EFFICIENCY OF A CYCLIC DESIGN AND A MULTIDIMENSIONAL SCALING SENSORY ANALYSIS TECHNIQUE IN THE STUDY OF SALT TASTE

Multidimensional scaling (MDS) procedures produce maps of stimulus points, the dimensions of which may be sensory and/or physicochemical attributes. Our objective was to test the efficiencies of a cyclic design and a MDS method in the study of saltiness perception. Two levels of NaCl (0.1% and 0.2%, w/v) were added to two nonionic and two ionic gum solutions prepared to two concentrations. Subjects scaled dissimilarities between pairs of solutions and rated each sample for flavor attributes. Solution pairs were selected using cyclic designs. Repeated measures ANOVA determined that added NaCl was the only significant factor affecting saltiness perception. In contrast, from KYST-2A MDS maps, we concluded that saltiness perception was influenced by gum property, gum concentration, presence of calcium and potassium, and related to binding of the sodium ion (Na⁺) as determined by ²³Na nuclear magnetic resonance (NMR) spectroscopy. The MDS cyclic design successfully reduced the number of samples and subjects when using experienced subjects.     

Isolation of methanogens from Arabian sea sediments and their salt tolerance

Abstract Sea sediments in tropical regions have been less studied for methanogenesis and methanogens present therein. Three species of methanogens viz. Methanobacterium bryantii, Methanococcus voltae and Methanosarcina barkeri were isolated from Arabian sea sediments collected near the west coast of India. Maximum methane was formed by M. voltae at 3.0% (w/v) NaCl and other two methanogens at 0.06% (w/v) NaCl. M. bryantii and M. barkeri tolerated 2.5 and 3.0% (w/v) NaCl respectively due to which these methanogens must have survived in salt conditions of the sea sediments.     

Effects of sodium chloride on biodegradation of crude oils by two species ofAeromonas

Summary The biodegradation of five weathered crude oils by two species ofAeromonas, (B59-4 and E. BOB) was investigated in varying concentrations of sodium chloride. A minimal salts medium whose NaCl concentration increased serially by 0.5% w/v up to 1.5% w/v was used to investigate the growth of these strains in glucose, and their biodegradation of the crude oils. The latter was also investigated in fresh and aged sea water. Strain B59-4 was more potent than E. BOB in the degradation of all five crude oils and at all four levels of salt concentration tested. The amount of oil degraded by each strain increased initially to a maximum level at 0.5% w/v NaCl, but thereafter decreased with increasing salt concentration, and the patterns were similar to those of aged and fresh sea water, respectively. The Forties and Nigerian crude oils with lower specific gravity, were more readily degraded than the Libyan and Venezuelan with higher specific gravity. The growth of the two strains ofAeromonas in glucose and their biodegradation of crude oils was optimal at 0.5% w/v NaCl, and thereafter decreased with increasing salt concentration of the basal medium.     

PERCEPTION OF FAT IN A MILK MODEL SYSTEM USING MULTIDIMENSIONAL SCALING

This study examined the contributions of stimulus fat content and flavor volatiles to the perception of fat in a milk model system. The model system was formulated by adding bland vegetable oil (0%, 5%, or 10% w/v) and natural cream flavor (0%, 0.5% or 1% w/v) to a skim milk base. Panelists judged pairs of samples for similarity on the basis of three attributes (fat content, mouthcoating and thickness) and the results were analyzed using a multidimensional scaling procedure. Two-dimensional solutions best represented the data. The stimulus spaces for fat content and mouthcoating were visually similar to each other and provided reasonable separation of the samples. Instrumental measures helped to define the underlying dimensions of the stimulus space for fat content. Dimension 1 related to texture and included the contributions of viscosity, and fat particle size and number distribution; dimension 2 related to flavor perception. The sample with a moderate fat content (5%) and the highest concentration of added cream flavor (1%) was perceived to be similar to the 10% fat samples with added flavor. Thus, the added flavor provided the sensation of higher fat content. These data suggest that flavor plays an important role in the preception of fat in dairy foods. A psychophysical model of fat perception in dairy foods is proposed which includes the contribution of viscosity, fat particle size and number distribution, and volatile flavor perceptions.     

