Completely noninvasive multi-analyte monitoring system for cell culture processes.

Although online monitoring of dissolved O2, pH, and dissolved CO2 is critical in bioprocesses, nearly all existing technologies require some level of direct contact with the cell culture environment, posing risks of contamination. This study addresses the need for an accurate, and completely noninvasive technique for simultaneous measurement of these analytes. A "non-contact" technique for simultaneous monitoring of dissolved O2, pH, and dissolved CO2 was developed. Instead of direct contact with the culture media, the measurements were made through permeable membranes via either a sampling port in the culture vessel wall or a flow cell. The efficacy of the "non-contact" technique was validated in Escherichia coli (E.coli), Chinese hamster ovary (CHO) culture processes, and dynamic environments created by sparging gases in cell culture medium. The measurements obtained through the developed techniques were comparable to those obtained through control methods. The noninvasive monitoring system can offer accurate, and contamination-minimized monitoring of critical process parameters including dissolved O2, pH, and dissolved CO2. These advancements will enhance the control and optimization of cell culture processes, promising improved cell culture performance.

Understanding the environmental drivers of summer dissolved carbon in lakes on the Qinghai-Tibetan Plateau.

How environmental factors affecting dissolved carbon remains unclear in lakes on the Qinghai-Tibetan Plateau (QTP), which limits the understanding of the lake carbon cycle. In this study, 60 lakes on the QTP in summer were investigated to clarify the variation in dissolved carbon, estimate dissolved carbon storage, and reveal how environmental factors affect the variation in dissolved carbon. The average dissolved organic carbon (DOC) and dissolved inorganic carbon (DIC) contents of 60 lakes on the QTP in summer were 12.78 mg/L and 103.66 mg/L, respectively. Salinity and total nitrogen were the important drivers of variations in DIC and DOC, respectively. Lake salinity and temperature were reduced only when precipitation was >50 mm, thus affecting the variations in lake dissolved carbon. Importantly, the elevation and area of the lake also significantly affected the variation in lake dissolved carbon. The total storage amounts of DOC and DIC in the 60 lakes on the QTP in summer were 58.94 Tg and 6.22 Tg, respectively. Lake area was the most direct factor influencing dissolved carbon storage in lakes on the QTP. Moreover, the TN and pH of the lake water also affected the DOC and DIC storage in the lakes, respectively. Interestingly, the lake pH at 9.1 was an important turning point that caused variations in lake DIC storage. Surprisingly, we found that rivers were able to transport 30 % of the DIC into QTP lakes and were the main source of DIC in the lakes in summer. The findings of this study clarify the sources of dissolved carbon and its drivers and improve our understanding of the carbon cycling processes in the lake system on the QTP.

Influences of carbonate weathering and hyporheic exchange on carbon fluxes in Pearl River Basin, China.

Deciphering riverine dissolved carbon dynamics is pivotal for a comprehensive picture of the global carbon cycle. Through rigorous in-situ sampling across the Pearl River Basin (PRB), our investigation reveals the Pearl River networks function as a significant carbon source, with the annual carbon dioxide (CO2) emission of 2.57 ± 1.94 Tg C, which offsets 10 ± 8 % of the forest carbon sequestration or 65 ± 49 % carbon sink via chemical weathering in the PRB. Based on the mass balance of 222Rn, we initially reveal that the contributions of water flux from the hyporheic zone increased with the river orders (Hack Order) across both dry and wet seasons. Conversely, the evasion rates of dissolved CO2 (CO2*) and dissolved inorganic carbon (DIC) from the hyporheic zone into river channels exhibited a decline with the increasing river orders. The hyporheic exchange contributes 4 - 11 % of the lateral and vertical DIC losses, thereby is a key mechanism in the riverine carbon cycle. Furthermore, CO2* derived from the hyporheic zone was ∼4 times of riverine CO2 emissions and this CO2* flux from the hyporheic zone was buffered into carbonates/bicarbonates in river channels, due to the high riverine pH resulted from carbonate weathering in the basin. These results not only highlight the substantial role of carbonates and hyporheic processes in modulating riverine carbon fluxes but also signify their broader implications on understanding riverine carbon dynamics at both regional and global scales.

Carbon export from submarine groundwater discharge in a semi-enclosed bay: Impact for the buffering capacity against coastal ocean acidification.

