Increase of ascorbic acid content and nutritional quality in spinach leaves during physiological acclimation to low temperature

The effect of acclimation to 10 °C on the leaf content of ascorbic and oxalic acids, was investigated in spinach (Spinacia oleracea L.). At 10 °C the content of ascorbic acid in leaves increased and after 7 days it was about 41% higher than in plants remaining under a 25 °C/20 °C day/night temperature regime. In contrast, the content of oxalate, remained unchanged. Transfer to 10 °C increased the ascorbic but not the oxalic acid content of the leaf intercellular washing fluid (IWF). Oxalate oxidase (OXO EC 1.2.3.4) activity was not detected in extracts of leaf blades. Therefore, oxalic acid degradation via OXO was not involved in the control of its content. Our results show that low temperature acclimation increases nutritional quality of spinach leaves via a physiological rise of ascorbic acid that does not feed-forward on the content of oxalic acid.     

A new approach to recycle oxalic acid during lignocellulose pretreatment for xylose production

Background

Dilute oxalic acid pretreatment has drawn much attention because it could selectively hydrolyse the hemicellulose fraction during lignocellulose pretreatment. However, there are few studies focusing on the recovery of oxalic acid. Here, we reported a new approach to recycle oxalic acid used in pretreatment via ethanol extraction.

Results

The highest xylose content in hydrolysate was 266.70 mg xylose per 1 g corncob (85.0% yield), which was achieved using 150 mmol/L oxalic acid under the optimized treatment condition (140 °C, 2.5 h). These pretreatment conditions were employed to the subsequent pretreatment using recycled oxalic acid. Oxalic acid in the hydrolysate could be recycled according to the following steps: (1) water was removed via evaporation and vacuum drying, (2) ethanol was used to extract oxalic acid in the remaining mixture, and (3) oxalic acid and ethanol were separated by reduced pressure evaporation. The total xylose yields could be stabilized by intermittent adding oxalic acid, and the yields were in range of 46.7–64.3% in this experiment.

Conclusions

This sustainable approach of recycling and reuse of oxalic acid has a significant potential application for replacing traditional dilute mineral acid pretreatment of lignocellulose, which could contribute to reduce CO₂ emissions and the cost of the pretreatment.

     

Modification of high lignin content kraft pulps with laccase to improve paper strength properties. 1. Laccase treatment in the presence of gallic acid

Laccase was reacted with gallic acid in the presence of a high-kappa (91) kraft pulp. The result was a modified pulp with 34%, 20%, and 72% improvements in burst, tensile, and wet tensile strength compared to untreated control samples. Fully bleached pulps were not responsive to the laccase treatment, indicating lignin was the major target for the fiber modification. The results indicate that the strength increases were a combined effect of improvements of hydrogen bonding between fibers and creation of phenoxy radical cross-links within the sheet.     

Optimization of organic acid pretreatment of wheat straw

The objective of this study was to determine the effectiveness of different organic acids (maleic, succinic, and oxalic acid) on enzymatic hydrolysis and fermentation yields of wheat straw. It was also aimed to optimize the process conditions (temperature, acid concentration, and pretreatment time) by using response surface methodology (RSM). In line with this objective, the wheat straw samples were pretreated at three different temperatures (170, 190, and 210°C), acid concentrations (1%, 3%, and 5%) and pretreatment time (10, 20, and 30 min). The findings show that at extreme pretreatment conditions, xylose was solubilized in liquid phase, causing an increase in cellulose and lignin content of biomass. Enzymatic hydrolysis experiments revealed that maleic and oxalic acids were quite effective at achieving high sugar yields (>90%) from wheat straw. In contrast, the highest sugar yields were 50–60%, when the samples were pretreated with succinic acid, indicating that succinic acid was not as effective. The optimum process conditions for maleic acid were, 210°C, 1.08% acid concentration, and 19.8 min; for succinic acid 210°C, 5% acid concentration, and 30 min; for oxalic acid 210°C, 3.6% acid concentration, and 16.3 min. The ethanol yields obtained at optimum conditions were 80, 79, and 59% for maleic, oxalic and succinic acid, respectively. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1487–1493, 2016     

