Green spaces and their biodiversity seem to improve the urban life quality. However, a wide range of descriptors were associated with multiple health outcomes, which limits the comparability of existing evidence. The objective of this systematic review was to organise this knowledge, investigating the relationship between biodiversity in urban green spaces and health outcomes in adults. Following PRISMA guidelines and a registered protocol (PROSPERO: CRD42025636281), a search of PubMed, Scopus, Web of Science identified studies published between January 2014 and July 2025. The search strategy (“biodiversity” AND “human health” AND “urban green space”) yielded 147 eligible studies. Most were published between 2017 and 2020 (57

Background Tobacco smoking is a leading cause of morbidity and mortality worldwide, responsible for about 61,300 deaths annually in Italy. In addition to tobacco cigarettes, the use of electronic cigarettes (e-cigs) and heated tobacco products (HTPs) has increased among young people, raising public health concerns. This study aimed to investigate smoking habits, exposure to passive smoke, and awareness of health risks among Italian university students. Methods A multicentre observational survey was carried out by 8 Italian Universities to promote smoke-free lifestyles among students. All students were sent an anonymous online questionnaire via email regarding their smoking habits, their exposure to second-hand smoke, and their knowledge of, and opinions on, smoking. Descriptive statistics and multinomial logistic regression models were applied to assess associations between tobacco and nicotine use with sex and areas of study. Results A total of 20,644 students from different study areas were included in the study: 62.3% did not smoke, 9.9% were former smokers, 14.4% were smokers of tobacco cigarettes exclusively, 6.5% of e-cigs or HTPs, and 6.9% were dual smokers (total smokers 27.8%). Smoking prevalence was highest among Law (35.9%) and Economics (33.7%) students, and lowest among Medicine and Sciences and Technology students (22%). Males were more likely to smoke tobacco or to be dual users, while females reported higher e-cig use; most smokers consumed fewer than 10 cigarettes/day with mild nicotine dependence. Among e-cig/HTP users, over 60% started as an alternative to cigarettes, but 22% started or resumed tobacco smoking after using them and 22% had never smoked tobacco cigarettes before. Passive smoke exposure was common: 26.5% lived with smokers and 58.9% reported peer exposure. While nearly all students recognized the harms of both active and passive tobacco smoking, only 60% considered passive exposure to e-cigs/HTPs as dangerous. About 70% were favourable to stricter enforcement of university smoking bans. Conclusions: Traditional cigarettes remain the most used product among Italian university students, but e-cigs and HTPs are increasingly widespread and socially accepted, even among never smokers. Awareness of the risks of alternative products is limited, and passive exposure remains underestimated. These findings underline the importance of targeted educational campaigns and stronger enforcement of smoke-free policies in academic settings.

The objective of this study was to use plant models, Allium cepa and Lepidium sativum, to assess the genotoxic effects of the urban particulate matter (PM) collected in a Northern Italian town. Aqueous extracts of different particle sizes (PM10-3, PM3-0.5, PM0.5) were tested alongside the organic extracts through the standard Ames test. The organic particulate matter extracts were subjected to mutagenicity testing in the Salmonella typhimurium strains TA98 and TA100 (without and with metabolic activation), whereas the aqueous extracts were evaluated for genotoxicity in the emerging seedlings of L. sativum and in the root tips of A. cepa bulbs using the comet test to detect the primary DNA damage. Furthermore, the micronuclei frequency was assessed in the bulbs of A. cepa. As expected, the organic extracts of PM3-0.5 and PM0.5 induced point mutations in bacteria. The aqueous extracts of the finest fractions caused a significant increase in genotoxic damage in both plant models. These findings indicate that the two plant models (L. sativum seeds and A. cepa bulbs) are able to detect the genotoxicity of aqueous extracts of air pollutants, with many potential advantages as screening-level tools to complement Ames testing for an easier assessment of urban air quality in terms of DNA toxicity.

