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1.

Introduction

The rising atmospheric CO2 level is continuously driving the dissolution of more CO2 into the oceans, and some emission scenarios project that the surface waters may reach 1000 μatm by the end of the century. It is not known if fish can detect moderately elevated CO2 levels, and if they avoid areas with high CO2. If so, avoidance behaviour to water with high CO2 could affect movement patterns and migrations of fish in the future. It is also being increasingly recognized that fish behaviour can be altered by exposure to CO2. Therefore this study investigated how long-term exposure to elevated pCO2 affects predator avoidance and CO2 avoidance in juvenile Atlantic cod (Gadus morhua). The fish were exposed to control water or CO2-enriched water (1000 μatm) for six weeks before being subjected to tests of behaviour.

Results

Despite long term exposure to elevated pCO2 the cod still strongly avoided the smell of a predator. These data are surprising because several coral reef fish have demonstrated reversal of olfactory responses after CO2 exposure, turning avoidance of predator cues into preference for predator cues. Fish from both treatment groups also demonstrated strong avoidance of CO2 when presented with the choice of control or CO2-acidified water, indicating that habituation to the CO2 sensory stimuli is negligible.

Conclusions

As Atlantic cod maintained normal behavioural responses to olfactory cues, they may be tolerant to CO2-induced behavioural changes. The results also suggest that despite the long-term exposure to CO2-acidified water, the fish still preferred the control water over CO2-acidified water. Therefore, in the future, fish may alter their movements and migrations in search of waters with a lower CO2 content.
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2.

Introduction

The aphid Rhopalosiphum padi L. is a vector of Barley yellow dwarf virus (BYDV) in wheat and other economically important cereal crops. Increased atmospheric CO2 has been shown to alter plant growth and metabolism, enhancing BYDV disease in wheat. However, the biochemical influences on aphid metabolism are not known.

Objectives

This work aims to determine whether altered host-plant quality, influenced by virus infection and elevated CO2, impacts aphid weight and metabolism.

Methods

Untargeted 1H NMR metabolomics coupled with multivariate statistics were employed to profile the metabolism of R. padi reared on virus-infected and non-infected (sham-inoculated) wheat grown under ambient CO2 (aCO2, 400 µmol mol?1) and future, predicted elevated CO2 (eCO2, 650 µmol mol?1) concentrations. Un-colonised wheat was also profiled to observe changes to host-plant quality (i.e., amino acids and sugars).

Results

The direct impacts of virus or eCO2 were compared. Virus presence increased aphid weight under aCO2 but decreased weight under eCO2; whilst eCO2 increased non-viruliferous (sham) aphid weight but decreased viruliferous aphid weight. Discriminatory metabolites due to eCO2 were succinate and sucrose (in sham wheat), glucose, choline and betaine (in infected wheat), and threonine, lactate, alanine, GABA, glutamine, glutamate and asparagine (in aphids), irrespective of virus presence. Discriminatory metabolites due to virus presence were alanine, GABA, succinate and betaine (in wheat) and threonine and lactate (in aphids), irrespective of CO2 treatment.

Conclusion

This study confirms that virus and eCO2 alter host-plant quality, and these differences are reflected by aphid weight and metabolism.
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3.

Background

Stress urinary incontinence (SUI) is a relatively common disorder that significantly affects the quality of life. Many conservative and surgical treatment methods have been recommended for SUI, but they have major limitations.

Aims

To assess the use of the CO2 fractional laser in the treatment of SUI.

Methods

This clinical trial included 55 patients with confirmed SUI. Patients underwent fractional CO2 laser treatment 3 times at 30-day intervals. Data on age, smoking history, sexual activity, menopause, and history of hormone replacement therapy (HRT) were collected. Response to treatment was assessed by SUI severity and the level of sexual satisfaction was assessed using the visual analog scale (VAS). Patients were evaluated at 3 different time points: before treatment, and 45 days and 6 months after the last laser treatment.

