Effects of components in culture medium on glutamate decarboxylase activity and γ-aminobutyric acid accumulation in foxtail millet (Setaria italica L.) during germination

The effects of glutamic acid (Glu), pyridoxal-5-phosphate (PLP) and calcium chloride (CaCl₂) in culture medium on glutamate decarboxylase (GAD) activity and γ-aminobutyric acid (GABA) accumulation in foxtail millet (Setaria italica L.) during germination were investigated in this study. The components in culture medium for GABA accumulation were optimised using response surface methodology (RSM). Results showed that GAD activity and GABA yield were dependent on the addition of Glu, PLP and CaCl₂ into the culture medium. Box–Behnken design indicated that the optimal culture components for GABA accumulation were: Glu at a concentration of 1.2 mg/ml, PLP at a concentration of 50 μM and CaCl₂ at a concentration of 2.5 mM. Under the optimal conditions, the maximal production of GABA (42.9 mg/100 g FW) was obtained. Analysis of variance for the regression model suggested that the model can quite exactly predict GABA accumulation in millet during germination.     

Metabolic fates of ammonia-N in ruminal epithelial and duodenal mucosal cells isolated from growing sheep

The objective of this experiment was to determine the capability of ruminant gut tissues to detoxify ammonia-N using short-term incubations of isolated cells in vitro. Ruminal epithelial cells (REC) and duodenal mucosal cells (DMC) were isolated from growing Texel-Polypay ram lambs (n = 4) fed a pelleted forage:concentrate-based diet. Immediately after isolation, primary cells were incubated for 60 min with glucose (1mM), glutamate (1mM), [¹⁵N]ammonium chloride (5, 10, 20, or 40 mM), and 1 of 4 combinations of substrates (1 mM each) that could support urea synthesis [control, N-carbamoylglutamate (NCG); NCG + ornithine (ONCG); and ONCG + aspartate (AONCG)]. Treatments were arranged in a 4 × 4 factorial design. Incorporation of ammonia-¹⁵N into alanine, citrulline, arginine, and urea was determined by gas chromatography-mass spectrometry. For both cell types, ammonia-N transfer to alanine was lower when incubation medium contained NCG compared with control, whereas use of ammonia-N for net alanine synthesis increased quadratically with ammonia concentration regardless of substrate treatment. For REC, ammonia-N was not incorporated into citrulline, arginine, or urea, nor into arginine or urea by DMC. Ammonia-N use for net citrulline synthesis exhibited an inverse relationship with ammonia concentration, decreasing linearly as media ammonia concentration increased. Thus, ala-nine synthesis may be a significant metabolic pathway for ruminant gut tissues to detoxify ammonia-N when it is presented luminally at high concentrations as compared with detoxification by the ornithine-urea cycle. Furthermore, DMC do exhibit a metabolic capability to incorporate ammonia-N into citrulline, but low or absent activity of downstream enzymes of the ornithine-urea cycle appears to limit ammonia-N transfers to urea.     

Amperometric assessment of glucose electrode behaviour in mixed solvents and determination of glucose in dairy products

An amperometric biosensor based on a ruthenium(III), nickel(II) and iron(II) hexacyanometallate (HCM)-modified graphite electrode and immobilized glucose oxidase has been used for the determination of glucose in water-miscible organic solvent/aqueous buffer mixtures. Although the specific activity of biochemically active molecule such as enzyme is reduced in organic environment, it was established that the presence of water soluble organic solvents such as methanol, ethanol and acetonitrile (φ = 10%) enhance the biosensor response. Hydrogen peroxide, produced by enzyme-catalysed reduction of glucose, was measured in phosphate buffer solution (pH = 6.86) at −50 mV against a reference Hg|Hg₂Cl₂|3 M KCl electrode to determine the concentration of glucose. The influence of the addition of different volume fractions (φ = 10–60%) of methanol, ethanol, acetone, acetonitrile and isopropanol on biosensor response was investigated. The obtained amperometric signals were fast, reproducible and linearly proportional to glucose concentrations in the range of 0.1–0.8 mM, with a squared correlation coefficient of 0.9994 for buffer solution. With the addition of ethanol (φ = 10% and 40%) the plateau on I/c curve was obtained for concentrations of glucose higher than 0.8 and 1.1 mM, respectively. The biosensor proved to be stable for several months. The recoveries of added glucose (0.200 and 0.300 mM) from aqueous solution and from solution with ethanol φ = 10% ranged from 96.0% to 108.0%. The biosensor was used for the determination of glucose in some food samples of dairy industry, and the results were consistent with those obtained with the commercially available glucose enzyme photometric kit.     

