Swedish University of Agricultural Sciences

Distinguishing the respective contributions of various microbes to methylmercury (MeHg) production is critical for predicting MeHg bioaccumulation and exposure risk. Metabolic inhibitors have been commonly used to block the activity of specific microbial groups and identify primary Hg methylating microbes. By reviewing literatures and our empirical data, we demonstrate how multiple factors, including (1) the addition of inappropriate amounts of inhibitors, (2) a tendency to overlook microbial syntrophy, and (3) the absence of comprehensive proxy systems of Hg methylation, would impact result interpretation of this approach. We thus suggest that the design of inhibition assays should consider the environmental properties, e.g., background levels of electron acceptors, concentrations of metabolic substrates, and abundances of methylating microbes. We also recommend that inhibitors should be added at multiple concentrations and that observed changes in Hg methylation should be assessed with comprehensive indicators. Revealing the key factors responsible for the improper usage of this method and inadequate interpretation of the results would help optimize inhibition assays for robust predictions of MeHg production in nature.

Rhizoctonia solani is a soilborne pathogen with a broad host range. An anastomosis group (AG) system based on hyphal fusions has been established to distinguish between different R. solani subgroups in this species complex. Members of the AG2-2IIIB subgroup can cause serious problems in sugar beet production, resulting in Rhizoctonia root and crown rot. In this review, we summarize the current molecular advances in the R. solani sugar beet pathosystem. The draft genome of R. solani AG2-2IIIB has an estimated size of 56.02 Mb, larger than any of the R. solani AGs sequenced to date. The genome of AG2-2IIIB has been predicted to harbor 11,897 protein-encoding genes, including a high number of carbohydrate-active enzymes (CAZymes). The highest number of CAZymes was observed for polysaccharide lyase family 1 (PL-1), glycoside hydrolase family 43 (GH-43), and carbohydrate esterase family 12 (CE-12). Eleven single-effector candidates were predicted based on AG2-2IIIB genome data. The RsLysM, RsRlpA, and RsCRP1 genes were highly induced upon early-stage infection of sugar beet seedlings, and heterologous expression in Cercospora beticola and model plant species demonstrated their involvement in virulence. However, despite the progress achieved thus far on the molecular interactions in this pathosystem, many aspects remain to be elucidated, including the development of efficient transformation systems, important for functional studies, and the silencing of undesirable traits in the sugar beet crop.

The escalating conflict between gillnet fisheries and the growing seal populations in Baltic Sea has increased the need to reduce direct effects in form of reduced or damaged catches. This study describes the design and catch rates of 20 different seal safe cod (Gadus morhua) pot designs suitable for the Baltic Sea. Catches in numbers of individuals and total catch weight were recorded in 2762 pot hauls with pots varying in terms of size, shape, number of entrances, entrance length, pot placement in relation to the sea floor and catch holding chamber. The aim was to investigate the how the different designs and soak times affected the catch rates under similar conditions. Volume and soak time was significantly positively correlated with catch rates and round bottom-standing pots with a fish holding chamber, funnels and 5 entrances had the highest catch rates. The findings are important for the development of cod pots, which can allow the fishers to continue fishing in areas of high seal density with static gear and hence low-carbon emission and minimum bycatch of marine mammals and sea birds.

