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At least 55 records · Page 3

Multiple sources of aerobic methane production in aquatic ecosystems include bacterial photosynthesis

Aquatic ecosystems are globally significant sources of the greenhouse gas methane to the atmosphere. Until recently, methane production was thought to be a strictly anaerobic process confined primarily to anoxic sediments. However, supersaturation of methane in oxygenated waters has been consistently observed in lakes and the ocean (termed the ‘methane paradox’), indicating that methane can be produced under oxic conditions through unclear mechanisms. Here we show aerobic methane production from multiple sources in freshwater incubation experiments under different treatments and based on biogeochemical, metagenomic, and metatranscriptomic data. We find that aerobic methane production appears to be associated with (bacterio)chlorophyll metabolism and photosynthesis, as well as with Proteobacterial degradation of methylphosphonate. Genes encoding pathways for putative photosynthetic- and methylphosphonate-based methane production also co-occur in Proteobacterial metagenome-assembled genomes. Our findings provide insight into known mechanisms of aerobic methane production, and suggest a potential co-occurring mechanism associated with bacterial photosynthesis in aquatic ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗

Genome-guided isolation of the hyperthermophilic aerobe Fervidibacter sacchari reveals conserved polysaccharide metabolism in the Armatimonadota

Few aerobic hyperthermophilic microorganisms degrade polysaccharides. Here, we describe the genome-enabled enrichment and optical tweezer-based isolation of an aerobic polysaccharide-degrading hyperthermophile, Fervidibacter sacchari, previously ascribed to candidate phylum Fervidibacteria. F. sacchari uses polysaccharides and monosaccharides for growth at 65–87.5°C and expresses 191 carbohydrate-active enzymes (CAZymes) according to RNA-Seq and proteomics, including 31 with unusual glycoside hydrolase domains (GH109, GH177, GH179). Fluorescence in-situ hybridization and nanoscale secondary ion mass spectrometry confirmed rapid assimilation of 13 C-starch in spring sediments. Purified GHs were optimally active at 80–100°C on ten different polysaccharides. Finally, we propose reassigning Fervidibacteria as a class within phylum Armatimonadota, along with 18 other species, and show that a high number and diversity of CAZymes is a hallmark of the phylum, in both aerobic and anaerobic lineages. Our study establishes Fervidibacteria as hyperthermophilic polysaccharide degraders in terrestrial geothermal springs and suggests a broad role for Armatimonadota in polysaccharide catabolism.

59 BASIC BIOLOGICAL SCIENCES↗

Metabolic multistability and hysteresis in a model aerobe-anaerobe microbiome community

Major changes in the microbiome are associated with health and disease. Some microbiome states persist despite seemingly unfavorable conditions, such as the proliferation of aerobe-anaerobe communities in oxygen-exposed environments in wound infections or small intestinal bacterial overgrowth. Mechanisms underlying transitions into and persistence of these states remain unclear. Using two microbial taxa relevant to the human microbiome, we combine genome-scale mathematical modeling, bioreactor experiments, transcriptomics, and dynamical systems theory to show that multistability and hysteresis (MSH) is a mechanism describing the shift from an aerobe-dominated state to a resilient, paradoxically persistent aerobe-anaerobe state. We examine the impact of changing oxygen and nutrient regimes and identify changes in metabolism and gene expression that lead to MSH and associated multi-stable states. In such systems, conceptual causation-correlation connections break and MSH must be used for analysis. Using MSH to analyze microbiome dynamics will improve our conceptual understanding of stability of microbiome states and transitions between states.

59 BASIC BIOLOGICAL SCIENCES↗

Convergent evolution of aerobic fermentation through divergent mechanisms acting on the same key glycolytic genes

Expression profiling by high throughput sequencing The model yeast Saccharomyces cerevisiae has evolved aerobic fermentation in part through a whole genome duplication and subsequent retention of glycolytic genes. To identify independent yeast lineages with a high glycolytic rate phenotype we developed an assay measuring extracellular acidification rates (ECAR) to evaluate the glycolytic rates across diverse yeast species. This assay identified a novel group of yeasts in the genus Saturnispora with rapid glycolytic rates. Through comparative approaches, we found that several glycolytic genes encoding hexokinases and enzymes in lower glycolysis had higher expression and modifications in promoter sequences in rapid ECAR species. Intriguingly, many of the upregulated genes are the same genes that are duplicated in S. cerevisiae. When the transcription factor required for their activation was deleted, the mutants had a slow glycolytic rate and more respiratory phenotype. This work shows how the aerobic fermentation phenotype has convergently evolved in Saturnispora through transcriptional rewiring. This divergent mechanism nevertheless impacts the same glycolytic genes, which suggests that there are evolutionary constraints on how aerobic fermentation can arise.

