True Ileal Amino Acid Digestibility and Protein Quality of 15N-Labeled Faba Bean in Healthy Humans.

BACKGROUND: The recommended transition toward more plant-based diets, particularly containing legumes, requires a wider knowledge of plant protein bioavailability. Faba beans are cultivated at different latitudes and are used increasingly in human nutrition. OBJECTIVES: We aimed to assess the nutritional quality of faba bean protein in healthy volunteers equipped with an intestinal tube to implement the ileal 15N balance method. METHODS: Nine volunteers completed the study (7 males, 2 females, aged 33 ± 10 y, BMI: 24.7 ± 2.6 kg/m2). They were equipped with a nasoileal tube. After fasting overnight, they ingested a test meal consisting of cooked mash of dehulled faba bean seeds (20 g protein per serving of approximately 250 g) intrinsically labeled with 15N. Samples of ileal contents, plasma, and urine were collected over an 8-h postprandial period. Undigested nitrogen (N) and amino acids (AAs) were determined using isotopic MS, and subsequently, ileal digestibility and digestible indispensable amino acid score (DIAAS) were calculated. The measurement of postprandial deamination allowed calculation of the net postprandial protein utilization (NPPU). RESULTS: The ileal N digestibility was 84.1% ± 7.7%. Postprandial deamination represented 19.2% ± 3.6% of ingested N, and the NPPU was 64.7% ± 9.7%. The ileal digestibility of individual AAs varied from 85.1% ± 13.7% for histidine to 94.2% ± 3.6% for glutamine + glutamate. The mean AA digestibility was ∼6 percentage points higher than the digestibility of N, reaching 89.8% ± 5.9%, whereas indispensable AA digestibility was 88.0% ± 7.3%. Histidine and tryptophan were the first limiting AAs [DIAAS = 0.77 (calculated by legume-specific N-to-protein conversion factor 5.4); 0.67 (by default factor 6.25)]. Sulfur AAs were limiting to a lesser extent [DIAA ratio = 0.94 (N × 5.4); 0.81 (N × 6.25)]. CONCLUSIONS: Protein ileal digestibility of cooked, dehulled faba beans in humans was moderate (<85%), but that of AAs was close to 90%. Overall protein quality was restricted by the limited histidine and tryptophan content. This trial was registered at clinicaltrials.gov as NCT05047757.

Organic matter lability modifies the vertical structure of methane-related microbial communities in lake sediments.

Eutrophication increases the input of labile, algae-derived, organic matter (OM) into lake sediments. This potentially increases methane (CH4) emissions from sediment to water through increased methane production rates and decreased methane oxidation efficiency in sediments. However, the effect of OM lability on the structure of methane oxidizing (methanotrophic) and methane producing (methanogenic) microbial communities in lake sediments is still understudied. We studied the vertical profiles of the sediment and porewater geochemistry and the microbial communities (16S rRNA gene amplicon sequencing) at five profundal stations of an oligo-mesotrophic, boreal lake (Lake Pääjärvi, Finland), varying in surface sediment OM sources (assessed via sediment C:N ratio). Porewater profiles of methane, dissolved inorganic carbon (DIC), acetate, iron, and sulfur suggested that sites with more autochthonous OM showed higher overall OM lability, which increased remineralization rates, leading to increased electron acceptor (EA) consumption and methane emissions from sediment to water. When OM lability increased, the abundance of anaerobic nitrite-reducing methanotrophs (Candidatus Methylomirabilis) relative to aerobic methanotrophs (Methylococcales) in the methane oxidation layer of sediment surface decreased, suggesting that Methylococcales were more competitive than Ca. Methylomirabilis under decreasing redox conditions and increasing methane availability due to their more diverse metabolism (fermentation and anaerobic respiration) and lower affinity for methane. Furthermore, when OM lability increased, the abundance of methanotrophic community in the sediment surface layer, especially Ca. Methylomirabilis, relative to the methanogenic community decreased. We conclude that increasing input of labile OM, subsequently affecting the redox zonation of sediments, significantly modifies the methane producing and consuming microbial community of lake sediments. IMPORTANCE Lakes are important natural emitters of the greenhouse gas methane (CH4). It has been shown that eutrophication, via increasing the input of labile organic matter (OM) into lake sediments and subsequently affecting the redox conditions, increases methane emissions from lake sediments through increased sediment methane production rates and decreased methane oxidation efficiency. However, the effect of organic matter lability on the structure of the methane-related microbial communities of lake sediments is not known. In this study, we show that, besides the activity, also the structure of lake sediment methane producing and consuming microbial community is significantly affected by changes in the sediment organic matter lability.