In hilly areas, agroforestry can be a more sustainable way of producing food and other products and services than agriculture based on sole-cropping. However, research is needed to evaluate and quantify formation of natural terraces in agroforestry and their contribution to soil conservation. This study quantified natural terrace formation and examined its role in reducing soil and nutrient losses during early stages of agroforestry with fruit trees, contour grass strips and maize or coffee in agroforestry systems on sloping land in northwest Vietnam. Two agroforestry systems, comprising longan (Dimocarpus longan L.)-mango (Mangifera indica L.)-maize (Zea mays L.)-guinea grass (Panicum maximum Jacq.) (fruit-maize-AF) and son tra (Docynia indica (Wall.) Decne.)-coffee (Coffea arabica L.)-guinea grass (fruit-coffee-AF) were compared with sole-cropped maize (sole-maize) and sole-cropped coffee (sole-coffee), respectively. Terrace formation was evaluated over five years using erosion pins placed above grass strips and the volume of terrace formed was estimated. Soil and nutrient losses were quantified using soil traps. The results showed that terraces formed as the systems developed, through gradual deposition of soil sediment above the living grass strips and trees. Accumulated soil sedimentation above the grass strips during the five-year study period raised the soil surface by 4.0 cm in fruit-maize-AF and 4.2 cm in fruit-coffee-AF, and the volume of terraces generated by the grass strips was 0.26 and 0.43 m³/m respectively. The fruit-maize-AF and fruit-coffee-AF systems significantly reduced losses of soil, soil organic carbon (SOC) and associated nutrients (N, P, K) compared with sole-maize and sole-coffee already in the first two years, while the reductions were greater from year 3 onwards. On average across experiments and years, the agroforestry systems reduced soil, SOC, N, P and K losses by 27–76%, 21–78%, 20–82%, 24–82% and 22–84%, respectively. These findings show that agroforestry with fruit trees, grass strips and crops could be a useful management practice and viable option for sustainable agricultural systems on sloping land, by reducing soil (and carbon and nutrient) losses through terrace formation.

The northern pike (Esox lucius) is an iconic predatory fish species of significant recreational value and ecological role in the Baltic Sea. Some earlier studies indicate local declines of pike in the region, but a thorough spatial evaluation of regional population trends of pike in the Baltic Sea is lacking. In this study, we collate data from 59 unique time-series from fisheries landings and fishery-independent monitoring programs to address temporal trends in pike populations since the mid-2000′s in eight countries surrounding the Baltic Sea. In a common analysis considering all time-series in concert, we found indications of an overall regional temporal decline of pike in the Baltic Sea, but trends differed among countries. Individual negative trends in time-series were moreover found in several regions of the Baltic Sea, but predominantly so in the central and southern parts, while positive trends were only found in Estonia and northern Finland. The mix of data used in this study is inherently noisy and to some extent of uncertain quality, but as a result of the overall negative trends, together with the socioeconomic and ecological importance of pike in coastal areas of the Baltic Sea, we suggest that actions should be taken to protect and restore pike populations. Management measures should be performed in combination with improved fishery-independent monitoring programs to provide data of better quality and development of citizen-science approaches as a data source for population estimates. Possible measures that could strengthen pike populations include harvest regulations (including size limits, no-take areas and spawning closures), habitat protection and restoration, and an ecosystem-based approach to management considering also the impact of natural predators.

Europe has experienced a substantial increase in non-indigenous crayfish species (NICS) since the mid-20th century due to their extensive use in fisheries, aquaculture and, more recently, pet trade. Despite relatively long invasion histories of some NICS and negative impacts on biodiversity and ecosystemfunctioning, large spatio-temporal analyses of their occurrences are lacking. Here, we used a large freshwater macroinvertebrate database to evaluate what information on NICS can be obtained from widely applied biomonitoring approaches and how usable such data is for descriptions of trends in identified NICS species. We found 160 time-series containing NICS between 1983 and 2019, to infer temporal patterns and environmental drivers of species and region-specific trends. Using a combination of metaregression and generalized linear models, we found no significant temporal trend for the abundance of any species (Procambarus clarkii, Pacifastacus leniusculus or Faxonius limosus) at the European scale, but identified species-specific predictors of abundances. While analysis of the spatial range expansion of NICS was positive (i.e. increasing spread) in England and negative (significant retreat) in northern Spain, no trend was detected in Hungary and the Dutch- German-Luxembourg region. The average invasion velocity varied among countries, ranging from 30 km/year in England to 90 km/year in Hungary. The average invasion velocity gradually decreased over time in the long term, with declines being fastest in the Dutch-German-Luxembourg region, and much slower in England. Considering that NICS pose a substantial threat to aquatic biodiversity across Europe, our study highlights the utility and importance of collecting high resolution (i.e. annual) biomonitoring data using a sampling protocol that is able to estimate crayfish abundance, enabling a more profound understanding of NICS impacts on biodiversity.