An extra-cellular alkaline metallolipase from Bacillus licheniformisMTCC 6824: Purification and biochemical characterization

An extra-cellular lipase produced by Bacillus licheniformis MTCC 6824 was purified to homogeneity by ammonium sulphate fractionation, ethanol/ether precipitation, dialysis, followed by anion-exchange chromatography on Amberlite IRA 410 (Cl⁻ form) and gel exclusion chromatography on Sephadex G 100 using Tris–HCl buffer (pH 8.0). The crude lipase extract had an activity of 41.7 LU/ml of culture medium when the bacterium was cultured for 48 h at 37 °C and pH 8.0 with nutrient broth supplemented with sardine oil as carbon source. The enzyme was purified 208-fold with 8.36% recovery and a specific activity of 520 LU/mg after gel exclusion chromatography. The pure enzyme is a monomeric protein and has an apparent molecular mass of 74.8 kDa. The lipase had a V_max and K_m of 0.64 mM/mg/min and 29 mM, respectively, with 4-nitro phenylpalmitate as a substrate, as calculated from the Lineweaver–Burk plot. The lipase exhibited optimum activity at 45 °C and pH 8.0, respectively. The enzyme had half-lives (T_1/2) of 82 min at 45 °C, and 48 min at 55 °C. The catalytic activity was enhanced by Ca²⁺ (18%) and Mg²⁺ (12%) at 30 mM. The lipase was inhibited by Co²⁺, Cu²⁺, Zn²⁺, Fe² even at low concentration (10 mM). EDTA, at 70 mM concentration, significantly inhibited the activity of lipase. Phenyl methyl sulfonyl fluoride (PMSF, 70 mM) completely inactivated the original lipase. A combination of Ca²⁺ and sorbitol induced a synergistic effect on the activity of lipase with a significantly high residual activity (100%), even after 45 min, as compared to 91.5% when incubated with Ca²⁺ alone. The lipase was found to be hydrolytically resistant toward triacylglycerols with more double bonds.     

Biochemical characteristics of Trametes multicolor pyranose oxidase and Aspergillus niger glucose oxidase and implications for their functionality in wheat flour dough

Similar to glucose oxidase (GO), pyranose oxidase (P₂O) may well have desired functionalities in some food applications in general, particularly breadmaking. As its name implies, P₂O oxidises a variety of monosaccharides. P₂O purified from a culture of Trametes multicolor (P₂O-Tm) had high affinity towards d-glucose (K_M = 3.1 mM) and lower affinity to other monosaccharides. GO from Aspergillusniger (GO-An) had a K_M value of 225 mM towards glucose, which points to a significant difference in glucose affinity between the two enzymes. Furthermore, P₂O-Tm had higher affinity towards O₂ (K_M = 0.46 mM) than GO-An (K_M = 2.9 mM). Dehydroascorbic acid did not accept electrons in the reactions catalysed by P₂O-Tm and GO-An. For the same activity towards glucose in saturating conditions, the rate of ferulic acid oxidation in a model system and of thiol oxidation in a wheat flour extract were higher with P₂O-Tm, than with GO-An. The demonstrated differences in properties and functional features between P₂O-Tm and GO-An allow prediction of differences in functional behaviour of the enzymes, in food applications.     