Development of resistance in Cronobacter sakazakii ATCC 29544 to thermal and nonthermal processes after exposure to stressing environmental conditions

Aims: The objective was to study the response of Cronobacter sakazakii ATCC 29544 cells to heat, pulsed electric fields (PEF), ultrasound under pressure (Manosonication, MS) and ultraviolet light (UV‐C) treatments after exposure to different sublethal stresses that may be encountered in food‐processing environments. Methods and Results: Cronobacter sakazakii stationary growth‐phase cells (30°C, 24 h) were exposed to acid (pH 4·5, 1 h), alkaline (pH 9·0, 1 h), osmotic (5% NaCl, 1 h), oxidative (0·5 mmol l^?1 H₂O₂, 1 h), heat (47·5°C, 1 h) and cold (4°C, 4 h) stress conditions and subjected to the subsequent challenges: heat (60°C), PEF (25 kV cm^?1, 35°C), MS (117 μm, 200 kPa, 35°C) and UV‐C light (88·55 mW cm^?2, 25°C) treatments. The inactivation kinetics of C. sakazakii by the different technologies did not change after exposure to any of the stresses. The combinations of sublethal stress and lethal treatment that were protective were: heat shock–heat, heat shock–PEF and acid pH–PEF. Conversely, the alkaline shock sensitized the cells to heat and UV‐C treatments, the osmotic shock to heat treatments and the oxidative shock to UV‐C treatments. The maximum adaptive response was observed when heat‐shocked cells were subjected to a heat treatment, increasing the time to inactivate 99·9% of the population by 1·6 times. Conclusions: Cronobacter sakazakii resistance to thermal and nonthermal preservation technologies can increase or decrease as a consequence of previous exposure to stressing conditions. Significance and Impact of the Study: The results help in understanding the physiology of the resistance of this emerging pathogen to traditional and novel preservation technologies.     

Combining Effect of Microbial Transglutaminase and Bambara Groundnut Protein Isolate on Gel Properties of Surimi from Sardine (Sardinella albella)

Effects of protein isolate from bambara groundnut (BGPI) at different levels (0–6 %, w/w) in combination with microbial transglutaminase (MTGase) at different concentrations (0, 0.3 and 0.6 U g^?1surimi) on gels properties of sardine (Sardinella albella) surimi were investigated. In the absence of MTGase, the increases in breaking force and deformation of gels were obtained when BGPI at levels of 1.5–3 % was incorporated (P?<?0.05). The further increases in BGPI levels (4.5–6 %) resulted in the decrease in breaking force and deformation (P?<?0.05). When MTGase (0.3 and 0.6 U g^?1surimi) was added, the increase in breaking force and deformation were noticed, regardless of BGPI levels, and the strengthening effect was in dose-dependent manner. The increases in hardness, gumminess and chewiness were also observed when surimi gel was added with BGPI and MTGase (P?<?0.05). Water-holding capacity of gels was improved with increasing level of BGPI, and MTGase incorporated (P?<?0.05). Whiteness of gels slightly decreased with increasing BGPI levels, however the addition of MTGase had no impact on whiteness (P?>?0.05). Based on electrophoretic study, myosin heavy chain decreased with addition of MTGase, indicating the formation of cross-links. More compact structure was observed in gel added with MTGase (0.6 U g^?1surimi) and 6 % BGPI, and was accompanied by an increased gel strength.     

Effect of temperature and pH on the yield of oxalic acid bySclerotium rolfsii

The yield of oxalic acid bySclerotium rolfsii was highest at 25 °C for the wild and X₁ strains and at 30 °C for the X₂ strain. The optimum pH for oxalic acid synthesis by wild X₁ and X₂ strains was 5.0, 6.0 and 4.5, respectively.     