Fine particulate matter (PM0.5) is an emerging public health concern due to its ability to penetrate deeply into tissues and induce oxidative stress–mediated damage, contributing to the development of chronic diseases. Despite increasing exposure in urban environments, its biological effects and potential mitigation strategies remain insufficiently explored. This study investigated whether selected foods typical of the Mediterranean diet (yoghurt, apples, lemons, peaches, kiwis, broccoli, onions, and tomatoes) can reduce PM0.5-induced DNA damage. The protective effects of these foods were evaluated using the Ames test on Salmonella typhimurium TA98 and the comet assay on human cell lines derived from lung (A549), liver (HepG2), and colorectal (HT-29) tissues. Exposure to PM0.5 organic extracts induced significant mutagenic and genotoxic effects. However, several foods demonstrated notable protective capacity. In bacterial assays, yoghurt, apple, onion, and tomato reduced mutagenicity by over 50

Many compounds of foods belonging to the Mediterranean diet were studied for their cardiovascular-protective and anticancer effects, as well as for antimutagenic activities. Among them, fermented foods as yoghurts and vegetables rich in antioxidants were frequently tested. However, the large majority of the studies have been carried out on single functional element of the food (such as bacterial strain or extracted compound). Differently, this study aimed to investigate whether a panel of commercially relevant foods belonging to Mediterranean diet (namely, yoghurt, apple, lemon, peach, kiwi, broccoli, onion, tomato) can counteract the effects on DNA of mutagens (2-nitrofluorene, sodium azide, 2-aminofluorene) by using the Ames test with preincubation on TA98 and TA100 strains of Salmonella typhimurium. The greatest inhibitory effect of the mutagens was achieved by yoghurt, followed, in decreasing order, by lemon = kiwi > peach = broccoli > onion = tomato > apple > tomato puree. The present findings provide a foundation for further research on the potential DNA protective activity of fermented foods and vegetables.

Given the considerable quantities of biogenic matrices employed in agricultural applications, there is growing concern about the potential negative effects resulting from the presence of harmful contaminants. The project "SLURP - SLUdge Recovery in Agriculture: Environment and Health Protection" planned a multi-stage approach in which the application of a wide battery of bioassays was proposed as an effective tool to measure the direct interaction of matrices with the different components of the ecosystem, from the molecular to the whole organism level. The aim of the present study, which is a part of the "SLURP" project, was to characterise the toxicological and genotoxicological properties of water extracts from biogenic wastes using several assays based on plant, bacterial and human cells. The aqueous extracts of four sewage sludges, a liming material, two manure slurries of swine and bovine origin, a digestate from bovine manure and a compost were chemically characterised for inorganic ions and heavy metals. Then the extracts were analysed using tests on A.cepa, C.sativus, L.sativum, S.typhimurium and human hepatoma cell line (HepG2) to assess toxicity (seed germination, root elongation, proliferation), mutagenicity and genotoxicity (primary DNA damage, chromosomal aberrations). The extracts exhibited chemical heterogeneity. Ammonia nitrogen, Ca2+, Fe and Zn were the most abundant elements. Toxic effects were caused on A.cepa and L.sativum by all extracts, while there were non-toxic effects on human cells. Genotoxic effects on A.cepa and L.sativum were instead caused by almost all the extracts, at least at the highest dose tested, while only four samples from one sewage sludge, liming material, digestate, and compost, caused DNA damage on human cells. None of the extracts induced mutagenic effects in S.typhimurium. A comprehensive interpretation of these results can only be achieved through the integrated evaluation of all eco-toxicological and chemical data obtained throughout the entire project.

Sewage sludge (biosolids) management represents a worldwide issue. Due to its valuable properties, approximately one half of the EU production is recovered in agriculture. Nevertheless, growing attention is given to potential negative effects deriving from the presence of harmful pollutants. It is recognized that a (even very detailed) chemical characterization is not able to predict ecotoxicity of a mixture. However, this can be directly measured by bioassays. Actually, the choice of the most suitable tests is still under debate. This paper presents a multilevel characterization protocol of sewage sludge and other organic residues, based on bioassays and chemical-physical-microbiological analyses. The detailed description of the experimental procedure includes all the involved steps: the criteria for selecting the organic matrices to be tested and compared; the sample pre-treatment required before the analyses execution; the chemical, physical and microbiological characterisation; the bioassays, grouped in three classes (baseline toxicity; specific mode of action; reactive mode of action); data processing. The novelty of this paper lies in the integrated use of advanced tools, and is based on three pillars: • the direct ecosafety assessment of the matrices to be reused. • the adoption of innovative bioassays and analytical procedures. • the original criteria for data normalization and processing.