Recycling eutrophic lake sediments into grass production: A four-year field experiment on agronomical and environmental implications.

Inefficient use of phosphorus (P) fertilizers leads to the transfer of P into water bodies, causing their eutrophication. Sediment removal is a promising lake restoration strategy that removes nutrients including P accumulated in lake sediments, and opens the opportunity to use removed nutrients in agriculture. In the present study, we investigated the effects of using a thick layer of sediment from the eutrophic Lake Mustijärv on plant growth, and estimated the environmental impacts of different sediment application methods by analyzing greenhouse gas emissions, N and P leaching, aggregate stability, and soil biota. The field experiment (2017-2020) was established on the lake shore with the following treatments: the agricultural control soil (Soil) surrounding the lake, pure sediment (Sed), biochar-treated sediment (SB), and biochar and soil mixed with sediment (SSB). The sediment-based treatments resulted in a similar grass growth performance to the Soil. The availability of most macro- and micronutrients including P (75 vs. 21 g m-3) were far greater in the Sed compared to the Soil. The sediment-based growing media emitted more CO2 than the Soil (579 vs. 400 mg CO2 - C m-2 h-1) presumably due to the high rate of organic matter decomposition. The bacterial and fungal community structures of the Sed were strongly differentiated from those of Soil. Also, Sed had lower bacterial diversity and a higher abundance of the bacterial phyla associated with solubilizing P including Proteobacteria and Chloroflexi. Sediment-based growing media increased more than seven times the risk of mineral N and P leaching, and the biochar treatment only had a short-lived beneficial effect on reduction of the sediment's leached P concentration. The sediment application rate should be adjusted to match the crop requirements to minimize greenhouse gas emissions and nutrient leaching when upscaling the case study to larger lakes with similar sediment properties.

Effects of liming on oxic and anoxic N2O and CO2 production in different horizons of boreal acid sulfate soil and non-acid soil under controlled conditions.

In acid sulfate (AS) soils, organic rich topsoil and subsoil horizons with highly variable acidity and moisture conditions and interconnected reactions of sulfur and nitrogen make them potential sources of greenhouse gases (GHGs). Subsoil liming can reduce the acidification of sulfidic subsoils in the field. However, the mitigation of GHG production in AS subsoils by liming, and the mechanisms involved, are still poorly known. We limed samples from different horizons of AS and non-AS soils to study the effects of liming on the N2O and CO2 production during a 56-day oxic and subsequent 72-h anoxic incubation. Liming to pH ≥ 7 decreased oxic N2O production by 97-98 % in the Ap1 horizon, 38-50 % in the Bg1 horizon, and 34-36 % in the BC horizon, but increased it by 136-208 % in the C horizon, respectively. Liming decreased anoxic N2O production by 86-94 % and 78-91 % in Ap1 and Bg1 horizons, but increased it by 100-500 % and 50-162 % in BC and C horizons, respectively. Liming decreased N2O/(N2O + N2) in anoxic denitrification in most horizons of both AS and non-AS soils. Liming significantly increased the cumulative oxic and anoxic CO2 production in AS soil, but less so in non-AS soil due to the initial high soil pH. Higher carbon and nitrogen contents in AS soil compared to non-AS soil agreed with the respectively higher cumulative oxic N2O production in all horizons, and the higher CO2 production in the subsoil horizons of all lime treatments. Overall, liming reduced the proportion of N2O in the GHGs produced in most soil horizons under oxic and anoxic conditions but reduced the total GHG production (as CO2 equivalents) only in the Ap1 horizon of both soils. The results suggest that liming of subsoils may not always effectively mitigate GHG emissions due to concurrently increased CO2 production and denitrification.

Recycling lake sediment to agriculture: Effects on plant growth, nutrient availability, and leaching.

Sediment removal from eutrophicated shallow lakes may not only be an effective method for lake restoration but also provides the potential for recycling nutrients from sediments to crop production. However, finding a suitable strategy for sustainably reusing the sediment remains a challenge. Therefore, current study focused on the best practices in applying the sediment from a shallow eutrophicated lake to the soil in terms of grass yield, nutrient uptake, and nutrient leaching. During a nine-month lysimeter experiment, 100-cm high columns were filled with six combinations of soil, sediment, and biochar, with or without meat bone meal organic fertilizer. Aboveground biomass, root mass distribution in soil, nutrient concentration, phosphorus (P) uptake of perennial ryegrass (Lolium perenne L.) along with easily soluble nutrients in the growing medium, and leached mineral nitrogen (N) and P levels were measured. Plant growth conditions were improved by sediment additions, as the yield and P uptake of ryegrass nearly doubled in treatments containing sediment compared to the control soil. While the sediment was richer in macro and micronutrients (e.g. P and N) compared to the soil, the leached N and P levels from both treatments were almost equivalent (N < 830 mg m-2 and P < 3 mg m-2). In addition, applying a 2-cm layer of biochar between the sediment and soil reduced P and N leaching by 50%. According to the results, applying a 75-cm thick layer of sediments on agricultural sandy loam soils surrounding the lake seems a promising practice for improving plant yield and soil nutrient status without increasing of P and N leaching from soil.

