Black pine (Pinus nigra Arn.) was chosen as bioindication species for several reasons. Black pine stands are located throughout Istria, at various distances from the above mentioned power plant. Black pine does not shed needles in the autumn, thus prolonging their exposure to sulphur dioxide emissions. Also neeedles stay on branches for several years, giving us the possibility to analyse the effect on older needles

Microorganisms in coastal sediments are fundamental for ecosystem functioning, and regulate processes relevant in global biogeochemical cycles. Still, our understanding of the effects anthropogenic perturbation and pollution can have on microbial communities in marine sediments is limited. We surveyed the microbial diversity, and the occurrence and abundance of metal and antibiotic resistance genes is sediments collected from the Pula Bay (Croatia), one of the most significantly polluted sites along the Croatian coast. With a collection of 14 samples from the bay area, we were able to generate a detailed status quo picture of a site that only recently started a cleaning and remediation process (closing of sewage pipes and reduction of industrial activity). The concentrations of heavy metals in Pula Bay sediments are significantly higher than in pristine sediments from the Adriatic Sea, and in some cases, manifold exceed international sediment quality guidelines. While the sedimentary concentrations of heavy metals did significantly influence the abundance of the tested metal resistance genes, no strong effect of heavy metal pollution on the overall microbial community composition was observed. Like in many other marine sediments, Gammaproteobacteria, Bacteroidota and Desulfobacterota dominated the microbial community composition in most samples, and community assembly was primarily driven by water column depth and nutrient (carbon and nitrogen) availability, regardless of the degree of heavy metal pollution.

The paper deals with the results of chemical analyses of the precipitation collected with the bulk method in and out of the pedunculate oak forest near Lipovljani in Croatia, for the period 1997-2001. A considerable higher total annual deposition of the majority of analysed determinants has been found in the forest. The results point to the importance and influence of the forest cover on the reception and distribution of dry and wet pollution in the forest ecosystem of pedunculate oak

This study focused on the short-term whole organism bioassays (WOBs) on fish (Danio rerio) and crustaceans (Gammarus fossarum and Daphnia magna) to assess the negative biological effects of water from the major European River Sava and the comparison of the obtained results with in vitro toxicity data (ToxCast database) and Risk Quotient (RQ) methodology. Pollution profiles of five sampling sites along the River Sava were assessed by simultaneous chemical analysis of 562 organic contaminants (OCs) of which 476 were detected. At each sampling site, pharmaceuticals/illicit drugs category was mostly represented by their cumulative concentration, followed by categories industrial chemicals, pesticides and hormones. An exposure-activity ratio (EAR) approach based on ToxCast data highlighted steroidal anti-inflammatory drugs, antibiotics, antiepileptics/neuroleptics, industrial chemicals and hormones as compounds with the highest biological potential. Summed EAR-based prediction of toxicity showed a good correlation with the estimated toxicity of assessed sampling sites using WOBs. WOBs did not exhibit increased mortality but caused various sub-lethal biological responses that were dependant relative to the sampling site pollution intensity as well as species sensitivity. Exposure of G. fossarum and D. magna to river water-induced lower feeding rates increased GST activity and TBARS levels. Zebrafish D. rerio embryo exhibited a significant decrease in heartbeat rate, failure in pigmentation formation, as well as inhibition of ABC transporters. Nuclear receptor activation was indicated as the biological target of greatest concern based on the EAR approach. A combined approach of short-term WOBs, with a special emphasis on sub-lethal endpoints, and chemical characterization of water samples compared against in vitro toxicity data from the ToxCast database and RQs can provide a comprehensive insight into the negative effect of pollutants on aquatic organisms.

Narrow city streets surrounded by tall buildings are favorable to inducing a general effect of a “canyon” in which pollutants strongly accumulate in a relatively small area because of weak or inexistent ventilation. In this study, levels of nitrogen-oxide (NO₂), elemental carbon (EC) and organic carbon (OC) mass concentrations in PM₁₀ particles were determined to compare between seasons and different years. Daily samples were collected at one such street canyon location in the center of Zagreb in 2011, 2012 and 2013. By applying machine learning methods we showed seasonal and yearly variations of mass concentrations for carbon species in PM₁₀ and NO₂, as well as their covariations and relationships. Furthermore, we compared the predictive capabilities of five regressors (Lasso, Random Forest, AdaBoost, Support Vector Machine and Partials Least squares) with Lasso regression being the overall best performing algorithm. By showing the feature importance for each model, we revealed true predictors per target. These measurements and application of machine learning of pollutants were done for the first time at a street canyon site in the city of Zagreb, Croatia.

Chemical analysis of plasma samples of wild fish from the Sava River (Croatia) revealed the presence of 90 different pharmaceuticals/illicit drugs and their metabolites (PhACs/IDrgs). The concentrations of these PhACs/IDrgs in plasma were 10 to 1000 times higher than their concentrations in river water. Antibiotics, allergy/cold medications and analgesics were categories with the highest plasma concentrations. Fifty PhACs/IDrgs were identified as chemicals of concern based on the fish plasma model (FPM) effect ratios (ER) and their potential to activate evolutionary conserved biological targets. Chemicals of concern were also prioritized by calculating exposure-activity ratios (EARs) where plasma concentrations of chemicals were compared to their bioactivities in comprehensive ToxCast suite of in vitro assays. Overall, the applied prioritization methods indicated stimulants (nicotine, cotinine) and allergy/cold medications (prednisolone, dexamethasone) as having the highest potential biological impact on fish. The FPM model pointed to psychoactive substances (hallucinogens/stimulants and opioids) and psychotropic substances in the cannabinoids category (i.e. CBD and THC). EAR confirmed above and singled out additional chemicals of concern - anticholesteremic simvastatin and antiepileptic haloperidol. Present study demonstrates how the use of a combination of chemical analyses, and bio-effects based risk predictions with multiple criteria can help identify priority contaminants in freshwaters. The results reveal a widespread exposure of fish to complex mixtures of PhACs/IDrgs, which may target common molecular targets. While many of the prioritized chemicals occurred at low concentrations, their adverse effect on aquatic communities, due to continuous chronic exposure and additive effects, should not be neglected.