The effect of volatile anaesthetics on levels of metabolites and on metabolic rate in brain

Anaesthesia with ether, halothane, methoxyflurane (Penthrane) and Ohio 347 (Ethrane) increased the energy stores in mouse brain as much as 1·7-fold above the control values. The greatest increases were observed in glucose and glycogen. Glucose-6-P was increased in some cases and UDP glucose was consistently lower in the anaesthetized animals. Hypothermia in conjunction with anaesthesia modified some of the observed changes. Hypothermia alone was associated with an increase in P-creatine and glucose and a decrease in UDPglucose in the brain. The cerebral metabolic rate was depressed by all the anaesthetic agents to about 50 per cent of the control value. When the body temperature was lowered to 25°, the cerebral metabolic rate fell to 73 per cent of the control rate. A temperature coefficient of 1·035 was calculated as the fractional change/degree between 25° and 34°.     

Oxalic acid production by Aspergillus niger

Aspergillus niger is able to produce a quite high concentration of oxalic acid using sucrose as carbon and energy source. Operating at pH higher than 6 and an enriched N and P medium is necessary in order to conduct the fermentation towards oxalic acid production. A pH?shift technique, operating at acid pH?in the first two days and then setting pH?to 6, allowed the productivity to slightly increase in shaking flasks cultures up to 3.0?kg/m³?·?d, with a final oxalic acid concentration of 29?kg/m³. When operating at more controlled conditions, in a stirred tank, both productivity and oxalic acid concentration were improved (4.1?kg/m³?·?d and 33.8?kg/m³, respectively). However the main drawback of this fermentation is the low yield attained (about 0.3?kg oxalic acid/kg sucrose) because most of glucose, resulting from the hydrolysis of sucrose by the extracellular enzymes secreted at the beginning of the fermentation, is very quickly oxidised to gluconic acid, a process which is favoured at a pH?close to 6. Milk whey was proved to be a very good substrate as it allows oxalic acid to be produced with a similar productivity (2.5?kg/m³?·?d in shaking flasks) giving excellent yields of almost 0.6?kg oxalic acid/kg lactose.     

Oxalic acid production by Aspergillus niger

Oxalic acid is formed by Aspergillus niger at nearly neutral pH values. In this study the applicability of milk whey as a carbon source was investigated, both in shaking flask experiments and in a stirred tank reactor. The influence of pH on oxalic acid formation showed that the maximum production rate and higher concentration of the product are observed at pH 6. At pH 7 the same production rate was obtained although at a lower oxalic acid concentration. The process was shown to be inhibited by product from an oxalic acid concentration of about 10?kg/m³ and its behaviour was fitted by Luong's equation. In a 10-dm³ strirred tank ferment the stirrer speed was varied in a range from 100 to 600 rpm. At values between 200 and 400 rpm, maximum production rates of oxalic acid of 6.8?kg/m³·d and 6.5?kg/m³·d were reached, respec-tively. A final concentration of 41.4?kg oxalic acid/m³ was reached operating at 400 rpm.     

The ecology of newt tadpoles

Food consumption of newt tadpoles caught during July and August was estimated as C=34W^1·11±0·12 where C is food consumption in milligrams wet weight/day and W is the live weight in grams. Well-fed tadpoles assimilated the food with an efficiency of 90–6%, but at low rates of feeding, the efficiency fell. Gross growth efficiency was about 35 % at maximum rations.     

Effect of environmental factors and precursors on oxalic acid production,mycelial biomass and virulence of a potential bioherbicide isolate of Sclerotium rolfsii SC64 produced in liquid culture

The fungus Sclerotium rolfsii is presently under development as a bioherbicide for broadleaf weed species using fungus-infested substrates as application material in this laboratory. The effect of environmental factors and three precursors (citric acid, ascorbic acid, and sodium succinate) on mycelial growth, oxalic acid production, and virulence by SC64 in liquid culture were investigated. The results showed that for mycelia growth the optimum liquid medium was Modified Richard's solution (MRS) among the five tested media, but potato dextrose broth (PDB) produced the maximum oxalic acid production and virulence on detached Solidago canadensis leaves. When PDB was used as the basic medium, the oxalic acid/mycelial dry weight (mg g^–1) ratio reached the peak 4 days after inoculation. The optimum temperature for oxalic acid production was at 27°C, but increased mycelial dry weight and virulence were observed at 30°C. The optimum range of initial pH value for oxalic acid accumulation was 4.0–6.0, with the optimal pH 5.0; highest mycelial growth was with an initial pH 3.5–6.0 (optimum pH 5.0) and subsequently pH 3.5–5.5 (maximum at pH 3.5). Both mycelial dry weight and oxalic acid production showed a decreasing trend as a result of the precursor of oxalic acid being added to PDB. Among the three precursors, the greatest decrease in mycelial dry weight, and oxalic acid production was caused by sodium succinate. This clarification of optimal conditions for production of mycelial biomass while insuring high concentrations of oxalic acid and high virulence should be useful for further development of this fungus as biocontrol agent.     