Submarine groundwater discharge (SGD) serves as an important pathway for the transport of dissolved carbon from land to ocean, significantly affecting the coastal biogeochemical cycles. However, the impact of SGD-derived dissolved carbon on the coastal carbon budget remains poorly understood. This study first quantified SGD and associated dissolved organic carbon (DOC), dissolved inorganic carbon (DIC) and total alkalinity (TA) fluxes in Daya Bay using mass balance models based on radium isotopes (223Ra, 224Ra, 226Ra and 228Ra). We then constructed carbon mass balance models to evaluate the impact of SGD-derived carbon on the buffering capacity against coastal ocean acidification. The estimated SGD fluxes ranged from 0.80 × 107 to 2.64 × 107 m3d-1. The DIC, DOC and TA fluxes from SGD were 17.90-36.44 mmol m-2d-1, 0.93-2.13 mmol m-2d-1, and 21.19-28.47 mmol m-2d-1, respectively. Based on carbon mass balances, the DIC flux from SGD was 19-39 times the riverine input, accounting for 27.16 % ∼ 37.64 % of the total carbon source. These results suggest that SGD is a major contributor to DIC, significantly affecting the coastal carbon budget. Furthermore, the average TA:DIC ratio of groundwater discharging into Daya Bay was approximately 1.13. High TA exports enhance the buffering capacity of the coastal ocean and contribute bicarbonate to the ocean, playing a significant role in the ocean carbon sequestration process. This study demonstrates the importance of SGD-derived dissolved carbon in the assessment of coastal carbon budgets.

Hydrochemical and isotopic characteristics of water sources for biological activity across a massive evaporite basin on the Tibetan Plateau: Implications for aquatic environments on early Mars.

Covered by vast eolian landforms, gravel deposits, and playas, the worldwide typical evaporite deposit land, Qaidam Basin, in northwestern China is analogous to early Mars when the aridification process had lasted for millions of years since the end of a wetter climate. This study aims to investigate the chemical and isotopic characteristics of waters in an evaporite-rich environment, as well as the habitable conditions therein, that have undergone a transformation similar to early Mars. In May 2023, a total of 26 water samples were collected across the representative central axis of a longitudinal aridity gradient in the Qaidam Basin, including categories of meteoric water, freshwater, standing water accumulated after precipitation, salty lacustrine water, and hypersaline brines to inspect compounds made up of carbon, nitrogen, phosphorus, sulfur, halogen, and metallic elements. As evaporation intensified, the salt types transformed from HCO3-Ca·Na to Cl·SO4-Na or ClMg. The dominance of carbonate will gradually be replaced by sulfate and chloride, leaving much more dilute and less detectable contents. The presence of trace ClO4-, ClO3-, ClO2-, and BrO3- was confirmed in a few of the sampled Qaidam waters, indicating the preservation of oxyhalides in waters within an arid region and possibly the presence of relevant microbial enzymes. The isotopes of water, carbonaceous, and nitrogenous compounds provide valuable references for either abiogenic or biogenic signatures. With undetectable amount, phosphorus was found to be the limiting nutrient in evaporative aquatic environments but not necessarily antibiosignatures. Overall, these results suggest that the paleo-lacustrine environments on Mars are more likely to preserve biosignatures if they feature the dominance of carbonate minerals, bioavailable nitrate, phosphorus, and organic carbon, the presence of thermodynamically unstable oxyhalides, and isotope ratios that point to the involvement of biological activity.

Connectivity of floodplain influences riverine carbon outgassing and dissolved carbon transport.