Evaluation of culture media for enumeration of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium animalis in the presence of Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus

The study compared the growth capability of probiotic (Lactobacillus acidophilus La05, Lactobacillus casei Lc01 and Bifidobacterium animalis Bb12) and non-probiotic (Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus) cultures on twenty-one culture media grouped according to selectivity: non-selective agars, selective agars without antibiotics and MRS agars containing different combinations of lithium chloride, cystein, bile salts and antibiotics. Four of these media were selected for quantitative enumeration of L. acidophilus La05, L. casei Lc01, and B. animalis Bb12. The best culture media and incubation conditions for enumeration of the probiotic cultures were: B. animalis: MRS agar with dicloxacillin, 37 °C or 42 °C, anaerobiosis; L. acidophilus: MRS agar with bile salts, 37 °C or 42 °C, aerobiosis; L. casei: MRS agar with lithium chloride and sodium propionate, 37 °C or 42 °C, aerobiosis or anaerobiosis. Plating on MRS with glucose replaced by maltose, 37 °C or 42 °C, anaerobiosis, will distinguish probiotic from non-probiotic cultures. For enumeration of each probiotic in a mixed culture, the following media and incubation conditions were recommended: B. animalis: 4ABC-MRS, 42 °C, anaerobiosis, L. acidophilus: LC medium, 42 °C, aerobiosis or anaerobiosis and L. casei: LP-MRS, 42 °C, aerobiosis or anaerobiosis. In all experiments, differences in counts using pour plating or surface plating were not significant (P ≤ 0.05).     

Enzymatic synthesis of primeverosides using transfer reaction by Trichoderma longibrachiatum xylanase

Enzymatic synthesis of two phenyl xylopyranosyl glucopyranosides, through transfer reaction by Trichoderma longibrachiatum endoxylanase, was achieved in the presence of n-hexane used as solvent, phenyl glucoside (10 mM) as acceptor and xylan (2 g/l) as donor. Kinetic study showed that only one compound, identified by ¹H and ¹³C NMR and heteronuclear 2D (¹H–¹³C) chemical shift correlation as phenyl primeveroside (phenyl 6-O-β-xylopyranosyl-1-β-d-glucopyranoside), was synthesized when the reaction time was beyond 1 h. Benzyl and hexyl primeverosides were obtained under the same conditions. When several phenyl glucoside concentrations, from 5 to 50 mM, were used with 2 g/l of xylan, a phenyl primeveroside isomer, identified as phenyl 4-O-β-xylopyranosyl-β-d-glucopyranoside, accumulated in the medium whereas the production of phenyl primeveroside decreased. Only phenyl primeveroside was produced when several xylan concentrations from 2 to 10 g/l were used with 10 mM of phenyl glucoside and its concentration in the reaction mixture increased with the increase of xylan concentration.     

Evaluation of ability of ferulic acid to control growth and fumonisin production of Fusarium verticillioides and Fusarium proliferatum on maize based media

The aim of this study was to evaluate the efficacy of ferulic acid (1, 10, 20 and 25 mM) at different water activity (a_w) values (0.99, 0.98, 0.96 and 0.93) at 25 °C on growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on maize based media. For both Fusarium species, the lag phase significantly decreased (p ≤ 0.001), and the growth rates increased (p ≤ 0.001) at the lowest ferulic acid concentration used (1 mM), regardless of the a_w. However, high doses of ferulic acid (10 to 25 mM) significantly reduced (p ≤ 0.001) the growth rate of both Fusarium species, regardless of the a_w. In general, growth rate inhibition increased as ferulic acid doses increased and as media a_w decreased. Fumonisin production profiles of both Fusarium species showed that low ferulic acid concentrations (1–10 mM) significantly increased (p ≤ 0.001) toxin production, regardless of the a_w. High doses of ferulic acid (20–25 mM) reduced fumonisin production, in comparison with the controls, by both Fusarium species but they were not statistically significant in most cases. The results show that the use of ferulic acid as a post-harvest strategy to reduce mycotoxin accumulation on maize needs to be discussed.     