Results

The mean patient age was 44.4±11.4 years (range: 28 to 68 years). Smoking history was positive in 6 patients (9.1%); 19 (54.3%) were menopausal on HRT. The SUI severity score at baseline (before treatment) was 8.56±0.62 and decreased to 2.28 6 months after treatment (p<0.0001). The sexual satisfaction score was 3±0.94 at baseline and increased to 7.87±0.93 6 months after treatment (day 180) (p<0.0001, slope = + 2.2)

Conclusion

Our findings are in line with a previous study that showed the value of transvaginal CO2 fractional laser treatment for alleviation of SUI symptoms and its potential as an alternative treatment. We also observed improved sexual satisfaction in SUI patients.
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4.

Background

The greenhouse gas (GHG) mitigation is one of the most important environmental benefits of using bioenergy replacing fossil fuels. Nitrous oxide (N2O) and methane (CH4) are important GHGs and have drawn extra attention for their roles in global warming. Although there have been many works of soil emissions of N2O and CH4 from bioenergy crops in the field scale, GHG emissions in large area of marginal lands are rather sparse and how soil temperature and moisture affect the emission potential remains unknown. Therefore, we sought to estimate the regional GHG emission based on N2O and CH4 releases from the energy crop fields.

Results

Here we sampled the top soils from two Miscanthus fields and incubated them using a short-term laboratory microcosm approach under different conditions of typical soil temperatures and moistures. Based on the emission measurements of N2O and CH4, we developed a model to estimate annual regional GHG emission of Miscanthus production in the infertile Loess Plateau of China. The results showed that the N2O emission potential was 0.27 kg N ha?1 year?1 and clearly lower than that of croplands and grasslands. The CH4 uptake potential was 1.06 kg C ha?1 year?1 and was slightly higher than that of croplands. Integrated with our previous study on the emission of CO2, the net greenhouse effect of three major GHGs (N2O, CH4 and CO2) from Miscanthus fields was 4.08 t CO2eq ha?1 year?1 in the Loess Plateau, which was lower than that of croplands, grasslands and shrub lands.

Conclusions

Our study revealed that Miscanthus production may hold a great potential for GHG mitigation in the vast infertile land in the Loess Plateau of China and could contribute to the sustainable energy utilization and have positive environmental impact on the region.
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5.

Aims

Despite extensive studies on effects of elevated CO2 concentration ([CO2]e) on plant growth, few studies have investigated the responses of native grassland plant species to [CO2]e in terms of nutrient acquisition.

Methods

The effects of [CO2]e (769 ± 23 ppm) on Artemisia frigida and Stipa krylovii, two dominant species in Inner Mongolia steppe were investigated by growing them for 7 weeks in Open-Top Chambers (OTC).

Results

Exposure to [CO2]e enhanced shoot and root growth of A. frigida and S. krylovii. Elevated [CO2] increased photosynthetic rates (Pn) by 34 % in A. frigida but decreased Pn by 52 % in S. krylovii. Moreover, root-secreted acid phosphatase activity in A. frigida was stimulated by [CO2]e, while exudation of malate from roots of S. krylovii was suppressed by [CO2]e. Exposure to [CO2]e led to a decrease in P concentration in shoots and roots of A. frigida and S. krylovii, but total amount of P accumulated in shoots and roots of both species was increased by [CO2]e.

Conclusions

The two dominant species in temperate steppes differed in their responses to [CO2]e, such that A. frigida was more adapted to [CO2]e than S. krylovii under low availability of soil P.
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6.

Background and aims

Saline and alkali soils severely impact plant growth. Endophyte and plant associations are known to significantly modify plant metabolism. This study reports the effects of a type of endophyte on organic acid (OA) accumulation and ionic balance in rice under Na2CO3 stress.

Methods

Rice seedlings with (E+) and without (E-) endophytic infection were subjected to different levels of Na2CO3 stress (0, 5, 10, 15, and 20 mM) for two weeks. Organic acids and mineral elements in the leaves and roots were determined.

Results

Seedlings with endophytic infection accumulated mainly citrate and fumarate, with some malate and succinate in the leaves. In the roots, accumulation of malate and fumarate was enhanced significantly by endophytic infection, while less citrate and succinate was accumulated under Na2CO3 stress, which suggested that leaves and roots use different mechanisms to control OA metabolism. Endophytes reduced the total Na and Na:K ratios, but increased ST values, the percent changes of other measured nutrients, Chl content, and dry weight per plant under Na2CO3 stress.

