only for the maximum production (parameter a), but also for the date on which it is reached (parameter b]. The use of these parameters can have an important practical implication when combined with the total production. For example, in 2013, two sites (SZGr and SZUngr] in the Zazid area had the same dry matter yield (about 1.5 t ha“). However, the first site (SZGr] reached its maximum production in mid-May, and had a growing period of 88 days (ND9O}, while the second (SZUngr] reached its peak production at the end of April and had a growing period of 108 days. Ine nore |NUr, AUr, AULJ and crude protein concentrations varied depending on the time of forage collection, moreover fibres did not show notable differences were found between sites. For all sites, and in both years an increase of protein concentration was observed from the beginning of the growing period until the end of May; this date occurred from 20 to 50 days (depending on the site] after the maximum production of period 1. The concentrations of structural carbohydrates (NDF, ADF and ADL] were similar for all pastures, with significant seasonal variation and higher percentages found in the more productive year (2013). It therefore follows that the sites differ more in their productivity than in the forage = Dolce, S., Stoch, F. & M. 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