Physiological performance of juvenile southern flounder, Paralichthys lethostigma (Jordan and Gilbert, 1884), in chronic and episodic hypoxia

Dissolved oxygen (DO) is proving to be one of the most important abiotic factors determining growth and survival of juvenile estuarine fish. In shallow, throughout estuarine systems, low DO can occur in two broad categories: a diel oscillating pattern resulting in repeated nocturnal hypoxia due to the photosynthesis-respiration cycle of algal populations, or as prolonged bottom water hypoxia or anoxia caused by stratification. A series of laboratory experiments was conducted to characterize the physiological performance of juvenile southern flounder, Paralichthys lethostigma, (55-65 mm TL) exposed to four treatments of DO: (1) constant normoxia (6.50+/-0.50 mg O(2) l(-1)), (2) constant hypoxia (2.79+/-0.19 mg O(2) l(-1)), (3) constant intermediate hypoxia (4.74+/-0.18 mg O(2) l(-1)), and (4) an oscillating oxygen environment cycling dielly between the normoxic and hypoxic levels (2.8-6.2 mg O(2) l(-1), daily mean=4.40 mg O(2) l(-1)). Routine respiration was positively correlated with DO level and increased significantly during the day in the oscillating treatment in response to increasing DO. Ventilation rates were negatively correlated with the DO level in the constant treatments and increased significantly at night in the oscillating treatment in response to nocturnal hypoxia. Similarly, hematocrit levels were negatively related to DO levels in the constant treatments after 5 and 26 days of exposure to the treatments. Hematocrit levels also increased significantly the oscillating treatment, apparently in response to the episodic nocturnal hypoxia. Growth was significantly reduced in the 2.8 mg O(2) l(-1) treatment and the oscillating treatment but not in the 4.7 mg O(2) l(-1) treatment. Acclimation was evident by an increase in growth rates from week 2 to week 3 and a decrease in hematocrit levels between 5 and 26 days of exposure in the 2.7 and 4.5 mg O(2) l(-1) treatments but was not evident in the normoxic or oscillating treatments. These results suggest that a juvenile fish must remain in even moderately low DO in order for acclimation to occur. The research presented demonstrates that correctly assessing habitat quality in terms of DO requires knowledge of a fish's physiological and environmental history.     

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.     

Survival,growth and physiological responses of juvenile Japanese flounder (Paralichthys olivaceus,Temminck & Schlegel, 1846) exposed to different dissolved oxygen concentrations and stocking densities

The aim of this study was to determine to what extent juvenile Japanese flounder can adapt to different stocking densities in captivity and to examine whether growth and some physiological parameters critical for welfare might be affected by different dissolved oxygen levels. Japanese flounder (Paralichthys olivaceus) juveniles (initial weight 1.27 ± 0.04 g/fish) were reared at five stocking densities (500, 1,000, 1,500, 2,000, 2,500 ind/m³) and two levels of dissolved oxygen (DO) concentration (5.5 ± 0.5 mg/L or 14 ± 2 mg/L) with duplicate tanks for each treatment in water‐recirculating systems for 40 days. Survival and activity of superoxide dismutase (SOD) were not affected either by stocking density or dissolved oxygen, whereas final body weight, specific growth rate (SGR) and feed conversion efficiency (FCE) of fish under low DO concentration decreased significantly with increasing stocking density. In contrast, growth of fish reared in high DO levels were unaffected by the stocking density. Furthermore, fish in this group had a higher feed intake and, consequently, grew faster (SGR) and achieved a higher final weight than fish reared at the low DO level. A significant reduction in hemoglobin (Hb) concentration and red blood cell (RBC) count of fish were recorded as DO concentration increased. Furthermore, the activity of protease decreased significantly with increasing stocking density and increased significantly with increasing DO concentration. The ventilation frequency results indicate that gill ventilation decreased significantly as DO levels increased. This study demonstrates that stocking density can affect the growth performance and physiological parameters critical for welfare of juvenile Japanese flounder. Also shown is that pure oxygen supplementation is an effective way to improve the growth of juvenile Japanese flounder when reared at a high stocking density.     