Conclusions

Endophytic infection plays a key role in maintaining plant growth by improving nutrient uptake and adjusting OA accumulation under Na2CO3 stress. The application of endophytes can enhance the resistance of rice to salinity.
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7.

Objectives

To investigate the feasibility of coupling carbonyl cyanide m-chlorophenylhydrazone-regulated photohydrogen production by Tetraselmis subcordiformis in a photobioreactor to an alkaline fuel cell (AFC).

Results

H2 evolution kinetics in the AFC integrated process was characterized. The duration of H2 evolution was prolonged and its yield was improved about 1.5-fold (to 78 ± 5 ml l?1) compared with that of the process without AFC. Improved H2 yield was possibly caused by removal of H2 feedback inhibition by H2 consumption in situ. Decreases in the H2 production rate correlated with the gradual deactivation of PSII and hydrogenase activities. The H2 yield was closely associated with catabolism of starch and protein.

Conclusion

A marine green algal CO2-supplemented culture integrated with in situ H2-consumption by an AFC system was developed as a viable protocol for the H2 production.
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8.

Objectives

To degrade enzymatically bisphenol A (BPA) that causes serious environmental concerns and is difficult to be degraded by chemical or physical methods.

Results

BPA (150 mg l?1) was completely degraded by chloroperoxidase (CPO)/H2O2 within 7 min at room temperature, atmospheric pressure with the enzyme at 6 μg CPO ml?1. The degradation products were identified by HPLC–MS, which suggested involvement of multiple steps. Enzymatic treatment followed by existing bioremediation technologies (activated sludge) enhanced removal of COD from 9 to 54 %. Using an ecotoxicity evaluation with Chlorella pyrenoidosa, the degradation products had a lower toxicity than BPA.

Conclusion

BPA can be degraded rapidly and efficiently under mild conditions with chloroperoxidase at 6 μg ml?1. The degradation products had a lower toxicity than BPA.
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9.

Background and Aims

The effects of Sb(V), alone or combined with Se, on the growth and root development of plants are unknown. The aim of this study is to investigate the interaction between selenite and different forms of Sb and the effects on their uptake in rice and on rice root morphology.

Methods

A hydroponic experiment was conducted that contained fourteen treatments. The treatment levels for Se were 0.5 and 1 mg L?1, and the treatment levels for Sb(III) and Sb(V) were 5 and 15 mg L?1.

Results

Sb(V) alone significantly reduced the surface area, mean diameter and volume of the roots, whereas Sb(III) alone reduced the values of most parameters of root morphology. The addition of 1 mg L?1 Se significantly enhanced the surface area, number of medium roots, and Sb concentration in the roots subjected to 15 mg L?1 Sb(V), but it decreased the number of root forks, the number and proportion of fine roots, and the shoot Sb concentration under exposure to 15 mg L?1 Sb(III). When the plants were subjected to 1 mg L?1 Se, the addition of 15 mg L?1 Sb(III) markedly reduced the shoot and root Se concentrations and the number of root tips, root forks, and fine roots and increased the mean root diameter. However, the addition of Sb(V) did not significantly affect the root and shoot Se concentrations but significantly decreased the number of root forks and fine roots and increased the proportion of medium roots.

Conclusions

Se and Sb(III) showed antagonistic effects on uptake in the shoots, but not in the roots, of paddy rice. A range of Se concentrations could stimulate the uptake of Sb in both the shoots and roots of paddy rice exposed to Sb(V).
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10.

Objectives

To evaluate the remediation efficiency of Mucor hiemalis by comparing media elimination, uptake, and biotransformation of microcystin-LR with exposure to pure toxin versus a crude bloom extract.

Results

With exposure to the extract, the elimination rate of microcystin-LR from the media, which was 0.28 ng MC-LR l?1 h?1, was significantly higher compared to that achieved with exposure to the pure toxin (0.16 ng MC-LR l?1 h?1) after 24 h. However, intracellular breakdown of microcystin-LR was significantly lower in the extract exposed pellets compared to the pure toxin treated fungal pellets over time. This coincided with reduced intracellular glutathione S-transferase activity with crude extract exposure which could be responsible for the detection of only the glutathione conjugate of microcystin-LR.

