Concentration of selected biogenic and risk elements in liver, kidneys and muscle of domestic rabbit and wild brown hare.

In the present study the concentration of selected elements in tissues of domestic rabbits and of wild brown-hares (kidneys, liver, and muscle - m. quadriceps femoris) in Slovakian habitats were determined. After mineralization the elements examined were detected using flame atomic absorption spectrophotometry/graphite furnace atomic absorption spectrophotometry. For rabbits, Fe in the liver was correlated with essential (Mn, Cu) (R2 = 0.94, p < 0.05; R2 = 0.96, p < 0.05 respectively) or toxic (Pb) elements (R2 = -0.93, p < 0.05). For hares, significant correlations were found between Cd and Cu or between Cd and Mn in the kidneys (R2 = -0.96, p < 0.05; R2 = 0.92, p < 0.05 respectively), which is the target organ for Cd. Higher concentrations of the elements were found in hare tissue, and this may be linked to pollution of their wild habitats. The xenobiotic elements as well as the essential elements were accumulated in the kidneys of the hares than rabbits. For liver, differences were less pronounced and significance was only for Fe and Cu. Muscle of hares was more contaminated than of rabbits for both biogenic and toxic elements. These results show that detectable concentrations of inorganic elements. These levels may be linked to contamination of the natural habitats of wild biota due to industry, traffic, agriculture, and urban sprawl.

A Machine Learning-Based Web Tool for the Severity Prediction of COVID-19.

Predictive tools provide a unique opportunity to explain the observed differences in outcome between patients of the COVID-19 pandemic. The aim of this study was to associate individual demographic and clinical characteristics with disease severity in COVID-19 patients and to highlight the importance of machine learning (ML) in disease prognosis. The study enrolled 344 unvaccinated patients with confirmed SARS-CoV-2 infection. Data collected by integrating questionnaires and medical records were imported into various classification machine learning algorithms, and the algorithm and the hyperparameters with the greatest predictive ability were selected for use in a disease outcome prediction web tool. Of 111 independent features, age, sex, hypertension, obesity, and cancer comorbidity were found to be associated with severe COVID-19. Our prognostic tool can contribute to a successful therapeutic approach via personalized treatment. Although at the present time vaccination is not considered mandatory, this algorithm could encourage vulnerable groups to be vaccinated.

Terrestrial bioassays for assessing the biochemical and toxicological impact of biosolids application derived from wastewater treatment plants.

Wastewater treatment plants (WWTP) are facilities where municipal wastewater undergoes treatment so that its organic load and its pathogenic potential are minimized. Sewage sludge is a by-product of this process and when properly treated is preferentially called "biosolids". These treatments may include some or most of the following: thickening, dewatering, drying, digestion, composting, liming. Nowadays it is almost impossible to landfill biosolids, which however can well be used as crop fertilizers. Continuous or superfluous biosolids fertilization may negatively affect non-target organisms such as soil macro-organisms or even plants. These effects can be depicted through bioassays on terrestrial animals and plants. It has been shown that earthworms have been affected to various degrees on the following endpoints: pollutants' bioaccumulation, viability, reproduction, avoidance behavior, burrowing behavior. Collembola have been affected on viability, reproduction, avoidance behavior. Other terrestrial organisms such as nematodes and diplopods have also shown adverse health effects. Phytotoxicity have been caused by some biosolids regimes as measured through the following endpoints: seed germination, root length, shoot length, shoot biomass, root biomass, chlorophyll content, antioxidant enzyme activity. Very limited statistical correlations between pollutant concentrations and toxicity endpoints have been established such as between juvenile mortality (earthworms) and As or Ba concentration in the biosolids, between juvenile mortality (collembola) and Cd or S concentration in the biosolids, or between phytotoxicity and some extractable metals in leachates or aquatic extracts from the biosolids; more correlations between physicochemical characteristics and toxicity endpoints have been found such as between phytotoxicity and ammonium N in biosolids or their liquid extracts, or between phytotoxicity and salinity. An inverse correlation between earthworm/collembola mortality and stable organic matter has also been found. Basing the appropriateness of biosolids only on chemical analyses for pollutants is not cost-effective. To enable risk characterization and subsequent risk mitigation it is important to apply a battery of bioassays on soil macro-organisms and on plants, utilizing a combination of endpoints and established protocols. Through combined analytical quantification and toxicity testing, safe use of biosolids in agriculture can be achieved.

