Robust multiple-input-multiple-output control of non-linear continuous fermentation processes

In this study, the removal of nitrate (NO₃^m) ions from aqueous streams with liquid membrane technique has been investigated. Among the other parameters (temperature, pH, acceptor phase type and medium concentration), the stirring speed was chosen as process parameter. From the experimental results, it has been determined that the reaction was diffusion controlled. The transport efficiency of nitrate ions increased with increasing stirring speed. The membrane entrance and exit rate constants (k_1d, k_2m and k_2a respectively) were linearly dependent on the stirring speed ratios of 100 to 250 rpm. Coupled transport of nitrate ions through a liquid membrane in 85% n-hexane-15% tricloromethane as diluent, containing tetraoctyl ammonium chloride (TOACl) as a carrier was examined at various stirring speeds. Membrane entrance (k_1d) and exit rates (k_2m and k_2a) increase with increasing the stirring speeds. The diffusion of the nitrate ion-carrier complex through the narrow stagnant layers was found to be rate determining step. The membrane was stable during the transport experiments. There is no leakage of carrier or nitrate ion-carrier complex to both aqueous phases and also, no supplementary water penetration into the membrane. This favours interfacial reaction of nitrate ion and carrier.     

Salt, nutrient uptake and transport, and ABA of Populus euphratica; a hybrid in response to increasing soil NaCl

The uptake and transport of salt ions (Na⁺, Cl^-), macronutrients (K⁺, Ca²⁺, Mg²⁺) and abscisic acid (ABA) response to increasing soil salinity were examined in 2-year-old seedlings of Populus euphratica and a hybrid, P. talassica Kom 2 (P. euphratica + Salix alba L.). Leaf burn symptoms appeared in the hybrid after 8 days of exposure to salinity when soil NaCl concentration increased to 206 mM, whereas P. euphratica exhibited leaf damage after day 21 when soil NaCl exceeded 354 mM. Leaf necrosis was the result of excess salt accumulation since the injury followed an abrupt increase of endogenous salt levels. Compared with the hybrid, P. euphratica exhibited a greater capacity to exclude salt ions from leaves under increasing salinity, especially Cl^-. Salt treatment altered nutrient balance of the hybrid, leaf K⁺, Ca²⁺ and Mg²⁺ concentrations significantly declined and the same trends were observed in roots with the exception of K⁺. Although K⁺ levels decreased in salinised P. euphratica, increasing salinity did not affect the levels of Ca²⁺ and Mg²⁺ in leaves, but did increase the uptake of these nutrients when salt stress was initiated. NaCl-induced increase of ABA concentration in xylem sap [ABA] was observed in the two tested genotypes, however xylem [ABA] increased more rapidly in P. euphratica and a fivefold increase of xylem [ABA] was recorded after the first day of exposure to salt stress. Therefore, we conclude that the increase of Ca²⁺ uptake may be associated with the rise of ABA, and thus contributes to membrane integrity maintenance, which enables P. euphratica to regulate uptake and transport of salt ions under high levels of external salinity in the longer term.     

Electrophysiological Measurements on the Root of Atriplex hastata

The ion contents and membrane potentials of the cells of young,hydroponically cultured seedlings of Atriplex hastata L. var.salina, Wallr. have been measured at several different NaClconcentrations. The total tissue concentrations of Na⁺ and Cl^–increase as external NaCl increases, but there is always a markedexcess of internal Na⁺ over Cl^–; this is balanced by endogenousorganic anion formation with a concomitant extrusion of H⁺ tothe bathing solution. Membrane potentials of the root cells remain essentially invariantwith changes in external NaCl at approx. –130 mV; thereis no evidence of a radial gradient of potential across theroot. The potential seems to contain a cyanide-sensitive electrogeniccomponent, also invariant with NaCl concentration, of about–70 mV, and a diffusion component. The electrogenic componentseems likely to be a H⁺ efflux, probably through a H⁺ uniportATPase.     

