Metabolic rate estimates for a eutrophic lake from diel dissolved oxygen signals

Estimates of net primary production, community respiration (R'), and gross primary production (P _g) are developed and presented for the productive layers of eutrophic Onondaga Lake, NY, U.S.A., for time scales ranging from diel to several months, based on 4 months of robotic diel profiles of dissolved oxygen (DO) and temperature. Metabolic rate calculations are made through application of a DO mass balance framework that also accommodates inputs and losses of DO mediated by exchange across the air–water interface and across the lower boundary of the productive layers. It is demonstrated that the dynamics of the flux across the air–water interface are important to the metabolic rate estimates, while vertical mixing-based losses to the underlying layers can be ignored. Study average estimates of R' (1.49 g O₂ m^–3 d^–1) and P _g (1.60 g O₂ m^–3 d^–1) obtained by this non-isolated community approach are consistent with levels reported in the literature for similar chlorophyll a concentrations, based on isolated community (bottle experiment) protocols to measure these metabolic rates. The non-isolated community approach is shown to have limited utility for quantifying day-to-day changes in these rates in this lake, apparently because of horizontal exchange with waters of different DO concentrations. However, this approach may support reliable estimates of metabolic rates at intermediate time scales; e.g., several days to a week. The DO mass balance framework is demonstrated to be valuable in resolving the relative roles of various physical and biological processes in regulating the DO pool of the productive layers.     

Climate-induced changes in the dissolved organic carbon budgets of boreal lakes

During 20 years of climatic warming, drought and increased forest firesbetween 1970 and 1990, DOC concentrations declined by 15--25%in lakesof the Experimental Lakes Area, northwestern Ontario, allowing increasedpenetration of both UV and photosynthetically-active radiation (PAR), andcausing deeper euphotic zones and thermoclines. Decreased input to thelakes of DOC from terrestrial catchments and upstream lakes was theprimary reason for the decline, although in-lake removal also increasedslightly. Decreased streamflow caused by drought was more important thanforest fires in affecting DOC exports from catchments. Experimentalacidification of lakes caused even greater losses in DOC, by enhancing ratesof in-lake removal. DOC in Lake 302S, acidified to pH 4.5 during the1980s, declined to less than 10% of preacidificationvalues.     

Relation between dissolved oxygen concentration and ajmalicine production rate in high-density cultures of Catharanthus roseus

The relation between dissolved oxygen (DO) and the ajmalicine production rate of Catharanthus roseus was investigated in 15-L tank reactors at constant stirrer speed and gas flow rate. Below a DO concentration of 29% of air saturation the ajmalicine production rate was less than 0.06 mumol/g/d. Above a DO of 43% the ajmalicine production rate was constant at 0.21 mumol/g/d. Between a DO of 29% and 43% there was a strong relation between the ajmalicine production rate and the DO concentration. After a period of at least 12 days at DO /=57%. A kinetic equation is proposed for the relation between DO and the specific ajmalicine production rate. (c) 1995 John Wiley & Sons, Inc.     

Neap-spring tidal effects on dissolved oxygen in two Malaysian estuaries

The longitudinal distributions of dissolved oxygen change dramatically during neap-spring tidal cycles in the lower Selangor and Klang Rivers. An oxygen deficit develops in both estuaries when the tidal range is high. The Selangor River inflow is nearly saturated with D.O., but during spring tides D.O. falls to as little as 15% of saturation in the middle reaches of the estuary. The Klang River has low D.O. freshwater input, an oxygen minimum develops during neap tides, and anoxic conditions are produced by spring tides. These oxygen minima are attributed to the high oxygen demand of sediment that rests on the bottom during neap tides and is resuspended during spring tides. The ecological effects of tidal cycling patterns of dissolved oxygen in these Malaysian estuaries need further study, within the context of land use patterns and other human activities.     

