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The worldwide decline and local extinctions of bumblebees have raised a need for fast and accurate tools for species identification. Morphological characters are often not sufficient, and molecular methods have been increasingly used for reliable identification of bumblebee species. Molecular methods often require high-quality DNA which makes them less suitable for analysis of low-quality or older samples. We modified the PCR- RFLP protocol for an efficient and cost-effective identification of four bumblebee species in the subgenus Bombus s. str. ( B. lucorum, B. terrestris, B. magnus and B. cryptarum). We used a short partial mitochondrial COI fragment (446 bp) and three diagnostic restriction enzymes (Hinf I, Hinc II and Hae III) to identify species from degraded DNA material. This approach allowed us to efficiently determine the correct species from all degraded DNA samples, while only a subset of samples 64.6% (31 of 48) resulted in successful amplification of a longer COI fragment (1064 bp) using the previously described method. This protocol can be applied for conservation and management of bumblebees within this subgenus and is especially useful for fast species identification from degraded samples. [ABSTRACT FROM AUTHOR]

Recognition of per- and polyfluoroalkyl substances (PFAS) as widespread environmental pollutants and a consequent risk to human health, has recently made the European Union (EU) adopt several regulatory measures for their management. The coherence of these measures is challenged by the diversity and the ubiquitous occurrence of PFAS, which also complicates the EU's endeavor to advance justified, harmonized, and transparent approaches in the regulatory assessment of chemical risks. Our study critically reviews the European approach for the risk assessment of PFAS, by applying a comparative analysis of the current and pending regulatory thresholds issued for these chemicals in water bodies, drinking water, and certain foodstuffs. Our study shows that the level of health protection embedded in the studied thresholds may differ by three orders of magnitude, even in similar exposure settings. This is likely to confuse the common understanding of the toxicity and health risks of PFAS and undermine reasonable decision-making and the equal treatment of different stakeholders. We also indicate that currently, no consensus exists on the appropriate level of required health protection regarding PFAS and that the recently adopted tolerable intake value in the EU is too cautious. Based on our analysis, we propose some simple solutions on how the studied regulations and their implicit PFAS thresholds or their application could be improved. We further conclude that instead of setting EU-wide PFAS thresholds for all the environmental compartments, providing the member states with the flexibility to consider case-specific factors, such as regional background concentrations or food consumption rates, in their national regulatory procedures would likely result in more sustainable management of environmental PFAS without compromising the scientific foundation of risk assessment, the legitimacy of the EU policy framework and public health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The considerable environmental burden of textiles is currently globally recognized. This burden can be mitigated by applying circular economy (CE) strategies to the commonly linear, short garment life cycles that end with incineration or landfill disposal. Even though all CE strategies strive to promote environmental sustainability, they might not be equally beneficial. Environmental data on different textile products is insufficiently available, which leads to complications when assessing and deciding on different CE strategies to be implemented. This paper studies the environmental impacts of a polyester T-shirt's linear life cycle through life cycle assessment (LCA) and evaluates the benefits attainable by adopting different CE strategies, and their order of priority, while noting uncertainty arising from poor data quality or unavailability. The LCA is complemented by assessing health and environmental risks related to the different options. Most of the linear life cycle's LCA-based impacts arise from use-phase washing. Hence, it is possible to reduce the environmental impact notably (37 %) by reducing the washing frequency. Adopting a CE strategy in which the shirt is reused by a second consumer, to double the number of uses, enables an 18 % impact reduction. Repurposing recycled materials to produce the T-shirt and recycling the T-shirt material itself emerged as the least impactful CE strategies. From the risk perspective, reusing the garment is the most efficient way to reduce environmental and health risks while washing frequency has a very limited effect. Combining different CE strategies offers the greatest potential for reducing both environmental impacts as well as risks. Data gaps and assumptions related to the use phase cause the highest uncertainty in the LCA results. To gain the maximum environmental benefits of utilizing CE strategies on polyester garments, consumer actions, design solutions, and transparent data sharing are needed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Red wood ants (RWAs) are a group of keystone species widespread in temperate and boreal forests of the Northern Hemisphere. Despite this, there is increasing evidence of local declines and extinctions. We reviewed the current protection status of RWAs throughout Europe and their International Union for the Conservation of Nature (IUCN) threat classification. Only some RWA species have been assessed at a global scale, and not all national red lists of the countries where RWAs are present include these species. Different assessment criteria, inventory approaches, and risk categories are used in different countries, and data deficiency is frequent. Legislative protection is even more complex, with some countries protecting RWAs implicitly together with the wildlife fauna and others explicitly protecting the whole group or particular species. This complexity often occurs within countries, for example, in Italy, where, outside of the Alps, only the introduced species are protected, whereas the native species, which are in decline, are not. Therefore, an international, coordinated framework is needed for the protection of RWAs. This first requires that the conservation target should be defined. Due to the similar morphology, complex taxonomy, and frequent hybridization, protecting the entire RWA group seems a more efficient strategy than protecting single species, although with a distinction between autochthonous and introduced species. Second, an update of the current distribution of RWA species is needed throughout Europe. Third, a protection law cannot be effective without the collaboration of forest managers, whose activity influences RWA habitat. Finally, RWA mounds offer a peculiar microhabitat, hosting a multitude of taxa, some of which are obligate myrmecophilous species on the IUCN Red List. Therefore, RWAs' role as umbrella species could facilitate their protection if they are considered not only as target species but also as providers of species-rich microhabitats., (© 2022 The Authors. Conservation Biology published by Wiley Periodicals LLC on behalf of Society for Conservation Biology.)