Rivers not only function as a conduit for the delivery of terrestrial constituents to oceans, but they also serve as an essential medium for biogeochemical processing of the constituents. While extensive research has been conducted on carbon transport in many rivers, little is known about carbon transformation in engineered rivers reconnected with their floodplain network. Being the largest distributary of the levee-confined Mississippi River (MR), the Atchafalaya River (AR) carries 25 % of the MR water, flowing through North America's largest freshwater swamp basin and emptying into the Gulf of Mexico. Previous studies reported that this 200-km long, 5-30-km wide river basin can remove a substantial amount of riverine nutrients and organic carbon. This study aimed to test the hypothesis that the AR emits significantly higher CO2 into the atmosphere as it flows through its extensive floodplain network than the levee-confined MR does. From January 2019 to December 2021, we conducted biweekly - monthly in-situ measurements in the lower AR at Morgan City and in the lower Mississippi River at Baton Rouge. Field measurements included partial pressure of dissolved CO2 (pCO2), water temperature, chlorophyll a, colored dissolved organic matter, dissolved oxygen, pH, and turbidity. During each field sampling, water samples were collected and analyzed for concentrations of dissolved organic and inorganic carbon (DOC and DIC). Mass transport of DOC and DIC and outgassing of CO2 were quantified for the two rivers. We found that pCO2 levels were significantly higher in the AR (mean: 3563 µatm; min-max: 1130-8650 µatm) than those in the MR (1931 µatm, 836-3501 µatm), resulting in a doubled CO2 outgassing rate in the AR (486 mmol m2 d-1) than in the MR (241 mmol m2 d-1). The AR had higher DOC (8.5 mg L-1) but lower chlorophyll a (153.9 AFU) when compared with the MR (7.5 mg L-1 and 164.0 AFU). Water temperature was constantly higher in the AR than in the MR, especially during the wintertime. Since the Mississippi-Atchafalaya River system is among the world's largest and most engineered river systems, our assessment offers a field case study to inform on the potential implications of reconnecting rivers with their floodplains networks.

Ecological water diversion activity changes the fate of carbon in a eutrophic lake.

Lake eutrophication mitigation measures have been implemented by ecological water diversion, however, the responses of carbon cycle to the human-derived hydrologic process still remains unclear. With a famous river-to-lake water diversion activity at eutrophic Lake Taihu, we attempted to fill the knowledge gap with integrative field measurements (2011-2017) of gas carbon (CO2 and CH4) flux, including CO2-equivalent, and dissolved carbon (DOC and DIC) at water-receiving zone and reference zone. Overall, results showed the artificial water diversion activity increased gas carbon emissions. At water-receiving zone, total gas carbon (expressed as CO2-equivalent) emissions increased significantly due to the occurring of water diversion, with CO2 flux increasing from 9.31 ± 16.28 to 18.16 ± 12.96 mmol C m-2 d-1. Meanwhile, CH4 emissions at water-receiving zone (0.06 ± 0.05 mmol C m-2 d-1) was double of that at reference zone. Water diversion decreased DOC but increased DIC especially at inflowing river mouth. Temporal variability of carbon emissions and dissolved carbon were linked to water temperature, chlorophyll a, and nutrient, but less or negligible dependency on these environment variables were found with diversion occurring. Water diversion may increase gas carbon production via stimulating DOC mineralization with nutrient enrichment, which potentially contribute to increasing carbon emissions and decreasing DOC at the same time and the significant correlation between CO2 flux and CH4 flux. Our study provided new insights into carbon biogeochemical processes, which may help to predict carbon fate under hydrologic changes of lakes.

Gas storage of peat in autumn and early winter in permafrost peatland.

The carbon (C) balance of permafrost peatlands in autumn and winter, which affects the annual C budget estimation, has become a hotspot of studies on peatlands C cycle. This study combined the static chamber method, in situ soil profile measurements, and incubation experiments to investigate release and storage of C during autumn and early winter in a permafrost peatland in the Da Xing'an Mountains, Northeast China. Our results showed that the peak values of CH4 fluxes (30 August 2016) lagged behind those of CO2 fluxes (24 July 2016). At the onset of soil freezing, CH4 fluxes slightly increased, while CO2 fluxes decreased. During soil freezing in autumn, gases were found to be mainly stored in the soil as dissolved CH4 and CO2 and dissolved C concentrations (CH4, CO2, and DOC (dissolved organic carbon)) increased with depth. DOC concentrations were closely related to dissolved C gases, implying that the stored dissolved C gases might be derived from DOC decomposition. The CO2: CH4 ratio decreased sharply from the freezing of the surface layer to the total freezing of the soil, indicating larger CH4 storage in totally frozen soil. The incubation experiments also showed larger CH4 storage in the frozen soils and the stored C gases could influence the assessment of C emissions during thawing. These findings have important implications for clarifying the gas storage of permafrost peatland in autumn and early winter. The results may also clarify the key link of C emissions between the growing season and the nongrowing season.