Drinking water treatment residual (DWTR) derived from flocculation and sedimentation of raw water using aluminum coagulants is a valuable environmental remediation byproduct capable of inactivating phosphorus (P). However, no generalizable model exists in the literature to describe reduction of releasable (mobile) P in lake sediment as a result of DWTR addition. The reduction of mobile P (sum of labile P and reductant soluble P) was investigated in over 100 sub-samples using five sediment samples from two lakes and three DWTRs from different water treatment plants. A consistent relationship was determined across a range of mobile P contents (0.23 g/ m2/cm to 0.92 g/m2/cm, or 15.8 to 186.1 μg/g DW) and DWTRs. The relationship was best described as a function of the mobile P content of the sediment and the oxalate-extractable aluminum content of the DWTR. An empirical model was developed to predict the immediate reduction in mobile P following the addition of DWTR containing aluminum. This model was validated using two additional lake sediments and one additional DWTR (R2 = 0.995). Thus, the immediate inactivation of P in lake sediment following DWTR addition can be predicted with this model, which can be used with internal P loading or other water quality goals to determine an appropriate DWTR dose. Further recommendations were made about dosing DWTRs for lake restoration, allowing practitioners to use DWTR to inactivate P in lake sediment without conducting individual sorption experiments.

Mitigating enteric methane (CH4) emissions is crucial as ruminants account for 5% of global greenhouse gas emissions. We hypothesised that less frequent harvesting, use of crops with lower WSC concentration, ensiling at low crop dry matter (DM) and extensive lactic acid fermentation would reduce in vitro CH4 production. Timothy (T), timothy + red clover mixture (T + RC) or perennial ryegrass (RG), cut either two or three times per season, was wilted to 22.5% or 37.5% DM and ensiled with or without formic acid-based additive. Silages were analysed for chemical composition and fermentation products. In vitro CH4 production was measured using an automated gas in vitro system. Methane production was, on average, 2.8 mL/g OM lower in the two-cut system than in the three-cut system (P < 0.001), and 1.9 mL/g OM lower in T than in RG (P < 0.001). Silage DM did not affect CH4 production (P = 0.235), but formic acid increased CH4 production by 1.2 mL/g OM compared to the untreated silage (P = 0.003). In conclusion, less frequent harvesting and extensive silage fermentation reduce in vitro CH4 production, while RG in comparison to T resulted in higher production of CH4.

Diversifying agriculture by rotating a greater number of crop species in sequence is a promising practice to reduce negative impacts of crop production on the environment and maintain yields. However, it is unclear to what extent cereal yields change with crop rotation diversity and external nitrogen fertilization level over time, and which functional groups of crops provide the most yield benefit. Here, using grain yield data of small grain cereals and maize from 32 long-term (10–63 years) experiments across Europe and North America, we show that crop rotational diversity, measured as crop species diversity and functional richness, enhanced grain yields. This yield benefit increased over time. Only the yields of winter-sown small grain cereals showed a decline at the highest level of species diversity. Diversification was beneficial to all cereals with a low external nitrogen input, particularly maize, enabling a lower dependence on nitrogen fertilisers and ultimately reducing greenhouse gas emissions and nitrogen pollution. The results suggest that increasing crop functional richness rather than species diversity can be a strategy for supporting grain yields across many environments.