Per- and polyfluoroalkyl substances (PFAS) comprise a wide group of persistent chemicals, whose ubiquitous occurrence in the environment, particularly due to their extensive use for fire suppression in aqueous film forming foams (AFFFs), has raised global attention. We evaluated the impacts of PFAS at three firefighting training sites and one industrial site in Finland, to highlight key elements to be considered in the retrospective risk assessment of these chemicals. The site assessments covered the occurrence and distribution of 23 PFAS in multiple environmental matrices, i.e., soil, sediment, surface water, groundwater, and biota, and the subsequent risks to human health and the environment owing to the migration of and exposure to the selected compounds. Our study showed that the extensive use of nowadays restricted or substituted PFAS, particularly PFOS, are still often the predominant compounds detected at AFFF-impacted sites and will continue to cause long-term risks to the environment. The most significant environmental or health risks at these sites are likely to concern aquatic ecosystems, fish consumption or groundwater usage due to the off-site migration of PFAS. Here, even a single fire extinguishing event can be a considerable contributor. We also illustrate that conventional procedures based on simple mass-balance, and exposure models, with a focus on PFOS and other site-specifically relevant PFAS may provide sufficient means to assess the risks. Moreover, we address that despite the exceedance of the very stringent regulatory threshold values issued recently for PFAS, the actual site-specific risks to human health and the environment may remain reasonably low., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Ants can influence ecological processes, such as the transfer of elements or radionuclides, in several ways. For example, they redistribute materials while foraging and maintaining their nests and have an important role in terrestrial food webs. Quantitative data of the transfer of elements into ants is needed, e.g., for developing improved radioecological models. In this study, samples of red wood ants (genus Formica), nest material, litter and soil were collected from a former uranium mining site in Eastern Finland. Concentrations of 33 elements were analyzed by Inductively Coupled Plasma-Mass Spectroscopy/Optical Emission Spectroscopy. Estimated element concentrations in spruce needles were used as a proxy for studying the transfer of elements into ants via aphids because spruces host the most important aphid farms in boreal forests. Empirically determined organism/medium concentration ratios (CRs) are commonly used in radioecological models. Ant/soil CRs were calculated and the validity of the fundamental assumption behind the of use of CRs (linear transfer) was evaluated. Elements that accumulated in ants in comparison to other compartments were cadmium, potassium, phosphorus, sulfur, and zinc. Ant uranium concentrations were low in comparison to soil, litter, or nest material but slightly elevated in comparison to spruce needles. Ant element concentrations were quite constant regardless of the soil concentrations. Non-linear transfer models could therefore describe the soil-to-ant transfer better than conventional CRs., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