The application of a battery of bioassays is widely recognized as a useful tool for assessing environmental hazard samples. However, the integration of different toxicity data is a key aspect of this assessment and remains a challenge. The evaluation of industrial waste leachates did not initially undergo any of the proposed integration procedures. This research addressed this knowledge gap. Twenty-five samples of waste foundry sands were subjected to a leaching test (UNI EN 12457-2) to evaluate waste recovery and landfill disposal. The leachates were evaluated using a battery of standardized toxicity bioassays composed of Aliivibrio fischeri (EN ISO 11348-3), Daphnia magna (UNI EN ISO 6341), and Pseudokirchneriella subcapitata (UNI EN ISO 8692), both undiluted and diluted. Daphnia magna and P. subcapitata were the most affected organisms, with significant effects caused by 68% and 64% of undiluted samples, respectively. The dilution of samples facilitates the calculation of EC50 values, which ranged from greater than the highest concentration tested to 2.5 g/L for P. subcapitata. The data on single-organism toxicity were integrated using three methods: the Toxicity Classification System, the toxicity test battery integrated index, and the EcoScore system. The three classifications were strongly similar. According to all applied systems, three samples were clearly nontoxic (from iron casting plants) and two were highly toxic (from steel casting plants). Moreover, the similar ranking between undiluted and diluted leachates suggests the possibility of using only undiluted leachates for a more cost-effective and time-efficient screening of waste materials. The findings of this study highlight the usefulness of integrating ecotoxicological waste assessment. Integr Environ Assess Manag 2024;20:2294-2311. © 2024 The Author(s). Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

The purposes of this review were to investigate the application of the comet assay in Allium cepa root cells to assess the genotoxicity of environmental samples and to analyse the experimental procedures employed. A literature search was performed selecting articles published between January 2000 and October 2023 from online databases using the combined search terms 'comet assay' and 'A. cepa'. Only 18 papers met the inclusion criteria. None of these were published in the first eight years (2000-2007), highlighting the increasing interest in using the comet assay on A. cepa to analyse environmental samples over the last decade. The majority of the selected studies (15/18, 83%) were performed on samples belonging to the water compartment on onion bulbs. Half of the selected studies (9/18) were conducted to demonstrate the DNA damaging effect of the sample, while the other half of the studies not only recognized the presence of genotoxic agents but also addressed possible remediation measures. Detailed analysis of the experimental procedures revealed heterogeneity in many key steps, such as exposure time, test controls, nuclei isolation solutions, duration of electrophoresis, and number of nuclei scored. This literature review has shown that the comet assay on A. cepa, although recognized as an appropriate tool, is underutilized in environmental toxicology. Greater standardization could lead to its more widespread use, providing valuable information on the genotoxicity of environmental samples and the ability of different processes to mitigate their negative effects on plants.

Steel slags, the main waste product from the steel industry, may have several reuse possibilities. Among others, building applications represent a crucial field. However, the potential impact of harmful substances on the environment should be assessed. The aim of this study was to assess the phytotoxicity of steel slags (SS) and concrete mixtures cast with a partial replacement of SS (CSS). Leaching tests were carried out on four SS and four CSS according to EN 12457-2 and UNI EN 15863, respectively. Each leachate was assayed using root elongation tests on 30 seeds of Allium cepa , Cucumis sativus , and Lepidium sativum , respectively, and on 12 bulbs of A. cepa . The latter also allowed the analysis of other macroscopic parameters of toxicity (turgidity, consistency, colour change and root tip shape) and the evaluation of the mitotic index on 20,000 root tip cells per sample. None of the samples induced phytotoxic effects on the organisms tested: all samples supported seedlings emergence, verified by root elongation comparable to, or even greater than, that of the negative controls, and did not affect cell division, as evidenced by mitotic index values. The absence of phytotoxicity demonstrated by the leachates allows SS and SS-derived concrete to be considered as reliable materials suitable for use in civil constructions or in other engineering applications, with economic and environmental advantages, such as the reduction of the final disposal in landfills as well as the consumption of natural resources.