Phytotoxicity of farm livestock manures in facultative heap composting using the seed germination index as indicator.

Static facultative heap composting of animal manure is widely used in China, but there is almost no systematic research on the phytotoxicity of the produced compost. Here, we evaluated the phytotoxic variation in compost produced by facultative heap composting of four types of animal manure (chicken manure, pig manure, sheep manure, and cattle manure) using different plant seeds (cucumber, radish, Chinese cabbage, and oilseed rape) to determine germination index (GI). The key factors that affected GI values were identified, including the dynamics of the phytotoxicity and microbial community during heap composting. Sensitivity to toxicity differed depending on the type of plant seed used. Phytotoxicity during facultative heap composting, evaluated by the GI, was in the order: chicken manure (0-6.6 %) < pig manure (14.4-90.5 %) < sheep manure (46.0-93.0 %) < cattle manure (50.2-105.8 %). Network analysis showed that the volatile fatty acid (VFA) concentration was positively correlated with Firmicutes abundance, and NH4+-N was correlated with Actinobacteria, Proteobacteria, and Bacteroidetes. More bacteria were stimulated to participate in conversions of dissolved organic carbon, dissolved nitrogen, VFA, and ammonia-nitrogen (NH4+-N) in sheep manure heap composting than that in other manure. The GI was most affected by VFA in chicken manure and cattle manure heap composting, while NH4+-N was the main factor affecting the GI in pig manure and sheep manure compost. The dissolved carbon and nitrogen content and composition, as well as the core and proprietary microbial communities, were the primary factors that affected the succession of phytotoxic substances in facultative heap composting, which in turn affected GI values. In this study, the key pathways of livestock manure composting that affected GI and phytotoxicity were found and evaluated, which provided new insights and theoretical support for the safe use of organic fertilizer.

Dynamics and internal links of dissolved carbon in a karst river system: Implications for composition, origin and fate.

Dissolved carbon (DC) deciphers biotic and abiotic processes in aquatic ecosystems, representing a critical component of global carbon cycling. However, underlying drivers of riverine DC dynamics and internal links have yet to be studied. Here, we investigated fluvial physicochemical characteristics, dissolved inorganic carbon (DIC) species, carbon dioxide (CO2) exchange, dissolved organic carbon (DOC) compositions and properties in a karst river system Qijiang, Southwest China. Carbonate dissolution combined with photosynthetic uptake could explain dynamics of DIC species. Carbon sequestration caused low-magnitude of partial pressure of aqueous CO2 (pCO2, 620.3 ± 1028.7 µatm) and water-air CO2 flux (F, 154.3 ± 772.6 mmol/m2/d), yielding an annual CO2 emission of 0.079 Tg CO2/y. Relatively high biological index (BIX, 0.77-0.96 on average) but low humification index (HIX, 0.67-0.78 on average) indicated notable autochthonous processes. Humic-like component was the predominant DOC, accounting for 39.0%-75.2% with a mean of 57.2% ± 6.17%. Meanwhile, tryptophan-like component (5.84% ± 2.31%) was also identified as collective DOC by parallel factor analysis (PARAFAC) across samples. Biological metabolism established internal linkages between DIC and DOC in the karst river system. Our findings highlighted biological process as a determinant for DC cycling in karst aquatic ecosystems.

Impact of Elevated Levels of Dissolved CO2 on Performance and Proteome Response of an Industrial 2'-Fucosyllactose Producing Escherichia coli Strain.

Large-scale microbial industrial fermentations have significantly higher absolute pressure and dissolved CO2 concentrations than otherwise comparable laboratory-scale processes. Yet the effect of increased dissolved CO2 (dCO2) levels is rarely addressed in the literature. In the current work, we have investigated the impact of industrial levels of dCO2 (measured as the partial pressure of CO2, pCO2) in an Escherichia coli-based fed-batch process producing the human milk oligosaccharide 2'-fucosyllactose (2'-FL). The study evaluated the effect of high pCO2 levels in both carbon-limited (C-limited) and carbon/nitrogen-limited (C/N-limited) fed-batch processes. High-cell density cultures were sparged with 10%, 15%, 20%, or 30% CO2 in the inlet air to cover and exceed the levels observed in the industrial scale process. While the 10% enrichment was estimated to achieve similar or higher pCO2 levels as the large-scale fermentation it did not impact the performance of the process. The product and biomass yields started being affected above 15% CO2 enrichment, while 30% impaired the cultures completely. Quantitative proteomics analysis of the C-limited process showed that 15% CO2 enrichment affected the culture on the protein level, but to a much smaller degree than expected. A more significant impact was seen in the dual C/N limited process, which likely stemmed from the effect pCO2 had on nitrogen availability. The results demonstrated that microbial cultures can be seriously affected by elevated CO2 levels, albeit at higher levels than expected.

