Measurement of k(L)a by dynamic pressure method in pilot-plant fermentor

The dynamic pressure method (DPM) is used for measurement of k(L)a in a 1-m(3) pilot scale fermentor in coalescing (distilled water) and noncoalescing (0.3 M Na(2)SO(4) aqueous solution) batches. The method consists in recording oxygen concentration in a batch after a small pressure change (20 kPa) in the fermentor. The upward pressure change is brought about by temporary closing and subsequent throttling of outlet gas stream and the downward change by full reopening of the gas outlet. Absorption of pure oxygen yields the same k(L)a values as absorption of air. In noncoalescing batch, the downward k(L)a values are always higher than the upward values owing to spontaneous nucleation of bubbles. The experiments performed in a stirred cell confirm this behavior. Thus, only upward pressure change should be used for measurement. The correlation of k(L)a data measured in small (18-L) and large (1000-L) vessels based on power dissipated and superficial gas velocity are in a good agreement. Unlike the DPM, the classical dynamic methods yield, under the same conditions, excessively low values of k(L)a (the dynamic startup method) or fail to produce data at all (the dynamic method with interchange of air for N(2)). (c) 1994 John Wiley & Sons, Inc.     

Combined sulfite method for the measurement of the oxygen transfer coefficient k(L)a in bio-reactors

The combined sulfite method is proposed for the measurement of oxygen transfer coefficients, k_La, in bioreactors. The method consists of a steady-state and a dynamic measurement which are carried out under the same experimental conditions and thus yield data for both methods during one experiment. The applied experimental conditions are shown to avoid chemical enhancement during the steady-state measurement. Moreover, no parallel sulfite oxidation occurs during the oxygen saturation phase of the dynamic measurement. Under the applied experimental conditions, no information about the sulfite oxidation kinetics is required and possible metal ion impurities in sulfite salts do not influence the measurement. The characterization of a laboratory-scale bioreactor aerated with pure oxygen yields k_La values during the steady-state and the dynamic measurements that are in good agreement with the dynamic pressure method, the correctness of which is generally accepted. When air is used for absorption, the steady-state measurement yields k_La values that correlate to the correct variant of the standard dynamic method. The dynamic measurement with air absorption yields a k_La value which considers the influence of the non-uniform bubble size distribution present in bubble-aerated bioreactors.     

Determination of the volumetric mass transfer coefficient (k(L)a) using the dynamic "gas out-gas in" method: Analysis of errors caused by dissolved oxygen probes

There are many dynamic methods for measuring the volumetric mass transfer coefficient. The "gas out-gas in" method can directly determine the volumetric mass transfer coefficient in a bioreactor system and provide estimates of the volumetric microbial oxygen uptake rate and the average oxygen saturation concentration at the gas-liquid interface. The errors on these parameters are large if the dissolved oxygen probe response time is not considered. For reliable measurements, deconvolution of the oxygen probe measurements must be made. (c) 1995 John Wiley & Sons, Inc.     

Infradian cycles of oxygen consumption in diapausing pupae of the flesh fly, Sarcophaga crassipalpis, monitored by a scanning microrespirographic method.

Fine details of the infradian O2 consumption cycles that characterize pupal diapause in flesh flies have been monitored by a newly designed microrespirographic method coupled with an electronically regulated O2 generator. During the 4-5 days between the peaks of elevated O2 consumption, the diapausing pupae maintained a very low and fairly constant respiratory rate (13 microl O2 x g-1.h-1). During the intercalated peaks of increased respiratory metabolism, which lasted an average of 33.6 h to 24-27 degrees C, the average maximum rate of O2 consumption was 86.9 microl.g-1.h-1, a value of 6.7 times higher than the interpeak values. The respiratory peaks started abruptly in some cases while the decline was consistently gradual. During the periods between the peaks there were no discontinuous bursts of CO2 release, a feature common to diapause in many other insects. Diapause was characteristically terminated during a peak of the O2 consumption cycle. At diapause termination O2 consumption remained at the maximum values of the peak for many hours and then gradually increased to levels characteristic of nondiapause development.     

Improving the dynamic response of a mediator-less microbial fuel cell as a biochemical oxygen demand (BOD) sensor

The dynamic behavior of a mediator-less, microbial fuel cell (MFC) was studied as a continuous biochemical oxygen demand (BOD) sensor. The response time and the sensitivity were analyzed through the step-change testing of the fuel concentration. The MFC of 25 ml had the shortest response time of 36± 2 min at the fuel-feeding rate of 0.53 ml min^–1 and the resistance of 10 A smaller MFC of 5 ml had a response time of 5± 1 min.     