Evaluation of the effect of carvacrol on the Escherichia coli 555 metabolome by using H-NMR spectroscopy

Cultures of Escherichia coli 555 were grown at four levels of carvacrol (0–2 mM) and the E. coli endo-metabolome was extracted and measured by ¹H NMR spectroscopy. The results show that glucose concentration is going up with concentration of carvacrol and so do formate until the highest concentration is reached, from which point it suddenly decreases. This is interpreted as if the bacteria are increasingly unable to further metabolize glucose and as if the bacteria increasingly shifts with higher levels of carvacrol toward sugar fermentation as carbon source, until the level of carvacrol reaches a level (2.00 mM), where the E. coli must give up. Additionally, the multivariate Principal Component Analysis suggests that the adaptation occurring at sub-lethal doses of carvacrol is different from that occurring at higher doses.     

Sucrose hydrolysis by gelatin-immobilized inulinase from Kluyveromyces marxianus var. bulgaricus

The crude cell-free medium from a culture of Kluyveromyces marxianus var. bulgaricus was immobilized in a gelatin-water support, with an immobilization yield of 82.60% for inulinase activity. The optimum pH for both free and immobilized inulinase was the same (3.5) and the optimum temperatures were 55 °C for the free and 60 °C for the immobilized enzyme. The Arrhenius plots were linear and activation energies were 56.20 (free enzyme) and 20.27 kJ/mol K (immobilized enzyme). The kinetic parameters were calculated by Lineweaver–Burk plots and the V_max and K_m were 37.60 IU/mg protein and 61.83 mM for the free inulinase and 31.45 IU/mg protein and 149.28 mM for the immobilized enzyme, respectively. The operational stability of the immobilized inulinase was studied in a continuous fixed-bed column reactor for 33 days, at the end of which the sucrose conversion was 58.12%.     

Effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and nutrient utilization in dairy does

The objective of this study was to evaluate effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and on feed intake and nutrient utilization in dairy does. Batch cultures of mixed rumen microorganisms were used to study effects of different concentrations of fumarate on fermentation with various N sources (ammonia as ammonium bicarbonate, casein amino acids, casein peptides, gelatin peptides) and feeds (bermudagrass hay, mixed diet of 60% bermudagrass hay plus 40% concentrate) for 6 and 24 h, respectively. Substrates were grouped into pairs for separate incubations. Monosodium fumarate was added to incubation tubes to achieve final concentrations of 0, 5, and 10 mM fumarate. More ammonia accumulated at the end of incubation with added ammonium bicarbonate. Ammonia concentration was higher for peptide compared with amino acid incubation, and for casein peptide compared with gelatin peptide. Addition of fumarate linearly decreased ammonia for all N sources and for feed substrates. For all substrate types, fumarate treatment increased acetate, propionate, and total volatile fatty acids (VFA), decreased acetate to propionate ratio, and tended to reduce branched-chain VFA. Digestion of feed neutral detergent fiber (NDF) by rumen microorganisms was improved by fumarate along with elevated endoglucanase and xylanase activities. In an animal metabolism experiment, 8 dairy does (4 per treatment) were used in a completely randomized design for 21 d. Does were fed a hay plus concentrate diet without (control) or with fumarate (6 g/head per day) supplementation to determine feed intake, whole-tract nutrient digestibility, and N utilization. Fumarate treatment did not affect weight change or feed intake but increased whole-tract digestion of gross energy, crude protein, and cellulose. Digested N was increased by fumarate supplementation; however, N retention was unaffected. Plasma glucose concentration was elevated with fumarate but urea N concentration remained unchanged. Fumarate addition had significant effects on rumen microbial fermentation by decreasing ammonia and branched-chain VFA, and by increasing acetate and propionate, and NDF digestion. These effects were reflected in the improvement in whole-tract gross energy, crude protein, and cellulose digestion and elevated plasma glucose concentration when dairy does were supplemented with fumarate.     