Dissolved carbon dioxide stripping while maintaining volatile components by feeding gaseous nitrogen using a microbubble generator.

Dissolved carbon dioxide (dCO2) stripping from a model solution containing sake flavor by feeding gaseous nitrogen (N2) using a microbubble (MB) generator was investigated. The effect of dCO2 stripping by N2MB increased significantly with increasing flow rate of gaseous N2 from 100 to 200 mL/min. dCO2 stripping from 3,000 mL of the model solution was achieved by feeding N2MB at a flow rate of 200 mL/min for 4 min. Volatile components from model solution containing sake flavor were hardly reduced even after feeding N2MB at a flow rate of 200 mL/min for 15 min by cooling to below 10 °C. On the other hand, non-microbubbled gaseous N2 at a flow rate of 200 mL/min was not very effective in stripping dCO2. Therefore, the use of N2MB with cooling to below 10 °C was effective in stripping dCO2 while maintaining the volatile components in model solution containing sake flavor.

Reclamation-induced tidal restriction increases dissolved carbon and greenhouse gases diffusive fluxes in salt marsh creeks.

Intertidal creeks play an important role in transporting nutrients between coastal ecosystems and ocean. Reclamation is a predominant anthropogenic disturbance in coastal regions; however, the influence of reclamation on carbon and nitrogen species and greenhouse gas (GHG) fluxes in creek remains unclear. In a subtropical salt marsh of eastern China, the seasonal patterns of dissolved carbon (DOC, DIC, CO2, and CH4) and inorganic nitrogen (NH4+-N, NO2--N, and NO3--N and N2O) species, and the diffusive fluxes of CO2, CH4, and N2O, were compared between the natural tidal creeks and the reclaimed creeks. Due to notably changed hydrological and biological conditions in the reclaimed creeks, concentrations of all dissolved carbon species, NH4+-N and NO2--N increased significantly by 60.2-288.2%, while NO3--N and N2O decreased slightly, compared to the natural tidal creeks. DIC and NO3--N were the primary components of the total dissolved carbon and inorganic nitrogen in both creek types; however, their proportions decreased as a result of elevated DOC, CO2, CH4, NH4+-N, and NO2--N following reclamation. Significantly higher global warming potential (0.58 ± 0.15 g CO2-eq m-2 d-1) was found in the reclaimed creeks, making them hotspot of greenhouse effects, compared to the natural tidal creeks. Our results indicated that changes in flow velocity, salinity, Chlorophyll a, and pH were the main factors controlling the dissolved carbon and nitrogen and consequent GHG emissions, due to reclamation. This study is helpful in understanding of carbon and nitrogen sink-source shifts resulting from land use changes in coastal wetlands.

The impact of alder litter on chemistry of Technosols developed from lignite combustion waste and natural sandy substrate: a laboratory experiment.

Alders, as an N-fixing species, are widely used as a phytomelioration species at post-industrial sites. The paper presents the effects of litter decomposition of different alder species-black alder, gray alder, and green alder-on changes to the soil solution chemistry of Technosols developed from two types of substrates: lignite combustion waste and sandy substrates from a former sand quarry. Under controlled conditions, the 13-week experiment investigated pH, electrical conductivity (EC), and the chemical parameters-dissolved organic carbon (DOC), dissolved nitrogen (DON), and macroelements S, Ca, Mg, K, and Na content-of filtrates in composites consisting of substrate and litter from alder species was investigated. Alder litter accelerated nutrient leaching processes from soil substrates and had the greatest impact on sandy substrates because fast leaching was possible at low absorption capacity characteristics. Higher leaching rates in sandy substrates were also observed, especially in the cases of DOC, DON, K, and P, in comparison to combustion waste substrates. In the first phase of the experiment in sandy soil substrates, the dominant process is the leaching of DOC from dead organic matter, while in the case of combustion waste, calcium leaching prevails in the first phase. Among the investigated species, black alder had the greatest impact on the chemistry of substrates and solutions, and the dynamic of these processes has a different path at various substrates.