大气CO2与吡虫啉对甘蓝土壤细菌与微生物生物量C的影响

采用田间开顶式CO2控制气室(OTC),研究了375μL/L、750μL/L两个CO2浓度和CK、LC50、LC903种吡虫啉浓度处理条件下,甘蓝根际土壤细菌与非根际土壤微生物生物量C的变化。750μL/L CO2处理对甘蓝根际细菌数量显著增加(P0.01),而在同一CO2水平下各农药处理间并无显著差异;根区土壤微生物生物量C只有在750μL/L CO2且无吡虫啉处理的条件下显著(P0.05)下降,在LC50、LC90处理的影响下并不显著。同一CO2水平下,根区土壤微生物生物量C受农药处理的影响不明显。     

Effects of elevated CO2 and O3 on the rate and duration of grain growth and harvest index in spring wheat (Triticum aestivum L.)

Wheat (Triticum aestivum L.) cv. Minaret was grown in open-top chambers (OTCs) in 1995 and 1996 under three carbon dioxide (CO₂) and two ozone (O₃) levels. Plants were harvested regularly between anthesis and maturity to examine the rate of grain growth (dG/dt; mg d^–1) and the rate of increase in harvest index (dHI/dt;% d^–1). The duration of grain filling was not affected by elevated CO₂ or O₃, but was 12 days shorter in 1995, when the daily mean temperature was over 3 °C higher than in 1996. Season-long exposure to elevated CO₂ (680 μmol mol^–1) significantly increased the rate of grain growth in both years and mean grain weight at maturity (MGW) was up to 11% higher than in the chambered ambient air control (chAA; 383 μmol mol^–1). However, the increase in final yield obtained under elevated CO₂ relative to the chAA control in 1996 resulted primarily from a 27% increase in grain number per unit ground area. dG/dt was significantly reduced by elevated O₃ under ambient CO₂ conditions in 1995, but final grain yield was not affected because of a concurrent increase in grain number. Neither dG/dt nor dHI/dt were affected by the higher mean O₃ concentrations applied in 1996 (77 vs. 66 nmol mol^–1); the differing effects of O₃ on grain growth in 1995 and 1996 observed in both the ambient and elevated CO₂ treatments may reflect the contrasting temperature environments experienced. Grain yield was nevetheless reduced under elevated O₃ in 1996, primarily because of a substantial decrease in grain number. The data obtained show that, although exposure to elevated CO₂ and O₃ individually or in combination may affect both dG/dt and dHI/dt, the presence of elevated CO₂ does not protect against substantial O₃-induced yield losses resulting from its direct deleterious impact on reproductive processes. The implications of these results for food production under future climatic conditions are considered.     

Influence of stem lacunar structure on gas transport: relation to the oxygen transport potential of submersed vascular plants

The influence of stem lacunar structure on the potential of diffusion and mass flow to meet estimated root O₂ demands was evaluated and compared in four submersed aquatic plant species. Internodal lacunae formed large continuous gas canals which were constricted at the nodes by thin, perforated diaphragms. Gas transport studies showed that nodes had little effect on diffusion, but significantly reduced mass flow. Measured diffusive resistances approximated those predicted by Fick's first law, ranged from 203 to 5107 × 10⁸ s m⁻⁴ and increased as lacunar area decreased in Potamogeton praelongus, two Myriophyllum species and Elodea canadensis. Our analysis suggested that diffusion could satisfy estimated root O₂ demands given the development of relatively steep O₂ gradients (0.15–0.35 mol O₂ mor⁻¹ per 0.5 m stem) between shoots and roots. Plants with high resistances (e.g. > 750 × 10⁸ s m⁻⁴) and long lacunar pathlengths may be unable, even during active photosynthesis, to support the O₂ demands of a large root system by diffusion alone. Measured nodal resistances to mass flow approximated those predicted by Hagen-Poiseuille law and ranged from 46 to 2029 × 10⁸ Pa s m⁻³. Our analysis suggested that these resistances were quite low and that relatively small pressure differentials (< 150 Pa per 0.5 m stem) could drive mass flow at rates which would support root O₂ demands. Possible mechanisms whereby plant architecture may serve to maintain these pressure differentials are proposed.     