Conclusion

This paper signifies the importance of using laboratory exposure scenarios which resemble conditions in nature to fully understand and evaluate remediation efficiency. There is merit in using M. hiemalis for mycoremediation of cyanotoxins in surface waters.
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11.

Purpose

Gold is one of the most significant metals in the world, with use in various sectors including the electronic, health, and fashion industries. The Philippines has the world’s third largest known Au deposits and is ranked 20th in global gold production. Of the country’s annual production, about 80% is from the small-scale gold mining (SSGM) sector. This work estimates the first location-specific life cycle energy use and CO2 emissions of SSGM establishments in the Philippines.

Methods

Process-based LCA was used with functional unit of 100 g Au and observed data from 2010 to 2011 for mining, comminution, recovery, and refining. Four gold production paths were observed in the provinces of Benguet and Camarines Norte, namely, amalgamation, cyanidation with carbon-in-leach (CIL), cyanidation with leaching with zinc, and combination of amalgamation and cyanidation with CIL.

Results and discussion

It was estimated that 3–18 g of Au was extracted for every ton of ore within 57–159 man-hours from mining to refining. Energy use estimates ranged from 3501 to 67,325 MJ/100 g Au, while CO2 emission estimates ranged from 398 to 5340 kg CO2/100 g Au. The combination of amalgamation and cyanidation with CIL processes was the least energy and carbon intensive, while cyanidation with CIL process was the most intensive. Electricity use accounted for 95–100% of total emissions, except in cyanidation with CIL where kerosene accounts for 77% of the total. Since SSGMs contributed 80% of the 40 tons of Au produced in the Philippines in 2014, the SSGM energy use was estimated to be between 1120 and 21,544 TJ and the CO2 emissions to be between 129 and 1726 ktons CO2. Energy estimates are most sensitive to refining process yield and electrical equipment efficiency.

Conclusions

The estimated life cycle emissions rate for SSGM in the Philippines is lower than available estimates of large-scale mining. Notwithstanding, given the sector’s reliance on fossil fuels for its energy needs and the Philippines’ pledge to reduce its CO2 footprint by 70% in 2030, every effort to mitigate energy use and CO2 emission counts. Three main recommendations toward energy consumption and CO2 emissions reduction in SSGMs are proposed: (1) policy to promote technologies that are energy-efficient and processes that maximize gold process yield, (2) effective Minahang Bayan (SSGM mining zone mandated by law) implementation to ensure use of higher-grade ores, and (3) adoption of renewable energy in Minahang Bayans to promote energy independence and mitigate CO2 emissions.
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12.

Aims

Hydro-biogeochemical processes in the rhizosphere regulate nutrient and water availability, and thus ecosystem productivity. We hypothesized that two such processes often neglected in rhizosphere models — diel plant water use and competitive cation exchange — could interact to enhance availability of K+ and NH4 +, both high-demand nutrients.

Methods

A rhizosphere model with competitive cation exchange was used to investigate how diel plant water use (i.e., daytime transpiration coupled with no nighttime water use, with nighttime root water release, and with nighttime transpiration) affects competitive ion interactions and availability of K+ and NH4 +.

Results

Competitive cation exchange enabled low-demand cations that accumulate against roots (Ca2+, Mg2+, Na+) to desorb NH4 + and K+ from soil, generating non-monotonic dissolved concentration profiles (i.e. ‘hotspots’ 0.1–1 cm from the root). Cation accumulation and competitive desorption increased with net root water uptake. Daytime transpiration rate controlled diel variation in NH4 + and K+ aqueous mass, nighttime water use controlled spatial locations of ‘hotspots’, and day-to-night differences in water use controlled diel differences in ‘hotspot’ concentrations.

Conclusions

Diel plant water use and competitive cation exchange enhanced NH4 + and K+ availability and influenced rhizosphere concentration dynamics. Demonstrated responses have implications for understanding rhizosphere nutrient cycling and plant nutrient uptake.
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13.

Objective

To investigate the inhibiting effect of formic acid on acetone/butanol/ethanol (ABE) fermentation and explain the mechanism of the alleviation in the inhibiting effect under CaCO3 supplementation condition.