The impact of clearance on mixing time for interface-added substrate.

This study was carried out to find the optimum clearance (impeller to bottom distance) for Rushton and pitch-blade turbine impellers in a stirred tank bioreactor for improved substrate mixing time added at interface, taking advantage of computational fluid dynamics. In this regard, the time needed for a thin layer of liquid, resembling substrate-rich or poor part, getting homogenously dispersed within the tank was calculated. The mixing time calculated in this way is called the surface aeration related mixing time (SARMT). SARMT was calculated using two approaches and was compared with each other. For the pitch-blade turbine impeller, a criterion which guarantees accurate mixing time by simulation was not satisfied, so the SARMT profile against clearance was not achieved. For the Rushton impeller, a general descending order of SARMT against impeller-bottom clearance was observed.

[Vertical Distribution of Soil Dissolved Carbon and Its Influencing Factors in the Artificial Shelterbelt Irrigated with Saline Water in an Extreme Drought Desert].

Dissolved carbon (DC) is the most active carbon fraction in soils. Vegetation restoration and reconstruction accelerate the carbon cycle in arid desert regions. Studying the DC distribution in soil profiles of artificial shelterbelt under saline irrigation can provide theoretical support and decision-making basis for artificial shelterbelt management, development, and utilization in arid desert areas. In this study, we took the artificial shelterbelts drip-irrigated with five different mineralization and one shifting sandy land (CK) along the Taklimakan Desert Highway as sampling plots, analyzed and discussed the vertical distribution characteristics of soil dissolved organic carbon (SDOC) and dissolved inorganic carbon (SDIC) in the 0-1 m profiles, and further analyzed their relationships among different factors. The results showed that SDOC and SDIC of CK and shelterbelts under 2.82 g·L-1 irrigation showed an "I" type distribution with a linear function relationship. The SDOC and SDIC of four other treatments showed a "Γ" type distribution with power function relationships. The fluctuation ability and contribution degree of SDOC and SDIC of different treatments in the surface layer were higher than those in the lower layers, and the fluctuation and contribution intensity of SDOC were higher than those of SDIC. Except for 2.82 g·L-1 treatment, the average SDOC contents of other treatments were 2-4 times those of SDIC. The average SDOC content of 2.82 g·L-1 treatment was lower than CK; other treatments were 3-5 times that of CK; and the average SDIC content of all treatments increased 15.0%-57.9% than CK. For the 0-5 cm layer, SDOC content increased with the irrigation water mineralization except the 2.82 g·L-1 treatment, but SDIC content firstly increased and then decreased with increasing mineralization, and that for the 4.82 g·L-1 treatment was highest. The SDOC and SDIC were positively correlated with EC, SOC, irrigation water mineralization, SIC, and soil moisture, for which they both showed a weak positive correlation with soil moisture; they were negatively correlated with soil depth. The SDOC and SDIC showed a weak negative correlation and a weak positive correlation with pH, respectively. In summary, the mineralization of irrigation water has an important impact on the vertical distribution of SDOC and SDIC, and their distribution also has close relationships with EC, SOC, SIC, soil moisture, and soil depth, which is of great significance for plantations in extremely drought deserts.

Nitrogen fertilizer application in the rice-growing season can stimulate methane emissions during the subsequent flooded fallow period.