Drinking water quality can be compromised at different stages, from raw water to treated one. This research aimed to evaluate the toxicity and genotoxicity of groundwater contaminated by fluorinated compounds treated in a drinking water treatment plant, through several bioassays. Water samples underwent chemical analyses and were assayed on Daphnia magna , Pseudokirchneriella subcapitata , Allium cepa , human leukocytes and Salmonella typhimurium . Physical–chemical parameters were always within the Italian legislation limits. Water after filtration and disinfection caused slight toxicity in D. magna ; the sample after filtration inhibited the proliferation of P. subcapitata . None of the water samples exerted toxicity in A. cepa . All the analysed samples had genotoxic effects on A. cepa and human leucocytes, while only disinfected water caused mutations in S. typhimurium . A battery composed of tests on D. magna , P. subcapitata , S. typhimurium and A. cepa could represent a useful tool to verify the toxicity/genotoxicity through the water treatment stages and to improve drinking water quality management.

Many fungal genera such as Aspergillus, Penicillium, Fusarium and Alternaria are able to produce, among many other metabolites, the aflatoxins, a group of toxic and carcinogenic compounds. To reduce their formation, synthetic fungicides are used as an effective way of intervention. However, the extensive use of such molecules generates long-term residues into the food and the environment. The need of new antifungal molecules, with high specificity and low off-target toxicity is worth. The aim of this study was to evaluate: i) the toxicity and genotoxicity of newly synthesized molecules with a good anti-mycotoxic activity, and ii) the suitability of the Allium cepa multi-endpoint assay as an early screening method for chemicals. Eight compounds were tested for toxicity by using the A. cepa bulb root elongation test and for genotoxicity using the A. cepa bulb mitotic index, micronuclei and chromosome aberrations tests. Three molecules showed no toxicity, while two induced mild toxic effects in roots exposed to the highest dose (100 µM). A more pronounced toxic effect was caused by the other three compounds for which the EC50 was approximately 50 μM. Furthermore, all molecules showed a clear genotoxic activity, both in terms of chromosomal aberrations and micronuclei. Albeit the known good antifungal activity, the different molecules caused strong toxic and genotoxic effects. The results indicate the suitability of experiments with A. cepa as a research model for the evaluation of the toxic and genotoxic activities of new molecules in plants before they are released into the environment.

The potential impact of concrete mixtures containing steel slag (SS) as a partial replacement of natural aggregates (NA) on the terrestrial ecosystem was assessed using a battery of plant‐based bioassays. Leaching tests were conducted on four concrete mixtures and one mixture containing only NA (reference concrete). Leachates were tested for phytotoxicity using seeds of Lepidium sativum , Cucumis sativus , and Allium cepa . Emerging seedlings of L. sativum and A. cepa were used to assess DNA damage (comet test). The genotoxicity of the leachates was also analyzed with bulbs of A. cepa using the comet and chromosome aberration tests. None of the samples caused phytotoxic effects. On the contrary, almost all the samples supported the seedlings; and two leachates, one from the SS‐containing concrete and the other from the reference concrete, promoted the growth of C. sativus and A. cepa . The DNA damage of L. sativum and A. cepa seedlings was significantly increased only by the reference concrete sample. In contrast, the DNA damage in A. cepa bulbs was significantly enhanced by the reference concrete but also by that of a concrete sample with SS. Furthermore, all leachates caused an increase in chromosomal aberrations in A. cepa bulbs. Despite some genotoxic effects of the concrete on plant cells, the partial replacement of SS does not seem to make the concrete more hazardous than the reference concrete, suggesting the potential use of SS as a reliable recycled material. Environ Toxicol Chem 2023;42:2193–2200. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The comet assay is a sensitive method for the evaluation of DNA damages and DNA repair capacity at single‐cell level. Allium cepa is a well‐established plant model for toxicological studies. The aim of this scoping review was to investigate the recent application of the comet assay in Allium cepa root cells to assess the genotoxicity. To explore the literature a search was performed selecting articles published between January 2015 and February 2023 from Web of Science, PubMed, and Scopus databases using the combined search terms “Comet assay” and “ Allium cepa ”. All the original articles that applied the comet assay to Allium cepa root cells were included. Of the 334 records initially found, 79 articles were identified as meeting the inclusion criteria. Some studies reported results for two or more toxicants. In these cases, the data for each toxicant were treated separately. Thus, the number of analyzed toxicants (such as chemicals, new materials, and environmental matrices) was higher than the number of selected papers and reached 90. The current use of the Allium ‐comet assay seems to be directed towards two types of approach: the direct study of the genotoxicity of compounds, mainly biocides (20% of analyzed compounds) and nano‐ and microparticles (17%), and assessing a treatment's ability to reduce or eliminate genotoxicity of known genotoxicants (19%). Although the genotoxicity identified by the Allium ‐comet assay is only one piece of a larger puzzle, this method could be considered a useful tool for screening the genotoxic potential of compounds released into the environment.

