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Effect of monensin on fermentation of hay and wheat bran investigated by the Rumen Simulation Technique (Rusitec). 2. End-products of fermentation and protein synthesis.

An experiment was performed with Rumen Simulation Technique (Rusitec) in which the fermentation of mixed ration of hay (12.8 g/d) and wheat bran (3.2 g/d) was compared with the fermentation of the same diet supplemented with 2.5 and 10 mg monensin. The duration of the experiment was 12 days. During the first six days the fermentation conditions in Rusitec were stabilised. The end products of fermentation and indices of protein synthesis were determined. The energy efficiency of volatile fatty acids (VFA), proportion of fermented hexose energy on VFA energy and on energy of bacterial matter were increased in the presence of monensin. The proportion of fermented hexose energy in methane energy was decreased. Utilization of glucose and production of adenosine triphosphate were not affected. The balance of metabolic H2 was reduced and this was manifested in decreasing production, utilization and recovery of metabolic H2. Microbial efficiency expressed per mol ATP (g/mol) was increased by monensin from 7.8 to 12.6. The indices of protein synthesis--protein conversion ratio, degradability of protein and microbial protein synthesis efficiency were increased and non protein utilization ratio decreased in the presence of monensin.

Animal Feed↗

[Fermentation process during the ensiling of green forage low in nitrate. 1. Fermentation process in untreated green forage].

The fermentation process of ensiling green forage, low in nitrate, was proved in 8 experiments with ryegrass, ochardgrass and grass-legumes mixture. It was demonstrated that also in easily fermentable green forage the absence of nitrate promote the occurrence of butyric acid simultaneously with the lactic acid fermentation even at an early stage of fermentation. It is supposed that the lack of nitrate as a natural inhibitor of clostridia leads to early formation of butyric acid. The evolution of clostridia in the beginning of the fermentation process was more rapid than expected. It seems that easily soluble carbohydrates are the substrates of butyric acid formation. That is why in green forage, low on nitrate, clostridia act as food competitors for lactic acid bacteria. Amino acids were not degraded in the early stage of fermentation. In spite of the high content of butyric acid the amount of ammonia was low. Apart from n-butyric acid volatile fatty acids > or = C4 did not occur. The lactic acid formation reached only a restricted extent, also in green forage with a high content of sucrose. It was assumed that there are differences in ensilability of carbohydrate fractions between forage plants.

Ammonia↗

Isolation and characterization of lactic acid bacteria from suan-tsai (fermented mustard), a traditional fermented food in Taiwan.

AIMS: To isolate, characterize, and identify lactic acid bacteria (LAB) in suan-tsai (fermented mustard), a traditional fermented food in Taiwan. METHODS AND RESULTS: Suan-tsai samples were collected at five time points from a fixed fermenting bucket. Fifty cultures were isolated from suan-tsai samples, and isolates were divided into classes by phenotype and then into groups by restriction-fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified two different bacterial groups (A and B), and the results showed that Pediococcus pentosaceus was the most abundant LAB during the initial fermentation time. However, the more NaCl-tolerant species Tetragenococcus halophilus took the place of P. pentosaceus and became the most abundant LAB later. All isolates were grown in de Man, Rogosa, and Sharpe (MRS) broth containing 6% NaCl, but T. halophilus could grow only in MRS broth containing 10% NaCl. CONCLUSIONS: These results suggest that the LAB P. pentosaceus and T. halophilus play roles in the fermentation of suan-tsai. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing the distribution and varieties of LAB that exist in the suan-tsai fermentation process.

Colony Count, Microbial↗

Isolation and characterization of acetic acid-tolerant galactose-fermenting strains of Saccharomyces cerevisiae from a spent sulfite liquor fermentation plant.

From a continuous spent sulfite liquor fermentation plant, two species of yeast were isolated, Saccharomyces cerevisiae and Pichia membranaefaciens. One of the isolates of S. cerevisiae, no. 3, was heavily flocculating and produced a higher ethanol yield from spent sulfite liquor than did commercial baker's yeast. The greatest difference between isolate 3 and baker's yeast was that of galactose fermentation, even when galactose utilization was induced, i.e., when they were grown in the presence of galactose, prior to fermentation. Without acetic acid present, both baker's yeast and isolate 3 fermented glucose and galactose sequentially. Galactose fermentation with baker's yeast was strongly inhibited by acetic acid at pH values below 6. Isolate 3 fermented galactose, glucose, and mannose without catabolite repression in the presence of acetic acid, even at pH 4.5. The xylose reductase (EC 1.1.1.21) and xylitol dehydrogenase (EC 1.1.1.9) activities were determined in some of the isolates as well as in two strains of S. cerevisiae (ATCC 24860 and baker's yeast) and Pichia stipitis CBS 6054. The S. cerevisiae strains manifested xylose reductase activity that was 2 orders of magnitude less than the corresponding P. stipitis value of 890 nmol/min/mg of protein. The xylose dehydrogenase activity was 1 order of magnitude less than the corresponding activity of P. stipitis (330 nmol/min/mg of protein).