Hypoxia affects root sodium and chloride concentrations and alters water conductance in salt-treated jack pine (<Emphasis Type="Italic">Pinus banksiana</Emphasis>) seedlings

The effects of NaCl were studied in 6-month-old jack pine (Pinus banksiana Lamb.) seedlings growing in solution culture under hypoxic (approximately 2 mg l^у O₂) and well-aerated (approximately 8 mg l^у O₂) conditions. The results showed that hypoxia led to further reduction of stomatal conductance (g_s) in plants treated with 45 mM NaCl. This effect was likely due to a reduction in root hydraulic conductance by both stresses. When applied individually or together, neither 45 mM NaCl nor hypoxia affected cell membrane integrity of needles as measured by tissue electrolyte leakage. Hypoxia did not alter shoot Na⁺ and Cl^m concentrations in NaCl-treated plants. However, root Na⁺ concentrations were lower in NaCl-treated hypoxic plants, suggesting that hypoxia affected the ability of roots to store Na⁺. Hypoxia also induced root electrolyte leakage from NaCl-treated and control plants. The higher root Cl^m concentrations compared with Na⁺ and the positive correlation between root Cl^m concentrations and electrolyte leakage suggest that Cl^m played a major role in salt injury observed in jack pine seedlings. Roots of well-aerated plants treated for 1 week with NaCl contained almost two-fold higher concentration of total non-structural carbohydrates compared with plants from other experimental treatments and these concentrations decreased in subsequent weeks. We suggest that under prolonged hypoxic conditions, roots lose the ability to prevent Cl^m uptake resulting in the increase in root Cl^m concentration, which has damaging effects on root cell membranes.     

Comparative studies on citric acid production by Aspergillus niger and Candida lipolytica using molasses and glucose

Citric acid production by Aspergillus niger NCIM 548 and Candida lipolytica NCIM 3472 has been studied in shake culture using glucose and molasses as carbon sources. Methanol addition (3% v/v) at 40 h of fermentation enhanced the production of citric acid by Aspergillus niger whereas a reduction in citric acid production by Candida lipolytica was observed with addition of methanol. Maximum citric acid concentration of 12 kg/m³ was obtained with Aspergillus niger using molasses in the presence of methanol, while maximum citric acid concentration of 8.4 kg/m³ was obtained with Candida lipolytica using glucose without methanol. It appears that product formation by Aspergillus niger is either non-growth associated or partially growth associated depending on the substrate. Methanol addition changes the nature of product formation in case of Candida lipolytica.     

A novel fiber optic probe for on-line monitoring of biomass concentrations

A fiber optic biomass probe for on-line measurement of biomass concentration was designed. Results of biomass concentration monitoring experiments with suspended cells of baker's yeast as well as an experimental cultivation of S. cerevisiae are presented. The device was able to observe biomass concentrations of 14 g l^у S. cerevisiae. By means of correlations the capability of estimating the biomass concentration from the probe signal is demonstrated.     

Effect of pH on Pb2+ accumulation in Saccharomyces cerevisiae and Aureobasidium pullulans

The optimum pH conditions of Pb²⁺ accumulation in Saccharomyces cerevisiae and Aureobasidium pullulans were 4~5 and 6~7, respectively. The initial Pb²⁺ accumulation rates according to the increase of initial Pb²⁺ concentration and pH were increased both in S. cerevisie and A. pullulans. And the initial Pb²⁺ accumulation rate of A. pullulans was much higher than that of S. cerevisiae because of the difference of Pb²⁺ accumulation mechanism. The Pb²⁺ accumulation isotherm of S. cerevisae obeyed a fully competitive inhibition, whereas that of A. pullulans showed a mixed inhibition of competition and non-competition associated with the proton (H⁺) as an accumulation inhibitor.     

Comparison of two membrane – photobioreactors, with free or immobilized cells, for the production of pigments by a marine diatom

The present study concerns the value enhancement of the microalga Haslea ostrearia. This marine diatom bears the peculiarity of synthesizing and secreting in the culture medium a blue-green pigment named "marennine". Anticancer research, cosmetics and aquaculture are the fields concerned with the utilizations of this hydrosoluble pigment. The aim of the study is to compare the pigment productivity obtained with two types of photobioreactors. In the first process, cells are free and recycled in a bioreactor combined with a membrane ultrafiltration equipment (external loop). In the second system, cells are entrapped in a tubular agar gel layer in a photobioreactor of original design. The influence of nitrate concentration and renewal rate is examined. Experiments, conducted on long term periods (up to 40 d) without any external contamination, revealed that marennine productivities of more than 5-7 mg 10⁹ cell^у d^у can be reached with both bioreactors. The advantages and drawbacks of each process design are also discussed.     