Anthropogenic Microparticles in Sea-Surface Microlayer in Osaka Bay, Japan.

The abundance, distribution, and composition of microparticles (MPs) in the sea-surface microlayer (S-SML, less than 100 µm of sea surface in this experiment) and in bulk water (1 m under the sea surface) were investigated to evaluate the pollution level of MPs in Osaka Bay in Japan. Both seawater fractions were collected at eight sites including ship navigation routes, the coastal area, and the center of Osaka Bay for 2021-2023. MPs were filtered for four size ranges (10-53, 53-125, 125-500, and >500 µm) and then digested with H2O2. MPs' abundance was microscopically assessed; and polymer types of MPs were identified by a Fourier transform infrared spectrometer (FTIR). For the 22 collections performed along eight sites, the average MPs' abundance was 903 ± 921 items/kg for S-SML, while for the 25 collections performed along the same sites, the average MPs' abundance was 55.9 ± 40.4 items/kg for bulk water, respectively. MPs in both S-SML and bulk water exhibited their highest abundance along the navigation routes. The smallest MPs (10-53 µm) accounted for 81.2% and for 62.2% of all MPs in S-SML and in bulk water among all sites, respectively. Polymethyl methacrylate (PMMA) was the major type of MPs identified while minor ones were polyethylene, polyesters, polystyrene, polypropylene, polyvinyl chloride, polyamide, etc. PMMA comprised 95.1% of total MPs in S-SML and 45.6% of total MPs in bulk water. In addition, PMMA accounted for 96.6% in S-SML and 49.5% in bulk water for the smallest MP category (10-53 µm). It can be assumed that the MP sources were marine paints-primarily APPs (antifouling paint particles)-as well as land coatings. Sea pollution due to microparticles from ship vessels should be given proper attention.

Floating particles with high copper concentration in the sea-surface microlayer.

This study sought to clarify whether suspended particles containing high Cu concentrations are present in the sea-surface microlayer (S-SML). For this reason, suspended particles (10-2000 µm) in the S-SML were collected periodically from a ship mooring pond during 2018-2020, and the acid-soluble Cu concentration in the suspended particles was measured as particulate Cu (P-Cu). The highest concentration of P-Cu in the S-SML of the pond was 75 µg L-1 with a 90th percentile value of 2.5 µg L-1. This is below P-Cu values reported for the S-SML in North American ports, but 140 times higher than this found in bulk seawater in the Atlantic Ocean. The highest P-Cu concentration in the S-SML of non-organism (abiotic) origin was 17 µg L-1, and the abiotic P-Cu to P-Cu ratio varied from 0.2 to 100%, likely depending on the quality and quantity of biogenic material in the S-SML samples. It is assumed that the S-SML particles examined here contain high Cu concentrations originating from ship antifouling paints.

Biosolid-Amended Soil Enhances Defense Responses in Tomato Based on Metagenomic Profile and Expression of Pathogenesis-Related Genes.

Biosolid application is an effective strategy, alternative to synthetic chemicals, for enhancing plant growth and performance and improving soil properties. In previous research, biosolid application has shown promising results with respect to tomato resistance against Fusarium oxysporum f. sp. radicis-lycopersici (Forl). Herein, we aimed at elucidating the effect of biosolid application on the plant-microbiome response mechanisms for tomato resistance against Forl at a molecular level. More specifically, plant-microbiome interactions in the presence of biosolid application and the biocontrol mechanism against Forl in tomato were investigated. We examined whether biosolids application in vitro could act as an inhibitor of growth and sporulation of Forl. The effect of biosolid application on the biocontrol of Forl was investigated based on the enhanced plant resistance, measured as expression of pathogen-response genes, and pathogen suppression in the context of soil microbiome diversity, abundance, and predicted functions. The expression of the pathogen-response genes was variably induced in tomato plants in different time points between 12 and 72 h post inoculation in the biosolid-enriched treatments, in the presence or absence of pathogens, indicating activation of defense responses in the plant. This further suggests that biosolid application resulted in a successful priming of tomato plants inducing resistance mechanisms against Forl. Our results have also demonstrated that biosolid application alters microbial diversity and the predicted soil functioning, along with the relative abundance of specific phyla and classes, as a proxy for disease suppression. Overall, the use of biosolid as a sustainable soil amendment had positive effects not only on plant health and protection, but also on growth of non-pathogenic antagonistic microorganisms against Forl in the tomato rhizosphere and thus, on plant-soil microbiome interactions, toward biocontrol of Forl.