High emission of tire rubber particles to the surrounding environment is an inevitable consequence of the current habits of transportation. Although most of the emissions stay within a close range of the sources, it has been proven that the smallest particles can be transported to remote locations through the atmosphere, including inland water bodies. It has been estimated that a relevant portion of the global emissions of tire rubber particles reach surface waters, but effects on aquatic life in the receiving water bodies are not completely understood. In the present study, we used the freshwater sediment dwellers Lumbriculus variegatus and Chironomus riparius to examine the toxicity of tire rubber particles at environmentally relevant concentrations, using different types of sediment and two particle sizes of tire rubber. Overall, the experiments were unable to discern any effects on the growth, survival or reproduction of the two animals tested. Significant differences were found among the animals dwelling on different sediments, but the effects were not attributable to the presence of tire rubber particles. This study provides important information regarding the lack of effect of tire rubber particles in laboratory experiments with model sediment dwellers and opens more questions about the potential effects of tire rubber particles in the real environment with longer durations and varying environmental factors. The influence of other factors such as the leaching of additives in the overall toxicity of tire rubber particles should be also considered., (© 2021. The Author(s).)

Background: Transplantation of human pluripotent stem cell-derived retinal pigment epithelium (RPE) is an urgently needed treatment for the cure of degenerative diseases of the retina. The transplanted cells must tolerate cellular stress caused by various sources such as retinal inflammation and regain their functions rapidly after the transplantation. We have previously shown the maturation level of the cultured human embryonic stem cell-derived RPE (hESC-RPE) cells to influence for example their calcium (Ca 2+ ) signaling properties. Yet, no comparison of the ability of hESC-RPE at different maturity levels to tolerate cellular stress has been reported., Methods: Here, we analyzed the ability of the hESC-RPE populations with early (3 weeks) and late (12 weeks) maturation status to tolerate cellular stress caused by chemical cell stressors protease inhibitor (MG132) or hydrogen peroxide (H 2 O 2 ). After the treatments, the functionality of the RPE cells was studied by transepithelial resistance, immunostainings of key RPE proteins, phagocytosis, mitochondrial membrane potential, Ca 2+ signaling, and cytokine secretion., Results: The hESC-RPE population with late maturation status consistently showed improved tolerance to cellular stress in comparison to the population with early maturity. After the treatments, the early maturation status of hESC-RPE monolayer showed impaired barrier properties. The hESC-RPE with early maturity status also exhibited reduced phagocytic and Ca 2+ signaling properties, especially after MG132 treatment., Conclusions: Our results suggest that due to better tolerance to cellular stress, the late maturation status of hESC-RPE population is superior compared to monolayers with early maturation status in the transplantation therapy settings., (© 2022. The Author(s).)

Information on transfer of elements and their radionuclides is essential for radioecological modeling. In the present study, we investigated the transfer of Cl, Co, Mo, Ni, Se, Sr, U and Zn in a boreal freshwater food chain. These elements were selected on the basis that they have important radionuclides that might be released into the biosphere from various stages of the nuclear fuel cycle. Water, sediment, chironomid larvae (Chironomus sp.), roach (Rutilus rutilus) and perch (Perca fluviatilis) were sampled from two ponds near a former uranium mine and one reference pond located further away from the mining area. Concentrations measured in water, sediment and the three animal species indicated the importance of sediment as a source of uptake for most of the elements (but not Cl). This should be considered in radioecological models, which conventionally predict concentration in aquatic organisms from concentration in water. The results also show that the assumption of linear transfer (constant concentration ratio) may not be valid for elements into fish. The results of this study show that further basic research is needed to understand the fundamental processes involved in transfer of elements into freshwater organisms in order to develop radioecological models., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Insects vary in the degree of their adaptability to environmental contamination. Determining the responses with phenotypic plasticity in ecologically important species in polluted environments will ease further conservation and control actions. Here, we investigated morphological characteristics such as body size, body mass, and color of the common wasp Vespula vulgaris in an industrially polluted environment, considering different levels of metal pollution, and we studied the localization of contaminants in the guts of wasps. We revealed some differences in morphological characteristics and melanization of wasps collected in habitats with high, moderate, and low levels of pollution. The results indicated that V. vulgaris from highly polluted environments had reduced melanin pigmentation on the face but increased melanin pigmentation on the 2nd tergite of the abdomen. In addition, with transmission electron microscopy (TEM) and energy dispersive X-ray analysis (EDX), we found metal particles from the midgut of wasps originating from the polluted environment. Most of the particles were encapsulated with melanin pigment. This finding confirmed that in wasps, ingested metal particles are accumulated in guts and covered by melanin layers. Our data suggest that wasps can tolerate metal contamination but respond phenotypically with modification of their size, coloration, and probably with the directions of the melanin investments (immunity or coloration). Thus, in industrially polluted areas, wasps might probably survive by engaging phenotypic plasticity with no significant or visible impact on the population.