Short communication: Interrelationship between butyrate and glucose supply on butyrate and glucose oxidation by ruminal epithelial preparations

The aim of this study was to determine whether dietary Na-butyrate supplementation affects butyrate and glucose oxidation by ruminal epithelial preparations and whether this effect can be acutely modulated by substrate (glucose and butyrate) supply. Eighteen Suffolk wether lambs (6 lambs/treatment) were blocked by body weight and, within block, randomly assigned to the control treatment (CON) or to diets containing differing Na-butyrate inclusion rates (1.58 or 3.16%) equating to 1.25 (B1.25), and 2.50% (B2.50) butyrate on a dry matter basis, respectively. All lambs received their diet for a period of 14 d. After dietary adaptation, lambs were killed and the ruminal epithelium was harvested from the ventral sac, minced finely, and used for in vitro incubations. Incubation medium contained either a constant concentration of glucose (4 mM) with increasing butyrate concentrations (0, 5, 15, 25, or 40 mM) or a constant butyrate concentration (15 mM) with increasing glucose concentrations (0, 1, 2, 4, or 8 mM) to allow for the evaluation of whether acute changes in the concentration of metabolic substrates affect the oxidation of glucose and butyrate. We observed no interactions between the in vivo and in vitro treatments. Increasing dietary butyrate supplementation linearly decreased glucose oxidation by ruminal epithelial preparations, but had no effect on butyrate oxidation. Increasing butyrate concentration in vitro decreased (cubic effect) glucose oxidation when butyrate concentration ranged between 5 and 15 mM; however, glucose oxidation was increased with a butyrate concentration of 40 mM. Butyrate oxidation decreased (cubic effect) as glucose concentration increased from 1 to 4 mM; however, butyrate oxidation increased when glucose was included at 8 mM. The results of this study demonstrate that dietary butyrate supplementation can decrease glucose oxidation by the ruminal epithelium, but the relative supply of glucose and butyrate has a pronounced effect on substrate oxidation.     

TBARs distillation method: Revision to minimize the interference from yellow pigments in meat products

The aim was to study the effect of the incubation method and TBA reagent (concentration/solvent) on yellow pigment interference in meat products. Distillates from red sausage, sucrose, malondialdehyde and a mixture of sucrose–malondialdehyde were reacted with four different TBA solutions at five different temperature/time relations. Two TBA solutions were prepared at 20 mM using 90% glacial acetic acid or 3.86% perchloric acid. In addition, an 80 mM TBA solution was prepared using distilled water adjusted to pH 4 and another using 0.8% TBA in distilled water. The temperature/time relations were: (1) 35 min in a boiling water bath; (2) 70 °C/30 min; (3) 40 °C/90 min; (4) room temperature (r.t.) (24 °C) in dark conditions for 20 h; and (5) 60 min in a boiling water bath. The results showed that aqueous or diluted acid solutions of TBA reagent and the application of 100 °C for less than 1 h provided the best conditions to minimize the presence of yellow pigments and maximize pink pigment formation in meat products.     

Hypoglycaemic effect of comatin,an antidiabetic substance separated from Coprinus comatus broth,on alloxan-induced-diabetic rats

Comatin, an inhibitor of the non-enzymatic glycosylation (NEG) reaction, was isolated from Coprinus comatus fermentation broth by macroporous resin separation, followed by chromatographic purification using a C18 reversed-phase column. The compound was identified as 4,5-dihydroxy-2-methoxy-benzaldehyde by high-resolution MS, IR, NMR and UV analyses. The hypoglycaemic effect of comatin, on both normal and alloxan-induced-diabetic rats, was investigated in the paper. The blood glucose concentration of normal rats treated by the comatin at 80 mg/kg body weight was reduced from 5.14 mM to 4.28 mM in 3 h. Also, the concentrations of fructosamine, triglycerides and total cholesterol in induced-diabetic rats were significantly decreased. These results indicated that comatin could maintain a low level of blood glucose and improve glucose tolerance.     