Influence of carbon dioxide enrichment, ozone and nitrogen fertilization on cotton (Gossypium hirsutum L.) leaf and root composition

The objective of this study was to test whether elevated [CO₂], [O₃] and nitrogen (N) fertility altered leaf mass per area (LMPA), non‐structural carbohydrate (TNC), N, lignin (LTGA) and proanthocyanidin (PA) concentrations in cotton (Gossypium hirsutum L.) leaves and roots. Cotton was grown in 14 dm³ pots with either sufficient (0·8 g N dm ^{? 3}) or deficient (0·4 and 0·2 g N dm ^{? 3}) N fertilization, and treated in open‐top chambers with either ambient or elevated ( + 175 and + 350 μ mol mol ^{? 1}) [CO₂] in combination with either charcoal‐filtered air (CF) or non‐filtered air plus 1·5 times ambient [O₃]. At about 50 d after planting, LMPA, starch and PA concentrations in canopy leaves were as much as 51–72% higher in plants treated with elevated [CO₂] compared with plants treated with ambient [CO₂], whereas leaf N concentration was 29% lower in elevated [CO₂]‐treated plants compared with controls. None of the treatments had a major effect on LTGA concentrations on a TNC‐free mass basis. LMPA and starch levels were up to 48% lower in plants treated with elevated [O₃] and ambient [CO₂] compared with CF controls, although the elevated [O₃] effect was diminished when plants were treated concurrently with elevated [CO₂]. On a total mass basis, leaf N and PA concentrations were higher in samples treated with elevated [O₃] in ambient [CO₂], but the difference was much reduced by elevated [CO₂]. On a TNC‐free basis, however, elevated [O₃] had little effect on tissue N and PA concentrations. Fertilization treatments resulted in higher PA and lower N concentrations in tissues from the deficient N fertility treatments. The experiment showed that suppression by elevated [O₃] of LMPA and starch was largely prevented by elevated [CO₂], and that interpretation of [CO₂] and [O₃] effects should include comparisons on a TNC‐free basis. Overall, the experiment indicated that allocation to starch and PA may be related to how environmental factors affect source–sink relationships in plants, although the effects of elevated [O₃] on secondary metabolites differed in this respect.     

Atmospheric impact of bioenergy based on perennial crop (reed canary grass,Phalaris arundinaceae,L.) cultivation on a drained boreal organic soil

Marginal organic soils, abundant in the boreal region, are being increasingly used for bioenergy crop cultivation. Using long‐term field experimental data on greenhouse gas (GHG) balance from a perennial bioenergy crop [reed canary grass (RCG), Phalaris arundinaceae L.] cultivated on a drained organic soil as an example, we show here for the first time that, with a proper cultivation and land‐use practice, environmentally sound bioenergy production is possible on these problematic soil types. We performed a life cycle assessment (LCA) for RCG on this organic soil. We found that, on an average, this system produces 40% less CO₂‐equivalents per MWh of energy in comparison with a conventional energy source such as coal. Climatic conditions regulating the RCG carbon exchange processes have a high impact on the benefits from this bioenergy production system. Under appropriate hydrological conditions, this system can even be carbon‐negative. An LCA sensitivity analysis revealed that net ecosystem CO₂ exchange and crop yield are the major LCA components, while non‐CO₂ GHG emissions and costs associated with crop production are the minor ones. Net bioenergy GHG emissions resulting from restricted net CO₂ uptake and low crop yields, due to climatic and moisture stress during dry years, were comparable with coal emissions. However, net bioenergy emissions during wet years with high net uptake and crop yield were only a third of the coal emissions. As long‐term experimental data on GHG balance of bioenergy production are scarce, scientific data stemming from field experiments are needed in shaping renewable energy source policies.     

Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix)

Hoflacher, H. and Bauer, H. 1982. Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix). – Physiol. Plant. 56: 177–182. Light acclimation was investigated during the juvenile and adult life phases of the whole-plant-development in Hedera helix L. For this purpose, cuttings of the juvenile and adult parts of one single parent plant were grown under low-light (PAR 30–50 μmol photons m^?2 s^?1) and high-light (PAR 300–500 μmol m^?2 s^?1) conditions: CO₂ exchange, chloroplast functions, and specific anatomy of fully developed leaves differentiated under these conditions were determined. In juvenile plants the leaves formed under low and high light had light-saturated rates of net photosynthesis of 6.5 and 11.1 mg CO₂ (dm leaf area)^?2 h^?1, respectively. In adult plants the rates were 9.4 and 22.2 mg dm^?2 h^?1, indicating a more pronounced capacity for acclimation to strong light in the adult life phase. Higher photosynthetic capacities were accompanied by higher conductances for the CO₂ transfer through the stomata, leading to almost the same CO₂ concentration in the intercellular spaces. Thus, stomatal conductances were not primarily responsible for the different photo-synthetic capacities. The higher rates in adult and high-light grown leaves were mainly the result of formation of thicker leaves with more chloroplasts per unit leaf area. Expressed per chloroplast, the photosynthetic capacity, the Hill reaction, and the activity of ribulose bisphosphate carboxylase were almost identical in plants grown in low-light and high-light. Measurements of photosynthetic capacity and thickness of leaves of Hedera sampled from field habitats with contrasting light regimes confirm the results of growth chamber studies. It is, therefore, concluded that both life phases of Hedera are capable of acclimating to strong light, but that during the juvenile phase this capacity is not fully developed.     