Comparison of oxygen consumption in freshwater mussels (Unionidae) from different habitats during declining dissolved oxygen concentration

The rate of oxygen consumption (OC) of 9 species of freshwater mussels was measured under declining dissolved oxygen (DO) concentrations. The effects of temperature for some species also was investigated. The pattern of the OC vs. DO curve for each species was used in a hyperbolic model to compare abilities to regulate OC under low oxygen conditions. At 24.5 °C, Pyganodon grandis (from lakes), Amblema plicata and Quadrula pustulosa (from mud or sand in large rivers), Elliptio complanata (from pool areas in rivers), and Elliptio fisheriana and Elliptio lanceolata (from bank margins of rivers) were better able to maintain OC under low DO than were Villosa iris and Villosa constricta (which inhabit riffles) and Pleurobema cordatum (found in rivers with moderate flow). Villosa iris was especially sensitive to low oxygen conditions. The ability to maintain normal OC at low DO was improved considerably at 16.5 °C for V. iris, P. grandis and E. complanata. It is concluded that oxygen regulation ability appears to be related to the degree of hypoxia a species normally experiences in its habitat type, and it is enhanced at low temperature. The measurement of OC vs. DO may be a useful technique for estimating DO water quality criteria for endangered species because it is noninvasive.     

Serum-independent modulation of hemicyst formation by dissolved oxygen in postconfluent epithelial monolayers

Summary Hemicyst formation is considered a manifestation of either transepithelial solute and fluid movement or secretory activity in culture. This study shows that hemicyst formation in postconfluent monolayers of rhesus monkey kidney (LLC-MK₂) cells is modulated by the dissolved oxygen concentration (P_O2) of the culture medium. Either daily replacement of serum-free medium or displacement of the gas phase with 18% vol/vol O₂ (initial medium P_O2=125 to 135 mm Hg) enhances formation of hemicysts. Use of 30% O₂ (medium P_O2≊175 mm Hg) does not further increase the incidence, but neither 10% O₂ (medium P_O2=90 to 95 mm Hg) nor 1% O₂ (medium P_O2=35 to 50 mm Hg), the approximate range of dissolved oxygen values in blood, supports hemicyst formation unless cultures are gently rocked to disrupt diffusion gradients. Phase photomicrography of living cultures shows that the surface of a turgid hemicyst is furrowed, and cinephoto-micrography reveals that the walls vibrate subtly. When hypoxic conditions (0 to 1% O₂) are introduced this vibration ceases within 2 to 3 h, whereas collapse and disappearance of turgid hemicysts requires 18 to 20 h, seems virtually synchronous, and is reversible. Hemicysts form in a broad osmotic range, and increased electrolyte concentration increases the incidence. Hemicysts persist in localyy dense areas when cell-free strips are etched in the postconfluent monolayer; no DNA synthesis is detected under these conditions, but two-dimensional cell spreading into the denuded area is seen along the edge of the wound. We conclude that the dissolved oxygen supply in the cellular microenvironment modulates functional expression by differentiated kidney epithelial cells in culture and that increased electrolyte concentration also enhances expression of this phenotypic marker.     

The effect of dissolved oxygen tension and the utility of oxygen uptake rate in insect cell culture