Effects of moderate hypothermia on O2 consumption at various O2 deliveries in a sheep model.

The effects of modest hypothermia on oxygen consumption (VO2) were studied at various levels of oxygen delivery (DO2) in six sheep. Each animal was placed on cardiopulmonary bypass by extrathoracic cannulations. DO2 was varied by changing blood flow through an extracorporeal circuit. VO2 was measured spirometrically across a membrane lung. VO2 was initially measured at various levels of DO2 at normothermic temperatures (39 degrees C). The animals were then cooled to 33 degrees C. DO2 was varied, and the corresponding VO2's were determined. The data at both temperatures demonstrated the biphasic relationship of VO2 to various levels of DO2. A critical level of DO2 (DO2 crit) was defined to reflect the transition area between the dependent and independent portions of the consumption-delivery curve. The average baseline VO2's on the delivery independent portion of the curve were calculated to be 5.33 and 3.17 ml O2.kg-1.min-1 at 39 and 33 degrees C, respectively (P less than 0.001). The corresponding DO2 crit's were 6.17 and 4.57 ml O2.kg-1.min-1 (P less than 0.05). The oxygen extraction ratios at DO2 crit for each of these temperatures did not differ significantly. We conclude that hypothermia, by lowering baseline VO2, reduces DO2 crit. Hypothermia may therefore reduce or eliminate the anaerobic metabolism and subsequent acidosis that would otherwise occur during normothermia at low levels of DO2.     

Hypoxia-induced growth rate reduction in two juvenile estuary-dependent fishes

As eutrophication of coastal waters increases, water quality issues such as hypoxia have come to the forefront of environmental concerns for many estuarine systems. Chronic hypoxia during the summer has become a common occurrence in numerous estuaries, degrading nursery habitat and increasing the potential for exposure of juvenile fish to low levels of dissolved oxygen (DO).We conducted a laboratory study to investigate how hypoxic conditions and temperature affect growth rates of two juvenile estuary-dependent fish: the Atlantic menhaden (Brevoortia tyrannus) and spot (Leiostomus xanthurus). For a 2-week period, we exposed the fish to one of four constant DO levels (6.0, 4.0, 2.0 or 1.5 mg O₂ l⁻¹), at one of two temperatures (25 or 30 °C). A fifth DO treatment, included for spot at 30 °C, allowed DO to fluctuate from 10.0 mg O₂ l⁻¹ during the day, to 2.0 mg O₂ l⁻¹ at night. This diel fluctuation approximated the natural DO cycle in tidal estuarine creeks. Size measurements were recorded at the beginning, middle and end of experiments.Growth rates were generally unaffected by low DO until concentrations dropped to 1.5 mg O₂ l⁻¹, resulting in 31-89% growth reductions. Our results suggest that DO levels must be severely depressed, and in fact, approaching lethal limits, to negatively impact growth of juvenile spot and Atlantic menhaden.     

Comparisons of the glycosylation of a monoclonal antibody produced under nominally identical cell culture conditions in two different bioreactors

The murine B-lymphocyte hybridoma cell line, CC9C10, was grown in serum-free continuous culture at steady-state dissolved oxygen (DO) concentrations of 10%, 50%, and 100% of air saturation in both LH Series 210 (LH) and New Brunswick Scientific (NBS) CelliGen bioreactors. All culture parameters were monitored and controlled and were nominally identical at steady state in the two bioreactors. The secreted monoclonal antibody (mAb), an immunoglobulin G(1), was purified and subjected to enzymatic deglycosylation using peptide N-glycosidase F (PNGase F). Asparagine-linked (N-linked) oligosaccharide pools released from mAb samples cultured in each bioreactor at each of the three DO setpoints were analyzed by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The predominant N-linked structures were core-fucosylated asialo biantennary chains with varying galactosylation. There were also minor amounts of monosialyl oligosaccharides and trace amounts of afucosyl oligosaccharides. The level of DO affects the glycosylation of this mAb. A definite reduction in the level of galactosylation of N-glycan chains was observed at lower DO in both bioreactors, as evidenced by prominent increases in the relative amounts of agalactosyl chains and decreases in the relative amounts of digalactosyl chains-with the relative amounts of monogalactosyl chains being comparatively constant. However, the quantitative results are not precise matches between the two bioreactors. The effect of DO on galactosylation is less pronounced in the NBS bioreactor than in the LH bioreactor, particularly the shift between the relative amounts of agalactosyl and digalactosyl chains in 10% and 50% DO. There are also perceptibly higher levels of sialylation of the mAb glycans in the NBS bioreactor than in the LH bioreactor at all three DO setpoints. The results indicate that the DO effect is not bioreactor specific and that nominally identical steady-state conditions in different chemostat bioreactors may still lead to some incongruities in glycosylation, possibly due to the particular architectures of the bioreactors and the design of their respective monitoring and control systems. The observed differences in N-linked glycosylation of the mAb secreted by the hybridoma grown in the LH and NBS bioreactors may be explained by the differences in oxygen supply and control strategies between the two bioreactors.     