Joint aerobic biodegradation of wastewater from table olive manufacturing industries and urban wastewater

Wastewater generated in the elaboration of table olives has been treated using activated sludge from a municipal wastewater plant after adequate acclimation. To avoid bactericide properties of some chemical structures present in this type of effluents, synthetic urban wastewater has been used to dilute the original wastewater. The main parameters affecting efficiency of biological processes have been studied. Thus, initial biomass concentration, temperature up to 303 K (upper working temperature limit = 313 K) and initial substrate concentration exerted a positive influence on COD degradation rate. The optimum pH was found to be around 7, experiencing a slight inhibition on cell activity at pH 4. Under the experimental conditions investigated other parameters like polyphenol content, absorbance at 254 nm and total organic carbon were also reduced to some extent. Only nitrates amount was increased after the biological process took place. A kinetic model based on Monod equation was proposed and applied to experimental results. The maximum specific growth rate was calculated by means of the aforementioned kinetic model. The value of this parameter as a function of temperature was fitted to an Arrhenius expression, w_max = 9.43 2 10¹⁰ exp(72021/RT) h^у (R in J mol^у K^у283 K < T < 303 K, pH , 7-10).     

Ion transport systems in the kidney and urinary bladder of two Antarctic teleosts, Chionodraco hamatus and Trematomus bernacchii

Na⁺-K⁺-Cl^- cotransport and Na⁺/K⁺ATPase were studied by immunohistochemistry in the kidney and urinary bladder of Trematomus bernacchii and Chionodraco hamatus. The activity was correlated to the density of mitochondria. The first segment of the renal proximal tubule was more active than the second one. In T. bernacchii and the temperate marine teleost Pagellus bogaraveo, the immunoreactivity for the antibody to cotransporters and to the !-subunit of the sodium pump was stronger than in the icefish. This difference indicates in the kidney of the icefish, a weaker secretory activity, a consequent lower osmolarity in the lumen and lower water loss, which correlates well with the need for a greater blood volume in the icefish. The epithelium of the urinary bladder in T. bernacchii, where intense immunostaining was observed, was composed of columnar cells. In C. hamatus the columnar cells, where the immunostaining was weaker, lined only a portion of the urinary bladder, the other region being composed of cuboidal cells.     

Water availability and carbon isotope discrimination in conifers

The stable C isotope composition ('¹³C) of leaf and wood tissue has been used as an index of water availability at both the species and landscape level. However, the generality of this relationship across species has received little attention. We compiled literature data for a range of conifers and examined relationships among landscape and environmental variables (altitude, precipitation, evaporation) and '¹³C. A significant component of the variation in '¹³C was related to altitude (discrimination decreased with altitude in stemwood, 2.53‰ km^-1 altitude, r²=0.49, and in foliage, 1.91‰ km^-1, r²=0.42), as has been noted previously. The decrease in discrimination with altitude was such that the gradient in CO₂ partial pressure into the leaf (P_a-P_i) and altitude were generally unrelated. The ratio of precipitation to evaporation (P/E) explained significant variation in P_a-P_i of stemwood (r²=0.45) and foliage (r²=0.27), but only at low (<0.8) P/E. At greater P/E there was little or no relationship, and other influences on '¹³C probably dominated the effect of water availability. We also examined the relationship between plant drought stress (O) and '¹³C within annual rings of stemwood from Pinus radiata and Pinus pinaster in south-western Australia. Differential thinning and fertiliser application produced large differences in the availability of water, nutrients and light to individual trees. At a density of 750 stems ha^-1, O and '¹³C were less (more negative) than at 250 stems ha^-1 indicating greater drought stress and less efficient water use, contrary to what was expected in light of the general relationship between discrimination and P/E. The greater '¹³C of trees from heavily thinned plots may well be related to an increased interception of radiation by individual trees and greater concentrations of nutrients in foliage - attributes that increase rates of photosynthesis, reduce P_i and increase '¹³C. '¹³C was thus modified to a greater extent by interception of radiation and by nutrient concentrations than by water availability and the '¹³C-O relationship varied between thinning treatments. Within treatments, the relationship between '¹³C and O was strong (0.38<r²<0.58). We conclude that '¹³C may well be a useful indicator of water availability or drought stress, but only in seasonally dry climates (P/E<1) and where variation in other environmental factors can be accounted for.     

Sperm extract and inositol 1,4,5-triphosphate induce cytosolic calcium rise in the central cell of Torenia fournieri

Microinjection of soluble sperm extract and Calcium Green-1 10 kDa-dextran conjugate (CG-1) into the mature central cell of Torenia fournieri induced a significant rise in cytosolic free calcium concentration ([Ca²⁺]_i). The rise reached a maximum at 20 min after injection and then steadily declined. Nevertheless, a relatively high level of [Ca²⁺]_i was maintained even 40 min after injection. Microinjection of sperm extract of maize into Torenia central cells, however, did not trigger any increase in [Ca²⁺]_i, suggesting the possibility of distinct triggers in different species. We also injected caged inositol 1,4,5-triphosphate (InsP₃) and caged cyclic ADP-ribose (cADPR) into Torenia central cells to compare the pattern of Ca²⁺ rise induced by the sperm extract. The results showed that [Ca²⁺]_i elevation triggered by the release of InsP₃ after photolysis appears much faster than that induced by sperm extract. The increase in [Ca²⁺]_i reached a maximum at 70-80 s and dropped to the resting level within 300 s after photolysis. Microinjection of cADPR, however, did not induce any changes in [Ca²⁺]_i. The results indicate that sperm extract might contain factors triggering the release of Ca²⁺ in the central cell.     