Horianopoulos, Linda C. [Department of Food Scienc↗

Frequency and duration of interval training programs and changes in aerobic power

The present study was designed to ascertain whether a training frequency of 2 days/wk for a 7- and 13-wk interval training program would produce improvement in maximal aerobic power comparable to that obtained from 7- and 13-wk programs of the same intensity consisting of 4 training days/wk. After training, there was a significant increase in maximal aerobic power that was independent of both training frequency and duration. Maximal heart rate was significantly decreased following training. Submaximal aerobic power did not change with training, but submaximal heart rate decreased significantly with greater decreases the more frequent and the longer the training.

Fox, E. L.↗

Considerations in prescribing preflight aerobic exercise for astronauts

The physiological effects of prolonged exposure to weightlessness are discussed together with the effects of aerobic exercise on human characteristics affected by weightlessness. It is noted that, although early data on orthostatic intolerance after spaceflight led to a belief that a high level of aerobic fitness for astronauts was detrimental to orthostatic tolerance on return to earth, most of the data available today do not suport this contention. Aerobic fitness was found to be beneficial to cardiovascular function and to mental performance; therefore, it may be important in performing extravehicular activities during flight.

Frey, Mary Anne Bassett↗

Aerobic fitness and orthostatic tolerance: Evidence against an association

This presentation will focus on only one side of the debate as to whether high levels of aerobic fitness have a deleterious effect on tolerance to gravitational stress. This issue was raised in the early 1970's as a result of two research publications. The first work investigated the carotid sinus baroreflex of humans with an airtight chamber that surrounded the head and neck. The steady-state reflex changes in blood pressure that were recorded 3 minutes after application of the head and neck stimuli, were attenuated in an athletic group compared to a sedentary group of volunteers. A second report in the NASA literature indicated that five endurance-trained runners were less tolerant to LBNP than five nonrunners. These early research findings have stimulated a considerable amount of interest that has lead to a growing number of research efforts seeking an association between aerobic fitness and orthostatic tolerance in humans. I will briefly review some of the more pertinent published research information which suggests that there is no relationship between aerobic fitness and orthostatic tolerance in humans.

Ebert, Thomas J.↗

Logistic Risk Model for the Unique Effects of Inherent Aerobic Capacity on (+)G(sub z) Tolerance Before and After Simulated Weightlessness

Small sample size (n less than 1O) and inappropriate analysis of multivariate data have hindered previous attempts to describe which physiologic and demographic variables are most important in determining how long humans can tolerate acceleration. Data from previous centrifuge studies conducted at NASA/Ames Research Center, utilizing a 7-14 d bed rest protocol to simulate weightlessness, were included in the current investigation. After review, data on 25 women and 22 men were available for analysis. Study variables included gender, age, weight, height, percent body fat, resting heart rate, mean arterial pressure, Vo(sub 2)max and plasma volume. Since the dependent variable was time to greyout (failure), two contemporary biostatistical modeling procedures (proportional hazard and logistic discriminant function) were used to estimate risk, given a particular subject's profile. After adjusting for pro-bed-rest tolerance time, none of the profile variables remained in the risk equation for post-bed-rest tolerance greyout. However, prior to bed rest, risk of greyout could be predicted with 91% accuracy. All of the profile variables except weight, MAP, and those related to inherent aerobic capacity (Vo(sub 2)max, percent body fat, resting heart rate) entered the risk equation for pro-bed-rest greyout. A cross-validation using 24 new subjects indicated a very stable model for risk prediction, accurate within 5% of the original equation. The result for the inherent fitness variables is significant in that a consensus as to whether an increased aerobic capacity is beneficial or detrimental has not been satisfactorily established. We conclude that tolerance to +Gz acceleration before and after simulated weightlessness is independent of inherent aerobic fitness.