Behaviour of rainbow trout Oncorhynchus mykiss presented with a choice of normoxia and stepwise progressive hypoxia

The objective of this study was to identify behavioural adjustments leading to avoidance of hypoxia. Using the oxygen-sensitive species rainbow trout Oncorhynchus mykiss as a model, individual fish were recorded while moving freely between two sides of a test arena: one with normoxia and one with stepwise progressive hypoxia [80-30% dissolved oxygen (DO) air saturation]. The results demonstrated a gradual decrease in the total time spent in hypoxia starting at 80% DO air saturation. At this DO level, the avoidance of hypoxia could not be attributed to changes in spontaneous swimming speed, neither in normoxia nor in hypoxia. Reducing the DO level to 60% air saturation resulted in decreased spontaneous swimming speed in normoxia, yet the number of trips to the hypoxic side of the test arena remained unchanged. Moreover, data revealed increased average residence time per trip in normoxia at DO levels ≤60% air saturation and decreased average residence time per trip in hypoxia at DO levels ≤50% air saturation. Finally, the spontaneous swimming speed in hypoxia increased at DO levels ≤40% air saturation and the number of trips to hypoxia decreased at the 30% DO air saturation level. Thus, avoidance of the deepest hypoxia was connected with a reduced number of trips to hypoxia as well as decreased and increased spontaneous swimming speed in normoxia and hypoxia, respectively. Collectively, the data support the conclusions that the mechanistic basis for avoidance of hypoxia may (1) not involve changes in swimming speed during mild hypoxia and (2) depend on the severity of hypoxia.     

Constant dissolved oxygen concentrations in cephalosporin C fermentation: Applicability of different controllers and effect on fermentation parameters

Summary The behaviour and applicability of several controllers for maintaining a constant dissolved oxygen concentration (DO) during the cephalosporin C production with Cephalosporium acremonium in a laboratory fermentor is described. The process controllers were realized on a MC 68000 based process computer using the real-time language PEARL. The discrete signum integral controller showed the best control action. In addition some derived fermentation data were calculated on-line by the process computer.The results obtained by comparison of fermentations carried out at DO between 10% and 40% saturation during ideophase indicate that high DO leads to a high specific production rate for cephalosporin C and a low specific production rate for penicillin N and vice versa. In the range of DO investigated the production of deacetyl and deacetoxy cephalosporin C is not affected by DO. A direct correlation between DO and the yield coefficients Y _P/S and Y _P/X could be established. The yield coefficient Y _P/O for cephalosporin C is constant in the DO range from 10%–40%.Dedicated to Prof. Dr. H. J. Rehm on the occasion of his 60th birthday     

Development of individual insulin infusion profiles for open loop infusion systems

A computer controlled syringe-type insulin infusion pump storing up to 254 different infusion rates, eight different meal programs and two different basal rates automatically changeable during 24 h in EPROM was used for insulin infusion applying a wavy step profile. This profile approaching the physiological postprandial insulin secretion of the B-cell was calculated by an algorithm following the biphasic insulin secretion model proposed by E. Cerasi . The computer program for the open loop infusion device simulated the feed-back structure of a closed loop insulin secretion control by an algorithm based upon a theoretical postprandial blood sugar profile. Fifteen unstable juvenile onset insulin requiring diabetics could be well controlled after two to three days of an intravenous open loop insulin infusion program. The programs consisted of two constant basal rates and superimposed wavy step profile programs activated at the beginning of each meal. The preabsorptive bolus or cephalic phase was an additional tool both for improved postprandial blood sugar control and further reduction of insulin consumption. The programmable insulin infusion device proved as a valuable tool for the study of a sophisticated insulin infusion profile suitable as well for open loop as for closed loop insulin infusion systems.     