Geographical Distribution and Long-Term Monitoring of Physokermes hellenicus (Hemiptera: Coccomorpha: Coccidae) on Abies spp. (Pinales: Pinaceae) in Greece.

The scale Physokermes hellenicus (Kozár & Gounari) (Hemiptera: Coccidae) has been recently included in the Greek entomofauna as a beneficial honeydew species. However, there are no adequate data about its geographical distribution and degree of infestation. Therefore, a study was conducted to examine these parameters in fifteen mountains of Greece. Furthermore, the monitoring of P. hellenicus infestation was carried out over a six-year period with regard to natural enemies and honeydew presence at three mountains (i.e., Menalon, Parnis and Tymfristos) that are traditional honeybee foraging areas. An extensive geographical distribution of the scale was negatively correlated with the latitude. Over the period of the study, P. hellenicus infestation exhibited a decreasing trend in the three mountains, which was more obvious at Menalon. The abundance of natural enemies of P. hellenicus, their effectiveness on honeydew excretion and the fecundity of P. hellenicus are discussed. The reduction in the honey produced at the Menalon mountain (a protected designation of origin product) could be attributed to the reduced presence of P. hellenicus in the fir forest. Among the other identified arthropods, Dreyfusia nordmannianae Eckstein (Hemiptera: Adelgidae) is reported for the first time infesting Abies cephalonica (Pinales: Pinaceae) in Greece. Furthermore, this species is reported for the first time as a co-parasite with P. hellenicus on A. cephalonica in Greece. Since D. nordmannianae is a serious pest, additional research is needed to determine its status in Greek fir forest ecosystems.

Evaluating toxic element contamination sources in groundwater bodies of two Mediterranean sites.

The Driver-Pressure-State-Impact-Response (DPSIR) framework is applied for assessing the pressures and impacts on groundwater bodies of two Mediterranean sites (Megara and Oropos-Kalamos basins). The study areas present joint driving forces (drivers) and pressures. The main driving forces in the areas studied mainly include geology, agricultural activities, and urban development, while the main pressures mainly include the weathering of ultramafic rock masses, application of agrochemicals, and groundwater abstractions for irrigation and drinking uses. Hexavalent chromium (Cr+6), chromium total (Crtotal), manganese (Mn), and nitrate (NO3-) contamination of groundwater bodies are attributed to both anthropogenic and lithological sources. Elevated Crtotal (up to 70.3 µg L-1), Mn (up to 87.7 µg L-1), and NO3- (up to 411 mg L-1) contents are recorded for groundwater samples in Megara basin. High Crtotal (up to 34.3 µg L-1), Cr6+ (up to 27.9 µg L-1), Mn (up to 132.5 µg L-1), and NO3- (up to 30 mg L-1) are also observed for groundwater samples in the Oropos-Kalamos basin. The major response actions needed for the management options of groundwater bodies are discussed. Among the proposed remedial measures, the installation of a continuous groundwater monitoring network and the control in the usage of nitrogen fertilizers seems to be the most effective and tangible for immediate action.

Determination of heavy metals in the territory of contaminated areas of Greece and their restoration through hyperaccumulators.