Ludwig, David A.↗

Stability Characteristics of "Aerobic" Acetyl-CoA Synthetase of Yeast

During the purification of the "aerobic" acetyl-CoA synthetase (ACS) of Saccharomyces cerevisiae, strain LK2Gl2, it was noted that stronge at 4 C resulted in the loss of enzyme activity within 24 hr. Similar losses were observed during column chromatography. Addition of boiled extracts from either aerobic or anerobic cells completely prevents this. The stabilizing factor (SF) in these extracts is non-dialyzable and organic in nature. SF is excluded on G-25 and G-50 Sephadex columns and is slightly retarded on G-75 columns. On G-100 columns, SF elutes as a peak exactly coincident with that of cytochrome c, indicating a molecular weight of 13,000. SF activity was not destroyed by Pronase treatment, was adsorbed onto Norite, and absorbed in the UV with a single maximum at 260 nm. The action of SF could be replaced by a number of nucleotides. At 0.01 M, the order of effectiveness was: ATP>ADP>AMP>GTP>CTP>/=UTP>XTP. Even at 2 x 10(exp -4) M, ATP and ADP, but not AMP, cyclic AMP, adenosine or adenine, were effective in stabilizing this ACS. The mechanism of stabilization by ATP and AMP appears to be the same, since AMP competitively inhibited the ACS with respect to ATP in in vitro assays, while ADP gave a mixed type of inhibition, thus indicating a different mechanism. ACS from nonaerobic cells is also unstable in the absence of SF but, unlike aerobic ACS, is not affected by ATP or other nucleotides.

Satyanarayana, T.↗

Aerobic growth of Anoxybacillus pushchinoensis K1(T): emended descriptions of A. pushchinoensis and the genus Anoxybacillus

In this work, corrections are made to the descriptions of the species Anoxybacillus pushchinoensis corrig. and the genus ANOXYBACILLUS: Experiments to determine the relationship of A. pushchinoensis K1(T) to oxygen showed that it was capable of aerobic growth, but preferred to grow anaerobically. During aerobic growth, the redox indicator resazurin was reduced as a result of hydrogen gas production. The facultatively anaerobic nature of K1(T) was ascertained by cultivation in aerobic liquid medium, where growth began at the bottom of the tube. The anaerobic nature of K1(T) was also indicated by a negative catalase reaction. This work is submitted to correct the description of the species A. pushchinoensis from obligate anaerobe to aerotolerant anaerobe and to emend the description of the genus Anoxybacillus from obligate anaerobes or facultative anaerobes to aerotolerant anaerobes or facultative anaerobes.

Bacillaceae/classification/growth & development/me↗

A two-year program of aerobics and weight training enhances bone mineral density of young women

Previous research suggests that physical activity may have a beneficial effect on bone mineral density (BMD) in women. This relationship was explored in a 2-year, randomized, intervention trial investigating the efficacy of exercise and calcium supplementation on increasing peak bone mass in young women. One hundred and twenty-seven subjects (ages of 20-35 years) were randomly assigned either to an exercise program that contained both aerobics and weight training components or to a stretching program. Calcium supplementation (up to 1500 mg/day including dietary intake) or placebo was given in a double-blinded design to all subjects. Spinal trabecular BMD was determined using quantitative computed tomography (QCT). Spinal integral, femoral neck, and trochanteric BMD were measured by dual X-ray absorptiometry (DXA) and calcaneal BMD by single photon absorptiometry (SPA). Fitness variables included maximal aerobic capacity (VO2max), and isokinetic muscle performance of the trunk and thigh. Measurements were made at baseline, 1 year, and 2 years. Sixty-three subjects (32 exercise, 31 stretching) completed the study, and all the measured bone parameters indicated a positive influence of the exercise intervention. There were significant positive differences in BMD between the exercise and stretching groups for spinal trabecular (2.5%), femoral neck (2.4%), femoral trochanteric (2.3%), and calcaneal (6.4%) measurements. The exercise group demonstrated a significant gain in BMD for spinal integral (1.3 +/- 2.8%, p < 0.02), femoral trochanteric (2.6 +/- 6.1%, p < 0.05), and calcaneal (5.6 +/- 5.1, p < 0.01) measurements. In contrast to exercise, the calcium intervention had no positive effect on any of the bone parameters. In regard to fitness parameters, the exercise group completed the study with significant gains in VO2max and isokinetic (peak torque) values for the knee flexion and extension and trunk extension. This study indicates that over a 2-year period, a combined regimen of aerobics and weight training has beneficial effects on BMD and fitness parameters in young women. However, the addition of daily calcium supplementation does not add significant benefit to the intervention.