In this study, the agricultural digestate from anaerobic biogas production mixed with food wastes was used as a substrate to grow Trichoderma reesei RUT-C30 and Trichoderma atroviride Ta13 in solid-state fermentation (SSF) and produce high-value bioproducts, such as bioactive molecules to be used as ingredients for biostimulants. The Trichoderma spp. reached their maximum growth after 6 and 3 SSF days, respectively. Both Trichoderma species were able to produce cellulase, esterase, and citric and malic acids, while T. atroviride also produced gibberellins and oxylipins as shown by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) profiling. Experimental evaluation of germination parameters highlighted a significant promotion of tomato seed germination and root elongation induced by T. atroviride crude extracts from SSF. This study suggests an innovative sustainable use of the whole digestate mixed with agro-food waste as a valuable substrate in fungal biorefineries. Here, it has been applied to produce plant growth-promoting fungi and bioactive molecules for sustainable agriculture.

This study aimed at valorizing digestate through Trichoderma spp. solid-state fermentation (SSF) to produce a potentially ameliorated fertilizer combined with fungal biomass as a value-added bioproduct. Plant-growth-promoting Trichoderma atroviride Ta13, T. reesei RUT-C30, T. asperellum R, and T. harzianum T-22 were tested on different SSF substrates: whole digestate (WD), digestate dried up with wood sawdust (SSF1), and digestate enriched with food waste and dried up with wood sawdust (SSF2). The fungal biomass was quantified by using a qPCR assay. The growth of the four Trichoderma spp. was only observed on the SSF2 substrate. The highest quantity of mycelium was produced by T. reesei RUT-30 (689.80 ± 80.53 mg/g substrate), followed by T. atroviride Ta13, and T. asperellum R (584.24 ± 13.36 and 444.79 ± 91.02 mg/g substrate). The germination of Lepidium sativum seeds was evaluated in order to assess the phytoxicity of the Trichoderma-enriched substrate. The treatments with 7.5% SSF2-R, 3.75% SSF2-T-22, and 1.8% SSF2-Ta13 equally enhanced the root elongation in comparison to the non-fermented SSF-2. This study demonstrated that digestate, mixed with agro-food waste, was able to support the cultivation of Trichoderma spp., paving the way to the valorization of fermented digestate as a proper biofertilizer.

From a circular economy perspective, the recovery and reuse of waste plays a fundamental role. Foundries purchase hundreds of millions of siliceous sands every year to create molds and cores that give shape to the casting. These sands, after several uses, become waste that must be properly recovered or disposed of; they are called waste foundry sands (WFS). The reuse of WFS leads to a reduction in: (i) the consumption of raw materials; (ii) the emissions into the atmosphere; and (iii) the amount of waste sent to landfill—on the other hand, the impact that their use generates on the environment and human health must be carefully assessed. Leaching tests are a fundamental tool for establishing the hazardousness of a waste and its release of contaminants into the environment. This paper presents an analysis of the scientific literature regarding the chemical characteristics of WFS and their release following leaching tests carried out in the laboratory; the environmental standards adopted by the countries that have issued guidelines regarding the reuse of WFS will also be presented.