Optimization of natural fermentative medium for selenium-enriched yeast by d-optimal mixture design

Natural fermentative medium for selenium-enriched yeast (Saccharomyces cerevisiae) culture was investigated using response surface methodology (RSM). The medium with a concentration of 15 μg/mL Na₂SeO₃, contained various ratios of juices from germinated brown rice (0.40 ∼ 0.80, 12 Brix), beerwort (0.10 ∼ 0.50, 12 Brix) and soybean sprout (0.10 ∼ 0.50, 12 Brix), which were optimized by applying d-optimal mixture design (DMD). The effects of their ratios on biomass yield and total selenium (Se) yield were analyzed. The results showed that when the mixed ratio of the components was 4:4:2 (v:v:v), the maximum value of biomass yield and total Se yield were 8.5 g/L and 3.53 mg/L, respectively. Verification experimental trials were performed for validating the models, and it indicated that the above mixture of these natural materials can be used as proper medium for the growth of selenium-enriched yeast and accumulation of Se.     

Sensory and physicochemical evolution of tropical cooked peeled shrimp inoculated by Brochothrix thermosphacta and Lactococcus piscium CNCM I-4031 during storage at 8°C

This study investigated the sensory quality and physicochemical evolution (pH, glucose, l-lactic acid, biogenic amine, free amino-acids and volatile compounds) during storage at 8 °C of cooked peeled shrimp inoculated with the specific spoilage bacteria Brochothrix thermosphacta alone or mixed with the protective strain Lactococcus piscium CNCM I-4031. Growth of both bacteria was monitored at regular intervals during storage by microbial counts and the thermal temperature gradient gel electrophoresis (TTGE) technique. Bacterial counts showed that L. piscium and B. thermosphacta inoculated at 7 log CFU/g and 3 log CFU/g were well adapted to shrimp, reaching a maximum level of 9 log CFU/g after 4 days and 10 days respectively. In mixed culture, the growth of B. thermosphacta was reduced by 3.2 ± 0.1 log CFU/g. The TTGE technique allowed monitoring the colonisation of the strains on the shrimp matrix and confirming the dominance of L. piscium in mixed culture throughout the experiment. Sensory analysis confirmed that B. thermosphacta spoiled the product after 11 days, when its cell number attained 8 log CFU/g with the emission of strong butter/caramel off-odours. This sensory profile could be linked to the production of 2,3 butanedione, cyclopentanol, 3-methylbutanol, 3-methylbutanal, 2-methylbutanal, 4-methyl-3-chloro-3-pentanol and ethanol, which were produced in more significant quantities in the B. thermosphacta batch than in the batches in which the protective strain was present. On the contrary, TVBN and TMA were not suitable as quality indicators for B. thermosphacta spoilage activity. In the products where the protective L. piscium strain was present, no adverse effect on sensory quality was noted by the sensory panels. Moreover, biogenic amine assessment did not show any histamine or tyramine production by this strain, underlining its safety profile. Both strains produced lactic acid (1850 mg/kg in L. piscium and B. thermosphacta batch on days 3 and 10 respectively; 3830 mg/kg on day 7 in mixed culture) and the pH decrease from 6.6 ± 0.0 to 5.9 ± 0.1 was similar in all batches. Lactic acid production or competition for free amino-acid was not involved in the inhibition mechanism; however rapid glucose consumption by L. piscium could partially explain the growth limitation of the spoilage micro-organism. This study demonstrated the spoilage characteristic of B. thermosphacta and the usefulness of L. piscium as a bioprotective culture for tropical cooked peeled shrimp without any adverse effect on the sensory quality of the product.     

Gold nanoparticle modified conducting polymer of 4-(2,5-di(thiophen-2-yl)-1H-pyrrole-1-l) benzenamine for potential use as a biosensing material

Gold nanoparticle (AuNP) modified conducting polymer of 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine (SNS-NH₂) was used as the biosensing platform for glucose analysis. Electrochemical measurements were carried out by following the consumed oxygen due to the enzymatic reaction of glucose oxidase (GOx) at −0.7 V vs Ag/AgCl. Optimisation of pH, enzyme loading, stability experiments were carried out. Effect of NP was investigated by monitoring the signal responses at different AuNP sizes and amounts. A linear relation of y = 1.597x + 0.264 (R² = 0.993) was found for glucose concentrations between 0.002 and 5.0 mM. The analytical characteristics of the system were also evaluated for glucose determination in flow injection analysis (FIA) mode. Finally, the system was checked for glucose detection on real samples.     