Winter-flooded rice paddy field (FR), characterized by water conserved in the field during the fallow period, is a typical cropping system in southwest China, leading to considerable methane (CH4) emissions. The effect of nitrogen (N) fertilization on CH4 emissions during rice-growing seasons is well studied in FR, further studies covering N fertilizer applied in the rice-growing seasons affects CH4 emissions during the subsequent fallow period is needed. Therefore, a field experiment was conducted in an FR of Sichuan province, China, with conventional N fertilized (CN) and N unfertilized (NN) treatments. The cumulative CH4 emission from CN treatment during the rice-growing season and the subsequent fallow period was 389 ± 29.4 and 158 ± 31.2 kg C ha-1, which were increased by 29.5% and 395% in comparison with the NN treatment, indicting N applied during the rice growing-season significantly facilitated CH4 emission during the subsequent fallow period. During the rice-growing season, higher CH4 emission from CN treatment could be attributed to elevated soil dissolved organic carbon (DOC) content that might have provided sufficient substrates for CH4 production. During the fallow period, as compared to NN treatment, higher CH4 emissions from CN treatment could be explained by greater linear regression slopes between CH4 fluxes, soil temperature and DOC to dissolved inorganic N (DIN) (DOC/DIN) ratio. Moreover, the structural equation model (SEM) described that the soil temperature exhibited the most significant effects on CH4 emissions for both treatments during the rice-growing season and subsequent fallow period. These findings are a major step forward to showing that N fertilizer applied in the rice-growing season could also affect CH4 emission during the subsequent fallow period, accompanying other soil parameters controlling CH4 emission.

Urbanization increases carbon concentration and pCO2 in subtropical streams.

Urbanization growth may alter the hydrologic conditions and processes driving carbon concentrations in aquatic systems through local changes in land use. Here, we explore dissolved carbon concentrations (DIC and DOC) along urbanization gradient in Santa Catarina Island to evaluate potential increase of CO2 in streams. Additionally, we assessed chemical, physical, and biotic variables to evaluate direct and indirect effects of urbanization in watersheds. We defined 3 specific urbanization levels: high (> 15% urbanized area), medium (15-5% urbanized area), and low (< 5% urbanized area) urbanization. The results showed that local changes due to growth of urban areas into watersheds altered the carbon concentrations in streams. DOC and DIC showed high concentrations in higher urbanization levels. The watersheds with an urban building area above 5% showed pCO2 predominantly above the equilibrium with the atmosphere. These findings reveal that local modifications in land use may contribute to changes in global climate by altering the regional carbon balance in streams.

Real-time dissolved carbon dioxide monitoring II: Surface aeration intensification for efficient CO2 removal in shake flasks and mini-bioreactors leads to superior growth and recombinant protein yields.

Mass transfer is known to play a critical role in bioprocess performance and henceforth monitoring dissolved O2 (DO) and dissolved CO2 (dCO2 ) is of paramount importance. At bioreactor level these parameters can be monitored online and can be controlled by sparging air/oxygen or stirrer speed. However, traditional small-scale systems such as shake flasks lack real time monitoring and also employ only surface aeration with additional diffusion limitations imposed by the culture plug. Here we present implementation of intensifying surface aeration by sparging air in the headspace of the reaction vessel and real-time monitoring of DO and dCO2 in the bioprocesses to evaluate the impact of intensified surface aeration. We observed that sparging air in the headspace allowed us to keep dCO2 at low level, which significantly improved not only biomass growth but also protein yield. We expect that implementing such controlled smart shake flasks can minimize the process development gap which currently exists in shake flask level and bioreactor level results.

Real-time dissolved carbon dioxide monitoring I: Application of a novel in situ sensor for CO2 monitoring and control.

Dissolved carbon dioxide (dCO2 ) is a well-known critical parameter in bioprocesses due to its significant impact on cell metabolism and on product quality attributes. Processes run at small-scale faces many challenges due to limited options for modular sensors for online monitoring and control. Traditional sensors are bulky, costly, and invasive in nature and do not fit in small-scale systems. In this study, we present the implementation of a novel, rate-based technique for real-time monitoring of dCO2 in bioprocesses. A silicone sampling probe that allows the diffusion of CO2 through its wall was inserted inside a shake flask/bioreactor and then flushed with air to remove the CO2 that had diffused into the probe from the culture broth (sensor was calibrated using air as zero-point calibration). The gas inside the probe was then allowed to recirculate through gas-impermeable tubing to a CO2 monitor. We have shown that by measuring the initial diffusion rate of CO2 into the sampling probe we were able to determine the partial pressure of the dCO2 in the culture. This technique can be readily automated, and measurements can be made in minutes. Demonstration experiments conducted with baker's yeast and Yarrowia lipolytica yeast cells in both shake flasks and mini bioreactors showed that it can monitor dCO2 in real-time. Using the proposed sensor, we successfully implemented a dCO2 -based control scheme, which resulted in significant improvement in process performance.