Chloride in Soils and its Uptake and Movement within the Plant: A Review

Natural inputs of chlorine (Cl) to soils come mainly from rainwater,sea spray, dust and air pollution. In addition, human practices,such as irrigation and fertilization, contribute significantlyto Cl deposition. In the soil solution, Cl occurs predominantlyas the chloride anion (Cl^-). The Cl^-anion does not form complexesreadily, and shows little affinity (or specificity) in its adsorptionto soil components. Thus, Cl^-movement within the soil is largelydetermined by water flows. Chlorine is an essential micronutrientfor higher plants. It is present mainly as Cl^-. Chloride isa major osmotically active solute in the vacuole and is involvedin both turgor- and osmoregulation. In the cytoplasm it mayregulate the activities of key enzymes. In addition, Cl^-alsoacts as a counter anion, and Cl^-fluxes are implicated in thestabilization of membrane potential, regulation of intracellularpH gradients and electrical excitability. Chloride enters plantsthrough the roots, and there is some concern over the uptakeof the long-lived radionuclide³⁶Cl, which enters into the foodchain through plants. Chloride is thought to traverse the rootby a symplastic pathway, and Cl^-fluxes across the plasma membraneand tonoplast of root cells have been estimated. These fluxesare regulated by the Cl^-content of the root. Chloride is mobilewithin the plant. The Cl^-concentrations of xylem and phloemsaps have been determined and Cl^-fluxes through the xylem andphloem have been modelled. Measurements of transmembrane voltagesand Cl^-activities in cellular compartments suggest (1) thatactive Cl^-transport across the plasma membrane dominates Cl^-influxto root cells at low Cl^-concentrations in the soil solutionand that passive Cl^-influx to root cells occurs under more salineconditions, and (2) that both active and passive Cl^-transportoccurs at the tonoplast. Electrophysiological studies have demonstratedthe presence of an electrogenic Cl^-/2H⁺symporter in the plasmamembrane of root-hair cells and Cl^-channels mediating eitherCl^-influx or Cl^-efflux across the plasma membrane. Similarly,there is both biochemical and electrophysiological evidencethat Cl^-channels mediate Cl^-fluxes in either direction acrossthe tonoplast and that a Cl^-/nH⁺antiport mediates Cl^-influxto the vacuole. This article reviews the availability of Cl^-inthe soil, the roles and distribution of Cl^-within the plant,the magnitude of Cl^-fluxes across membranes and between tissues,the mechanisms of Cl^-transport across membranes and the electricalcharacteristics and molecular biology of Cl^-channels. Copyright2001 Annals of Botany Company Review, Arabidopsis thaliana, channel, chloride (Cl^-), influx, phloem, plasma membrane, radiochlorine (³⁶Cl), soil, tonoplast, transport, uptake, xylem     

Optimized fed-batch fermentation of L-β-hydroxy isobutyric acid by Yarrowia lipolytica

Yarrowia lipolytica KCCM50506, which transforms isobutyric acid to L-#-hydroxy isobutyric acid (L-#-HIBA), was screened. Chemostat cultures were carried out in jar fermentors at dilution rates of 0.02 h^у to 0.12 h^у. L-#-HIBA fermentation-regulating factors were determined to be specific growth rate, and concentrations of glucose and isobutyric acid in fermentor from analysis of steady-state data. The specific productivity of L-#-HIBA increased as the specific growth rate increased, apparently as a growth-associated type of product formation. A fed-batch culture was carried out under optimum conditions where the concentrations of glucose and isobutyric acid in the fermentor were maintained at 23 g l^у and 9 g l^у, respectively. The concentrations of cells and L-#-HIBA obtained at the end of fermentation were 20 g l^у and 49 g l^у, respectively, corresponding to 2.0 and 2.7 times more than concentrations in batch culture.     