Acetates↗

Potential rates of fermentation in digesta from the gastrointestinal tract of pigs: effect of feeding fermented liquid feed.

Microbial catabolic capacity in digesta from the gastrointestinal tract of pigs fed either dry feed or fermented liquid feed (FLF) was determined with the PhenePlate multisubstrate system. The in vitro technique was modified to analyze the kinetics of substrate catabolism mediated by the standing stock of enzymes (potential rates of fermentation), allowing a quantitative evaluation of the dietary effect on the catabolic capacity of the microbiota. In total, the potential rates of fermentation were significantly reduced in digesta from the large intestine (cecum, P < 0.1; colon, P < 0.01; and rectum, P < 0.0001) of pigs fed FLF compared to pigs fed dry feed. No effect of diet was observed in the stomach (P = 0.71) or the distal part of the small intestine (P = 0.97). The highest rates of fermentation and the most significant effect of diet were observed for readily fermentable carbohydrates like maltose, sucrose, and lactose. Feeding FLF to pigs also led to a reduction in the large intestine of the total counts of anaerobic bacteria in general and lactic acid bacteria specifically, as well as of microbial activity, as determined by the concentration of ATP and short-chain fatty acids. The low-molecular-weight carbohydrates were fermented mainly to lactic acid in the FLF before being fed to the animals. This may have limited microbial nutrient availability in the digesta reaching the large intestine of pigs fed FLF and may have caused the observed reduction in activity and density of the cecal and colonic microbial population. On the other hand, feeding FLF to pigs reduced the viable counts of coliform bacteria (indicator of Escherichia coli and Salmonella spp.) most profoundly in the stomach and the distal part of the small intestine, probably due to the bactericidal effect of lactic acid and low pH. The results presented clearly demonstrate that feeding FLF to pigs had a great impact on the indigenous microbiota, as reflected in bacterial numbers, short-chain fatty acid concentration, and substrate utilization. However, completely different mechanisms may be involved in the proximal and the distal parts of the gastrointestinal tract. The present study illustrates the utility of the PhenePlate system for quantifying the catabolic capacity of the indigenous gastrointestinal tract microbiota.

Animal Feed↗

Effects of Aspergillus oryzae fermentation extract on in vitro equine cecal fermentation.

The objective of this study was to examine the effects of Aspergillus oryzae fermentation extract on the in vitro equine cecal fermentation of soluble starch, amino acids/peptides, coastal bermudagrass hay, and alfalfa hay. Cecal contents were obtained from a cecally fistulated Quarter Horse gelding fed coastal bermudagrass and grain (70:30) either unadapted or adapted to dietary A. oryzae supplementation (2 g/d). Mixed cecal microorganisms were incubated in anaerobic media for either 24 h (soluble starch, amino acids) or 48 h (bermudagrass hay, alfalfa hay). A. oryzae was added to the incubation bottles (n = 4) at concentrations of 0, .07, or .7 g/L. Fermentation of soluble starch in the presence of .7 g/L of A. oryzae resulted in increased concentrations of acetate, propionate, NH3, and L-lactate and decreased final pH. Addition of .7 g/L of A. oryzae to amino acid/peptide fermentations decreased final pH and increased concentrations of H2, acetate, propionate, butyrate, and total VFA. When alfalfa hay or bermudagrass hay was fermented with .7 g/L of A. oryzae, CH4, IVDMD, and digestion of NDF and ADF decreased. When adapted mixed cecal microorganisms were used, .7 g/L of A. oryzae did not inhibit methanogenesis or fiber digestion and L-lactate concentrations were not increased with soluble starch as the substrate. These results suggest that cecal microorganisms exposed to A. oryzae via the diet may adapt to the product. Incorporation of A. oryzae into in vitro incubations at concentrations similar to current recommended usage levels resulted in little change in final pH and fermentation products.

Animal Feed↗

Effect of inclusion of fermentable carbohydrates in the diet on fermentation end-product profile in feces of weanling piglets.

An in vivo experiment was conducted to monitor the changes in fermentation end products in the feces of weaning piglets due to the inclusion of selected fermentable carbohydrates in the diet. The experiment involved 3 groups of 16 piglets each. Specially raised piglets (neither antibiotics nor creep feeding) were weaned abruptly at 4 wk of age. The piglets were offered 1 of 2 dietary treatments [a control diet (CON), or a fermentable carbohydrate-enriched diet (CHO)] and were subjected to 1 of the 2 fasting treatments (fasting for 2 d at the beginning of the experimental period or nonfasting). Fecal samples were collected per rectum every day during the experimental period. Piglets were slaughtered at the end of the 10-d experimental period, and digesta samples were collected from different parts of the gastrointestinal tract (GIT): the first half of the small intestine, the second half of the small intestine, the cecum, and colon. The DM, VFA profile, and ammonia concentrations were analyzed from the fecal and digesta samples. Daily feed intake was also recorded. There was no difference in concentrations of VFA in feces between the treatment groups. Ammonia concentration was lower (P < 0.05) in piglets fed the CHO diet compared with those fed the CON diet in both feces and digesta from different parts of GIT. Fasting had no effect on fermentation end products in feces. This study demonstrated that the inclusion of fermentable carbohydrates in weanling diets reduces protein fermentation along the GIT and also reduced the fecal concentration of ammonia.