Randomized Controlled Trial↗

Comparing the effects of two in-flight aerobic exercise protocols on standing heart rates and VO(2peak) before and after space flight

The effects of regular aerobic exercise on orthostatic tolerance have been the subject of a long-standing controversy that will influence the use of exercise during space flight. To examine these effects, astronauts performed continuous (CE) aerobic exercise (n = 8), interval (IE) aerobic exercise (n = 4), or no (NE) exercise (n = 5) during flights of 7 to 11 days. Heart rate (HR) responses to an orthostatic challenge (stand test) were measured 10 days before flight and on landing day. VO(2peak) (graded treadmill exercise) was measured 7 to 21 days before and 2 days after flight. No significant differences across the groups were observed in standing HRs before or after flight. However, the within-group mean HRs significantly increased in the NE (71-89 beats/min) and CE (60-85 beats/min) groups after space flight. The HRs for the IE group did not significantly increase (75-86 beats/min) after space flight. VO(2peak) decreased (P < .05) in the NE (-9.5%) group, but did not change in the CE (-2.4%) and IE (1%) groups. The relationship (r = 0.237) between the delta HR and delta VO(2peak) was not significant. These preliminary results indicate that: (1) continuous exercise does not affect the orthostatic HR response after space flight; (2) interval exercise may minimize an increase in the postflight orthostatic HR; and (3) both exercise protocols can maintain VO(2peak).

NASA Discipline Musculoskeletal↗

WIse-2005: Combined Aerobic and Resistive Exercise May Help Mitigate Bone Loss During 60-D Simulated Microgravity in Women

Exercise can attenuate bone loss associated with disuse during bed rest (BR), an analog of space flight. Previous studies have examined the efficacy of aerobic or resistive exercise countermeasures, but not in combination. We sought to determine the effect of a combined resistive and aerobic exercise regimen on bone metabolism during BR. After a 20-d ambulatory adaptation to confinement and diet, 16 women participated in a 60-d head-down-tilt BR. Control subjects (CN, n=8) performed no countermeasures. Exercise subjects, (EX, n=8) participated in exercise alternating daily between supine treadmill exercise within lower body negative pressure and resistive fly-wheel exercise (6-d wk(sup -1)). In the last week of BR, bone resorption was greater (p less than 79 plus or minus 44%, mean plus or minus SD) and EX groups (64 50%). N-telopeptide also increased (CN: 51 plus or minus 34%; EX: 43 plus or minus 56%). However, bone-specific alkaline phosphatase, a bone formation marker, tended to be higher in EX (26 plus or minus 18%) than in CN (8 plus or minus 33%) groups. The combination of resistive and aerobic exercise does not prevent bone resorption, but may promote formation, potentially mitigating the net bone loss associated with simulated microgravity. This study was supported by CNES, CSA, ESA, NASA, and NASA grant NNJ04HF71G to ARH. MEDES (French Institute for Space Medicine and Physiology) organized the study.

Smith, Scott M.↗

WISE-2005: LBNP/Treadmill and Resistive Exercise Countermeasures Maintain Aerobic Capacity during a 60-d Bed Rest