Bioaccumulation of organic contaminants from contaminated food sources, might pose an underestimated risk towards shredding invertebrates. This assumption is substantiated by monitoring studies observing discrepancies of predicted tissue concentrations determined from lab-based experiments compared to measured concentrations of systemic pesticides in gammarids. To elucidate the role of dietary uptake in bioaccumulation, gammarids were exposed to leaf material from trees treated with a systemic fungicide mixture (azoxystrobin, cyprodinil, fluopyram and tebuconazole), simulating leaves entering surface waters in autumn. Leaf concentrations, spatial distribution, and leaching behaviour of fungicides were characterised using LC-HRMS/MS and MALDI-MS imaging. The contribution of leached fungicides and fungicides taken up from feeding was assessed by assembling caged (no access) and uncaged (access to leaves) gammarids. The fungicide dynamics in the test system were analysed using LC-HRMS/MS and toxicokinetic modelling. Additionally, a summer scenario was simulated where water was the initial source of contamination and leaves contaminated by sorption. The uptake, translocation and biotransformation of systemic fungicides by trees was compound dependent. Internal fungicide concentrations of gammarids with access to leaves were much higher than in caged gammarids of the autumn scenario, but the difference was minimal in the summer scenario. In food choice and dissectioning experiments gammarids did not avoid contaminated leaves and efficiently assimilated contaminants from leaves indicating the relevance of this exposure pathway in the field. The present study demonstrates the potential impact of dietary uptake on in situ bioaccumulation for shredders in autumn, outside the main application period. Obtained toxicokinetic parameters facilitate modelling of environmental exposure scenarios. The uncovered significance of the dietary uptake for detritivores warrants further consideration from scientific but also regulatory perspectives. This article is protected by copyright. All rights reserved. Environ Toxicol Chem 2023;00:0-0. © 2023 SETAC.

Purpose The purpose of using circularity indicators is to show the effect of changes from linear to more circular systems. This paper contributes to highlighting the importance of methodological aspects of circularity indicators in the agricultural sector when using a life cycle thinking approach. Selected circularity indicators have been explored and compared with LCA impact categories by using them to evaluate the circularity of a livestock system. Methods Circularity indicators were tested on a theoretical pig production system where several circularity strategies and associated mitigation actions were applied. The strategies and mitigation actions were as follows: anaerobic digestion of manure (closing resource loops), anaerobic digestion of bread waste (closing resource loops), precision fertilization (narrowing resource loops), use of cover crops in feed production (regenerating resource flows), and use of bread waste as feed (slowing resource loops). The functional unit was 1 kg pork as carcass weight, and the treatment of 1.1 kg bread waste for all impact categories and indicators. For each mitigation action, relevant circularity indicators were tested. Based on this, the functionality and suitability of these indicators were discussed. Results and discussion Four of the circularity indicators were based on nitrogen (N) or phosphorus (P) substances: N recycling index, partial N balance, consumption of fossil-P fertilizers, and emissions to water bodies (P). Even if the indicators do not capture the impact of emissions of N and P as the eutrophication impact categories, they provide a useful indication of the circularity of a system. The other three circularity indicators tested were as follows: renewable energy production, soil organic carbon, and land use ratio. The renewable energy production indicator is easy to understand and communicate and provides unique information. Soil organic carbon presents a potential for soil carbon sequestration. Land use ratio is based on the same data as land occupation but provides an assessment of whether feed production competes for the suitable area for food production by including production of human-digestible protein. Conclusions Circularity indicators provide valuable information about the circularity of an agricultural product system. The circularity indicators and LCA impact categories can be used either separately or together, or to complement each other. The choice of indicators depends on the questions raised, i.e., goals and scope, and it is therefore important to have a number of circular indicators to choose from in order to achieve a comprehensive assessment.

