Electrode System for the Determination of Microbial Populations

Determinations of microbial populations were carried out by using a new electrode system composed of two electrodes. Each electrode was constructed from a platinum anode and a silver peroxide cathode. The anode of the reference electrode was covered with cellulose dialysis membrane. The response time of the electrode system was 15 min in culture broth, and current differences between the two electrodes were proportional to populations of microbial cells in cultures of Saccharomyces cerevisiae and Lactobacillus fermentum. Current differences were reproducible; the average relative error was 5%. Furthermore, cell populations of S. cerevisiae in a fermentor could be continuously estimated by using this electrochemical method.     

Lignocellulose Mineralization by Arctic Lake Sediments in Response to Nutrient Manipulation

Mineralization of specifically labeled ¹⁴C-cellulose- and ¹⁴C-lignin-labeled lignocelluloses by Toolik Lake, Alaska, sediments was examined in response to manipulation of various environmental factors. Mineralization was measured by quantifying the amount of labeled CO₂ released from the specifically labeled substrates. Nitrogen (NH₄NO₃) and, to a greater degree, phosphorus (PO₄⁻³) additions enhanced the mineralization of white pine (Pinus strobus) cellulose during the summer of 1978. Nitrogen and phosphorus together had no cumulative effect. During the summer of 1979, nitrogen or phosphorus alone had only a slight stimulatory effect on the mineralization of a sedge (Carex aquatilis) cellulose; however, together, they had a dramatic effect. This variable response of mineralization to nutrient addition between 1978 and 1979 was probably attributable to year-to-year variation in nutrient availability within the lake. Cellobiose addition and oxygen depletion inhibited the amount of pine cellulose mineralized. Whereas addition of nitrogen to oxygen-depleted treatments had limited effect, addition of phosphorus resulted in mineralizations equal to or greater than that of the controls. Nitrogen had no effect on mineralization of pine or Carex lignins. Phosphorus, however, inhibited mineralization of both lignins. With Carex lignin, the phosphorus inhibition occurred at a concentration as low as 0.1 μM. The antagonistic role of phosphorus in cellulose and lignin mineralizations may be of significance in understanding the increased proportion of lignin relative to cellulose in decomposing litter.     

Embryonic rotational behaviour in the pond snail Lymnaea stagnalis: influences of environmental oxygen and development stage

Responses of freshwater organisms to environmental oxygen tensions (PO₂) have focused on adult (i.e. late developmental) stages, yet responses of embryonic stages to changes in environmental PO₂ must also have implications for organismal biology. Here we assess how the rotational behaviour of the freshwater snail Lymnaea stagnalis changes during development in response to conditions of hypoxia and hyperoxia. As rotation rate is linked to gas mixing in the fluid surrounding the embryo, we predicted that it would increase under hypoxic conditions but decrease under hyperoxia. Contrary to predictions, early, veliger stage embryos showed no change in their rotation rate under hyperoxia, and later, hippo stage embryos showed only a marginally significant increase in rotation under these conditions. Predictions for hypoxia were broadly supported, however, with both veliger and hippo stages showing a marked hypoxia-related increase in their rotation rates. There were also subtle differences between developmental stages, with hippos responding at PO₂s (50% air saturation) greater than those required to elicit a similar response in veligers (20% air saturation). Differences between developmental stages also occurred on return to normoxic conditions following hypoxia: rotation in veligers returned to pre-exposure levels, whereas there was a virtual cessation in embryos at the hippo stage, likely the result of overstimulation of oxygen sensors driving ciliary movement in later, more developed embryos. Together, these findings suggest that the spinning activity of L. stagnalis embryos varies depending on environmental PO₂s and developmental stage, increasing during hypoxia to mix capsular contents and maintain a diffusive gradient for oxygen entry into the capsule from the external environment (“stir-bar” theory of embryonic rotational behaviour).     

The design,construction, and characteristics of a new long-lived steam sterilizable oxygen electrode

A steam sterilizable oxygen electrode for fermentor use is described. The electrode has a silver cathode, lead anode, phosphate electrolyte, and a membrane of a fluorinated ethylene-propylene copolymer film (FEP.). The electrode has a linear response to partial pressure of oxygen from 1.5 × 10^?2 to 10³ mm Hg.     

