Introduction of Participatory Conservation in Croatia, Residents’ Perceptions: A Case Study from the Istrian Peninsula

Croatia, like many other transition countries has undergone radical changes in its nature protection models. This paper discusses a historical overview, present situation and future possibilities for nature conservation in Croatia. A conservative top-down approach to nature protection was applied in the past in Croatia and is now being replaced by a prevalent bottom-up approach. Social context is crucial to introducing participatory conservation, therefore special concern is given to the perception of the local population towards protected area management in Istria as a case study in Croatia. Survey data were used to assess the conservation knowledge of local populations and their perception towards Protected Areas (PAs), leadership activities and management authorities in Istria County. This paper examines the perceptions of 313 residents living in and around six natural PAs located in Istria. The results revealed a moderate general knowledge about PAs in Istria and environmental issues, and a low awareness of institutions managing PAs, eagerness to participate in the activities of PAs and general support for the conservation cause. Understanding the perception of local residents enables the creation of feasible, long-term strategies for the implementation of participatory conservation. The research identifies the need for greater human, technical and financial efforts to strengthen the management capabilities of local agencies responsible for PAs. The process of participatory conservation optimization in Croatia is underway and world experiences must be observed in order to create a congruent, site-specific model with the best possible results.     

Determination of C(5)-C(12) perfluoroalkyl carboxylic acids in river water samples in the Czech Republic by GC-MS after SPE preconcentration

A method employing solid phase extraction followed by gas chromatography-mass spectrometry with negative chemical ionization has been developed for determination of ultratrace concentrations of perfluoroalkyl carboxylic acids in river water. The effects of the experimental parameters, such as the pH, additions of NaCl and an ion-pairing agent (tetraethylammonium bromide) and the kind of the elution agent, on the efficiency of the test acid extraction have been studied. The analyte extraction recoveries and the limits of detection and determination have been found. The method developed has been tested on determinations of perfluoroalkyl carboxylic acids in the waters of the two largest Czech rivers, Vltava and Labe (Elbe). The best extraction results have been attained without any alteration of the sample pH, with an addition of tetrabutylammonium bromide (a concentration of 50 μg mL⁻¹ in the sample) and using methanol as the elution agent. Under these conditions, the recoveries of the test acids in the spiked real samples are within ranges from 60% to 104% and 53% to 111% for analyte concentration levels of 1.40 ng mL⁻¹ and 0.14 ng mL⁻¹, respectively, depending on the lengths of the perfluorinated chains of the acids. In general, the recovery decreases with increasing length of this chain. The method developed exhibits very low limits of detection and determination and the results are fully comparable with those obtained when using more expensive HPLC-MS/MS instrumentation. Typical values amount to tenths to tens of pg mL⁻¹ and units to one hundred pg mL⁻¹ for the limits of detection and determination, respectively; the measuring sensitivity increases with increasing length of the analyte chain. The analyzes of real samples from the Vltava and Elbe rivers have demonstrated that the results obtained are similar to the values published for contamination of the Elbe and other rivers in western and central Europe. The concentrations determined are of the order of units to tens of pg mL⁻¹ and the C₈-C₁₀ acids occur most often.     

The electronic properties of trimethylnaphthalenes as properties for the prediction of biodegradation rates: ab initio and DFT study

There is little information on trimethylnaphthalenes (TMNs) which are constituents of diesel fuel and bitumen emissions. In this study, a theoretical investigation of the electronic properties of all trimethylnaphthalene (TMN) isomers and their relation to biodegradation are presented. Equilibrium geometries, ionization potentials (IPs), electron affinities (EAs), dipole moments and electronic dipole polarizabilities of TMN isomers calculated by ab initio and Density Functional Theory (DFT) methods are reported. Polarizability and dipole moment computations have been performed in gas and in water solution using the polarizable continuum model (PCM). The results obtained show that the IP value varies little along the series of isomers while averaged static dipole polarizabilities (〈α〉) increase on passing from α,α,α-TMN to β,β,β-TMN isomers. This indicates that the binding affinity between TMNs and active site of bacterial enzymes is mainly determined by dispersive and inductive effects. Therefore, the computed polarizability values of TMNs can be used as predictors of the rates of biodegradation of TMNs.     

Survey of cyanobacterial toxins in Czech water reservoirs—the first observation of neurotoxic saxitoxins