Internal phosphorus loading in a small shallow Lake: Response after sediment removal.

Mankind is taking advantage of numerous services by small shallow lakes such as drinking water supply, irrigation, and recreational function; however, many of these lakes suffer from eutrophication. Given the key role of phosphorus (P) in eutrophication process, one of the effective restoration methods especially for small shallow lakes is removal of sediments enriched with nutrients. In our study, we used interannual, seasonal, and spatial data to examine the changes in sediment P mobility after removal of sediments in 2016 from a 1-ha highly eutrophic lake. We measured the sediment redox potential, analyzed soluble reactive P (SRP) in the pore water and P fractional composition of the surface sediments, and calculated the diffusive flux of P in three locations in two continuous years (2017 and 2018) after the excavation. Similar measurements were done before sediment removal at central site of the lake in 2015. Removing nutrient-rich sediment also removed 6400 kg of P, and thus the potential for release of P from sediments decreased on a long-term scale. However, a large pool of releasable P was rebuilt soon after the sediment removal due to high external P loading, resulting in extensive anoxia of sediment surface and associated internal P loading as high as 1450 mg m-2 summer-1. Moreover, the Fe-P and labile P fractions were the most important sources of P release, as evidenced by their considerable seasonal and interannual changes after the sediment removal. The sediment total Fe negatively correlated with diffusive flux of P, pore water SRP, and near-bottom water total P and SRP concentrations which indicated a strong linkage between sediment P dynamics and Fe after the restoration. Sediment removal could be a beneficial restoration approach, but the effects on lake water quality remain only short-term unless there is an adequate control on external loading to the lake.

Strains of the toxic and bloom-forming Nodularia spumigena (cyanobacteria) can degrade methylphosphonate and release methane.

Nodularia spumigena is a nitrogen-fixing cyanobacterium that forms toxic blooms in the Baltic Sea each summer and the availability of phosphorous is an important factor limiting the formation of these blooms. Bioinformatic analysis identified a phosphonate degrading (phn) gene cluster in the genome of N. spumigena suggesting that this bacterium may use phosphonates as a phosphorus source. Our results show that strains of N. spumigena could grow in medium containing methylphosphonic acid (MPn) as the sole source of phosphorous and released methane when growing in medium containing MPn. We analyzed the total transcriptomes of N. spumigena UHCC 0039 grown using MPn and compared them with cultures growing in Pi-replete medium. The phnJ, phosphonate lyase gene, was upregulated when MPn was the sole source of phosphorus, suggesting that the expression of this gene could be used to indicate the presence of bioavailable phosphonates. Otherwise, growth on MPn resulted in only a minor reconstruction of the transcriptome and enabled good growth. However, N. spumigena strains were not able to utilize any of the anthropogenic phosphonates tested. The phosphonate utilizing pathway may offer N. spumigena a competitive advantage in the Pi-limited cyanobacterial blooms of the Baltic Sea.

Newtonian boreal forest ecology: The Scots pine ecosystem as an example.

Isaac Newton's approach to developing theories in his book Principia Mathematica proceeds in four steps. First, he defines various concepts, second, he formulates axioms utilising the concepts, third, he mathematically analyses the behaviour of the system defined by the concepts and axioms obtaining predictions and fourth, he tests the predictions with measurements. In this study, we formulated our theory of boreal forest ecosystems, called NewtonForest, following the four steps introduced by Newton. The forest ecosystem is a complicated entity and hence we needed altogether 27 concepts to describe the material and energy flows in the metabolism of trees, ground vegetation and microbes in the soil, and to describe the regularities in tree structure. Thirtyfour axioms described the most important features in the behaviour of the forest ecosystem. We utilised numerical simulations in the analysis of the behaviour of the system resulting in clear predictions that could be tested with field data. We collected retrospective time series of diameters and heights for test material from 6 stands in southern Finland and five stands in Estonia. The numerical simulations succeeded to predict the measured diameters and heights, providing clear corroboration with our theory.