Compensation for measuring error produced by finite response time in determination of rates of oxygen consumption with a Clark-type polarographic electrode

In the determination of the rates of oxygen consumption with a Clark-type oxygen electrode, and experimental error is caused by finite response time of the oxygen electrode for a rapid oxidation reaction. A theoretical equation between the observed pseudo first-order rate constant (k_obs) and the true rate constant (k)

$\text{1}\text{k}{}_{\mathrm{obs}}\text{=}\text{1}\text{k}\text{+T}$

where T is a time constant for a first-order response of the oxygen electrode, was derived and found to hold up to k = 23 min^?1 in oxidation of hydroquinone at pH 7.60–8.63.     

Amperometric method for monitoring anaerobic respiration with nitrous oxide as the terminal electron acceptor

A commercially available Clark-type electrode with a platinum cathode, commonly used for monitoring oxygen uptake, may also be used to monitor respiration with N₂O. The disadvantages of this system include a high sensitivity to O₂, which may be overcome by excluding O₂, and sensitivity to acetylene. The advantage of the method is that it may be used to monitor directly the reduction of N₂O by respiring cells.     

Critical oxygen partial pressures and maximal tracheal conductances for Drosophila melanogaster reared for multiple generations in hypoxia or hyperoxia

In Drosophila melanogaster and other insects, increases in atmospheric oxygen partial pressure (aPO₂) tend to increase adult body size and decrease tracheal diameters and tracheolar proliferation. If changes in tracheal morphology allow for functional compensation for aPO₂, we would predict that higher aPO₂ would be associated with higher critical PO₂ values (CritPO₂) and lower maximal tracheal conductances (G_max). We measured CritPO₂ and G_max for adult and larval vinegar flies reared for 7-9 generations in 10, 21 or 40 kPa O₂. The CritPO₂, CO₂ emission rates and G_max values were generally independent of the rearing PO₂ these flies had experienced, suggesting that minimal functional changes in tracheal capacities occurred in response to rearing PO₂. Larvae were able to continue activity during 20 min of anoxia. The lack of multigenerational rearing PO₂ effects on tracheal function suggests that the functional compensation at the whole-body level due to tracheal morphological changes in response to aPO₂ may be minimal; alternatively the benefits of such compensation may occur in specific tissues or during processes not assessed by these methods. In larvae, the CritPO₂ and the capacity for movement in anoxia suggest adaptations for life in hypoxic organic matter.     

Design and application of electrodes for the determination of dissolved oxygen

The design and circuitry of solid electrodes of the Clark type for the determination of oxygen dissolved in microbial cultures and the suitable techniques are described. For the electrodes a portable battery instrument with an amplifier and recorder was developed. The electrode materials tested were: Au, Pt, Ag (cathode) and Ag/Ag₂O and Ag/AgCl (anode). The effect of the presence of carbon dioxide on the course of the polarographic curves was determined. The temperature effect was determined for various membrane materials and its compensation with a thermistor was investigated. The effect of the circuit and design of the electrodes on stability, response time, reproducibility of measurement, durability of the electrodes and applicability in microbiology are discussed.     

Oxygen poisoning of NAD biosynthesis: a proposed site of cellular oxygen toxicity.

Quinolinate phosphoribosyl transferase was rapidly inactivated in $\text{Escherichia}\text{coli}$ exposed to hyperbaric oxygen. The enzyme is essential for de novo biosynthesis of NAD in $\text{E.}\text{coli}$ and man. Because of its sensitivity and essentiality, inactivation of this enzyme is proposed as a significant mechanism of cellular oxygen toxicity. Niacin which enters the NAD biosynthetic pathway below the oxygen-poisoned enzyme provided significant protection against the decrease in pyridine nucleotides and the growth inhibition from hyperoxia in $\text{E.}\text{coli}$ and could be useful in cases of human oxygen poisoning.     

Oxygen-dependence of upper thermal limits in crustaceans from different thermal habitats

The critical thermal maximum (CT_MAX) is the temperature at which animals exhibit loss of motor response because of a temperature-induced collapse of vital physiological systems. A central mechanism hypothesised to underlie the CT_MAX of water-breathing ectotherms is insufficient tissue oxygen supply for vital maintenance functions because of a temperature-induced collapse of the cardiorespiratory system. The CT_MAX of species conforming to this hypothesis should decrease with declining water oxygen tension (PO₂) because they have oxygen-dependent upper thermal limits. However, recent studies have identified a number of fishes and crustaceans with oxygen-independent upper thermal limits, their CT_MAX unchanged in progressive aquatic hypoxia. The previous studies, which were performed separately on cold-water, temperate and tropical species, suggest the oxygen-dependence of upper thermal limits and the acute thermal sensitivity of the cardiorespiratory system increases with decreasing habitat temperature. Here we directly test this hypothesis by assessing the oxygen-dependence of CT_MAX in the polar Antarctic krill (Euphausia superba), as well as the temperate Baltic prawn (Palaemon adspersus) and brown shrimp (Crangon crangon). We found that P. adspersus and C. crangon maintain CT_MAX in progressive hypoxia down to 40 mmHg, and that only E. superba have oxygen-dependent upper thermal limits at normoxia. In E. superba, the observed decline in CT_MAX with water PO₂ is further supported by heart-rate measurements showing a plateauing, and subsequent decline and collapse of heart performance at CT_MAX. Our results support the hypothesis that the oxygen-dependence of upper thermal limits in water-breathing ectotherms and the acute thermal sensitivity of their cardiorespiratory system increases with decreasing habitat temperature.     