The extent until which plastics are present in our surrounding environment completely exceeds our expectations. Plastic materials in the form of microplastics have been found in terrestrial, freshwater and marine environments and are transported through the atmosphere even to remote locations. However, we are still far from understanding the effects that they may have caused and are causing to biota. In the present study, we investigated the alterations in the expression of twelve genes in the aquatic insect Chironomus riparius after 36 h exposures to polystyrene and tire rubber microplastics at nominal concentrations of 1 and 10 mg L -1 . The results indicated that several genes encoding for heat shock proteins (hsp90, Glycoprotein 93 (Gp93), hsc70, hsp60, hsp40, and the small HSP hsp17) were overexpressed respect to the control. In addition, the genes coding for manganese superoxide dismutase (SOD Mn, related to alleviation of oxidative stress) and for the FK506-binding protein of 39 kDa. (FKBP39, related to development and pupation) showed altered expression. Most of the alterations on gene expression level occurred at a concentration of 10 mg L -1 of tire rubber microplastics, although specific modifications arose at other concentrations of both rubber and polystyrene. On the contrary, one hsp gene (hsp10) and genes related to biotransformation and detoxification (Cyp9f2, Cyp12a2, and ABCB6) did not alter their expression in any of the treatments. Overall, the results of the gene expression indicated that microplastics (especially tire rubber) or their additives caused cellular stress that led to some alterations in the normal gene expression but did not cause any mortality after 36 h. These results highlight the need for more studies that describe the alterations caused by microplastics at the molecular level. Additionally, it opens questions about the effects caused to aquatic fauna in environmental realistic situations, especially in hot spots of microplastic contamination (e.g., tire rubber released in storm water runoff discharge points)., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Human pluripotent stem cell-derived retinal pigment epithelium (RPE) transplantation is currently under evaluation as treatment for macular degeneration. For therapeutic applications, cryostorage during cell production is typically needed with potential consequences to cell functionality. We have previously shown that the culture substrate affects human embryonic stem cell-derived RPE (hESC-RPE) properties in fresh cultures. Here, we aimed to further identify the role of RPE basement membrane proteins type IV collagen (Col-IV), laminin (LN), and nidogen-1 in the maturation and functionality of hESC-RPE after cryopreservation. In addition to cell attachment and morphology, transepithelial electrical resistance, expression of key RPE proteins, phagocytosis capacity and Ca 2+ signalling were analysed. After cryostorage, attachment of hESC-RPE on culture surfaces coated with Col-IV alone was poor. Combining Col-IV and LN with or without nidogen-1 significantly improved cell attachment and barrier properties of the epithelium. Furthermore, functional homogeneity of the hESC-RPE monolayer was enhanced in the presence of nidogen-1. Our results suggest that the choice of coating proteins for the cell culture may have implications to the functional properties of these cells after cryostorage cell banking.

Bird feces are commonly used as a proxy for measuring dietary metal exposure levels in wild populations. Our study aims to improve the reliability and repeatability of fecal metal measurements and gives some recommendations for sampling. First, we studied levels of variation in metallic element (arsenic, calcium, cadmium, cobalt, copper, nickel, lead) concentrations: temporal variation within an individual, among siblings in a brood and among-brood/spatial variation. Second, we explored the variation caused by dual composition (urate vs. feces) of bird droppings. Two sets of fresh fecal samples were collected from pied flycatcher ( Ficedula hypoleuca ) nestlings living in a metal polluted area in summers 2017 (dataset 1) and 2018 (dataset 2). We found a great deal of temporal intra-individual variation in metal levels, suggesting that dietary exposure varied markedly in a short time scale (within a day). A sample from only one nestling per brood did not well describe the brood mean value, and we recommend that at least four siblings should be sampled. Brood level samples give relatively good temporal repeatability for most metals. For all the metals, the levels in the fecal portion were more than double to those in the urate portion. Since the mass proportion of urate in the bird droppings varied a great deal among samples, standardizing sampling, e.g., by collecting only the fecal part, would markedly reduce the variation due to composition. Alternatively, urate portion could be used for biomonitoring of internally circulated bioavailable metal.