Climate change, biodiversity loss and chemical pollution are planetary-scale emergencies requiring urgent mitigation actions. As these "triple crises" are deeply interlinked, they need to be tackled in an integrative manner. However, while climate change and biodiversity are often studied together, chemical pollution as a global change factor contributing to worldwide biodiversity loss has received much less attention in biodiversity research so far. Here, we review evidence showing that the multifaceted effects of anthropogenic chemicals in the environment are posing a growing threat to biodiversity and ecosystems. Therefore, failure to account for pollution effects may significantly undermine the success of biodiversity protection efforts. We argue that progress in understanding and counteracting the negative impact of chemical pollution on biodiversity requires collective efforts of scientists from different disciplines, including but not limited to ecology, ecotoxicology and environmental chemistry. Importantly, recent developments in these fields have now enabled comprehensive studies that could efficiently address the manifold interactions between chemicals and ecosystems. Based on their experience with intricate studies of biodiversity, ecologists are well equipped to embrace the additional challenge of chemical complexity through interdisciplinary collaborations. This offers a unique opportunity to jointly advance a seminal frontier in pollution ecology and facilitate the development of innovative solutions for environmental protection.

Background: Zoonotic livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is widely spread in pig herds in many countries. However, the knowledge regarding the survival of LA-MRSA in the pig farm environment is currently limited. The aim of this study was to assess the survival of LA-MRSA on different surface materials found in the farm environment. The study investigated the survival of two different LA-MRSA strains belonging to the clonal complex (CC) 398 on four different surfaces: stainless steel, polypropylene plastic, K30 concrete and commercial concrete disk coupons. The survival of the bacteria over time was determined by the viable count method and, where possible, fitting a model to the observed data by using nonlinear least squares method to calculate the half-life ([Formula: see text]) for different strain and material combinations. Results: The study showed that the half-life of the bacteria was longer on polypropylene plastic ([Formula: see text]=11.08-15.78 days) than on stainless steel ([Formula: see text]=2.45-7.83 days). On these materials, both LA-MRSA strains survived through the 14 week observation period. The bacterial decay was fastest on the concrete surfaces, where LA-MRSA became undetectable after 3-9 weeks. Conclusions: The survival of LA-MRSA in the pig farm environment may be affected by different surface materials. A more frequent sampling protocol (< 7 days) is needed to determine the half-life on concrete surfaces.

Small rodents are prevalent and functionally important across the world's biomes, making their monitoring salient for ecosystem management, conservation, forestry, and agriculture. There is a growing need for cost-effective and noninvasive methods for large-scale, intensive sampling. Fecal pellet counts readily provide relative abundance indices, and given suitable analytical methods, feces could also allow for the determination of multiple ecological and physiological variables, including community composition. In this context, we developed calibration models for rodent taxonomic determination using fecal near-infrared reflectance spectroscopy (fNIRS). Our results demonstrate fNIRS as an accurate and robust method for predicting genus and species identity of five coexisting subarctic microtine rodent species. We show that sample exposure to weathering increases the method's accuracy, indicating its suitability for samples collected from the field. Diet was not a major determinant of species prediction accuracy in our samples, as diet exhibited large variation and overlap between species. fNIRS could also be applied across regions, as calibration models including samples from two regions provided a good prediction accuracy for both regions. We show fNIRS as a fast and cost-efficient high-throughput method for rodent taxonomic determination, with the potential for cross-regional calibrations and the use on field-collected samples. Importantly, appeal lies in the versatility of fNIRS. In addition to rodent population censuses, fNIRS can provide information on demography, fecal nutrients, stress hormones, and even disease. Given the development of such calibration models, fNIRS analytics could complement novel genetic methods and greatly support ecosystem- and interaction-based approaches to monitoring.