Treating cattle with antibiotics affects greenhouse gas emissions, and microbiota in dung and dung beetles.

Antibiotics are routinely used to improve livestock health and growth. However, this practice may have unintended environmental impacts mediated by interactions among the wide range of micro- and macroorganisms found in agroecosystems. For example, antibiotics may alter microbial emissions of greenhouse gases by affecting livestock gut microbiota. Furthermore, antibiotics may affect the microbiota of non-target animals that rely on dung, such as dung beetles, and the ecosystem services they provide. To examine these interactions, we treated cattle with a commonly used broad-spectrum antibiotic and assessed downstream effects on microbiota in dung and dung beetles, greenhouse gas fluxes from dung, and beetle size, survival and reproduction. We found that antibiotic treatment restructured microbiota in dung beetles, which harboured a microbial community distinct from those in the dung they were consuming. The antibiotic effect on beetle microbiota was not associated with smaller size or lower numbers. Unexpectedly, antibiotic treatment raised methane fluxes from dung, possibly by altering the interactions between methanogenic archaea and bacteria in rumen and dung environments. Our findings that antibiotics restructure dung beetle microbiota and modify greenhouse gas emissions from dung indicate that antibiotic treatment may have unintended, cascading ecological effects that extend beyond the target animal.

Perennial crop growth in oil-contaminated soil in a boreal climate.

Soil contamination by petroleum hydrocarbons is a global problem. Phytoremediation by plants and their associated microorganisms is a cost-effective strategy to degrade soil contaminants. In boreal regions the cool climate limits the efficiency of phytoremediation. The planting of oil-tolerant perennial crops, especially legumes, in oil-contaminated soil holds promise for great economic benefits for bioenergy and bio-fertilizer production while accelerating the oil degradation process. We established a multi-year field experiment to study the ecological and agronomic feasibility of phytoremediation by a legume (fodder galega) and a grass (smooth brome) in a boreal climate. In 40 months, soil oil content decreased by 73%-92%, depending on the crop type. The oil degradation followed first-order kinetics with the reduction rates decreasing as follows: bare fallow > galega-brome grass mixture > brome grass > galega. Surprisingly, the presence of oil enhanced crop dry matter and nitrogen yield, particularly in the fourth year. The unfertilized galega-brome grass mixture out-yielded the N-fertilized pure grass swards over years by an average of 33%. Thus, a perennial legume-grass mixture is both ecologically and agronomically sustainable as a cropping system to alleviate soil contamination in the boreal zone, with considerable potential for bioenergy and bio-fertilizer production.

A multi-scale comparison of dissolved Al, Fe and S in a boreal acid sulphate soil.

Acid sulphate (AS) soils are most prevalent in the tropics, but the acidic discharge from cultivated AS soils also threatens water bodies under boreal conditions. Feasible options to reduce the acid load are needed. In this study, the groundwater of an AS field was monitored for 3.5 years, and the efficiency of waterlogging in mitigating the environmental risks caused by acidic discharge was investigated in a 2.5-year experiment with 10 monolithic lysimeters taken from the same field. In order to unravel the transferability of the results from lysimeters to the field scale, the Al, Fe and S concentrations in discharge water from the lysimeters were compared with those in the groundwater of the AS field (pedon and field scale), and in pore water (pedon and horizon scale). In the waterlogged bare lysimeters (HWB), the Al, Fe and S concentrations in discharge waters were broadly similar to those measured in the groundwater and followed the changes in the pore water. In the waterlogged cropped (reed canary grass, Phalaris arundinacea) lysimeters (HWC), in contrast, the discharge waters were markedly higher in Fe and lower in Al than the groundwater in the field. This outcome was attributable to the reduction of Fe(3+) to the more soluble Fe(2+) and the reduction-induced increase in pH, which enhanced the formation of Al(3+) hydroxy species. Lowering of the water table (LWC) caused soil ripening, which resulted in increased saturated hydraulic conductivity and porosity and enhanced the oxidation of sulphidic materials and acid formation. The responses of Al, Fe and S in drainage waters from HWC and LWC lysimeters resembled previous findings in AS soils. Based on this and the similarity between dissolved element concentrations in the discharge water of HWB lysimeters and groundwater in the field, we conclude that our monolithic lysimeters yielded realistic results concerning the efficiency of various methods in mitigating environmental risks related to cultivated AS soils.

Response of pore water Al, Fe and S concentrations to waterlogging in a boreal acid sulphate soil.