The environmental occurrence and concentrations of cyanobacterial toxins (cyanotoxins) were investigated in the Czech Republic. Concentrations of microcystins (MCs), cylindrospermopsin (CYN) or saxitoxins (STXs) were determined immunochemically by ELISA assays in 30 water samples collected from the surface layers of 19 reservoirs during the summer season of 2010. MCs were detected in 18 reservoirs and 83 % of samples, with median and maximal concentration being 1.5 and 18.6 μg/L, respectively. The high frequency of MC occurrence coincided with prevalence of cyanobacterium Microcystis sp., which was detected in 87 % samples, followed by Dolichospermum (Anabaena) sp. observed in 33 % samples. CYN was detected by ELISA only in one sample at a concentration of 1.2 μg/L. STXs presence was indicated for the first time in Czech water reservoirs when the toxins were found at low concentrations (0.03–0.04 μg/L) in two samples (7 %) collected from two different reservoirs, where STXs co-occurred with MCs and eventually also with CYN. In both STX-positive samples, the phytoplankton community was dominated by Microcystis sp., but Dolichospermum sp. and/or Aphanizomenon sp. were also present as putative producers of STX and/or CYN. Cyanotoxins commonly occurred in Czech water reservoirs, and MCs frequently at concentrations possibly associated with human health risks. MCs were the most prevalent and abundant cyanotoxins, but also other cyanotoxins were detected, though sporadically. Further research and regulatory monitoring of cyanotoxins other than MCs is therefore required.     

Photodynamic effects of 31 different phthalocyanines on a human keratinocyte cell line

Phthalocyanines (Pcs, colored macromolecular compounds with the ability to generate singlet oxygen) represent a promising group of photosensitizers due to their intense absorption in the red and UV portion of the spectrum which leads to their excitation. In order to characterize possible toxic effects associated with eventual practical use and application of these chemicals, we employed an in vitro cell culture model to evaluate cytotoxic effects of 31 different phthalocyanines using neutral red uptake assay. An immortalized human keratinocyte cell line HaCaT was exposed to the tested chemicals for 2 or 24 h, either with or without illumination in the last 60 min of the exposure period. After 2- or 24-h exposure without illumination, no cytotoxic effects or weak cytotoxic effects were induced by any Pc under the study and EC50 values could not be obtained within the tested concentration ranges (1.25–20 mg L⁻¹ or 0.625–10 mg L⁻¹). On the other hand, exposure to phthalocyanines under illumination induced a significant cytotoxic effect. The most pronounced cytotoxicity was elicited by Pcs previously shown to have high positive charge densities at peripheral parts of substituent groups, which is most likely the factor responsible for the binding of Pc to negatively charged membranes on the cell surface and thus guaranteeing the tight connection necessary for the singlet oxygen attack on the cell surface.     

Metallophyte status of violets of the section Melanium

Violets from metal-enriched soils have controversially been described as both heavy-metal accumulators and excluders in the literature. The present study solves the issue for violets of the section Melanium (zinc violets, Viola lutea ssp. calaminaria and V. lutea ssp. westfalica; hartsease or wild pansy, Viola tricolor; and mountain pansy, V. lutea). The aims were to determine the concentrations of heavy metals in the soil and in the roots and shoots of field-collected plants, to evaluate the potential impact of colonisation by arbuscular mycorrhizal fungi on heavy-metal concentrations in the plant tissues, and to quantitatively define the localisation of the elements in root cross-sections. When these violets grow in low-metal soils, higher concentrations of the heavy metals were found in the roots and shoots than in the soil, whereas the opposite was seen in samples from high-metal soils. Under all field conditions examined, the roots of all of these species were colonised by arbuscular mycorrhizal fungi. However, V. tricolor was marginally colonised when the concentrations of Zn and P were higher in the soil. Determination of the spatial distribution of the elements in root cross-sections of these violets indicates tissue-specific deposition of elements within the vascular tissue, the cortex, and the rhizodermis. These data indicate that violets of the section Melanium are heavy-metal excluders.     

Seasonal and spatial variations of Po and Pb activity concentrations in Mytilus galloprovincialis from Croatian coast of the Adriatic Sea

Results of 2 years monitoring of ²¹⁰Po and ²¹⁰Pb activity concentrations in soft tissue of the species Mytilus galloprovincialis from Croatian part of the Adriatic coast are presented. The samples were collected at thirteen coastal stations (some of which are also a part of the Mediterranean Mussel Watch Project) in spring and autumn of 2010 and 2011. The collected mussels were ranging between 4 cm and 6 cm in shell length. After sample pre-treatment lead and polonium were radiochemically separated on Sr resin. ²¹⁰Po was determined by alpha-particle spectrometry and ²¹⁰Pb was determined, via ²¹⁰Bi, by a low-level gas proportional counter. The results of ²¹⁰Po activity concentrations were found to vary between (104 ± 11) and (1421 ± 81) Bq kg⁻¹ dry weight while ²¹⁰Pb activity concentrations were much lower and in range (8.2 ± 5.3)–(94.1 ± 29.8) Bq kg⁻¹ dry weight. Higher ²¹⁰Po and ²¹⁰Pb activity concentrations were determined in spring period. The inter-site differences seen in their activity concentrations can be due to natural background levels of sites. The ²¹⁰Po/²¹⁰Pb activity concentration ratios in all cases exceeded unity for all mussel samples and ranged between 4.0 and 47.9.     