Warming and hypoxia reduce the performance and survival of northern bay scallops (Argopecten irradians irradians) amid a fishery collapse

Warming temperatures and diminishing dissolved oxygen (DO) concentrations are among the most pervasive drivers of global coastal change. While regions of the Northwest Atlantic Ocean are experiencing greater than average warming, the combined effects of thermal and hypoxic stress on marine life in this region are poorly understood. Populations of the northern bay scallop, Argopecten irradians irradians across the northeast United States have experienced severe declines in recent decades. This study used a combination of high-resolution (~1 km) satellite-based temperature records, long-term temperature and DO records, field and laboratory experiments, and high-frequency measures of scallop cardiac activity in an ecosystem setting to quantify decadal summer warming and assess the vulnerability of northern bay scallops to thermal and hypoxic stress across their geographic distribution. From 2003 to 2020, significant summer warming (up to ~0.2°C year⁻¹) occurred across most of the bay scallop range. At a New York field site in 2020, all individuals perished during an 8-day estuarine heatwave that coincided with severe diel-cycling hypoxia. Yet at a Massachusetts site with comparable DO levels but lower daily mean temperatures, mortality was not observed. A 96-h laboratory experiment recreating observed daily temperatures of 25 or 29°C, and normoxia or hypoxia (22.2% air saturation), revealed a 120-fold increased likelihood of mortality in the 29°C-hypoxic treatment compared with control conditions, with scallop clearance rates also reduced by 97%. Cardiac activity measurements during a field deployment indicated that low DO and elevated daily temperatures modulate oxygen consumption rates and likely impact aerobic scope. Collectively, these findings suggest that concomitant thermal and hypoxic stress can have detrimental effects on scallop physiology and survival and potentially disrupt entire fisheries. Recovery of hypoxic systems may benefit vulnerable fisheries under continued warming.     

A simple and rapid method for monitoring dissolved oxygen in water with a submersible microbial fuel cell (SBMFC)

A submersible microbial fuel cell (SBMFC) was developed as a biosensor for in situ and real time monitoring of dissolved oxygen (DO) in environmental waters. Domestic wastewater was utilized as a sole fuel for powering the sensor. The sensor performance was firstly examined with tap water at varying DO levels. With an external resistance of 1000?, the current density produced by the sensor (5.6±0.5-462.2±0.5mA/m(2)) increased linearly with DO level up to 8.8±0.3mg/L (regression coefficient, R(2)=0.9912), while the maximum response time for each measurement was less than 4min. The current density showed different response to DO levels when different external resistances were applied, but a linear relationship was always observed. Investigation of the sensor performance at different substrate concentrations indicates that the organic matter contained in the domestic wastewater was sufficient to power the sensing activities. The sensor ability was further explored under different environmental conditions (e.g. pH, temperature, conductivity, and alternative electron acceptor), and the results indicated that a calibration would be required before field application. Lastly, the sensor was tested with different environmental waters and the results showed no significant difference (p>0.05) with that measured by DO meter. The simple, compact SBMFC sensor showed promising potential for direct, inexpensive and rapid DO monitoring in various environmental waters.     

