Fractions of barley spent grain as media for growth of probiotic bacteria

The application of the protein and polysaccharide fractions of barley spent grain as a basis of growth media for probiotic bacteria was studied. High values of biomass yield, cell viability, and organic acid production were observed in the variants of media containing the barley spent grain supplemented with lactose, ascorbic acid, yeast extract, and mineral salts. Cells of lactic acid bacteria and bifidobacteria had the typical rod-shaped morphology.     

A rapid procedure for detection of bacterial amino acid decarboxylases

The detection of decarboxylases of arginine, glutamic acid, histidine, lysine, ornithine, phenylalanine, tryptophan, and tyrosine in bacteria by thin-layer chromatography on polyamide sheets is described. The bacteria were grown on agar medium plates supplemented with eight amino acids at pH 5.5 for induction of amino acid decarboxylases, then transferred to amine-production media. The decarboxylation products in the spent media (amines and/or γ-amino-n-butyric acid) were dansylated and the dansyl derivatives were separated by thin-layer chromatography on polyamide sheets. This method requires only two separate incubations of the decarboxylase-induced bacteria in amine-production media for 1 h at 37°C for simultaneous detection of eight bacterial amino acid decarboxylases using 0.4 μl of the spent media.     

Production of Propionibacterium shermanii biomass and vitamin B12 on spent media

AIMS: The propionibacteria are commercially important due to their use in the cheese industry, and there is a growing interest for their probiotic effects. Stimulatory effects of lactic acid bacteria (LAB) on propionic acid bacteria have been observed. This study was designed to examine the possibility of using spent media previously used to grow LAB for the production of biomass and metabolites of Propionibacterium freudenreichii subsp. shermanii. METHODS AND RESULTS: Seventeen MRS and vegetable juice media were prefermented by various LAB and evaluated for their ability to subsequently support the growth of Propionibacterium, using automated spectrophotometry (AS). Growth of Propionibacterium in spent media was strongly affected by the LAB strain used to produce the spent medium. The native MRS medium (not prefermented) yielded the highest optical density values followed by prefermented media by Lactobacillus acidophilus, Bifidobacterium longum and Lactococcus lactis. Prefermented cabbage juice enabled good growth of Propionibacterium. For the production of organic acids and vitamin B12, cells of Propionibacterium were concentrated and immobilized in alginate beads in the aim of accelerating the bioconversions. More propionic acid was obtained in spent media than in native MRS. The concentration of vitamin B12 was higher in media fermented with free cells than those with immobilized cultures; with the free cells, its concentration varied from 900 to 1800 ng ml(-1) of media. CONCLUSIONS: It was demonstrated that spent media could be recycled for the production of Propionibacterium and metabolites, depending on the LAB strain that was previously grown. Media remediation is needed to improve the production of vitamin B12, especially with immobilized cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents an option for recycling of spent media generated by producers of LAB or producers of fermented vegetables. The propionic fermentation may result in three commercial products: biomass, vitamin B12 or organic acids, which may be used as starters, supplements or food preservatives. It is an attractive process from economical and environmental standpoints.     

Influence of barley grain particle size and treatment with citric acid on digestibility,ruminal fermentation and microbial protein synthesis in Holstein calves

Chemical and physical treatments of barley grain increase ruminally resistant starch and can improve the rumen fermentation pattern. The objective of the present study was to evaluate the effects of chemical (addition of citric acid, CA) and physical (grinding to two different particle sizes, PS) treatment of barley grain on performance, rumen fermentation, microbial protein yield in the rumen and selected blood metabolites in growing calves. In all, 28 male Holstein calves (172±5.1 kg initial BW) were used in a complete randomised design with a factorial arrangement of 2 barley grain particle sizes×2 levels of citric acid. The diets were as follows: (i) small PS (average 1200 µm) barley grain soaked in water (no CA addition); (ii) small PS barley grain soaked in a CA solution (adding 20 g CA/kg barley); (iii) large PS (average 2400 µm) barley grain soaked in water (no citric acid addition) and (iv) large PS barley grain soaked in a citric acid solution (adding 20 g CA/kg barley). Barley grain was then incorporated at 35% in a total mixed ration and fed to the calves for 11 weeks. Feeding small PS barley decreased feed intake (P=0.02) and average daily weight gain (P=0.01). The addition of CA to barley grain did not affect intake but increased weight gain (P<0.01) and improved feed to gain ratio (P=0.03). Digestibility of organic matter and NDF tended (P<0.10) to increase, whereas faecal scoring was improved (P=0.03) and the presence of undigested grain particles in faeces was reduced (P<0.01) with CA-treated barley grain. Glucose and urea concentrations were increased (P<0.01) in the blood of calves fed the CA-treated barley grain. Ruminal pH tended (P=0.08) to be decreased with more finely ground barley and was increased when barley grain was treated with CA. Total volatile fatty acid concentrations in the rumen did not differ among treatments (P>0.05). However, the molar proportion of propionate was increased (P=0.03) when barley was more finely ground, and that of acetate was increased (P=0.04) when CA was added to barley grain. The ruminal concentration of ammonia nitrogen was increased (P<0.01) and microbial nitrogen synthesis in the rumen tended to decrease by adding CA to barley. Treating barley grain with citric acid increased fibre digestibility of total mixed rations, attenuated the decrease in ruminal pH, and improved weight gain and feed efficiency in male Holstein growing calves fed a high-cereal diet (550 g cereal grain/kg diet).     