Biodegradation of PCBs by ligninolytic fungi and characterization of the degradation products

The aim of the present study was to compare the degrading capabilities of eight ligninolytic fungal representatives towards a technical mixture of polychlorinated biphenyls (Delor 103). Axenic cultures of the fungi, either in complex or N-limited liquid media, were spiked with the technical mixture of Delor 103. All of the fungal strains were able to degrade the pollutant significantly after 6weeks of incubation in both media. Outstanding results were achieved by the treatment with Pleurotus ostreatus, which removed 98.4% and 99.6% of the PCB mixture in complex and mineral media, respectively. This fungus was the only one capable of breaking down penta- and hexachlorinated biphenyls in the complex medium. Ecotoxicological assays performed with the luminescent bacterium Vibrio fischeri demonstrated that all of the fungal strains employed in this study were able to remove the toxicity only temporarily (e.g., after 28d of incubation), while P. ostreatus was capable of suppressing the toxicity associated to PCBs along the whole incubation period in both media. We also performed an extensive set of qualitative GC/MS analyses and chlorinated derivatives of hydroxy- and methoxy-biphenyls were detected along with monoaromatic structures, i.e. chlorobenzoic acids, chlorobenzaldehydes and chlorobenzyl alcohols. This results indicate that both intracellular (cytochrome P-450 monooxigenase, aryl-alcohol dehydrogenase and aryl-aldehyde dehydrogenase) and extracellular (ligninolytic enzymes) enzymatic systems could be involved in the biotransformation of PCB by ligninolytic fungi. The data from this work also document that the fungi are able to degrade further the main metabolites on the PCB pathway (i.e. chlorobenzoic acids) simultaneously with PCBs.     

Chemical and morphological characterization of aerosol particles at Mt. Krvavec, Slovenia, during the Eyjafjallajökull Icelandic volcanic eruption

Objective

In this work, continuous and size-segregated aerosol measurements at Mt. Krvavec, Slovenia, during the Eyjafjallajökull volcanic eruption were performed. Based on chemical and morphological characteristics of size-segregated particles, the presence of the volcanic aerosols after long-range transport to Slovenia was to be confirmed.

Results and conclusions

Continuous measurements with the aethalometer and SMPS indicated the suspected volcanic ash plume passing over the sampling site. The aerosols collected by discrete sampling showed a chemical signature similar to the known elemental signature of the Icelandic volcanic ash. Coarse particles showed a composition typical for silicates rich in metals; in many cases also S was present. Morphological analysis showed particles with features indicative of an explosive volcanic eruption, e.g., pumice and pumice shards, glass shards, minerals, evidence of steam condensation, etc. The high sulfate concentration associated with the fine particles resulted in sulfate crystallization within the cascade impactor leading to the formation of large structures resembling a “fern”. Mass size distributions for Fe, Ti, Mn, Ca, Na, and Mg showed one primary peak (for Fe, Mn, and Ti at 2.8 μm; for Ca, Na, and Mg at ca. 4 μm), which supports the fact that most of the particles in the coarse sizes were silicates rich in metals. The size distribution of the water-soluble SO ₄ ^2? showed a maximum peak at 0.75 μm, which also confirms the high sulfate concentration in the fine particles. Chemical and morphological characterization of aerosols collected at Mt. Krvavec indeed confirmed that volcanic ash plume passed over Slovenia.     

Determination of wine microbiota using classical method, polymerase chain method and Step One Real-Time PCR during fermentation process

The aim of our study was the identification of grape, must and wine microbiota during the fermentation process using a classical microbiological method and Real-Time PCR. The changes in different groups of microorganisms were monitored in total counts of bacteria, lactobacilli and yeasts. Microbiological parameters were observed during the current collection and processing of grapes in 2009. Samples were taken during the fermentation process in wine enterprises and a private vineyard. During this period 30 samples of wine among Müller Thurgau, Cabernet Sauvignon, Chardonnay, Tramin and Red Bio-wine were examined. Samples were collected from stages of grape-must unfiltered, grape-must filtered, the beginning of fermentation, fermentation, late fermentation and young wine. The highest total counts of bacteria ranged from 0.00 to 176 ± 15 CFU.mL(-1) in the wine of Müller Thurgau, the highest number of yeast ranged from 0.00 to 150 ± 9 CFU.mL(-1) in the wine of Müller Thurgau and the number of Lactobacillus spp. ranged from 0.00 to 92 ± 5 CFU.mL(-1) in the sample of Cabernet Sauvignon wine. The presence and sensitivity of Gram-positive and Gram-negative bacterial species Enterococcus faecium, Lactobacillus acidophilus, Lactobacillus crispatus and Lactobacillus salivarius were detected using Real-Time PCR (RTQ PCR). Susceptibility of Enterococcus faecium varied in different isolates from 1 to 10(6) CFU.mL(-1), the sensitivity of the species Lactobacillus acidophilus in different isolates of the wine samples ranged from 1 to 10(5) CFU.mL(-1). We also monitored representation of species Lactobacillus crispatus, which were captured by RTQ PCR sensitivity and ranged from 1 to 10(5) CFU.mL(-1). Identification of the species Lactobacillus salivarius in each of isolates by RTQ PCR method showed the presence of these bacteria in the range of 1 to 10(4) CFU.mL(-1).