Improvement of Kefir yeast by mutation with N-methyl-N- nitrosoguanidine

A commercial culture of Kefir grew well at 30°C and 37°C with NaCl up to 4% (w/v) but at 42°C and 4% (w/v) NaCl it grew very slowly. These conditions were selected for the mutation with N-methyl-N-nitrosoguanidine in order to improve cell yield at high temperatures and salt concentration. The mutated cells had higher biomass and growth rate compared with the original culture. The improvement of Kefir at high temperatures and salt concentrations offers advantages for uses in single cell protein production and alcohol production from whey containing high salt.     

Microflora of the aerobic preservation of directly brined green olives from Hojiblanca cultivar

New procedures for the preservation stage of ripe olives from Hojiblanca cultivar were studied. An aerobic fermentative process was used with initial pH correction (0.3% acetic acid) and various NaCl concentrations: 6, 3 and 0% (w/v) in tap water. Treatments were carried out at industrial level and the spontaneous changes monitored. At initial salt concentrations of 6 and 3% (w/v) NaCl, pH rose progressively, reaching 4.3 at equilibrium maintaining during this period a constant free lactic acidity of around 0.4% (w/v). When the initial solution was tap waste, however, the pH decreased rapidly to stabilize at about 3.7, and lactic acidity increased continuously to reach values over 1% (w/v) at the end of the preservation process. In all treatments aeration effectively purged the carbon dioxide from the preservation brines, preventing shrivelling of olives. The microbial growth was strongly influenced by the initial NaCl concentration. At 6 and 3%, only yeasts grew, the most abundant being Pichia membranaefaciens, P. vini, P. fermentans and Hansenula polymorpha. However, when there was no NaCl, lactic acid bacteria colonized the solution. Lactobacillus plantarum and Pediococcus inopinatus were the only species found. In this case there was a co-existence between yeasts and lactic acid bacteria. As the treatment that supported lactic acid bacteria achieved the best final pH and acidity for olive stability, it may help to overcome the obstacles to a lactic fermentative process during the preservation stage of ripe olives from the Hojiblanca cultivar.     

Effect of carbohydrates and inhibitors of GA3 biosynthesis on embryogenenic potential of salt tolerant and non-tolerant callus lines of orange (Citrus sinensis osbeck)

Embryogenesis in callus lines of Citrus sinensis (orange) selected for tolerance to NaCl stress was compared to an unselected line. Addition of inhibitors of gibberellic acid (GA₃) synthesis, 2.6 × 10⁻⁶M paclobutrazol or 6.2 × 10⁻⁶M daminozide (butanedioic acid mono-(2-2-dimethylhydrazide) to the medium enhanced embryogenesis in the unselected line and two of the three salt tolerant lines, and also overcame embryo browning in the salt tolerant line R13. Sucrose as a carbon source was generally ineffective for embryogenesis. Two percent (w/v) glycerol was more effective than 2% (w/v) galactose, especially in older cultures. Formation of embryos was quicker in the non-selected line. Only a slight improvement in embryogenic potential was noted upon removal of salt tolerant lines for three passages from NaCl containing medium to basal medium without salt.     

Defense against lethal treatments and de novo protein synthesis induced by NaCl in Enterococcus faecalis ATCC 19433

Enterococcus faecalis was strongly resistant to high osmotic pressure in complex medium; however, when it was subjected to a moderate osmotic stress [6.5% (w/v) NaCl or 52% (w/v) sucrose] for 2 h, it showed cross-protection against ethanol (22%), detergents stresses [bile salts (0.3%) and SDS (0.017%)], hydrogen peroxide challenge (45 mM), and to a minor extent against lethal temperature (62° C). In response to salt stress [6.5% (w/v) NaCl], E. faecalis induced a large number of stress proteins. In addition, NaCl strongly induced the synthesis of many proteins more than tenfold. Although the acquired thermotolerance was inhibited markedly by chloramphenicol, the other NaCl-induced cross-tolerances seemed not to be correlated with de novo protein synthesis. The relationship between the stress protein synthesis and the induction of different types of cross-protection is discussed. Received: 7 September 1995 / Accepted: 29 January 1996     

The effect of sodium chloride and NADH on the growth of six strains of haemophilus species pathogenic to chickens.

Six strains of Haemophilus species, pathogenic to chickens, required 1-0 to 1-5% (w/v) NaCl for optimum growth. The requirement was for Na+ rather than NaCl. A sodium salt buffer influenced the optimum NaCl requirement and enhanced growth. Each strain required a different concentration of NADH for an optimum rate of growth.     