Forest fragmentation increases the amount of edges in the landscape. Differences in wind, radiation and vegetation structure create edge-to-interior gradients in forest microclimate, and these gradients are likely to be more pronounced during droughts and heatwaves. Although the effects of climate extremes on edge influences have potentially strong and long-lasting impacts on forest understory biodiversity, they are not well understood and are not often considered in management and landscape planning. Here we used a novel method of retrospectively quantifying growth to assess biologically relevant edge influences likely caused by microclimate using Hylocomium splendens, a moss with annual segments. We examined how spatio-temporal variation in drought across three years and 46 sites in central Sweden, affected the depth and magnitude of edge influences. We also investigated whether edge effects during drought are influenced by differences in forest structure. Edge effects were almost twice as strong in the drought year as in the non-drought years, but we did not find clear evidence that they penetrated deeper into the forest in the drought year. Edge influences were also greater in areas that had fewer days with rain during the drought year. Higher levels of forest canopy cover and tree height buffered the magnitude of edge influence in times of drought. Our results demonstrate that edge effects are amplified by drought, suggesting that fragmentation effects are aggravated when droughts become more frequent and severe. Our results suggest that dense edges and buffer zones with high canopy cover can be important ways to mitigate negative drought impacts in forest edges.

Internal, slow-release implants can be an effective way to manipulate animal physiology or deliver a chemical exposure over long periods of time without the need for an exogenous exposure route. Slow-release implants involve dissolving a compound in a lipid-based carrier, which is inserted into the body of an organism. However, the release kinetics of the compound from the implant to body tissues also requires careful validation. Here, we tested and validated a slow-release implant methodology for exposing fish to a pharmaceutical pollutant, fluoxetine. We tested two lipid-based carriers (coconut oil or vegetable shortening) in the common roach (Rutilus rutilus). The implants contained either a high (50 μg/g), low (25 μg/g), or control (0 μg/g) concentration of fluoxetine, and we measured tissue uptake in the brain, muscle, and plasma of implanted fish over 25 days. The two carriers released fluoxetine differently over time: coconut oil released fluoxetine in an accelerating manner (tissue uptake displayed positive quadratic curvature), while vegetable shortening released fluoxetine in a decelerating manner (negative quadradic curvature). For both carrier types, fluoxetine was measured at the highest concentration in the brain, followed by muscle and plasma. We showed that the implants delivered an internal exposure that would be similar to if fish were exposed in surface waters containing effluents by comparing the implant exposures to waterborne exposures in the published literature. Overall, we show that slow-release internal implants are an effective method for delivering chronic exposures of fluoxetine over at least one-month timescales. Internal exposures can be an especially powerful experimental tool when coupled with field-based study designs to assess the impacts of pharmaceutical pollutants in complex, natural environments. This article is protected by copyright. All rights reserved. Environ Toxicol Chem 2023;00:0-0. © 2023 SETAC.

Ammonia (NH3) volatilization, nitrous oxide (N2O) emissions, and nitrate (NO3-) leaching from agriculture cause severe environmental hazards. Research studies and mitigation strategies have mostly focused on one of these nitrogen (N) losses at a time, often without an integrated view of the agro-food system. Yet, at the regional scale, N2O, NH3, and NO3- loss patterns reflect the structure of the whole agro-food system. Here, we analyzed at the resolution of NUTS2 administrative European Union (EU) regions, N fluxes through the agro-food systems of a Temperate-Mediterranean gradient (France, Spain, and Portugal) experiencing contrasting climate and soil conditions. We assessed the atmospheric and hydrological N emissions from soils and livestock systems. Expressed per ha agricultural land, NH3 volatilization varied in the range 6.2-44.4 kg N ha-1 yr-1, N2O emission and NO3 leaching 0.3-4.9 kg N ha-1 yr-1 and 5.4-154 kg N ha-1 yr-1 respectively. Overall, lowest N2O emission was found in the Mediterranean regions, where NO3- leaching was greater. NH3 volatilization in both temperate and Mediterranean regions roughly follows the distribution of livestock density. We showed that these losses are also closely correlated with the level of fertilization intensity and agriculture system specialization into either stockless crop farming or intensive livestock farming in each region. Moreover, we explored two possible future scenarios at the 2050 horizon: (1) a scenario based on the prescriptions of the EU-Farm-to-Fork (F2F) strategy, with 25% of organic farming, 10% of land set aside for biodiversity, 20% reduction in N fertilizers, and no diet change; and (2) a hypothetical agro-ecological (AE) scenario with generalized organic farming, reconnection of crop and livestock farming, and a healthier human diet with an increase in the share of vegetal protein to 65% (i.e., the Mediterranean diet). Results showed that the AE scenario, owing to its profound reconfiguration of the entire agro-food system would have the potential for much greater reductions in NH3, N2O, and NO3- emissions, namely, 60-81% reduction, while the F2F scenario would only reach 24-35% reduction of N losses.