Wastewater treatment plant (WWTP) effluents are pointed as hotspots for the introduction of both commensal and pathogenic bacteria as well as their antibiotic resistance genes (ARGs) in receiving water bodies. For the first time, the effect of partially treated submarine effluents was explored at the bottom and surface of the water column to provide a comprehensive overview of the structure of the microbiome and associated AR, and to assess environmental factors leading to their alteration. Seawater samples were collected over a 5-month period from submarine outfalls in central Adriatic Sea, Croatia. 16S rRNA amplicon sequencing was used to establish taxonomic and resistome profiles of the bacterial communities. The community differences observed between the two discharge areas, especially in the abundance of Proteobacteria and Firmicutes, could be due to the origin of wastewaters treated in WWTPs and the limiting environmental conditions such as temperature and nutrients. PICRUSt2 analysis inferred the total content of ARGs in the studied microbiomes and showed the highest abundance of resistance genes encoding multidrug efflux pumps, such as MexAB-OprM, AcrEF-TolC and MdtEF-TolC, followed by the modified peptidoglycan precursors, transporter genes encoding tetracycline, macrolide and phenicol resistance, and the bla operon conferring β-lactam resistance. A number of pathogenic genera introduced by effluents, including Acinetobacter, Arcobacter, Bacteroides, Escherichia-Shigella, Klebsiella, Pseudomonas, and Salmonella, were predicted to account for the majority of efflux pump-driven multidrug resistance, while Acinetobacter, Salmonella, Bacteroides and Pseudomonas were also shown to be the predominant carriers of non-efflux ARGs conferring resistance to most of nine antibiotic classes. Taken together, we evidenced the negative impact of submarine discharges of treated effluents via alteration of physico-chemical characteristics of the water column and enrichment of bacterial community with nonindigenous taxa carrying an arsenal of ARGs, which could contribute to the further propagation of the AR in the natural environment.

During the last 70 years 1, 2, 3, 4, 5, 6-Hexachlorocyclohexane (HCH) has been one of the most extensively used pesticides. Only the gamma-isomer has insecticidal properties. For the marketing of gamma-HCH (lindane) the other 85% HCH isomers which are formed as by-products during HCH production had to be separated and became finally hazardous waste. For each tonne of lindane 8–12 tonnes of waste HCH isomers were produced and production of the approximately 600,000 t of lindane has therefore generated 4.8 to 7.2 million tonnes of HCH/POPs waste. These waste isomers were mostly buried in uncontrolled dumps at many sites around the world. The stockpiles and the large contaminated sites can be categorized as “mega-sites”. Countries with HCH legacy problems include Albania, Argentina, Austria, Azerbaijan, Brazil, China, Croatia, Czech Republic, France, Germany, Hungary, India, Italy, Japan, Macedonia, Nigeria, Poland, Romania, Russia, Slovakia, South Africa, Spain, Switzerland, Turkey, The Netherlands, UK, Ukraine and the USA.As lindane and alpha- and beta-HCH have been listed as POPs in the Stockholm Convention since August 2010, the problem of stockpiles of HCH waste is now documented and globally acknowledged.This article describes briefly the legacy of HCH and lindane that has been created. Three of the mega-sites are being discussed and demonstrate the increase in pollution footprint over time. Recent developments in the EU (including the Sabinanigo project in Aragon/Spain) and on a global level are presented. A short overview is given on lack of activities and on actions of countries within their obligations as Parties of the Stockholm Convention. Furthermore, current country activities supported by the Global Environment Facility (GEF), the “financing mechanism” of the convention, are listed. Finally, conclusions and recommendations are formulated that will contribute to the solution of this problem over the next 25 years.

Marine bioinvasions require integrating monitoring tools with other complementary strategies. In this study, we collected information about the invasive alien crab Callinectes sapidus in Italy, Croatia and Montenegro, by means of online questionnaires administered to recreational fishers (n = 797). Our records matched the current distribution of the species: C. sapidus resulted far more common in the Adriatic/Ionian than in the Tyrrhenian/Ligurian sector. Most respondents rated the species as ‘occasional’ or ‘rare’. Moreover, the more C. sapidus was considered to be abundant, the more fishers tended to perceive it as a negative disturbance over fisheries and the environment. Our findings suggest that C. sapidus is more common than previously thought in most of the study area, and it could have reached the levels of a true invasions in the south-eastern Adriatic Sea. This experience demonstrates that online questionnaires can be appropriate tools to effectively engage stakeholders in alien species monitoring.

The present paper is a contribution to the first initiative of the Port Baseline Survey (PBS) for Non-indigenous species (NIS) in the Mediterranean Sea. It presents a report on the soft-bottom macrobenthos from the five Adriatic ports: Bari, Ancona (Italy), Koper (Slovenia), Pula, Rijeka (Croatia), with a focus on the presence and contribution of NIS to native assemblages. Out of 451 species identified, only four were common to all ports. A total of eight NIS were recorded, five in surveyed ports and three in the lagoon connected to the Port of Koper. The highest number of NIS was recorded in Bari, and the highest abundance in Ancona and Bari. Generally, the number, abundance and contribution of NIS seems too low to cause a substantial impact on native communities in surveyed ports. The suitability of methods adopted for PBS for soft-bottom NIS was discussed and suggestion for methodological improvement is provided.