Malt Extract as Protective Medium for Lactic Acid Bacteria in Cryopreservation

S ummary : Malt extracts prepared from barley and sorghum malts were used as media for preserving bacteria at -196°. The results are compared with those obtained with other protective substances such as dimethylsulphoxide, glycerol, malic acid, sucrose and sodium glutamate. The malt extract medium is recommended for use in the food industry.     

Impact of barley form on equine total tract fibre digestibility and colonic microbiota

This study aimed at assessing the impact of four barley forms on total tract apparent digestibility of dietary fibre in horses fed a large amount of starch in the morning meal (0.27% BW). Processed barley forms had a greater pre-caecal starch digestibility than the whole form. Based on this result, we hypothesised that using barley-processing methods would limit the potential dumping of undegraded starch in the hindgut of horses and, consequently, the potential negative effect on fibre degradation in the hindgut. In a 4×4 latin square design, four mature geldings fitted with a right ventral colon-fistula were fed a meadow hay : concentrate (62 : 38; dry matter (DM) basis) diet at 1.7% BW. The concentrate was made of 80% barley distributed either as whole grain or as processed forms: 2.5 mm ground, pelleted or steam-flaked. For each period, total tract apparent digestibilities of DM, NDF and ADF were determined over 3 consecutive days by total faecal collection, whereas pH, volatile fatty acids (VFA) concentrations and cultural functional bacteria counts (total anaerobic, cellulolytic bacteria, lactic acid producers, amylolytic bacteria and lactic acid utilisers) in colonic content were evaluated on 1 day 4 h after the morning meal. Total tract apparent digestibility of DM and dietary fibre was influenced (P<0.05) by barley form. Diets including thermo-mechanically treated barley forms led to a higher (P<0.05) total tract apparent digestibility of NDF than those constituted of ground barley and also led to a greater (P<0.05) total tract apparent digestibility of ADF than those made of whole or ground barley forms. However, no significant difference was observed in colonic pH, VFA concentrations and cultural bacteria concentrations. Owing to a high starch supply in the morning meal, the concentration of the functional bacteria in the colonic content averaged 7.8 log CFU/ml, 5.9 NPM/ml, 6.9 and 7.3 CFU/ml for total anaerobic, cellulolytic, amylolytic and lactic acid-utilising bacteria, respectively. Consequently, providing horses with pelleted or steam-flaked instead of ground barley forms may limit the negative impact of starch on fibre digestibility in horses fed a high level of starch in the morning meal (0.27% BW). Moreover, the fibre-to-starch ratio fed in this experiment did not cause any digestive upset.     

Manufacture and prebiotic potential of oligosaccharides derived from industrial solid wastes

The solid waste obtained in malting industries when dehulling barley grains, which was mainly made up of barley husks, spent grains and grain fragments, was subjected to a double hydrothermal processing under selected conditions. The liquor from the second stage (containing xylooligosaccharides, XOS) was refined by membrane and ion exchange processing (with or without a previous endoxylanase treatment to reduce the XOS molecular weight). Three XOS concentrates with different purity and/or molecular weight distribution were fermented in vitro with faecal inocula to assess their prebiotic potential. Succinate, lactate, formiate, acetate, propionate and butyrate were generated in fermentations, confirming the prebiotic potential of the various products assayed. The purity of XOS concentrates did not play a significant role in fermentation, whereas the sample with shorter average degree of polymerization presented a faster fermentation kinetics and led to the highest concentration of lactic acid.     