Isolation of haloarchaea that grow at low salinities

Summary Archaea, the third domain of life, were long thought to be limited to environmental extremes. However, the discovery of archaeal 16S rRNA gene sequences in water, sediment and soil samples has called into question the notion of Archaea as obligate extremophiles. Until now, none of these novel Archaea has been brought into culture, a critical step for discovering their ecological roles. We have cultivated three novel halophilic Archaea (haloarchaea) genotypes from sediments in which the pore-water salinity was close to that of sea water. All previously reported haloarchaeal isolates are obligate extreme halophiles requiring at least 9% (w/v) NaCl for growth and are typically the dominant heterotrophic organisms in salt and soda lakes, salt deposits and salterns. Two of these three newly isolated genotypes have lower requirements for salt than previously cultured haloarchaea and are capable of slow growth at sea-water salinity (2.5% w/v NaCl). Our data reveal the existence of Archaea that can grow in non-extreme conditions and of a diverse community of haloarchaea existing in coastal salt marsh sediments. Our findings suggest that the ecological range of these physiologically versatile prokaryotes is much wider than previously supposed.     

Effects of Culture Age, Washing and Storage Conditions on Desiccation Tolerance of Colletotrichum truncatum Conidia

Colletotrichum truncatum conidia produced from a one week-old culture in a liquid semi-defined medium with a C:N ratio of 5:1 were more tolerant of desiccation than those harvested from two or three week-old cultures. Conidia washed with 20% (w/v) sucrose germinated better than unwashed conidia or those washed in 10% (w/v) sucrose, 10 and 20% (w/v) glucose or fructose, 0.1% (w/v) soluble starch, 0.9% (w/v) NaCl or deionized water. Washing with sucrose (20% w/v) also resulted in significantly longer germ tubes than those produced by unwashed conidia or conidia washed with deionized water or NaCl (0.9% w/v). Conidia washed twice in sucrose showed greater desiccation tolerance during storage at 15% relative humidity (RH) and 15°C than at 30% RH and 15 or 25°C or at 15% RH and 25, 5 or -10°C.     

Cytochrome P-450 arachidonic acid epoxygenase. Regulatory control of the renal epoxygenase by dietary salt loading.

The rat kidney microsomal epoxygenase catalyzed the asymmetric epoxidation of arachidonic acid to generate as major products: 8(R),9(S)-, 11(R),12(S)- and 14(S),15(R)-epoxyeicosatrienoic acids with optical purities of 97, 88, and 70%, respectively. Inhibition studies utilizing a panel of polyclonal antibodies to several rat liver cytochrome P-450 isoforms, indicated that the renal epoxygenase(s) belongs to the cytochrome P-450 2C gene family. Dietary salt, administered either as a 2-2.5% (w/v) solution in the drinking water or as a modified solid diet containing 8% NaCl (w/w), resulted in marked and selective increases in the renal microsomal epoxygenase activity (416 and 260% of controls, for the liquid and solid forms of NaCl, respectively) with no significant changes in the microsomal omega/omega-1 oxygenase or in the hepatic arachidonic acid monooxygenase reaction. Immunoblotting studies demonstrated that dietary salt induced marked increases in the concentration of a cytochrome P-450 isoform(s) recognized by polyclonal antibodies raised against human liver cytochrome P-450 2C10 or rat liver cytochrome P-450 2C11. Comparisons of the stereochemical selectivity of the induced and non-induced microsomal epoxygenase(s) with that of purified rat liver cytochrome P-450 2C11 suggest that the salt-induced protein(s) is catalytically and structurally different from liver cytochrome P-450 2C11. The in vivo significance of dietary salt in regulating the activities of the kidney endogenous arachidonic acid epoxygenase was established by the demonstration of a salt-induced 10-20-fold increase in the urinary output of epoxygenase metabolites. These results, in conjunction with published evidence demonstrating the potent biological activities of its metabolites, suggest a role for the epoxygenase in the renal response to dietary salt.     

Combined solvent and water activity stresses on turgor regulation and membrane adaptation in Oceanimonas baumannii ATCC 700832