Results

From the medium containing 50 g sugars l?1 and 0.5 g formic acid l?1, only 0.75 g ABE l?1 was produced when pH was adjusted by KOH and fermentation ended prematurely before the transformation from acidogenesis to solventogenesis. In contrast, 11.4 g ABE l?1 was produced when pH was adjusted by 4 g CaCO3 l?1. The beneficial effect can be ascribed to the buffering capacity of CaCO3. Comparative analysis results showed that the undissociated formic acid concentration and acid production coupled with ATP and NADH was affected by the pH buffering capacity of CaCO3. Four millimole undissociated formic acid was the threshold at which the transformation to solventogenesis occurred.

Conclusion

The inhibiting effect of formic acid on ABE fermentation can be alleviated by CaCO3 supplementation due to its buffering capacity.
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14.

Purpose

CD4+CD25+ regulatory T (Treg) cells are present in increased numbers in patients with advanced cancer and CD25+ T cell depletion potentiates tumour immunity in animal models. The aim of this study was to assess the feasibility and safety of adoptive transfer of CD25+ depleted autologous T cells in patients with advanced renal cell carcinoma and to examine resulting changes in lymphocyte subsets.

Patients and methods

Six patients with advanced renal cell carcinoma underwent leukapheresis followed by conditioning chemotherapy with cyclophosphamide and fludarabine. The autologous leukapheresis product was depleted of CD25+ cells using CliniMACS® System then re-infused into the patient.

Results

Efficient CD25+ depletion from all leukapheresis products was achieved and 0.55–5.87 × 107/kg CD3+ cells were re-infused. Chemotherapy related haematological toxicity was observed, but blood counts recovered in all patients allowing discharge after a mean inpatient stay of 21 days. One patient subsequently developed a rapidly progressive neurological syndrome. A transient reduction in CD25+ subset was noted in the peripheral blood of 5 out of 6 patients with evidence of increased T cell responses to PHA in 4 out of 6 patients. One patient showed increased specific proliferative responses to the tumour associated antigen h5T4 coinciding with the nadir of Treg cells.

Conclusions

Given the transient nature of the reduction in CD25+ subset and the observed toxicity there is a need to explore further strategies to improve the safety and efficacy of this approach. Nevertheless, the results provide proof of concept in potentiation of tumour antigen T cell responses when Treg cell levels are depleted.
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15.

Background

The understanding of bicarbonate kinetics and CO2 retention in the body is necessary to conduct amino acid tracer oxidation studies in both humans and laboratory animals. Significant metabolic activity is associated with eating which can affect bicarbonate steady state kinetics. A study was conducted to assess the impact of feeding regimen on the recovery of labelled bicarbonate and energy expenditure in adult female pigs (sows). Five catheterized sows (235 ± 5 kg) were fed semi-synthetic diets as: a single meal 2 h into the infusion after an overnight fast, or in eight hourly meals starting 2 h before the infusion. Oxygen consumption, CO2 production and 14CO2 recovery (ie fraction not retained) were determined during primed, constant intravenous infusions of NaH14CO3.

Results

The 14CO2 recovery (%) after fasting (58.1 ± 4.8) was lower than that after single meal feeding (78.8 ± 5.9) or hourly meal feeding (81.0 ± 2.6, P = 0.03). CO2 production correlated with 14CO2 recovery during hourly feeding (r = 0.40, P = 0.01); this relationship was not significant after single meal feeding (P = 0.30), probably due to physical activity-associated CO2 production.

Conclusions

The correlation of CO2 retention factors with CO2 production during hourly feeding suggests that this regimen should be preferred for future amino acid kinetics studies.
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16.

Background and aims

Elemental uptake in serpentine floras in eastern North America is largely unknown. The objective of this study was to determine major and trace element concentrations in soil and leaves of three native pseudo-metallophyte C4 grasses in situ at five sites with three very different soil types, including three serpentine sites, in eastern USA.

Methods

Pseudo-total and extractible concentrations of 15 elements were measured and correlated from the soils and leaves of three species at the five sites.

Results

Element concentrations in soils of pseudo-metallophytes varied up to five orders of magnitude. Soils from metalliferous sites exhibited higher concentrations of their characteristic elements than non-metalliferous. In metallicolous populations, elemental concentrations depended on the element. Concentrations of major elements (Ca, Mg, K) in leaves were lower than typical toxicity thresholds, whereas concentrations of Zn were higher.