Fluxes and composition of settling particles during summer in an Antarctic shallow bay of Livingston Island, South Shetlands

A moored experiment using a sediment trap was conducted at Johnson's Dock, Livingston Island from 11 December 1997 to 24 February 1998, as part of the EASIZ Programme activities carried out at the Juan Carlos I Spanish Antarctic base. Total mass vertical fluxes ranged from 23,235 mg m^-2 day^-1 to 89,073 mg m^-2 day^-1 during the experiment, with a mean value of 42,857 mg m^-2 day^-1. Lithogenic components were the major contributors to the settling particulate flux. Organic components accounted for a low fraction of the settling particulate matter, showing an inverse relation to total mass flux. Nevertheless, the fluxes of organic components at Johnson's Dock are as high as in the open sea. The increases in chlorophyll a in water were related to increases in the organic carbon content, which dominated over inorganic carbon during the whole experiment. Calcium carbonate particles settle without being significantly altered in the water column and are dissolved in the upper centimetres of the bottom sediments, once they are buried. The settling material consisted of fine particles, with coarse clasts transported by icebergs. Antarctic shallow environments receive important sediment fluxes from the erosion and transport action of ice.     

Electrical properties of intact pollen grains of Lilium longiflorum: characteristics of the non-germinatingpollen grain

As a first step to characterizing membrane function during theprocess of germination in pollen, the transport characteristicsof Lilium longiflorum Thunbpollen were examined in the quiescentgrains.Membrane voltages were recorded, and conventional voltage-clampmeasurements were carried out with double-barrelled microelectrodes.The resting membrane voltage (V_m) was found to depend on theexternal K⁺; concentration and generally followed the equilibriumpotential for K^plus; (E_k). In some cells a more negative V_mwas measured indicating the presence of an electrogenic pump.The presence of a pump current was also detected by a depolarizationof the plasma membrane after inhibition of ATP synthesis byaddition of CN^– and SHAM. Besides this pump, two othercurrent components were found in the plasma membrane of ungerminatedpollen grains: an outward potassium current (l_k,out) and aninward K+ current (l_k,in). Outward K⁺ currents were detectedat membrane voltages more positive than the resting voltageand were blocked by externally applied TEA⁺ and Ba²⁺. The voltageat which l_k,out was first detected shifted in parallel withthe equilibrium potential for K⁺. By contrast, activation ofl_k,in was less affected by the external K⁺ concentration, exceptthat the magnitude of the inward current increased with K⁺ concentration.The detected current components may be involved in initiationof osmotic water influx for germination by allowing a K⁺ influxafter the membrane voltage has been driven more negative thanE_k by an electrogenic pump. Key words: Pollen, germination, potassium channel, proton pump, voltage-clamp     

Effect of extracellular polymeric substances (EPS) on Pb2+ accumulation by Aureobasidium pullulans

In Pb²⁺ accumulation by Aureobasidium pullulans, the time to reach an equilibrium state was not dependent on the initial cell dry weight. The Pb²⁺ accumulation capacity was increased from 56.9 to 215.6 mg Pb²⁺/g cell dry weight as the biomass was stored from 1 to 53 days, and correlated with the amount of excreted extracellular polymeric substances (EPS). It was observed that Pb²⁺ accumulated only on the surface of the intact cells of A. pullulans due to the existence of EPS, whereas Pb²⁺ penetrated into the inner cellular parts of the EPS-extracted cells.     

Spatial distribution of benthic microalgae on coral reefs determined by remote sensing

Understanding the ecological role of benthic microalgae, a highly productive component of coral reef ecosystems, requires information on their spatial distribution. The spatial extent of benthic microalgae on Heron Reef (southern Great Barrier Reef, Australia) was mapped using data from the Landsat 5 Thematic Mapper sensor, integrated with field measurements of sediment chlorophyll concentration and reflectance. Field-measured sediment chlorophyll concentrations, ranging from 23-1,153 mg chl a m^-2, were classified into low, medium, and high concentration classes (1-170, 171-290, and >291 mg chl a m^-2) using a K-means clustering algorithm. The mapping process assumed that areas in the Thematic Mapper image exhibiting similar reflectance levels in red and blue bands would correspond to areas of similar chlorophyll a levels. Regions of homogenous reflectance values corresponding to low, medium, and high chlorophyll levels were identified over the reef sediment zone by applying a standard image classification algorithm to the Thematic Mapper image. The resulting distribution map revealed large-scale (>1 km²) patterns in chlorophyll a levels throughout the sediment zone of Heron Reef. Reef-wide estimates of chlorophyll a distribution indicate that benthic microalgae may constitute up to 20% of the total benthic chlorophyll a at Heron Reef, and thus contribute significantly to total primary productivity on the reef.