Oceanimonas baumannii ATCC 700832 is a Gram negative marine bacterium capable of utilising phenol as a sole carbon source. The ability of the bacterium to tolerate low water activity when utilising either succinate or phenol as a substrate in minimal medium was studied. The membrane lipid and protein composition showed two discreet adaptive phases as salinity increased. Firstly, when NaCl concentration was increased from 0.15% (w/v), the minimum at which growth was observed, to 1% NaCl (w/v), the ratio of zwitterionic to anionic phospholipids in the membrane increased significantly. At the same time the ratio of saturated to unsaturated fatty acids and the total membrane protein decreased significantly. The second phase was observed when salinity was increased from 1% to 7% NaCl (w/v) as the ratio of zwitterionic to anionic phospholipids decreased and membrane protein increased. However, the ratio of saturated to unsaturated fatty acids was unaffected. Salinity also affected the tolerance of cultures to elevated levels of phenol. Cultures grown in 0.15% NaCl (w/v) could tolerate 12 mM phenol, whereas in the presence of 1% NaCl (w/v) cultures continued to grow in up to 20 mM phenol and in 7% NaCl (w/v) cultures 8 mM phenol could be tolerated. Changes to the composition of the membrane phospholipids and fatty acids were also observed when phenol concentrations were at the maximum that could be tolerated. Under such conditions the ratio of zwitterionic to anionic phospholipids decreased twofold compared to cultures utilising 4 mM phenol as the substrate, in all salinities except in 7% NaCl (w/v) cultures, where there was no significant effect. The ratio of saturated to unsaturated fatty acids increased significantly in all salinities compared to cultures grown with 4 mM phenol. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of salinity increase on carotenoid accumulation in the green alga Dunaliella salina

The effect of sudden salinity increases on the kinetics of growth and carotenogenesis was studied in three geographically diverse isolates of Dunaliella saliva. A sudden increase in salinity results in a lag phase in growth and the length of this lag phase is dependent on the final salinity and the magnitude of the salinity change (no lag at 10–15% w/v NaCl, 4-day lag at 30% NaCl). There is also a lag before an increase in the total carotenoid content can be measured following the salinity up-shock, and the length of the lag depends largely on the initial salinity and the magnitude of the salinity up-shock, whereas the rate of carotenogenesis and the final carotenoid content reached depend on the final salinity. The increase in total carotenoid content is mainly due to β-carotene. Following the salinity up-shock (especially from 10% to 20% NaCl) the proportion of lutein as a percentage of total carotenoids decreases, whereas zeaxanthin increases. This suggests that the pathway synthesising lutein is more sensitive to salt or osmotic stress and is inhibited at higher salinities, thus leading to β-carotene formation. The proportion of α-carotene does not change.     

Stimulation of intermolecular ligation with E. coli DNA ligase by high concentrations of monovalent cations in polyethylene glycol solutions.

In the presence of high concentrations of the nonspecific polymer polyethylene glycol (PEG), intermolecular cohesive-end ligation with the DNA ligase from Escherichia coli was stimulated by high salt concentrations: 200 mM NaCl or 300 mM KCl in 10% (w/v) PEG 6000 solutions, and 100-200 mM NaCl or 150-300 mM KCl in 15% PEG 6000 solutions. Intermolecular blunt-end ligation with this ligase was also stimulated at 100-150 mM NaCl or 150-250 mM KCl in 15% PEG 6000 solutions. The extent of such intermolecular ligation increased and the salt concentrations at which ligation was stimulated extended to lower concentrations when we raised the temperature from 10 to 37 degrees C.     

Synthesis of the compatible solutes glucosylglycerol and trehalose by salt-stressed cells of Stenotrophomonas strains

In this study, physiological processes were analysed, which are involved in salt acclimation of two Stenotrophomonas species, Stenotrophomonas maltophilia strain DSM 50170 and Stenotrophomonas rhizophila strain DSM 14405. S. maltophilia accumulated trehalose as the only osmolyte, whereas S. rhizophila produced additionally to trehalose glucosylglycerol (GG). The different spectrum and amounts of compatible solutes in these two strains led to differences in terms of their salt tolerance. The human-associated S. maltophilia was able to grow in media containing up to 3% NaCl (w/v). In contrast, S. rhizophila propagated in salinities up to 5% NaCl (w/v). The strain was isolated from the rhizosphere, a microenvironment which is characterised by high and changing salinities. Light microscopic analysis of S. rhizophila cells showed a significant increase in cell length of salt-treated cells in comparison to control cells. Cells of S. rhizophila exposed to more than 2% NaCl excreted GG into the medium during the transition from exponential to stationary growth phase, while the internal trehalose pool remained constant. This feature offers a high potential for the biotechnological production of GG.     