Studies on chitin. 2. Preparation and properties of chitin membranes

A chitin membrane was prepared by a new procedure involving coagulation of the chitin solution in N,N-dimethyl acetamide, N-methyl 2-pyrrolidone and lithium chloride (DMA-NMP-LiCl) with 2-propanol. The solute permeability, water sorption and mechanical properties were compared with membranes prepared by two previously reported methods (coagulation of a formic acid and dichloroacetic acid (FA-DCA) solution of chitin with 2-propanol; and coagulation of a trichloroacetic acid and dichloroethane (TCA-DCE) solution of chitin with acetone). The permeability coefficients of the three chitin membranes were higher than a regenerated cellulose membrane (Cuprophane®). The membrane prepared from DMA-NMP-LiCl solution had a higher tensile strength (3·3 Mpa) in the wet state than the others. The membrane obtained from TCA-DCE solution absorbed more water (360%) and the membrane prepared from FA-DCA solution was relatively weak (1·8 MPa) in the wet state. It was suggested that 2-propanol was a favourable coagulant for membrane production. In addition, the effect of the origin of chitin on molecular weight and tensile properties of the membranes was studied.     

Effects of partial acid hydrolysis on physical and chemical properties of agar from a newly reported Japanese agarophyte (Gracilariopsis lemaneiformis)

Physical and chemical properties of alkali-treated agar polymers extracted from Gracilariopsis lemaneiformis, newly reported Japanese agarophyte, were investigated after partial acid hydrolysis. The alkali-treated agar was hydrolyzed in boiling 0.1 N, 0.01 N, and 0.001 N sulfuric acid, oxalic acid, acetic acid, and citric acid solutions, for 1, 2, and 3 h at 100 °C. Partial acid hydrolysis of the agar polymers indicated strong effects on the physical properties. Different kinds of acid used for hydrolysis gave different agar properties. Gelling polymers were obtained from the agar hydrolysed in boiling 0.001 N acetic acid, oxalic acid, and citric acid solutions, and in 0.01 N acetic acid solution. High gel strength (715 ± 74.6 g cm-2) with low viscosity (2.47 cP) was obtained from 1 h treatment by 0.001 N acetic acid on hydrolysed agar. The results indicated that partial hydrolysis of agar under appropriate conditions probably improve agar quality and produce good grade agar from the Japanese agarophyte. This revised version was published online in August 2006 with corrections to the Cover Date.     

High potency of bombesin for stimulation of human gastrin release and gastric acid secretion

Dose-response studies were performed in 6 human volunteer subjects to determine the threshold and optimal doses of intravenous bombesin for stimulation of gastric acid secretion and gastrin release. A significant stimulation of both acid and gastrin was obtained with a very low dose, 3 pmol · kg^?1 · h^?1. Peak stimulation of acid secretion (67% of pentagastrin PAO) was obtained at 12.5 pmol · kg^?1 · h^?1. Serum gastrin response to this dose of bombesinn was similar to that obtained after a high protein meal. Higher doses of bombesin caused further increases in serum gastrin but not in acid secretion. Since very low doses of bombesin, too small to produce detectable increases in immunoreactive serum bombesim, caused parallel increases in gastrin and acid secretion, it is possible that the bombesin-like peptides present in human gastrointestinal tissues contribute to regulation of human gastric secretion.     