Environmental hazards caused by acid sulphate (AS) soils are of worldwide concern. Among various mitigation measures, waterlogging has mainly been studied in subtropical and tropical conditions. To assess the environmental relevance of waterlogging as a mitigation option in boreal AS soils, we arranged a 2.5-year experiment with monolithic lysimeters to monitor changes in the soil redox potential, pH and the concentrations of aluminium (Al), iron (Fe) and sulphur (S) in pore water in response to low and high groundwater levels in four AS soil horizons. The monoliths consisted of acidic oxidized B horizons and a reduced C horizon containing sulphidic material. Eight lysimeters were cropped (reed canary grass, Phalaris arundinacea) and two were bare without a crop. Waterlogging was conducive to reduction reactions causing a slight rise in pH, a substantial increase in Fe (Fepw) and a decrease in Al (Alpw) in the pore water. The increase in Fepw was decisively higher in the cropped waterlogged lysimeters than in the bare ones, which was attributable to the microbiologically catalysed reductive dissolution of poorly ordered iron oxides and secondary minerals. In contrast to warmer climates, Fepw concentrations remained high throughout the experiment, indicating that the reduction was poised in the iron range, while sulphate was not reduced to sulphide. Therefore, the precipitation of iron sulphide was negligible in the environment with a low pH and abundant with poorly ordered Fe oxides. Increased Fe in pore water counteracts the positive effects of waterlogging, when water is flushed from fields to watercourses, where re-oxidation of Fe causes acidity and oxygen depletion. However, waterlogging prevented further oxidation of sulphidic materials and decreased Alpw to one-tenth of the initial concentrations, and even to one-hundredth of the levels in the low water table lysimeters.

The microbial communities and potential greenhouse gas production in boreal acid sulphate, non-acid sulphate, and reedy sulphidic soils.

Acid sulphate (AS) soils along the Baltic coasts contain significant amounts of organic carbon and nitrogen in their subsoils. The abundance, composition, and activity of microbial communities throughout the AS soil profile were analysed. The data from a drained AS soil were compared with those from a drained non-AS soil and a pristine wetland soil from the same region. Moreover, the potential production of methane, carbon dioxide, and nitrous oxide from the soils was determined under laboratory conditions. Direct microscopic counting, glucose-induced respiration (GIR), whole cell hybridisation, and extended phospholipid fatty acid (PLFA) analysis confirmed the presence of abundant microbial communities in the topsoil and also in the deepest Cg2 horizon of the AS soil. The patterns of microbial counts, biomass and activity in the profile of the AS soil and partly also in the non-AS soil therefore differed from the general tendency of gradual decreases in soil profiles. High respiration in the deepest Cg2 horizon of the AS soil (5.66 µg Cg(-1)h(-1), as compared to 2.71 µg Cg(-1)h(-1) in a top Ap horizon) is unusual but reasonable given the large amount of organic carbon in this horizon. Nitrous oxide production peaked in the BCgc horizon of the AS and in the BC horizon of the non-AS soil, but the peak value was ten-fold higher in the AS soil than in the non-AS soil (82.3 vs. 8.6 ng Ng(-1)d(-1)). The data suggest that boreal AS soils on the Baltic coast contain high microbial abundance and activity. This, together with the abundant carbon and total and mineral nitrogen in the deep layers of AS soils, may result in substantial gas production. Consequently, high GHG emissions could occur, for example, when the generally high water table is lowered because of arable farming.

Quantifying beetle-mediated effects on gas fluxes from dung pats.

Agriculture is one of the largest contributors of the anthropogenic greenhouse gases (GHGs) responsible for global warming. Measurements of gas fluxes from dung pats suggest that dung is a source of GHGs, but whether these emissions are modified by arthropods has not been studied. A closed chamber system was used to measure the fluxes of carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O) from dung pats with and without dung beetles on a grass sward. The presence of dung beetles significantly affected the fluxes of GHGs from dung pats. Most importantly, fresh dung pats emitted higher amounts of CO2 and lower amounts of CH4 per day in the presence than absence of beetles. Emissions of N2O showed a distinct peak three weeks after the start of the experiment--a pattern detected only in the presence of beetles. When summed over the main grazing season (June-July), total emissions of CH4 proved significantly lower, and total emissions of N2O significantly higher in the presence than absence of beetles. While clearly conditional on the experimental conditions, the patterns observed here reveal a potential impact of dung beetles on gas fluxes realized at a small spatial scale, and thereby suggest that arthropods may have an overall effect on gas fluxes from agriculture. Dissecting the exact mechanisms behind these effects, mapping out the range of conditions under which they occur, and quantifying effect sizes under variable environmental conditions emerge as key priorities for further research.