A novel approach to controlling dissolved oxygen levels in laboratory experiments

Hypoxia is a widespread environmental stressor that affects marine, estuarine and freshwater systems worldwide. Investigating the effects of hypoxia on aquatic animals in the natural environment is difficult and expensive. Laboratory experiments provide an alternative that allows manipulation of environmental variables and simulation of altered conditions that are expected in the future. However, controlling dissolved oxygen (DO) levels precisely in laboratory test situations is challenging and generally costly and time intensive. In this paper, we describe a novel chamber design that is capable of maintaining low DO levels precisely for a period of at least 4 days with minimal re-adjustment and nitrogen gas requirement. The system is simple, inexpensive and easy to use, and offers the additional benefit of being portable. The utility of this design is demonstrated with an experiment to test the effects of hypoxia and high salinity on hatch rates and yolk-sac larvae survival in the estuarine fish black bream Acanthopagrus butcheri (Munro, 1949). The results show that early life-stage black bream are sensitive to hypoxia and increased salinity, indicating that deterioration in estuarine environmental conditions may have negative effects on spawning success and recruitment to fish populations. This new laboratory technique will be useful for testing future scenarios for the estuaries of southern Australia as predicted by climate change models. Other potential applications and modifications for further development of this new technique are discussed.     

Control of dissolved oxygen concentration in mammalian cell culture using a computer-coupled mass flow controller

Summary A simple proportional control system for dissolved oxygen (DO) concentration in cell culture medium was developed by using a computer-coupled mass flow controller. The DO levels were very stable during the cultivation of Vero-6, while flow rates of air and/or oxygen enriched air were gradually changed depending on the DO concentration and the preset DO level. Vero-6 cells could grow normally to the confluence in the range of 30% and 50% of DO. Growth of Vero-6 at 10% of DO was markedly retarded.     

The effect of dissolved oxygen,sediment, and sewage treatment plant discharges upon growth,survival and density of Asiatic clams

The biology of Corbicula fluminea, the Asiatic clam, in the Vermilion River, Louisiana, as affected by sediment, dissolved oxygen (DO) levels, and sewage treatment plant (STP) effluents was investigated. A point source of high DO water to the Vermilion River established a gradient of DO that decreased as the river moved towards the Gulf of Mexico. Lowering DO levels were exacerbated by municipal sewage treatment plant discharges in the 20 km reach studied. Low dissolved oxygen was associated with reduced Corbicula density in the river and 30-day in-stream growth studies (weight and length) demonstrated that low DO inhibited growth. Generally, if DO was < 1.0 mg l^–1 in sediment pore water and/or < 3.0 mg l^–1 at the sediment-water interface, growth was significantly impaired (p < 0.05). Corbicula experienced substantial mortality near the STP discharges (up to 70% in 30 days) and laboratory toxicity tests with Ceriodaphnia dubia, a sensitive cladoceran, also strongly suggested discharges were chronically toxic at 6.25–25.0% effluent. Respiration experiments along with environmental measurements of DO, temperature, and STP discharge chemistry support a hypothesis that clam populations are adversely affected by the suite of environmental conditions present in the Vermilion River. Further, growth studies were consistent with observed population densities in situ.     

Information processing in molecular systems

Information processing, or selective dissipation, is mediated by switching elements in classical systems and by enzyme catalysis in biochemical systems. There are important differences in the character of this dissipation (from the standpoint of energy and control) in self-reproducing systems based on molecular interactions and those based on conventional computers. Conventional computers process information in a single level mode, i.e., the state of each unit of the system is accessed independently. This includes the manipulable memory units which store the computer program. In contrast, molecular self-reproducing systems process information in a hierarchical mode, based on the fact of hierarchy in molecular structure. In particular, the enzyme is described genetically at the primary level of structure (amino acid sequence) but functions at the higher, tertiary level on the basis of the interactions of many manipulable units. As a consequence it is not possible to program a biochemical system in any conventional sense. However, this is compensated by an increased capacity for accumulating appropriate information through evolution by variation and natural selection. This is possible because systems operating in the hierarchical mode are amenable to gradual modification of function. The degree of gradualness is itself an evolved property of biological molecules.     