Author index

Binding of dexamethasone · receptors with isolated nuclei, DNA-cellulose and cellulose has been compared with respect to dependence on salt concentration and resistance to KCl extraction and DNAase I digestion. A solution of cytoplasmic dexamethasone-receptor complexes was prepared by the incubation of rat thymus cells with steroid at 3°C and breaking the cells by hypotonic lysis. Activation of the complexes was accomplished by warming the solution at 25°C for 15 min. Activation significantly increased the ability of dexamethasone · receptors to bind to nuclei and DNA-cellulose but not to cellulose. Dexamethasone-receptor complexes bound to nuclei at 3°C are completely resistant to extraction with 0.1 M KCl, 76% resistant to 0.2 M KCl and 20% resistant to 0.4 M KCl. Dexamethasone · receptors bound to DNA-cellulose are 45% resistant to extraction with 0.1 M and 0.2 M KCl and 29% resistant to 0.4 M KCl extraction. Cellulose-bound dexamethasone · receptors are not resistant to any of these extractions. DNAase I treatment releases 60% of the dexamethasone · receptors bound to DNA-cellulose but only 13% of those bound to nuclei, though at least 60% of the nuclear DNA is solubilized. The presence of 0.15 M KCl decreases binding of activated dexamethasone · receptors to nuclei by 73% but to DNA-cellulose by only 17%. Pretreatment of nuclei with 0.1–0.4 M KCl reduces their capacity to bind activated dexamethasone · receptors by 90% whereas similar treatment reduces the capacity of DNA-cellulose to bind dexamethasone · receptors by only 29%. Nuclei extracted with 0.1 M KCl appear to have a limited capacity to accept dexamethasone · receptors. These studies demonstrate that binding of dexamethasone · receptors to nuclei and DNA-cellulose differs by (a) the higher resistance of nuclear complexes to KCl and DNAase I treatment; (b) the much greater sensitivity of nuclei to KCl treatment.     

Post-harvest age-induced seed dormancy of Acer ginnala and its alleviation by growth regulator and low temperature treatments

One-month-old fruits of Acer ginnala with winged pericarp attached gave 44% germination and this was not increased by cold treatment at 4°C for 0, 10, 20, or 30 days, gibberellic acid treatment at 0, 1, 10, 100 or 1000 mg litre^-1, or ethephon treatment at 0, 2, 20, 200 or 2000 mg litre^-1. After 6 months of storage at 20–25 °C, germination of untreated fruits fell to 5% but could be restored to that of 1-month-old fruits by incubation at 4 °C for 30 days. After 9 months storage, no germination occurred in untreated fruits. Cold treatment (30 days at 4 °C partially restored germination (26%). Treatment with either gibberellic acid (1000 mg litre^-1) and 30 days at 4 °C (40%) or ethephon (100 mg litre^-] and 30 days at 4 °C improved germination (69%). The combination of all three treatments, i.e. 100 mg litre^-1 gibberellic acid, 100 mg litre^-1 ethephon and 30 days at 4 °C, optimised germination (86%). Thus, dormancy of A. ginnala developed during storage but could be reversed by a combination of treatment with low temperature and growth regulators. The highest germination (86%) was achieved after low temperature and growth regulator treatment of stored fruit.     

Effects of ambient temperature and methionine supplementation of a low protein diet on the performance of laying hens