Dissolved oxygen tension and oxygen uptake rate are critical parameters in animal cell culture. However, only scarce information of such variables is available for insect cell culture. In this work, the effect of dissolved oxygen tension (DOT) and the utility of on-line oxygen uptake rate (OUR) measurements in monitoring Spodoptera frugiperda (Sf9) cultures were determined. Sf9 cells were grown at constant dissolved oxygen tensions in the range of 0 to 30%. Sf9 metabolism was affected only at DOT below 10%, as no significant differences on specific growth rate, cell concentration, amino acid consumption/production nor carbohydrates consumption rates were found at DOT between 10 and 30%. The specific growth rate and specific oxygen uptake rate followed typical Monod kinetics with respect to DOT. The calculated _max and max were 0.033 h^-1 and 3.82×10^-10 mole cell^-1h^-1, respectively, and the corresponding saturation constants were 1.91 and 1.57%, respectively. In all aerated cultures, lactate was consumed only after glucose and fructose had been exhausted. The yield of lactate increased with decreasing DOT. It is proposed, that an apparent DOT in non-instrumented cultures can be inferred from the lactate yield of bioreactors as a function of DOT. Such a concept, can be a useful and important tool for determining the average dissolved oxygen tension in non-instrumented cultures. It was shown that the dynamic behavior of OUR can be correlated with monosaccharide (fructose and glucose) depletion and viable cell concentration. Accordingly, OUR can have two important applications in insect cell culture: for on-line estimation of viable cells, and as a possible feed-back control variable in automatic strategies of nutrient addition.Abbreviations DOT Dissolved oxygen tension - OUR Oxygen uptake rate - specific oxygen uptake rate - specific growth rate - X_v viable cell concentration - C_L, C^*, and oxygen concentrations in liquid phase, in equilibrium with gas phase, and medium molar concentration, respectively - H Henry's constant - K_La volumetric oxygen transfer coefficient - P_T total pressure - oxygen partial pressure - oxygen molar fraction - i discrete element     

溶氧水平对鲫鱼代谢模式的影响

为了探讨水体溶氧水平对鲫幼鱼(Carassius carassius)运动、消化能力及其交互作用的影响,在(25.0±0.5)℃温度条件下,测定了8(饱和溶氧水平)、2和1mg/L溶氧水平下摄食(饱足摄食)和空腹组(空腹2 d)鲫鱼的临界游泳速度(Ucrit)、运动前耗氧率(MO2pre-exercise)、活跃耗氧率(MO2active)和代谢范围(MS)。摄食诱导的耗氧率上升在各溶氧水平下无显著差异。在饱和溶氧水平下,摄食组和空腹组的Ucrit没有显著差异,但在1和2 mg/L条件下,摄食组的Ucrit显著低于空腹组(P<0.05)。在饱和溶氧水平条件下,消化和运动诱导的耗氧率上升在各个游泳水平均能完全叠加,且摄食组鱼类与空腹组鱼类具有相似的MS和Ucrit和更高的MO2active,提示鲫鱼在常氧下为添加代谢模式。随着溶氧水平下降至2和1mg/L,呼吸能力(摄食组的MO2active)对溶氧水平下降较运动耗氧率更为敏感,消化诱导的耗氧率增加只能在较低游泳速度叠加,与空腹组鱼类比较,摄食组鱼类的MS和Ucrit显著下降,MO2active无显著差异,提示低氧下消化和运动对氧气需求竞争的加剧使其代谢模式转化为消化优先。     

Relationship between production of carrot somatic embryos and dissolved oxygen concentration in liquid culture

To evaluate the relationship between somatic embryogenesis and dissolved oxygen concentration, somatic embryo cultures of carrot (Daucus carota L.) were cultured under various dissolved oxygen concentration levels (bubble free aeration with 4%, 7%, 20%, 30%, and 40% oxygen in flasks). The system used allows dissolved oxygen concentration control without bubble aeration or mixing speed modification. The total number of somatic embryos was not affected by the dissolved oxygen (DO) concentration tested. Even if globular-stage embryos were induced at a low level of oxygen aeration, heart-stage embryo formation was still repressed. Oxygen enrichment (20%, 30% and 40% oxygen) enhanced torpedo and cotyledonary-stage embryo production. The oxygen-enriched aeration was effective in promoting the growth of the late developmental stages. Sugar consumption did not increase when the oxygen concentration was enriched above the ambient level. The number of heart-stage embryos increased as oxygen concentration increased up to the 7% level, while above the 20% level no change in production was observed. The production of cotyledonary-stage embryos was directly related to oxygen concentration. These results support that oxygen-enriched aeration provides oxygen to the low oxygen areas in somatic embryo. After the heat-stage embryos, which were grown at the 7% level were transferred to a flask with ambient, they developed an elongated root part and eventually grew to normal plantlets. This revised version was published online in June 2006 with corrections to the Cover Date.     