Utilization of renewable agricultural residues for the production of extracellular halostable cellulase from newly isolated Halomonas sp. strain PS47

A newly isolated biopolymer-degrading halophilic bacterium, Halomonas sp. strain PS47, yielded higher cellulase activity (0.0076 U/ml) in mineral salt medium (MM63). Activity increased to 0.029 U/ml when carboxymethyl cellulose (0.5 % w/v) was used as carbon source and further to 0.138 U/ml when a combination of yeast extract and peptone was used as nitrogen source. Enzyme secretion was maximal during late exponential and stationary phases (0.15 U/ml, 48 h). Among different agro-residues (1 % w/v), wheat bran gave the highest activity (0.12 U/ml) at pH 7.5, 30 °C and 6 % (w/v) NaCl. The cellulase exhibited higher activity at pH 7.1 and 50 °C. The enzyme exhibited activity over a wide range of NaCl concentrations (0–4 M). Optimum activity was at 0–1 M NaCl. At 4 M NaCl, activity was reduced to 65 % of the initial value. The present investigation thus contributes to the limited information available on halostable cellulases.     

Biochemical changes induced by salt stress in halotolerant bacterial isolates are media dependent as well as species specific

Halophilic bacteria respond to salt stress by regulating the cytosolic pools of organic solutes to achieve osmotic equilibrium. In order to understand the metabolic regulation of these organic solutes, for the first time, we have investigated the effect of salt on growth and biochemical changes in four major moderately halophilic bacterial strains isolated from a saltern region of the Kumta coast, India. The strains under study were Halomonas hydrothermalis VITP9, Bacillus aquimaris VITP4, Planococcus maritimus VITP21, and Virgibacillus dokdonensis VITP14, which exhibited similar salt tolerance (0% to 10% w/v NaCl) with optimal growth at 5% w/v NaCl. Biochemical analysis showed that the total intracellular organic solutes increased significantly with increasing NaCl concentration in the growth medium, and the compositions of the solutes were dependent on the type of strain and also on the nutrient richness of the growth medium. Glutamic acid levels increased in all the strains under salt stress, indicating the significance of glutamic acid as the anionic counterpart of K⁺/Na⁺ ions and precursor for other synthesized nitrogenous osmolytes. Though initial studies were performed with thin-layer chromatography, mass spectrometry was used to identify the major solutes accumulated by the strains under salt stress, such as proline (VITP4), ectoine (VITP14 and VITP9), and sugars (VITP21) under minimal medium and glycine betaine (by all the strains under study) under complex growth medium conditions. Such comparative study on the stress-dependent metabolic differences of different microbes, under identical experimental condition, helps to identify possible bacterial sources for the production of industrially important solutes.     

Effect of salt on aerobic biodegradation of petroleum hydrocarbons in contaminated groundwater

Hydrocarbon-contaminated soil and groundwater at oil and gas production sites may be additionally impacted by salts due to release of produced waters. However, little is known about the effect of salt on the in-situ biodegradation of hydrocarbons by terrestrial microbes, especially at low temperatures. To study this effect, we prepared a groundwater-soil slurry from two sites in Canada: a former flare pit site contaminated with flare pit residue (Site A), and a natural gas processing facility contaminated with natural gas condensate (Site B). The slurry with its indigenous microbes was amended with radiolabeled hydrocarbons dissolved in free product plus nutrients and/or NaCl, and incubated in aerobic biometer flasks with gyrotory shaking at either 25 or 10°C for up to 5 weeks. Cumulative production of ¹⁴CO₂ was measured and the lag time, rate and extent of mineralization were calculated. For Site A, concentrations of NaCl ≥1% (w/v) delayed the onset of mineralization of both ¹⁴C-hexadecane and ¹⁴C-phenanthrene under nutrient-amended conditions, but once biodegradation began the degradation rates were similar over the range of salt concentrations tested (0–5% NaCl). For Site B, increasing concentrations of NaCl ≥1% (w/v) increased the lag time and decreased the rate and extent of mineralization of aliphatic and aromatic substrates. Of particular interest is the observation that low concentrations of salt (≤1% NaCl) slightly stimulated mineralization in some cases.     

Thermokinetic and energetic profiles of the yeast Debaryomyces hansenii in the presence of sodium chloride

Summary Sodium chloride decreased the growth yield with respect to glucose by 60% (at 17% NaCl, w/v), and narrowed the temperature span of growth of Debaryomyces hansenii from 6.8–40.5°C without the salt to 13.2–34.5°C with 16% NaCl. The thermokinetic profile (conjunct display of the Arrhenius plots of the specific rates of growth and thermal death) was dissociative either without or with NaCl, the minimum temperature of thermal death increasing from 42 to 45°C with 10% NaCl.