Ability of ectomycorrhizal fungi to utilize starch and related substrates

Basidiomycetous fungi of 55 strains of 33 species in 15 genera which are thought to be ectomycorrhizal were grown on starch and related substrates as a sole carbon source, and their ability to utilize these substrates was determined. Mycellial weights of the fungi grown on agar media containing starch and amylose varied between the strains from 1.1 to 94.9 mg/flask and from 0.4 to 93.3mg/flask, respectively. Mycelial growth rates ranged from 0 to 1.17 mm/d on barley grain medium and from 0 to 2.03 mm/d on rice grain medium; the highest rate corresponded to about half of the average of reference wood-rotting fungi. Most of the mycorrhizal fungi that grew well on amylose gave higher growth rates on barley. Several strains inLyophyllum, Hebeloma, Sarcodon, andTricholoma grew well on both glucose and starch media.     

Fatty acid content and composition in relation to grain size of barley

The relationship between grain size and total fatty acid (TFA) content and composition, as well as the contribution of corn size distribution to the variation of fatty acid contents between barley types, were studied in nine barley varieties (six-rowed and two-rowed winter barleys, and two-rowed spring barleys) fractionated according to grain size. Smaller grains contained as much TFA as the bigger ones. On average, the proportions of saturated fatty acids (16:0 and 18:0) and of oleic acid (18:1) are higher in large grains than in small kernels, whereas the percentages of linoleic (18:2) and linolenic (18:3) acids are lower. The higher 18:1 and the lower 18:3 proportions in two-rowed barleys compared to their six-rowed counterparts, can be explained by the indirect effect of the different grain size distribution between the barley types.     

THE COMPETITION BETWEEN BARLEY AND CERTAIN WEEDS UNDER CONTROLLED CONDITIONS

Chickweed (Stellaria media) is one of the commonest annual weeds on almost all soils, and forms 20–30 % of the weed herbage on the lower greensand soils of Woburn, Beds. It occurs in all crops in this area except on very acid soils.
When barley and chickweed are planted together, with abundance of water and nutrients for both, it is found that increasing the density of planting of the barley reduces the loss due to chickweed competition, but even with very close-planted barley, the loss caused by the weed amounts to nearly two-thirds of the total fresh weight and to four-fifths of the grain yields. This is much greater than with the two annual weeds previously studied (spurrey and mayweed). In a sparse crop of barley, increase in chickweed growth may reduce the barley to less than 10 % of its growth without the weed, while the chickweed itself is relatively little affected by the presence of the barley.
There is no evidence of any specific effect of the roots of the one plant on the other; they intertwine without any sign of attraction or repulsion between them.
Comparisons are made with other annual weeds previously studied, and it is suggested that the method adopted furnishes a means of assessing the relative effectiveness of these weeds in competing with barley.     

Grain source and marginal changes in forage particle size modulate digestive processes and nutrient intake of dairy cows

This study investigated the effects of, and interactions between, dietary grain source and marginal changes in alfalfa hay (AH) particle size (PS) on digestive processes of dairy cows. A total of eight Holstein dairy cows (175 days in milk) were allocated in a replicated 4 × 4 Latin square design with four 21-day periods. The experiment was a 2 × 2 factorial arrangement with two levels of theoretical PS of AH (fine = 15 mm or long = 30 mm) each combined with two different sources of cereal grains (barley grain alone or barley plus corn grain in a 50 : 50 ratio). Results showed that cows consuming diets supplemented with corn had greater dry matter and nutrient intakes (P < 0.01), independent of forage PS. In addition, the apparent digestibility of fiber fractions was greater for diets supplemented with corn (P = 0.01). The feeding of barley grain-based diets was associated with greater apparent digestibility of non-fiber carbohydrates, and this variable was even greater when long AH was fed (P = 0.04). Moreover, the feeding of long AH resulted in longer time spent eating (P = 0.03) and higher pH (P < 0.01), as well as a tendency for higher acetate-to-propionate ratio in the rumen fluid (P = 0.06) at 3 h post feeding. In conclusion, the results indicated that the marginal increase of PS of AH may prolong eating time and improve rumen fermentation, particularly in diets based on barley grain. Partial substitution of barley grain by corn can improve feed intake and fiber digestibility in mid-lactation dairy cows.     

Effect of molybdate ions on methanation of simulated and natural waste-waters

Summary Sulphate in concentrations of 500 and 1000 mg SO₄-S/l did not inhibit methanation of synthetic waste-water (acetate + methanol + glucose) by sludge from a digester treating neutral spent sulphite process effluents. The role of sulphate reducers in the conversion of those substrates was minor although sulphate-reducing bacteria were present with a viable count similar to that of methane-producing bacteria in the sludge. Neutral spent sulphite liquor was partially converted to methane (40% of chemical oxygen demand) under these conditions.Molybdate (20 mM) inhibited methanation of both synthetic waste-water and neutral spent sulphite liquor. Acetate accumulated in glucose plus molybdate media. Molybdate had a direct inhibitory effect on enriched acetoclastic methane-producing bacteria. Molybdate was bacteriocidic to sulphate-reducing bacteria and bacteriostatic to methane-producing bacteria.     