Conclusions

In grasses, species can maintain relatively low metal concentrations in their leaves even when soil concentrations are richer. However, in highly Zn-contaminated soil, we found evidence of a threshold concentration above which Zn uptake increases drastically. Finally, absence of main characteristics of serpentine soil at one site indicated the importance of soil survey and restoration to maintain serpentinophytes communities and avoid soil encroachment.
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17.

Introduction

Despite the use of buffering agents the 1H NMR spectra of biofluid samples in metabolic profiling investigations typically suffer from extensive peak frequency shifting between spectra. These chemical shift changes are mainly due to differences in pH and divalent metal ion concentrations between the samples. This frequency shifting results in a correspondence problem: it can be hard to register the same peak as belonging to the same molecule across multiple samples. The problem is especially acute for urine, which can have a wide range of ionic concentrations between different samples.

Objectives

To investigate the acid, base and metal ion dependent 1H NMR chemical shift variations and limits of the main metabolites in a complex biological mixture.

Methods

Urine samples from five different individuals were collected and pooled, and pre-treated with Chelex-100 ion exchange resin. Urine samples were either treated with either HCl or NaOH, or were supplemented with various concentrations of CaCl2, MgCl2, NaCl or KCl, and their 1H NMR spectra were acquired.

Results

Nonlinear fitting was used to derive acid dissociation constants and acid and base chemical shift limits for peaks from 33 identified metabolites. Peak pH titration curves for a further 65 unidentified peaks were also obtained for future reference. Furthermore, the peak variations induced by the main metal ions present in urine, Na+, K+, Ca2+ and Mg2+, were also measured.

Conclusion

These data will be a valuable resource for 1H NMR metabolite profiling experiments and for the development of automated metabolite alignment and identification algorithms for 1H NMR spectra.
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18.

Objectives

To improve the production and activity of an alkaline zinc metalloprotease from Salinivibrio proteolyticus in response to ZnSO4 (ionic and nanoparticle forms) and low intensity direct electric current (LIDC).

Results

A DC of 50 µA for 10 min increased enzyme production from 35 to 53 U ml?1 when applied to the stationary phase bacterial cells. Zn2+ improved enzyme production better than zinc nanoparticles (52 vs. 43.5 U ml?1). Zinc nanoparticles (0.5 mM) added to an enzyme reaction mixture containing casein (0.65 %) and 20 mM Tris/HCl buffer (pH 8) improved enzyme activity more than Zn2+ (42 vs. 36 U ml?1).

Conclusion

LIDC exposure (50 µA, 10 min) to the stationary phase bacterial cells increases metalloprotease production in Salinivibrio. A low concentration of zinc nanoparticles (0.5 mM) increases maximum enzyme activity.
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19.
20.

Aims

Slow or failed tree regeneration after forest disturbance is increasingly observed in the central European Alps, potentially amplifying the carbon (C) loss from disturbance. We aimed at quantifying C dynamics of a poorly regenerating disturbance site with a special focus on the role of non-woody ground vegetation.

Methods

Soil CO2 efflux, fine root biomass, ground vegetation biomass, tree increment and litter input were assessed in (i) an undisturbed section of a ~ 110 years old Norway spruce stand, (ii) in a disturbed section which was clear-cut six years ago (no tree regeneration), and (iii) in a disturbed section which was clear-cut three years ago (no tree regeneration).

Results

Total soil CO2 efflux was similar across all stand sections (8.5 ± 0.2 to 8.9 ± 0.3 t C ha?1 yr.?1). The undisturbed forest served as atmospheric C sink (2.1 t C ha?1 yr.?1), whereas both clearings were C sources to the atmosphere. The source strength three years after disturbance (?5.5 t C ha?1 yr.?1) was almost twice as high as six years after disturbance (?2.9 t C ha?1 yr.?1), with declining heterotrophic soil respiration and the high productivity of dense graminoid ground vegetation mitigating C loss.

Conclusions

C loss after disturbance decreases with time and ground vegetation growth. Dense non-woody ground vegetation cover can hamper tree regeneration but simultaneously decrease the ecosystem C loss. The role of ground vegetation should be more explicitly taken into account in forest C budgets assessing disturbance effects.
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