Steel slags (SS) are the major waste produced by iron and steel industry. Slags may be reused as recycled materials, instead of natural aggregates (NA), to reduce the final disposal in a landfill and the exploitation of raw materials. However, the reuse of SS may generate a potential release of toxic compounds for the environment and humans. The purpose of this study was to evaluate the toxicity and genotoxicity of SS, in comparison with NA, by using an integrated chemical-biological approach to enable their safe reuse in engineering applications. Leaching solutions from samples were obtained by using short-term leaching tests (CEN EN 12457-2, 2004) usually adopted for the evaluation of waste recovery and final disposal. Chemical analyses of leachates were performed according to the Italian legislation on waste recovery (Ministerial Decree 186/2006). The leaching solutions were assayed by using toxicity test on Daphnia magna. Moreover, mutagenicity/genotoxicity tests on Salmonella typhimurium , Allium cepa , and human leucocytes and fibroblasts were carried out. The releases of pollutants from all samples were within the limits of the Italian legislation for waste recovery. Despite the effects that SS and NA could have on different cells, in terms of toxicity and genotoxicity, globally, SS do not seem to be any more hazardous than NA. This ecotoxicological assessment, never studied before, is important for promoting further studies that may support the decision-making process regarding the use of such types of materials.

A low-energy paradigm was adopted for sustainable, affordable, and effective urban waste valorization. Here a new, eco-designed, solid-state fermentation process is presented to obtain some useful bio-products by recycling of different wastes. Urban food waste and scraps from trimmings were used as a substrate for the production of citric acid (CA) by solid state fermentation of Aspergillus niger NRRL 334, with a yield of 20.50 mg of CA per gram of substrate. The acid solution was used to extract metals from waste printed circuit boards (WPCBs), one of the most common electronic waste. The leaching activity of the biological solution is comparable to a commercial CA one. Sn and Fe were the most leached metals (404.09 and 67.99 mg/L, respectively), followed by Ni and Zn (4.55 and 1.92 mg/L) without any pre-treatments as usually performed. Commercial CA extracted Fe more efficiently than the organic one (123.46 vs. 67.99 mg/L); vice versa, biological organic CA recovered Ni better than commercial CA (4.55 vs. 1.54 mg/L). This is the first approach that allows the extraction of metals from WPCBs through CA produced by A. niger directly grown on waste material without any sugar supplement. This "green" process could be an alternative for the recovery of valuable metals such as Fe, Pb, and Ni from electronic waste.

Steel slags (SS) are the major waste produced by iron and steel industry. Slags may be reused as recycled materials, instead of natural aggregates (NA), to reduce the final disposal in a landfill and the exploitation of raw materials. However, the reuse of SS may generate a potential release of toxic compounds for the environment and humans. The purpose of this study was to evaluate the toxicity and genotoxicity of SS, in comparison with NA, by using an integrated chemical-biological approach to enable their safe reuse in engineering applications. Leaching solutions from samples were obtained by using short-term leaching tests (CEN EN 12457-2, 2004) usually adopted for the evaluation of waste recovery and final disposal. Chemical analyses of leachates were performed according to the Italian legislation on waste recovery (Ministerial Decree 186/2006). The leaching solutions were assayed by using toxicity test on Daphnia magna. Moreover, mutagenicity/genotoxicity tests on Salmonella typhimurium, Allium cepa, and human leucocytes and fibroblasts were carried out. The releases of pollutants from all samples were within the limits of the Italian legislation for waste recovery. Despite the effects that SS and NA could have on different cells, in terms of toxicity and genotoxicity, globally, SS do not seem to be any more hazardous than NA. This ecotoxicological assessment, never studied before, is important for promoting further studies that may support the decision-making process regarding the use of such types of materials.