Influence of the surface temperature of packaging specimens on the inactivation of Bacillus spores by means of gaseous H(2) O(2)

Aims: To determine the influence of condensation as a function of the surface temperature of aseptic packaging, on the inactivation of Bacillus spores [Bacillus subtilis (DSM 347), B. subtilis SA22, Bacillus atrophaeus] having different surface properties by means of vaporized H₂O₂. Methods and Results: The packaging specimens inoculated with Bacillus spores were tempered and subsequently exposed to H₂O₂‐vapour. During the exposure, surface temperature curves were measured and the spore survival was determined. Results showed that decreasing the initial surface temperature of the packaging specimens had a positive effect on the sporicidal activity of H₂O₂‐vapour, where the effect was less pronounced for less hydrophilic spores. The surfaces of spores were characterized by means of the water contact angle. Conclusions: For starting surface temperatures below the dew point temperature of the sterilant gas, the condensation of highly concentrated liquid H₂O₂ on the packaging surface accelerates the killing of the spores, while the inferior wettability of more hydrophobic spores compared to more hydrophilic ones diminishes the effect. Significance and Impact of the Study: Regarding industrial packaging sterilization, a mixed microflora has to be inactivated. Promoting the condensation of H₂O₂ improves in general the killing of different species of spores, however, at various degrees depending on the wettability of spores.     

Affinity, capacity and oxygen sensitivity of two different mechanisms for bicarbonate utilization in Ulva lactuca L. (Chlorophyta)

Ulva lactuca, collected on the west coast of Sweden at the end of May, was able to utilize the HCO₃ ^? pool of seawater only through extracellular dehydration via carbonic anhydrase, followed by uptake of the CO₂ formed. A decrease in the CO₂ supply via this mechanism resulted in the gradual development of an additional method of HCO₃ ^? utilization, namely a direct uptake of HCO₃ ^? . Photosynthesis could then be supported by either a ‘HCO₃ ^? dehydration mechanism’ or a ‘HCO₃ ^? uptake mechanism’. Through selective inhibition of either of these mechanisms, the physiological properties of the other could be assessed. These properties suggest that the HCO₃ ^? uptake mechanism of U. lactuca is important under conditions when low concentrations of inorganic C, high pH and high external O₂ concentrations would limit photosynthesis supported by the HCO₃ ^? dehydration mechanism. Such conditions may occur during intense irradiation of the alga in rockpools or in shallow bays with low rates of water exchange. The results are discussed in relation to a possible coupling between mechanisms for inorganic C acquisition and cell structure (or even morphology) of green macroalgae. They also illustrate some necessary precautions when using Michaelis–Menten kinetics for estimations of V_max and K_1/2 values.     

Influence of mass transfer limitations on determination of the half saturation constant for hydrogen uptake in a mixed-culture CH(4)-producing enrichment

There is strong evidence in the literature supporting the existence of significant mass transfer limitations on the kinetics of exogenous H(2) consumption by methanogens. The half saturation constant for H (2) uptake by a mixed-culture, CH(4) producing enrichment was measured using an experimental protocol that avoided internal mass transfer limitations. The value obtained was two orders of magnitude smaller than any other previously reported. A mathematical model for acetogenic syntrophic associations was developed to check the capacity of H(2) as electron transporter between syntrophic partners. It was found that H(2) diffusion could account for the rate of transport of electrons between the syntrophic microorganisms and that formate is not a necessary intermediate. The possibility that formate may be an intermediate in this system was not ruled out. A Monod-type kinetic equation was modified to include the observed H(2) threshold effect. This modified equation was used to predict the CH(4)-production rate in a batch-fed digester. The results show that the external and internal H(2) pools are kinetically coupled. (c) 1992 John Wiley & Sons, Inc.     