Metabolic responses of the mussels Perna viridis and Perna indica to declining oxygen tension at different salinities

• 1.1. Oxygen uptake and ammonia loss were monitored during responses to reductions of both salinity and oxygen tension (PO₂) in the marine mussels Perna viridis and Perna indica from southern India.
• 2.2. The proportional contribution of protein to total catabolic substrates under natural environmental conditions was as much as 96% in P. viridis, relative to only 19% in P. indica.
• 3.3. Normoxic oxygen consumption remained statistically unchanged in P. viridis conditioned to salinities between 32 and 15‰, with no obvious signs of distress. Although equally unaffected at salinities between 32 and 20‰, P. indica showed significantly reduced oxygen uptake following transfer from 32 to 15‰, and had died within the next 7 days.
• 4.4. At salinities greater than 20‰, P. viridis was better able than P. indica to regulate oxygen consumption independent of PO₂.
• 5.5. P. indica showed a compensatory increase in oxyregulatory capacity at 15‰. This exceeded unstressed abilities, helping to maintain albeit reduced oxygen uptake throughout wider ranges of PO₂.
• 6.6. Different responses recorded in each of these tropical and often intertidal species were in accordance with their natural distributions. Nevertheless, the oxyregulatory capacity in both species was higher than in bivalves from temperate and/or subtidally restricted habitats.

     

Respiratory rate,haemolymph oxygen tension and haemocyanin level in the shrimp Palaemon adspersus: Rathke

The marine shrimp Palaemon adspersus Rathke, frequenting Zostera meadows, shows an oxygen consumption rate (MO₂) that is independent of water oxygen tension (P_wO₂) as PO₂ decreases to a critical point (P_cr) near 70 mm Hg. This respiratory independence is associated with maintenance of a relatively constant “arterial” (post-branchial) haemolymph tension (P_aO₂) at 70–80 mm Hg. At lower P_wO₂ values, both MO₂ and P_aO₂ fall, reflecting, in contrast to the above independence, a direct dependence of MO₂ on internal PO₂.Haemolymph copper measurements demonstrate relatively high haemocyanin concentrations and oxygen-carrying capacities, but MO₂ levels reflect an insignificant rôle for the pigment as an oxygen store.The data are discussed as regards adaptations for aerobic metabolism.     

Respiration and oxygen transport functions of the blood from an intertidal fish,Helcogramma medium (Tripterygiidae)

Synopsis Aquatic and aerial oxygen uptake (̇O₂), ventilation frequency, and oxygen transport properties of the blood were determined for the intertidal fish Helcogramma medium. Ventilation frequency increased in response to decreased environmental PO₂ and aquatic ̇O₂ was maintained down to a critical PO₂ of 30–40 mm Hg. Below PO₂ 30 mm Hg fish intermittently gulped air and finally emerged into air at PO₂ 18 mm Hg. After 1 h exposure to air ̇O₂ decreased to 60% of the aquatic rate and this was accompanied by an increase in blood lactate. Aerobic expansibility was reduced in air (×1.2) compared to water (× 5.5). The Hb concentration was 0.47 ± 0.13 mmol 1^–1 and hematocrit 11.55 ± 3.61% indicating a moderate O₂-carrying capacity. Oxygen affinity was not especially high (P₅₀ = 19 mm Hg at pH 7.7 and 15°C) and ATP was the predominant acid-soluble phosphate regulating P₅₀. The equilibrium curve was essentially hyperbolic (Hill's n = 1.2) with a marked Bohr effect = –1.06) and Root effect (saturation depressed by 50% at pH7.1). The pattern of respiration and the respiratory properties of the blood together with observations of the behaviour of the fish during aerial exposure indicated that Helcogramma is adapted to living in a well-aerated environment yet can adequately tolerate short term exposure to low aquatic PO₂ or air.     