Abiotic influences on dissolved oxygen in a riverine environment

Dissolved oxygen (DO) is an important parameter in riverine health. Periphyton and/or macrophytes are frequently the drivers behind the fluctuation of DO levels in aquatic environments; however, the effects of abiotic and biotic factors on biomass and in turn on DO may be variable from river to river. The objective of this paper is to understand which factors govern DO levels in terms of daily minimum DO (DO_min) and daily DO variation (ΔDO) in a major wastewater-impacted river using statistical data analysis and modeling. Both climatic conditions (reflected in water temperature) and hydrometric conditions (flow) were major factors influencing DO_min and ΔDO. The effect of flow on DO_min and ΔDO was discontinuous, depending on the flow magnitude. Nutrient loading from wastewater effluent was not identified to have a significant impact on a daily basis; however, their role over large time scales is unclear. In the data-driven modeling approaches, the non-linear approach using multiple-layer perceptron neural network, which has very flexible architecture, was superior to the linear approach used (multiple linear regression). Although nutrients are likely related to the DO_min and ΔDO, flow and temperature were sufficient to obtain robust prediction in the data-driven modeling. This approach is useful to model and understand complicated processes when their governing mechanisms are not well presented in conceptual- or physically based models.     

On-line gas analysis in animal cell cultivation: I. Control of dissolved oxygen and pH

To monitor gas reaction rates in animal cell culture at constant dissolved oxygen concentration (DO) and constant pH it was necessary to develop improved control methods. Decoupling of both controllrs was obtained by manipulation of molar fractions of oxygen and carbon dioxide in the gas phase. Two pairs of DO and pH controllers were designed and tested both in simulation and exprimental runs. The first controller pair was developed for headspace aeration only, whereas the second controller pair was designed for bubble aeration using a microsparger and flushing the headspace with helium. pH was controlled by a conventional discrete PID controller in its velocity form. For DO control two linear state space feedback controllers with parameter adaptation were established. In these controllers the oxygen uptake rate (OUR) was considered as a disturbance and was not included in the mathematical model. The feedback gain adaptation was based on the difference between the actual molar fraction of oxygen at time step n and the initial molar fraction. This difference is related to OUR and was used to increase or decrease the state feedback controller gain (k and k(1), respectively) in a slow manner. With these controllers it was possible to get an excellent online estimate of OUR. In the case of bubble aeration a simple gas phase mass balance was sufficient, whereas during the headspace aeration a liquid phase balance was required. It has been shown that determination of OUR using gas balance requires a significantly better controller performance compared to just keeping DO and pH within reasonable limits. (c) 1995 John Wiley & Sons, Inc.     

Effect of dissolved oxygen concentration on the metabolism of glucose inPseudomonas putida BM014

The effect of dissolved oxygen concentration on the metabolism of glucose inPseudomonas putida BM014 was investigated. Glucose was completely converted to 2-ketogluconatevia extracellular oxdative pathway and then taken up for cell growth under the condition of sufficient dissolved oxygen concentration. On the other hand, oxygen limitation below dissolved oxygen tension (DOT) value of 20% of air saturation caused the shift of glucose metabolism from the extracellular oxidative pathway to the intracellular phosphorylative pathway. Specific activities of hexokinase and gluconate kinase in intracellular phosphorylation pathway decreased as the DOT increased, while 2-ketogluconokinase activity in extracellular oxidative pathway increased under the same condition. This result can be usefully applied to microbial transformation of glucose to 2-ketogluconate, the synthetic precursor for iso-vitamine C, with almost 100% yieldvia extracellular oxidation by simple DOT control.     