We have previously documented that supine treadmill exercise within lower body negative pressure (LBNPex) performed 6 sessions (raised dot) wk(sup -1) during 15- and 30-day bed rests (BR) maintained upright aerobic capacity (VO2pk). In the present study, ure are evaluating whether aerobic capacity is maintained during a 60-d BR when the LBNPex frequency is reduced to 2-4 sessions (raised dot) wk(sup -1) and resistance exercise (REX) is added 2-3 sessions (raised dot) wk(sup -1). Eight healthy women (32 plus or minus 4 yrs; 56.4 plus or minus 3.6 kg; 164 plus or minus 8 cm; mean plus or minus SD) performed maximal-exertion, graded treadmill tests before and 3 days after a 60-d, 6 deg. head-down tilt BR. (Earliest day the medical monitors would permit a maximal exercise test post-BR). During BR, four subjects performed no exercise (CON), while four other subjects (EX) performed LBNPex and REX on separate days. The LBNPex countermeasure employed an intermittent (40-80% pre-BR VO2pk), 40-min protocol against an LBNP pressure (-49 plus or minus 3 mmHg) applied to provide a footward force equivalent to 1.0-1.2 body weight. REX consisted of maximal concentric and eccentric supine leg press and heel raise exercises using a gravity-independent flywheel ergometer. Comparisons were performed using paired (within-group) or non-paired (between-group) t-tests. Three days post-BR, VO2pk of the CON group was reduced significantly from pre-BR (Pre:37.2 plus or minus 1.2, Post: 29.4 plus or minus 2 ml (raised dot) kg(sup -1) (raised dot) min(sup -1), P less than 0.05), while the VO2pk of the EX group was not significantly reduced (Pre: 39.6 plus or minus 1.9, Post: 38.0 plus or minus 0.6 ml (raised dot) kg(sup -1) (raised dot) min(sup -1)). Peak heart rate, ventilation, rating of perceived exertion, and respiratory exchange ratio were not significantly different between the two groups pre- and post-BR. These preliminary results suggest that the combined LBNPex and REX countermeasures may be sufficient to maintain upright aerobic capacity after long-duration space flights.

Schneider, Suzanne M.↗

Cu‐Catalyzed Aerobic Oxidative C─C Cleavage in Lignin‐Derived Oligomers and Biological Funneling of the Monomeric Products

Existing methods for lignin deconstruction to aromatic monomers primarily cleave carbon–oxygen bonds within the polymer, resulting in sub-optimal monomer yields and formation of oligomers that retain intact carbon–carbon bonds. Here, we demonstrate that copper-catalyzed aerobic oxidation under aqueous alkaline conditions promotes oxidative cleavage of carbon–carbon bonds in lignin oligomers derived from reductive catalytic fractionation (RCF) of pine and poplar biomass. Fundamental insights are gained from reactions of model compounds that resemble subunits present in RCF oligomers. Optimal results are achieved in a flow reactor that provides precise control over O2 delivery, temperature, and reaction residence time. The Cu-catalyzed aerobic oxidation conditions access aromatic monomers in 19 and 34 wt% monomer yields, respectively, from pine- and poplar-derived RCF oligomers. Overall, the sequence consisting of biomass RCF into monomers and oligomers followed by oxidative deconstruction of the RCF oligomers generates substantially higher yields of aromatic monomers from lignin. Engineered strains of Pseudomonas putida support biological funneling of the oligomer-derived oxygenated aromatic compounds into cis,cis-muconic acid from pine or 2-pyrone-4,6-dicarboxylic acid from poplar.

09 BIOMASS FUELS↗

Mechanistic investigation of the aerobic oxidation of 2-pyridylacetate coordinated to a Ru(ii) polypyridyl complex

A new ruthenium polypyridyl complex, [Ru(bpy) 2 (acpy)] + (acpy = 2-pyridylacetate, bpy = 2,2'-bipyridine), was synthesized and fully characterized. Distinct from the previously reported analog, [Ru(bpy) 2 (pic)] + (pic = 2-pyridylcarboxylate), the new complex is unstable under aerobic conditions and undergoes oxidation to yield the corresponding α-keto-2-pyridyl-acetate (acpyoxi) coordinated to the Ru II center. The reaction is one of the few examples of C–H activation at mild conditions using O 2 as the primary oxidant and can provide mechanistic insights with important implications for catalysis. Theoretical and experimental investigations of this aerobic oxidative transformation indicate that it takes place in two steps, first producing the α-hydroxo-2-pyridyl-acetate analog and then the final product. The observed rate constant for the first oxidation was in the order of 10 -2 h -1 . The reaction is hindered in the presence of coordinating solvents indicating the role of the metal center in the process. Theoretical calculations at the M06-L level of theory were performed for multiple reaction pathways in order to gain insights into the most probable mechanism. Our results= indicate that O 2 binding to [Ru(bpy) 2 (acpy)] + is favored by the relative instability of the six-ring chelate formed by the acpy ligand and the resulting Ru III -OO˙- superoxo is stabilized by the carboxylate group in the coordination sphere. C–H activation by this species involves high activation free energies (ΔG ‡ = 41.1 kcal mol -1 ), thus the formation of a diruthenium μ-peroxo intermediate, [(Ru III (bpy) 2 (O-acpy)) 2 O 2 ] 2+ via interaction of a second [Ru(bpy) 2 (acpy)] + was examined as an alternative pathway. The dimer yields two Ru IV =O centers with a low ΔG ‡ of 2.3 kcal mol -1 . The resulting Ru IV =O species can activate C–H bonds in acpy (ΔG ‡ = 23.1 kcal mol -1 ) to produce the coordinated α-hydroxo-2-pyridylacetate. Further oxidation of this intermediate leads to the α-keto-2-pyridyl-acetate product. The findings provide new insights into the mechanism of C–H activation catalyzed by transition-metal complexes using O 2 as the sole oxygen source.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Metabolic Model of the Nitrogen-Fixing Obligate Aerobe Azotobacter vinelandii Predicts Its Adaptation to Oxygen Concentration and Metal Availability