Influence of the pH on (open) mixed culture fermentation of glucose: a chemostat study

Catabolic products from anaerobic fermentation processes are potentially of industrial interest. The volatile fatty acids and alcohols produced can be used as building blocks in chemical processes or applied directly as substrates in a mixed culture process to produce bioplastics. Development of such applications requires a predictable and controllable product spectrum of the fermentation process. The aim of the research described in this paper was (i) to investigate the product spectrum of an open mixed culture fermentation (MCF) process as a function of the pH, using glucose as substrate, and (ii) to relate the product spectrum obtained to generalized biochemical and thermodynamic considerations. A chemostat was operated under carbon and energy limitation in order to investigate the pH effect on the product spectrum in a MCF process. A transition from CO(2)/H(2) production at lower pH values to formate production at higher pH values was observed. The ratio of CO(2)/H(2) versus formate production was found to be related to the thermodynamics of formate dehydrogenation to CO(2)/H(2). This transition was associated with a shift in the catabolic products, from butyrate and acetate to ethanol and acetate, likely due to a decrease in the oxidation state of the electron carriers in the cell. The product spectrum of the MCF process as a function of the pH could largely be explained using general biochemical considerations.     

The P140K mutant of human O(6)-methylguanine-DNA-methyltransferase (MGMT) confers resistance in vitro and in vivo to temozolomide in combination with the novel MGMT inactivator O(6)-(4-bromothenyl)guanine

The O(6)-methylguanine-DNA-methyltransferase (MGMT) inactivator O(6)-benzylguanine (O(6)-beG) is currently under clinical investigation as a potential tumour-sensitising agent. In clinical trials its use has been associated with increased myelotoxicity and a reduced maximum tolerated dose (MTD) for BCNU. Thus the concept of myeloprotection by gene therapy with an O(6)-beG-insensitive mutant of MGMT is soon to be tested. Recently, an alternative inactivator has been described (O(6)-(4-bromothenyl)guanine, PaTrin-2), which shows potential advantages over O(6)-beG in terms of higher activity against wild-type MGMT and oral formulation. The use of PaTrin-2 has also been associated with increased myelotoxicity in clinical trials and thus PaTrin-2 may also be a candidate for use in conjunction with mutant MGMT gene transfer in genetic chemoprotective strategies. However, its activity against mutant MGMTs has not been reported. We show here that the P(140)K mutant of MGMT is highly resistant to inactivation by PaTrin-2. Furthermore, we show that a human haemopoietic cell line (K562) transduced with a retroviral vector encoding MGMT(P140K) is highly resistant to the cytotoxic effects of PaTrin-2 in combination with the methylating agent temozolomide, and that cells expressing MGMT(P140K) can be effectively enriched in vitro following challenge with this drug combination. Finally, we show that animals reconstituted with bone marrow expressing MGMT(P140K) exhibit haemopoietic resistance to PaTrin-2/temozolomide, which results in in vivo selection of gene-modified cells. All of these effects were comparable to those also achieved using O(6)-beG/temozolomide. Thus our data show that MGMT(P140K) is a suitable candidate for chemoprotective gene therapy where PaTrin-2 is being used in conjunction with temozolomide.     

The effects of elevated CO2 (0.5%) on chloroplasts in the tetraploid black locust (Robinia pseudoacacia L.)

Some ploidy plants demonstrate environmental stress tolerance. Tetraploid (4×) black locust (Robinia pseudoacacia L.) exhibits less chlorosis in response to high CO₂ than do the corresponding diploid (2×) plants of this species. We investigated the plant growth, anatomy, photosynthetic ability, chlorophyll (chl) fluorescence, and antioxidase activities in 2× and 4× black locusts cultivated under high CO₂ (0.5%). Elevated CO₂ (0.5%) induced a global decrease in the contents of total chl, chl a, and chl b in 2× leaves, while few changes were found in the chl content of 4× leaves. Analyses of the chl fluorescence intensity, maximum quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), K‐step (V_k), and J‐step (V_J) revealed that 0.5% CO₂ had a negative effect on the photosynthetic capacity and growth of the 2× plants, especially the performance of PSII. In contrast, there was no significant effect of high CO₂ on the growth of the 4× plants. These analyses indicate that the decreased inhibition of the growth of 4× plants by high CO₂ (0.5%) may be attributed to an improved photosynthetic capacity, pigment content, and ultrastructure of the chloroplast compared to 2× plants.