Phytoremediation is an effective technique for the processing of contaminated soil and for sequestering environmental contaminants such as heavy metals. This kind of pollution is ubiquitous in mines and similar excavation sites. This mini-review aims to provide an overview of the alternative forms of restoring contaminated soils (phytoremediation) highlighting the areas of Kozani (Agios Dimitrios) and Chalkidiki (Stratoni) in Greece characterized by exorbitant metallurgical activities. Firstly, a literature search in databases such as ScienceDirect, SciFinder, PubMed, Google Scholar, and Scopus was conducted, using the following keywords to identify relevant articles: "utilization", "plants", "phytoremediation", "sustainability", "environment", and "restoration". Two case studies have subsequently been chosen in Northern Greece, one characterized by metal mining (Pb, Zn, Ag) and hot summer Mediterranean climate and another one involving lignite mining and concomitant Cr, Zn, Cu, and Ni pollution under subtropical climate. Based on the literature search some possible, non-invasive plant species were proposed, focusing on the most efficient hyperaccumulators.

Chemical and ecotoxicological assessment of sludge-based biosolids used for corn field fertilization.

Sludge-based biosolids can be used for crop fertilization and for soil enrichment with organic matter. The prerequisites for their application are laid out in 86/278/EEC which sets limits for metals of toxicological concern in the biosolid and in the recipient soil. In this context, three kinds of sludge-based biosolids from a municipal wastewater treatment plant were submitted to the leaching tests EN 12457-2 and ΝΕΝ 7341. The leachates were examined for their physicochemical, chemical, and ecotoxicological characteristics four times within a year. Even in the more aggressive metal leaching method (ΝΕΝ 7341), results were much lower than the limits set in Council Directive 86/278/EEC. Correlations were noted between numerous parameters and principal component analysis grouped them in 3 components explaining 76% of total variation. These leaching tests were also performed on soil-biosolid samples from another investigative research in which two of the three kinds of biosolids had been incorporated in corn fields at doses 0, 20, and 40 tn/ha. EN 12457-2 leaching tests indicated that Ni and Cd leaching was affected by biosolid incorporation in soil, in relation to the type of the receiving soil (clay or sand). The leachates from the soil-biosolid mixtures were practically not toxic for the organisms tested (D. magna, V. fischeri, higher plants). In summary the biosolids were considered of low environmental risk when utilized in the field. A detailed risk assessment encompassing both chemical and ecotoxicological analysis is necessary for integrated evaluation of biosolids.

Organic pollution and its effects in the marine mussel Mytilus galloprovincialis in Eastern Mediterranean coasts.

Persistent chemicals and emerging pollutants are continuously detected in marine waters and biota. Out of these, polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCs) are significant contaminants with decades of presence in the marine environment. The Mediterranean Sea is an ecosystem directly affected by a variety of anthropogenic activities including industry, municipal, touristic, commercial and agricultural. The Mediterranean mussel (Mytilus galloprovincialis) is a filter feeder, which presents wide distribution. In this regard, the specific organism was used as a biological indicator for the monitoring and evaluation of pollution in the studied areas with focus on the mentioned chemical groups. Pristine Turkish sites with minimum effect from anthropogenic activities, in contrast with Greek sites which were subjected to heavy industrial and shipping activity, were selected. A gas chromatographic tandem mass spectrometric method (GC-MS/MS) was developed and validated to monitor 34 compounds (16 EPA priority PAHs and 18 OCs). Analyses of mussel samples in 2011 from sites with the limited anthropogenic pollution shores have shown the occurrence of 11 pollutants (6 PAHs, 5 OCs), while in the samples from sites with intensive activity and expected pollution, 12 PAHs and 6 OCs were detected. Biochemical and biological responses studied only in mussels samples from the sites with the highest contamination showed a situation that was under strong seasonal influence. The intensity of the response was also influenced by deployment duration. Noteworthy correlations were detected among biochemical/biological effects and between mussel body burden and these effects. Continuous monitoring of priority pollutants of East Mediterranean Sea is vital both for ecological and human risk assessment purposes.

Modulation of CYP1A1 and CYP1A2 hepatic enzymes after oral administration of Chios mastic gum to male Wistar rats.