Overcoming challenges in WAVE Bioreactors without feedback controls for pH and dissolved oxygen

The biopharmaceutical industry is increasing its use of the WAVE Bioreactor for culturing cells. Although this disposable bioreactor can be equipped to provide real-time pH and dissolved oxygen (DO) monitoring and control, our goal was to develop a process for culturing CHO cells in this system without relying on pH and DO feedback controls. After identifying challenges in culturing cells without controlling for pH and DO in the WAVE Bioreactor, we characterized O(2) and CO(2) transfer in the system. From these cell-free studies, we identified rock rate and rock angle as key parameters affecting O(2) transfer. We also identified the concentration of CO(2) in the incoming gas and the rate of gas flow into the headspace as key parameters affecting CO(2) transfer--and therefore pH--in the disposable culture chamber. Using a full-factorial design to evaluate the rock rate, rock angle, and gas flow rate defined for this WAVE Bioreactor process, we found comparable cell growth and pH profiles in the ranges tested for these three parameters in two CHO cell lines. This process supported cell growth, and maintained pH and DO within our desired range--pH 6.8-7.2 and DO exceeding 20% of air saturation--for six CHO cell lines, and it also demonstrated comparable cell growth and viability with the stirred-tank bioreactor process with online pH and DO control. By eliminating the use of pH and DO probes, this process provides a simple and more cost-effective method for culturing cells in the WAVE Bioreactor.     

Spatial and temporal variation in dissolved oxygen in natural egg pockets of chum salmon, in Kennedy Creek, Washington

The effects of dissolved oxygen (DO) on incubating salmonid embryos have been studied extensively in the laboratory but there is little information on levels experienced by salmon embryos in complex natural channels. We monitored 33 natural egg pockets of chum salmon Oncorhynchus keta , from shortly after spawning until emergence, and found that DO levels varied considerably among and within egg pockets over time. Egg pocket DO levels varied from 2–10 mg l⁻¹ at the time of spawning and generally declined during incubation but the declines did not occur in all pockets and were not always steady. Much of the variability may be attributed to local channel topography. Pool tailouts had the highest and least variable DO levels whereas levels in lateral bars were generally lower and more variable. Levels in glides and riffles tended to be intermediate between those in pool tailouts and lateral bars. In spite of the variation in DO levels and habitats used by chum salmon, DO levels were not correlated with egg pocket sediment composition (per cent of the sample<1.0 or 4.0 mm diameter) or with the egg pocket's depth.     

A model for predicting the temporal evolution of dissolved oxygen concentration in shallow estuaries

The design of sewage discharge systems in estuaries needs to consider the dissolved oxygen concentration among other water quality indicators. Due to the great number of factors affecting the dissolved oxygen, the prediction of the temporal evolution of this element requires the use of mathematical tools. In the case of shallow estuaries with extensive intertidal zones, the complexity of this task increases since the water domain varies continuously. This work describes a numerical model which solves the vertical integrated transport equation including the effect of extensive tidal flats. The model makes use of the tidal velocities computed by a previous run of a hydrodynamic model. This procedure allows the dissolved oxygen modelling to be performed using a larger time step than that used for the velocity field calculation. The numerical scheme developed for the model guarantees that even in the drying and wetting of intertidal areas the water column dissolved oxygen concentration is not modified by numerical errors. The representations of the processes included in the model (advection, dispersion and reaction) have been validated successfully in several theoretical cases. An application to the Urdaibai Estuary, a shallow estuary in the Basque Country, Northern Spain, is also presented.     

Distribution of fingerling brook trout, Salvelinus fontinalis (Mitchill), in dissolved oxygen concentration gradients

A self-recording linear gradient tank and procedures are described in which individual brook trout fingerlings unstressed by recent transfer, unaccustomed surroundings or the presence of an observer could move freely in 16 oxygen concentration gradients within the limits of 1 and 8.9 mg O₂1⁻¹. They avoided oxygen concentrations below 4 mg 1⁻¹ most of the time and preferred 5 mg 1⁻¹ or higher more than half the time, which supports the field-derived belief that fish avoid oxygen concentrations below 5 mg l⁻¹ in the natural environment if they can.     