Self-cycling fermentation in a stirred tank reactor

Self-cycling fermentations (SCFs) were conducted in a stirred tank apparatus using Bacillus subtilis and Acinetobacter calcoaceticus. The systems were very stable and the experiments lasted through many cycles. The variation of parameters such as biomass and doubling time from cycle to cycle was small. The stirred tank reactor (STR) allowed a much better control of the working volume in the fermentor from cycle to cycle, compared to the cyclone column, and it was not necessary to make periodic corrections.The production of surfactin from B. subtilis was achieved without extending the cycle time. The harvested broth at the end of each cycle was allowed to remain in a secondary vessel, at ambient temperature, before being collected. It is exhaustion of the limiting nutrient which causes an increase in dissolved oxygen (DO). At this point, the computer, which constantly monitors the DO, triggered the harvesting sequence to end the cycle. Thus, the mature culture in the secondary vessel experienced appropriate conditions for the production of the secondary metabolite. Meanwhile, the next batch of cells was being grown in the primary reactor.The response of a gas analyzer on the effluent paralleled that of the DO measurements in the fermentor. These data for oxygen and carbon dioxide exhibited less noise than the DO readings. Either would be a more reliable parameter for feedback control of the SCF because the problem of fouling of the DO probe after extended runs of many cycles would be eliminated. (c) 1993 John Wiley & Sons, Inc.     

Feedforward-feedback control of dissolved oxygen concentration in a predenitrification system

As the largest single energy-consuming component in most biological wastewater treatment systems, aeration control is of great interest from the point of view of saving energy and improving wastewater treatment plant efficiency. In this paper, three different strategies, including conventional constant dissolved oxygen (DO) set-point control, cascade DO set-point control, and feedforward-feedback DO set-point control were evaluated using the denitrification layout of the IWA simulation benchmark. Simulation studies showed that the feedforward-feedback DO set-point control strategy was better than the other control strategies at meeting the effluent standards and reducing operational costs. The control strategy works primarily by feedforward control based on an ammonium sensor located at the head of the aerobic process. It has an important advantage over effluent measurements in that there is no (or only a very short) time delay for information; feedforward control was combined with slow feedback control to compensate for model approximations. The feedforward-feedback DO control was implemented in a lab-scale wastewater treatment plant for a period of 60 days. Compared to operation with constant DO concentration, the required airflow could be reduced by up to 8–15% by employing the feedforward-feedback DO-control strategy, and the effluent ammonia concentration could be reduced by up to 15–25%. This control strategy can be expected to be accepted by the operating personnel in wastewater treatment plants.     

Computer-assisted method for multi-exponential curve fitting for determining cerebral blood flow.

A computer program has been developed for use in determining cerebral blood flow using an inert radioactive gas. The basic algorithm involves the determination of multiple exponential coefficients from the complex concentration-time function. The exponential coefficients are determined by 'peeling' away slower exponentials complex function one at a time. The procedure involves the use of a small laboratory computer in the interactive graphics mode. The method is currently in use analyzing data in a cerebral vascular research laboratory.     

Economic weights of maternal and direct traits of pigs calculated by applying gene flow methods

Multiple trait selection indexes in pig breeding programmes should take into account the population structure and time delay between parent selection and expressions of traits in all production levels next to the trait impacts on economic efficiency of production systems. Gene flow procedures could be used for the correct evaluation of maternal and direct traits of pig breeds involved in breeding or crossbreeding systems. Therefore, the aim of this study was to expand a previously developed bioeconomic model and computer program to calculate the marginal economic values by including a gene flow procedure to calculate the economic weights for maternal and direct traits in pig breeds. The new program was then applied to the three-way crossbreeding system of the Czech National Programme for Pig Breeding. Using this program, the marginal economic values of traits for dam breeds Czech Large White in the dam position and Czech Landrace in the sire position, and for the sire breed Pietrain were weighted by the number of discounted gene expressions of selected parents of each breed summarised within all links of the crossbreeding system during the 8-year investment period. Economic weights calculated in this way were compared with the approximate economic weights calculated previously without a gene flow procedure. Taking into account the time delay between parent selection and trait expression (using discounting with half-year discount rates of 2% or 5%) and including more than one generation of parent progeny had little impact on the relative economic importance of maternal and direct traits of breeds involved in the evaluated three-way crossbreeding system. These results indicated that this gene-flow method could be foregone when estimating the relative economic weights of traits in pig crossbreeding systems applying artificial insemination at all production levels.