The effect of calcium and sodium hydroxides and of sodium acrylate on the fermentation and digestibility in vitro of ensiled whole-crop wheat and barley harvested at different stages of maturity

Whole-crop wheat and barley were each harvested at the soft-, medium- and hard-dough stages of grain development. Material from each harvest was ensiled in polythene bag silos without additive or after the addition of calcium hydroxide, sodium hydroxide or sodium acrylate at 50, 50 and 12.5 g kg_?1 of the crop dry matter (DM), respectively. All silages were opened after 60 days.With advancing maturity there was an increase in the content of DM, starch and insoluble-nitrogen, but a reduction in water-soluble carbohydrates (WSC) and ash.When crops were ensiled without additives, only medium-dough barley fermented to give butyric acid. However, the addition of calcium hydroxide to crops of low DM (soft-dough) and medium DM (medium-dough) promoted the activity of clostridial bacteria giving rise to the production of butyric acid, but this did not occur with crops of high DM (hard-dough). Sodium hydroxide gave rise to butyric acid only at low DM, and to restricted fermentation at high DM content. Sodium acrylate restricted fermentation and prevented butyric acid production in all silages.Ensiling led to an average reduction of 5 percentage units in the digestible organic matter (DOM) of the control silages compared to that of the crops. Addition of calcium hydroxide and sodium acrylate gave values similar to the control silages. Only sodium hydroxide consistently increased DOM, the effect becoming more marked as the crops matured. The increases over the control silages were 10, 18 and 26 units for wheat and 15, 21 and 20 units for barley at low, medium and high DM, respectively.     

Biological efficiency and nutritional value of Pleurotus ostreatus cultivated on spent beer grain

Unpretreated spent beer grains were successfully used as a basic substrate material for the cultivation of Pleurotus ostreatus. The effects of spent grain types, additives, substrate moisture content, and substrate packing density on the yield and nutrition of fruit bodies were investigated. The cultivation results showed that few fruit bodies were formed on spent grain alone; however, a significantly high biological efficiency (19.1%) was obtained with the addition of wheat bran to (45%). The chemical analysis of fruit bodies indicated that P. ostreatus cultivated on spent grain substrate had a higher nutritional value than those grown on other reported types of substrates. The total amino acid content in the fruit bodies was 347.5 mg/g dry matter, and the crude protein content was as high as 53.3% on a dry weight basis. It was also found that the cultivation of P. ostreatus increased the crude protein content, while it decreased the ratio of lignin to cellulose, of the spent grain substrate.     

Production of polyunsaturated fatty acids by Pythium ultimum in solid-state cultivation

The oleaginous fungus Pythium ultimum was cultivated on various solid substrates in order to achieve fungal oil enriched in the polyunsaturated fatty acids arachidonic acid and eicosapentaenoic acid. Cultivation parameters, such as incubation temperature and time, substrate composition, and moisture were optimized, so as to obtain as much as 3.5 mg eicosapentaenoic acid and 2.6 mg arachidonic acid/g of wet substrate consisting of 28.5% pearled barley, 5.75% spent malt grains, 5.75% linseed oil, and 60% nutrient solution.     

Programmed cell death in cereal aleurone

Progress in understanding programmed cell death (PCD) in the cereal aleurone is described. Cereal aleurone cells are specialized endosperm cells that function to synthesize and secrete hydrolytic enzymes that break down reserves in the starchy endosperm. Unlike the cells of the starchy endosperm, aleurone cells are viable in mature grain but undergo PCD when germination is triggered or when isolated aleurone layers or protoplasts are incubated in gibberellic acid (GA). Abscisic acid (ABA) slows down the process of aleurone cell death and isolated aleurone protoplasts can be kept alive in media containing ABA for up to 6 months. Cell death in barley aleurone occurs only after cells become highly vacuolated and is manifested in an abrupt loss of plasma membrane integrity. Aleurone cell death does not follow the apoptotic pathway found in many animal cells. The hallmarks of apoptosis, including internucleosomal DNA cleavage, plasma membrane and nuclear blebbing and formation of apoptotic bodies, are not observed in dying aleurone cells. PCD in barley aleurone cells is accompanied by the accumulation of a spectrum of nuclease and protease activities and the loss of organelles as a result of cellular autolysis.     