The effects of ambient temperature and the supplementation of methionine to a low protein diet on egg production, egg quality, blood constituents and nitrogen excretion of laying hens were studied. The objective was to derive an environmental friendly feed formulation for warm climate. Seventy-two 29-week-old commercial White Leghorn hens of Babcock ISA white strain were used in this trial. The design is a completely randomized design with a 2×3 factors arrangement of treatments. Two constant ambient temperatures were 24±1°C and 34±1°C with 85% relative humidity. The three dietary treatments were 170 g kg⁻¹ crude protein, 140 g kg⁻¹ crude protein and 140 g kg⁻¹ crude protein supplemented with methionine 1.4 g kg⁻¹. Hens were allotted into six groups according to egg production and body weight. Birds were raised in individual wire cages for the experimental feeding period of five weeks. At the end of the feeding trial, one replicate of laying hens (four birds) from each treatment were selected for a four-day metabolic study for the daily collection of the excreta. The blood samples were withdrawn from the wing vein for analysis of hematocrit, blood glucose, sodium, potassium, magnesium, calcium, phosphorus and uric acid. Experimental results indicate that increases in ambient temperature significantly depress feed intake, egg production, egg weight and live weight of laying hens. High ambient temperature also caused inferior egg quality, including shell weight, shell thickness, shell breaking strength and specific gravity. Ambient temperature also changes the egg components with heavier egg albumin and yolk in the low-temperature group. Increasing ambient temperature also caused an increase in pH value in the plasma. This increase revealed a trend of depressed glucose (P<0.05) in the plasma of the laying hens. The dietary treatments, however, did not significantly influence feed intakes. Except egg weight, laying hens that were fed with the low protein (140 g kg⁻¹) with methionine supplemented diet produced similar numbers of egg and feed conversion as the layers fed with the high protein diet. The low protein with methionine supplemented diet produced significantly lighter eggs than the high dietary protein diet under the high ambient temperature, but produced heavier egg under the low ambient temperature. The concentration of uric acid in the plasma and nitrogen in the excreta of the high protein group was significantly higher than the other two low protein dietary groups (P<0.05).     

Seed treatments with Vitavax for the control of loose smut of wheat and barley

Seed dressings with Vitavax—75 % w.p.—eliminated Ustilago nuda in spring barley seeds and greatly reduced infection due to U. tritici in winter wheat. Emergence and yield of these crops were not adversely affected. Seed soak treatments including 0·2 % aqueous Vitavax for 6 h at 30 °C (wheat and barley), 0·2% thiram for 24 h at 30 °C (barley) and 0·2% Vitavax for 1 h at 30 °C (barley) also rid the seeds of infection. In other tests with barley 2 h soaks in 0·2 % aqueous suspensions of Vitavax at 30 °C gave equivalent control to 12 h soaks in 0·2% thiram at 30 °C.     

Effect of incubation temperature on muscle growth of barramundi Lates calcarifer at hatch and post-exogenous feeding

Muscle morphology was investigated in newly hatched barramundi Lates calcarifer larvae incubated at set temperatures (26, 29 and 31° C) prior to hatching. Three days after hatching (the start of exogenous feeding), larvae from the 26 and 31° C treatments were each divided into two groups and reared at that temperature or transferred over the period of several hours to 29° C (control temperature). Incubation temperature significantly affected muscle cellularity in the developing embryo, with larvae incubated at 26° C (mean ±s .e . 223·3 ± 7·9) having on average 14·4% more inner muscle fibres than those incubated at 31° C (195·2 ± 8·8) and 4·8% more than those incubated at 29° C (213·5 ± 4·7). Conversely, inner muscle fibre cross‐sectional area significantly increased at the warm incubation temperature in L. calcarifer, so that the total cross‐sectional muscle area was not different between treatment groups. The total cross‐sectional area of superficial muscle fibres and the proportion of superficial to total fibre cross‐sectional area in just hatched L. calcarifer were also affected by incubation temperature, with incubation at the cool temperature (26° C) increasing both the total cross‐sectional area and proportion of superficial muscle fibres. By 9 days post‐hatch, the aforementioned differences were no longer significant. Similarly, there was no difference in total superficial fibre cross‐sectional area between any treatment groups of L. calcarifer, whereas incubation temperature still significantly affected the proportion of superficial to total muscle fibre cross‐sectional area. Larvae hatched and grown at 31° C had a significantly reduced percentage of superficial muscle cross‐sectional area (mean ±s .e . 5·11 ± 0·66%) compared with those incubated and grown at 29° C (8·04 ± 0·77%) and 26° C (9·32 ± 0·56%) and those incubated at 26° C and transferred to 29° C (7·52 ± 0·53%), and incubated at 31° C and transferred to 29° C (6·28 ± 0·69%). These results indicate that changes in muscle cellularity induced by raising or lowering the incubation temperature of L. calcarifer display varying degrees of persistence over developmental time. The significance of these findings to the culture of L. calcarifer is discussed.