Construction and operation of freshwater sediment microbial fuel cell for electricity generation

In this work, sediment microbial fuel cell (SMFC) with granule activated carbon (GAC) cathode and stainless steel anode was constructed in laboratory tests and various factors on SMFC power output were investigated. The maximum power densities for the SMFC with GAC cathode was 3.5 mW m⁻², it was much higher than SMFC with round stainless steel cathode. Addition of cellulose reduced the output power from SMFC at the beginning of experiments, while the output power was found to increase after adding cellulose to sediments on day 90 of operation. On 160 day, maximum power density from the SMFC with adding 0.2% cellulose reached to 11.2 mW m⁻². In addition, the surface morphology of stainless steel anode on day 90 was analyzed by scanning electron microscope. It was found that the protection layer of the stainless steel as electrode in SMFCs was destroyed to some extent.     

Tissue oxygen partial pressure in organs of chickens in the second half of embryogenesis and first days after hatching

The aim of this study is to measure the oxygen partial pressure (pO₂) in developing chicken tissues, namely, in the cerebral hemispheres, liver, m. pectoralis, and m. gastrocnemius, and to estimate the correlation of pO₂ with the earlier measured values (laser Doppler flowmetry) of volume blood flow (BF) in these organs. We have studied 10-, 15-, and 19-day-old embryos and 4-day-old chickens anesthetized with urethane. The pO₂ has been measured in the surface layers of organs with a membrane amperometric Clark-type O₂ electrode (cathode diameter of approximately 50 μm) placed in the center of the sensor unit (outer diameter of 3.4 mm). Noticeable distinctions between both the tissue pO₂ values in different organs and the dynamics of their changes during the observation time have been recorded. The following differences are the most important: (1) the lowest pO₂} {cm(and BF) is observed in the brain and, especially, in the liver of 10-day-old embryos; (2) in the subsequent period of embryogenesis, the pO₂ in the brain increases 1.9-fold (BF also increases), falls 1.7-fold in m. pectoralis, and displays minor changes in the liver and m. gastrocnemius on the background of constant BF value in the liver and both muscles; and (3) after hatching, pO₂ in the liver and m. pectoralis increases severalfold (BF increases too) but does not change in a statistically significant manner in the brain and m. gastrocnemius despite an increase in BF (more pronouncedly in the muscle). Two possible mechanisms underlying the changes in the tissue pO₂ in developing chicken organs have been proposed: one is determined by the specific features of intracardiac blood flows and the other is associated with the oxyhemoglobin dissociation pattern in the blood capillary circulation in the organs, determined by the specific features in its oxidative metabolism.     

Ductus arteriosus responses to prostaglandin E1 at high and low oxygen concentrations

It previously has been suggested that prostaglandin E₁ (PGE₁) relaxes the ductus arteriosus in a low but not in an elevated oxygen environment. However, in the experiments reported here PGE₁ relaxed rings on fetal lamb ductus arteriosus at both low (14 to 20 torr) and high (680 to 720 torr) oxygen tensions. The threshold concentration for PGE₁ was 10⁻¹⁰ M in either PO₂ and the ED50's of PGE₁ relaxation in high and low oxygen were 8.5 ± 3.4 × 10⁻¹⁰ M and 5.5 ± 0.7 × 10⁻¹⁰ M respectively. The magnitude of the relaxation was greater for the oxygen contracted ductus arteriosus than for that exposed to low oxygen. It is suggested that earlier reports of the lack of response of the ductus arteriosus to PGE₁ in a high oxygen environment following relaxation in a low oxygen environment may be related to loss of response of the ductus arteriosus to repeated doses of PGE₁ rather than to differences in PO₂. Prostaglandin E₁ therefore may play a significant role in the regulation of ductus arteriosus tone in the elevated oxygen environment of the newborn as well as the low oxygen environment of the fetus.     

A novel oxyconforming response in the freshwater fish Galaxias maculatus

How fish oxygen consumption is modulated by external PO₂ has long been a matter of interest, yet is an experimentally complicated question to answer. In this study closed and semi-closed respirometry were used to evaluate the oxygen consumption rate of the scaleless galaxiid fish, inanga (Galaxias maculatus) as a function of decreasing external PO₂. Both respirometry techniques showed that as environmental oxygen levels declined, oxygen consumption rates also decreased. At no point did inanga regulate oxygen consumption. This is strong evidence that inanga is an oxyconformer. Partitioned respirometry experiments showed that skin plays an important role in oxygen uptake in this fish species, and cutaneous oxygen uptake may have an important role in shaping the oxygen consumption response to hypoxia.