Chios mastic gum (CMG), a resin derived from Pistacia lentiscus var. chia, is known since ancient times for its pharmacological activities. CYP1A1 and CYP1A2 enzymes are among the most involved in the biotransformation of chemicals and the metabolic activation of pro-carcinogens. Previous studies referring to the modulation of these enzymes by CMG have revealed findings of unclear biological and toxicological significance. For this purpose, the modulation of CYP1A1 and CYP1A2 enzymes in the liver of male Wistar rats following oral administration of CMG extract (CMGE), at the levels of mRNA and CYP1A1 enzyme activity, was compared to respective enzyme modulation following oral administration of a well-known bioactive natural product, caffeine, as control compound known to involve hepatic enzymes in its metabolism. mRNA levels of Cyp1a1 and Cyp1a2 were measured by reverse transcription real-time polymerase chain reaction and their relative quantification was calculated. CYP1A1 enzyme induction was measured through the activity of ethoxyresorufin-O-deethylase (EROD). The results indicated that administration of CMGE at the recommended pharmaceutical dose does not induce significant transcriptional modulation of Cyp1a1/2 and subsequent enzyme activity induction of CYP1A1 while effects of the same order of magnitude were observed in the same test system following the administration of caffeine at the mean daily consumed levels. The outcome of this study further confirms the lack of any toxicological or biological significance of the specific findings on liver following the administration of CMGE.

Monitoring of systemic exposure to plant protection products and DNA damage in orchard workers.

The systemic exposure of plum tree growers and operators to plant protection products (PPPs) and effects on DNA were assessed. Specifically, a GC-MS/MS method was developed and validated for the analysis of serum samples for the presence of seven active substances of PPPs. The analytical results verified the presence of myclobutanil, propargite, cypermethrin and deltamethrin in 7 out of 19 serum samples. The incidence of DNA damage was monitored using the single cell electrophoresis assay (comet assay). A paired Student's t-test revealed a statistically significant increase of SSBs in the blood samples collected at the end of the cropping period as compared to the samples collected from the same subjects before the start of PPPs application period. Moreover, the group of seven subjects with detectable serum pesticides levels revealed statistically significant increase of SSBs as compared to the group of subjects with no detectable PPP levels. The results of the present study demonstrate that the agriculture workers may exhibit detectable level of systemic exposure to the applied PPPs which are correlated to increased DNA damage during the cultivation period.

Excision of an oxidative DNA lesion (8-oxo-deoxyguanosine) in Mytilus edulis populations from two sites in the eastern Irish Sea.

Mussels (Mytilus edulis) from the urban/industrialised site of New Brighton, Merseyside, UK and the reference site of Llandudno, North Wales, UK were collected at various time-points in 2005-2006. The excision capacity of mussel gill towards a radio-labelled probe containing the oxidative lesion 8-oxo-dG (8-oxo-deoxyguanosine) was quantified for both populations within 24 h of collection. Fpg (formamidopyrimidine glycosylase)-sensitive sites (which are proportional to 8-oxo-dG levels) were also quantified via the Fpg-modified Comet assay. Generally, excision capacity in New Brighton mussels was lower compared to Llandudno mussels, with the exception of May when activities from both groups were at their lowest. No substantial differences between Fpg-sensitive sites were noted but the ratio "mean Fpg-sensitive sites/mean 8-oxo-dG excision capacity" was elevated for New Brighton mussels. It is possible that the evident oxidative stress these mussels undergo is linked to pollutants found in their natural environment. These pollutants may act not only by direct oxidative DNA damage but also by down-regulation of DNA repair pathways.

Oxidative damage in gill of Mytilus edulis from Merseyside, UK, and reversibility after depuration.

Mussels were collected from the urban/industrialized site of New Brighton, Merseyside and the relatively non-industrial site of Llandudno, North Wales. All mussels were identified as Mytilus edulis by PCR amplification of Mefp1. DNA single strand breaks and 8-oxo-7,8-dihydro-2'-deoxyguanosine were measured in gill within 24h of collection, using the COMET assay, both with and without formamidopyrimidine glycosylase. Gill lipid peroxidation was also measured within 24h. No difference between sites was found for frank SSB and malonaldehyde levels, however 8-oxo-dG and 4-hydroxynonenal were significantly greater in New Brighton mussels compared to Llandudno mussels. After 1-month laboratory maintenance, lipid peroxidation and 8-oxo-dG levels were lower. In contrast, frank SSB were higher. This could reflect enhanced DNA repair excision, though we cannot exclude the possibility of other non-oxidative DNA damage. The results suggest that laboratory maintenance allows recovery from environmentally induced oxidative damage, which was more extensive at Merseyside.