Exchange of viral segments between one or more influenza virus subtypes can contribute to a shift in virulence and adaptation to new hosts. Among several influenza subtypes, H9N2 is widely circulating in poultry populations worldwide and has the ability to infect humans. Here, we studied the reassortant compatibility between chicken H9N2 with N1–N9 gene segments of wild bird origin, either with an intact or truncated stalk. Naturally occurring amino acid deletions in the NA stalk of the influenza virus can lead to increased virulence in both mallard ducks and chickens. Our findings show extended genetic compatibility between chicken H9Nx gene segments and the wild-bird NA with and without 20 amino acid stalk deletion. Replication kinetics in avian, mammalian and human cell lines revealed that parental chH9N2 and rH9N6 viruses with intact NA-stalk replicated significantly better in avian DF1 cells compared to human A549 cells. After introducing a stalk deletion, an enhanced preference for replication in mammalian and human cell lines could be observed for rH9N2 Δ (H6), rH9N6 Δ and rH9N9 Δ compared to the parental chH9N2 virus. This highlights the potential emergence of novel viruses with variable phenotypic traits, warranting the continuous monitoring of H9N2 and co-circulating subtypes in avian hosts.

Several mass spectrometry and spectroscopic techniques have been used in the search for molecular biomarkers on Mars. A major constraint is their capability to detect and identify large and complex compounds such as peptides or other biopolymers. Multiplex immunoassays can detect these compounds, but antibodies must be produced for a large number of sequence-dependent molecular targets. Ancestral Sequence Reconstruction (ASR) followed by protein "resurrection" in the lab can help to narrow the selection of targets. Herein, we propose an immunoanalytical method to identify ancient and universally conserved protein/peptide sequences as targets for identifying ancestral biomarkers in nature. We have developed, tested, and validated this approach by producing antibodies to eight previously described ancestral resurrected proteins (three β-lactamases, three thioredoxins, one Elongation Factor Tu, and one RuBisCO, all of them theoretically dated as Precambrian), and used them as a proxy to search for any potential feature of them that could be present in current natural environments. By fluorescent sandwich microarray immunoassays (FSMI), we have detected positive immunoreactions with antibodies to the oldest β-lactamase and thioredoxin proteins (ca. 4 Ga) in samples from a hydrothermal environment. Fine epitope mapping and inhibitory immunoassays allowed the identification of well-conserved epitope peptide sequences that resulted from ASR and were present in the sample. We corroborated these results by metagenomic sequencing and found several genes encoding analogue proteins with significant matches to the peptide epitopes identified with the antibodies. The results demonstrated that peptides inferred from ASR studies have true counterpart analogues in Nature, which validates and strengthens the well-known ASR/protein resurrection technique and our immunoanalytical approach for investigating ancient environments and metabolisms on Earth and elsewhere.

The benefits and costs of wildlife are contingent on the spatial overlap of animal populations with economic and recreational human activities. By using a production function approach with dynamic spatial panel data models, we analyze the effects of human hunting and carnivore predation pressure on the value of ungulate game harvests. The results show evidence of dynamic spatial dependence in the harvests of roe deer and wild boar, but not in those of moose, which is likely explained by the presence of harvesting quotas for the latter. Results suggest the impact of lynx on roe deer harvesting values is reduced by 75% when spatial effects are taken into account. The spatial analysis confirms that policymakers’ aim to reduce wild boar populations through increased hunting has been successful, an effect that was only visible when considering spatial effects.