Effect of water activity on stress tolerance and biocontrol activity in antagonistic yeast Rhodosporidium paludigenum

The aims of this study were to optimize the salt-adaptation conditions of the marine antagonistic yeast Rhodosporidium paludigenum and investigate the biocontrol activity of salt-induced cell suspensions of R. paludigenum on postharvest pathogens in fruits. Low water activity (a_w = 0.98, 0.97, 0.96, and 0.95) inhibited the growth of R. paludigenum in nutrient yeast dextrose broth, but the yeast grew better in the medium modified with NaCl solute than other nonionic solutes. R. paludigenum grown in 6.6% NaCl-modified medium had higher viabilities (92.1%) at low water activity (a_w = 0.95) than control (81.1%) after 48 h incubation. The salt-adapted R. paludigenum also showed better viability than the un-adapted cells after being frozen, which may be related to the accumulation of intracellular trehalose. Moreover, the best biocontrol inhibition in pears and Chinese winter jujubes was obtained when R. paludigenum was grown in NaCl-modified medium. Therefore, this study implies that improving physiological inducement methods may be a promising strategy for accelerating commercialization of biocontrol agents.     

Short communication: Eicosatrienoic acid and docosatrienoic acid do not promote vaccenic acid accumulation in mixed ruminal cultures

Previous research found that docosahexaenoic acid (C22:6n-3) was a component of fish oil that promotes trans-C18:1 accumulation in ruminal cultures when incubated with linoleic acid. The objective of this study was to determine if eicosatrienoic acid (C20:3n-3) and docosatrienoic acid (C22:3n-3), n-3 fatty acids in fish oil, promote accumulation of trans-C18:1, vaccenic acid (VA) in particular, using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of C20:3n-3 (ETA), control plus 5 mg of C22:3n-3 (DTA), control plus 15 mg of linoleic acid (LA), control plus 5 mg of C20:3n-3 and 15 mg of linoleic acid (ETALA), and control plus 5 mg of C22:3n-3 and 15 mg of linoleic acid (DTALA). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the concentrations of trans-C18:1 (0.87, 0.88, and 0.99 mg/culture), and VA (0.52, 0.56, and 0.62 mg/culture) were similar for the control, ETA, and DTA cultures, respectively. The concentrations of trans-C18:1 (5.51, 5.41, and 5.36 mg/culture), and VA (4.78, 4.62, and 4.59 mg/culture) were also similar between LA, ETALA, and DTALA cultures, respectively. These data suggest that C20:3n-3 and C22:3n-3 are not the active components in fish oil that promote VA accumulation when incubated with linoleic acid.     

Structural characterization and antioxidant activity of an extracellular polysaccharide isolated from Brevibacterium otitidis BTS 44

A water-soluble acidic extracellular polysaccharide reaching a maximum concentration of 23.4 g/l growth medium, coded as BSMA, was isolated from the non-pathogenic soil bacteria Brevibacterium otitidis BTS44, by precipitating with two volumes of ethanol. BSMA consisted of arabinose, mannose, glucose and mannouronic acid in ratios of 2.7:3.6:2.1:1.0. No protein was detected in the BSMA fraction, and its molecular weight was about 127 kDa. It has a backbone composed of (1 → 5)-linked arabinose, (1 → 6)-linked mannose with three branches attached to O-3 of (1 → 6)-linked mannose and terminated with either mannose, or mannose and glucose; all the glucose and most of the mannouronic acid are distributed in branches. Partial acid hydrolysis of BSMA gave four sub-fractions termed BSMA-1, BSMA-2, BSMA-3 and BSMA-4. BSMA-1 was composed of arabinose, mannose and trace amounts of mannouronic acid; BSMA-2 was only composed of arabinose and mannose; BSMA-3 was composed of mannose, mannouronic acid and glucose, and BSMA-4 was only composed of mannose and glucose. In the in vitro antioxidant assay, BSMA was found to possess DPPH radical-scavenging activity, with an IC₅₀ value of 120 μg/ml.