Online detection of feed demand in high cell density cultures of Escherichia coli by measurement of changes in dissolved oxygen transients in complex media

A starvation-based dissolved oxygen (DO) transient controller was developed to supply growth-limiting substrate to high cell density fed-batch cultures of recombinant Escherichia coli. The algorithm adjusted a preexisting feed rate in proportion to the culture's oxygen demand, which was estimated from transients in the DO concentration after short periods of feed interruption. In this manner, the addition of glucose feed was precisely controlled at a rate that did not exceed the acetate production threshold, thus preventing acetate accumulation. In comparison to exponential feed algorithms commonly used in industry, the implementation of the new feeding strategy increased the final cell density from 32 to 44 g (dry cell weight).L(-1), with less than 16 mM acetate accumulated, producing an ideal culture for subsequent induction. Despite a constant starvation level and relatively low levels of acetate, experimental cultivations still tended to produce acetate towards the end of the process. The use of a simple Monod model provided an explanation as to why this may occur in high cell density cultivations and suggests how it may be overcome.     

Daily cycles of dissolved oxygen and nutrient content in a shallow fishpond: The impact of water renewal

This study was conducted in a shallow fishpond located on the eastern side of Arcachon Bay, France.Dissolved oxygen, pH and nutrient content (NH₄, NO₂, NO₃ and PO₄) showed pronounced diel variations.These phenomena seemed to be directly controlled by biological processes. Microbially mediated degradation of organic matter constitutes the main source of nutrients in the water column and primary producers consume the major part of these products. These processes are closely coupled to the light-dark cycle.Man-induced water renewal caused low modifications in nutrient concentration and did not alter its temporal dynamics.     

Temporal and spatial variations of dissolved oxygen concentrations in some Dutch polder ditches

This study deals with the temporal and spatial variations of dissolved oxygen concentration, temperature and occasionally pH in three polder ditches in the Netherlands. A routine programme including different environmental parameters served to characterize the ditch environment. Conclusions concerning water quality criteria are presented briefly.     

Long-term continuous monitoring of dissolved oxygen in cell culture medium for perfused bioreactors using optical oxygen sensors

For long-term growth of mammalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to ascertain the health of the cells. An optical oxygen sensor based on dynamic fluorescent quenching was developed for long-term continuous measurement of DO for NASA-designed rotating perfused bioreactors. Tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II) chloride is employed as the fluorescent dye indicator. A pulsed, blue LED was chosen as the excitation light source. The sensor can be sterilized using an autoclave. The sensors were tested in a perfused rotating bioreactor supporting a BHK-21 (baby hamster kidney) cell culture over one 28-day, one 43-day, and one 180-day cell runs. The sensors were initially calibrated in sterile phosphate-buffered saline (PBS) against a blood-gas analyzer (BGA), and then used continuously during the entire cell culture without recalibration. In the 180-day cell run, two oxygen sensors were employed; one interfaced at the outlet of the bioreactor and the other at the inlet of the bioreactor. The DO concentrations determined by both sensors were compared with those sampled and measured regularly with the BGA reference. The sensor outputs were found to correlate well with the BGA data throughout the experiment using a single calibration, where the DO of the culture medium varied between 25 and 60 mm Hg at the bioreactor outlet and 80-116 mm Hg at the bioreactor inlet. During all 180 days of culture, the precision and the bias were +/-5.1 mm Hg and -3.8 mm Hg at the bioreactor outlet, and +/- 19 mm Hg and -18 mm Hg at inlet. The sensor dynamic range is between 0 and 200 mm Hg and the response time is less than 1 minute. The resolution of the sensor is 0.1 mm Hg at 50 mm Hg, and 0.25 mm Hg at 130 mm Hg.