There is considerable interest in promoting biological nitrogen fixation (BNF) as a mechanism to reduce the inputs of nitrogenous fertilizers in agriculture, but considerable fundamental knowledge gaps still need to be addressed. BNF is catalyzed by nitrogenase, which requires a large input of energy in the form of ATP and low potential electrons. Diazotrophs that respire aerobically have an advantage in meeting the ATP demands of BNF but face challenges in protecting nitrogenase from inactivation by oxygen. Here, we constructed a genome-scale metabolic model of the nitrogen-fixing bacterium Azotobacter vinelandii, which uses a complex respiratory protection mechanism to consume oxygen at a high rate to keep intracellular conditions microaerobic. Our model accurately predicts growth rate under high oxygen and substrate concentrations, consistent with a large electron flux directed to the respiratory protection mechanism. While a partially decoupled electron transport chain compensates for some of the energy imbalance under high-oxygen conditions, it does not account for all substrate intake, leading to increased maintenance rates. Interestingly, the respiratory protection mechanism is required for accurate predictions even when ammonia is supplemented during growth, suggesting that the respiratory protection mechanism might be a core principle of metabolism and not just used for nitrogenase protection. We have also shown that rearrangement of flux through the electron transport system allows A. vinelandii to adapt to different oxygen concentrations, metal availability, and genetic disruption, which cause an ammonia excretion phenotype. Accurately determining the energy balance in an aerobic nitrogen-fixing metabolic model is required for future engineering approaches.

59 BASIC BIOLOGICAL SCIENCES↗

Aligned FTICR-MS, Respiration, and Biochemical Transformation Data from Columbia River Sediment Incubations Associated with: “Carbon Limitation Leads to Thermodynamic Regulation of Aerobic Metabolism”

Columbia River hyporheic zone sediments from the Hanford Site 300 Area in eastern Washington were incubated in a full factorial design with different organic matter (OM) amendments. The aim of the experiment was to test the influence of OM thermodynamics on aerobic respiration across different organic carbon pools and concentrations. This dataset is comprised of (1) a csv with high resolution characterization of dissolved organic matter via 12 Tesla Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) analyzed through the Environmental Molecular Sciences Laboratory (EMSL; https://www.pnnl.gov/environmental-molecular-sciences-laboratory), (2) a csv with sample IDs, replicates, amendments, dissolved oxygen (DO) measurements, and sampling time points, (3) a csv with sample IDs, replicates, respiration rates, FTICR summary metrics, and relative abundance of chemical classes; and (4) a csv with the number of chemical transformations in each sample replicate (microcosm). The data are associated with the publication “Carbon Limitation Leads to Thermodynamic Regulation of Aerobic Metabolism” published in ES&T Letters (Garayburu-Caruso et al., 2020). Please use the data package’s DOI to cite the data package. We ask that you acknowledge the U.S. Department of Energy (DOE) Biological and Environmental Research (BER) Subsurface Biogeochemical Research (SBR) program when using the data. All data are free to be used for any purpose, such as for manuscripts, presentations, and grant proposals. There is no obligation to include data package authors as co-authors.

54 ENVIRONMENTAL SCIENCES↗