Interspecies interaction based on transfer of a thioredoxin-like compound in anaerobic chitin-degrading mixed cultures

Abstract Fermentation of chitin by mixed cultures of the chitinolytic Clostridium sp. strain 9.1 and various non-chitinolytic bacteria proceeded up to eight times faster than in pure cultures. The addition of spent media of such mixed cultures also resulted in a marked stimulation of chitinolysis in pure cultures of strain 9.1. Pure cultures fermented chitin much faster if supplemented with either spent media or cell-free extracts of the non-chitinolytic bacteria. The compound responsible for this stimulation was thermostable (10 min at 85° C) and could not be removed by passage over Sephadex G-25, indicating a molecular weight of more than 1500. The heat stable enzyme thioredoxin (from Saccharomyces cerevisiae ) was shown to stimulate the chitin fermentation in a similar manner. Alkylation of this enzyme reduced its stimulatory action significantly indicating its (di)thiol: disulfide interchanging activity.
It is hypothesized that essential sulfhydryl groups in the chitinolytic system of strain 9.1 are reduced by thioredoxin and/or similar thiol: disulfide transhydrogenases present in the cell-free extracts and spent media, resulting in an acceleration of chitin hydrolysis and fermentation. This stimulation may thus be the result of a new type of interspecies interaction in anaerobic mixed cultures.     

Distinct developmental defense activations in barley embryos identified by transcriptome profiling

Proper embryo development is crucial for normal growth and development of barley. Numerous related aspects of this process--for example how the embryo establishes and sustains disease resistance for extended periods during dormancy--remain largely unknown. Here we report the results of microarray analyses of >22,000 genes, which together with measurements of jasmonic acid and salicylic acid during embryo development provide new information on the initiation in the developing barley embryo of at least two distinct types of developmental defense activation (DDA). Early DDA is characterized by the up-regulation of a specific set of genes around 20 days after flowering, including co-regulation of those for encoding 9-lipoxygenase and several oxylipin-generating enzymes, possibly leading to the formation of alpha-ketols. The same developmental phase includes an up-regulation of several defense genes, and indications of co-regulation of those for enzymes involved in the generation of phenylpropanoid phytoalexins. Late DDA is initiated prior to grain desiccation, around 37 days after flowering, with up-regulation of several genes encoding proteins with roles in antioxidant responses as well as a simultaneous up-regulation of several PR genes is notable. Throughout barley embryo development, there are no indications of an increased biosynthesis of either jasmonic acid or salicylic acid. Collectively, the results help explain how the proposed DDA enables protection of the developing barley embryo and grain for purposes of disease resistance.     

Salicylic Acid-Mediated Regulation of Morpho-Physiological and Yield Attributes of Wheat and Barley Plants in Deferring Salinity Stress

Salinity has been observed to be a global problem that impede the physiological characteristics of plants. Salicylic acid (SA) as a phytohormone play multifaceted role in plants in terms of development as well as stress management. The current study was conducted to evaluate the effect of salinity and salicylic acid on the performance of wheat and barley plants under field experimentation followed by on-farm study to validate the results. This research was firstly conducted in a 4-year research barley field (2012–2013 and 2013–2014) and wheat (2014–2015 and 2015–2016) and subsequently in an on-farm research in four places (2017–2018). Results depicted that salinity decreased plant yield components and altered ion concentrations (Na⁺/K⁺) causing reduced grain and biological yield. However, SA foliar application induced yield components, especially grain number of plants in both years in non-saline and saline conditions. Exogenously SA application not only led to higher grain yield of barley and wheat but also significantly improved their salt tolerance. Our findings revealed that optimum SA concentrations for achieving highest barley yield were 0.85 and 0.78 mM under saline and non-saline conditions, respectively, while on-farm scale studies observed that foliar application of SA increased grain and biological yield of wheat in Ardakan, Ashkzar (saline soil and water) and Mehrabad (non-saline field) regions. There was no significant effect in Tijerd, a completely non-saline field. The grain yields were higher in SA-treated Ardakan, Ashkzar, and Mehrabad plants in field by 19, 16, and 15%, respectively. Based on present detailed studies, it was concluded that SA improved salinity tolerance and increased crop yield. So, optimum concentration (1.0–1.5 mM) with proper time application (double ridges), SA increased wheat and barley yields up to 20%. Therefore, SA priming could be used as a potent strategy to cope up salinity stress from plants.