Acetic Acid↗

Influence of the amount of dietary fiber on the available energy from hindgut fermentation in growing pigs: use of cannulated pigs and in vitro fermentation.

Available energy from hindgut fermentation to pigs fed various amounts of dietary fiber was investigated using an in vivo-in vitro methodology. Six growing pigs fitted with a simple T-shaped cannula at the terminal ileum, and following a Latin-square design, were fed 3 diets differing in the content of non-starch polysaccharides (NSP): a low fiber diet (LFD, 77 g/kg of DM), a standard fiber diet (SFD, 160 g/kg of DM), and a high fiber diet (HFD, 240 g/kg of DM). After adaptation to the diet for 10 d, samples from feces and ileum were collected and analyzed for DM, energy, NSP, and chromic oxide; feces were also analyzed for short chain fatty acids (SCFA). Freeze-dried ileal samples (10 g/L) were fermented in vitro in a fecal slurry consisting of an anaerobic mineral salt medium and feces (50 g/L) from cannulated pigs fed the same diets. Available energy was calculated from the amount of SCFA produced in vitro after 48 h of incubation. Nonstarch polysaccharide content in the fermented material was measured to assess the in vitro degradation of this fraction. Increasing dietary NSP from 77 to 240 g/kg of feed DM increased (P < 0.001) ileal flow from 199 to 468 g/kg of feed, leading to a reduction in the energy digested at the terminal ileum, from 15 to 11 MJ/kg of feed DM and an increment in energy digested in the hindgut, from 1.6 to 3.5 MJ/kg of feed DM. Total in vitro production of SCFA/kg of feed DM was dependent on the amount of ileal substrate available for fermentation; that is, increased concentrations of NSP in the diet led to an increase in the SCFA that may be available to the animal (P < 0.001). The molar ratio of SCFA produced in vitro was affected by diet; the high fiber diet showed the greatest (P = 0.004) proportion of acetic acid, and the low fiber diet showed a tendency (P = 0.081) to an increased butyric acid proportion compared with the other 2 diets. Net disappearance of NSP during fermentation in vivo and in vitro were compared and showed a close relationship (P < 0.001, slope = 0.906, r = 0.960). In our experimental conditions, available energy as SCFA to the animal from hindgut fermentation increased with the concentration of dietary NSP (P < 0.001) and provided between 7.1 and 17.6% of the total available energy.

Animal Feed↗

Effects of pearl millet (Pennisetum typhoides), and fermented and processed fermented millet on Nubian goats.

The effects of Pearl millet, fermented millet, or processed fermented millet were investigated in 15 goats assigned as untreated controls, Pearl millet-fed (166.6g/animal/d), fermented millet-fed (166.6 g/animal/d or 83.3g/animal/d), and processed fermented millet-fed (166.6g/ animal/d). Pearl millet caused goiter within 72-108 d, and changes in thyroid follicles were correlated with clinical manifestations and alterations in serum and tissue iodine and selenium concentrations. Neither fermented nor processed fermented millet produced goiter, but their toxicity was characterized by ruminal bloat, dyspnea, diarrhea, ataxia, and enterohepatonephrotoxicity with death within 5-15 d. Organ lesions correlated with changes in serum enzymes and other serum constituents, hematology, and tissue and serum iodine and selenium concentrations.

Animals↗

Competitive growth experiments with related pairs of tartrate-fermenting and tartrate-non-fermenting strains of Salmonella typhimurium: relevance to biotyping studies.

For each of the isomers of tartaric acid, meso- or d- or l-, a pair of strains of Salmonella typhimurium was obtained, the one, a naturally occurring, non-fermenting strain and the other a spontaneous, tartrate-fermenting mutant derived from it. For each of the pairs of strains, competitive mixed cultures were grown from inocula of the tartrate-fermenting and tartrate-non-fermenting strains in peptone medium without or with the appropriate tartrate isomer. In each experiment, small numbers of tartrate-fermenting bacteria outgrew small or large numbers of tartrate-non-fermenting bacteria in 24 hours in tartrate-containing but not in tartrate-free peptone medium. The results of the experiments are discussed with reference to the choice of the definitive time of reading for tartrate-utilisation tests in the biotyping of S. typhimurium.

Animals↗

Comparative fermentation behaviour and chemical characteristics of Saccharomyces and Zymomonas fermented culled apple juice.

Ethanol production from culled apple juice showed that fermentability of the juice could be enhanced by addition of DAHP or ammonium sulphate in Saccharomyces and DAHP in Zymomonas fermentation. Addition of trace elements inhibited both the fermentations and ethanol, consequently. With respect to by-products of fermentation, no clear advantage of Zymomnas fermentation of culled apple juice could be observed. Differences in physico-chemical characteristics of the fermented apple juice were also noted. Saccharomyces cerevisiae proved to be better than Zymomonas in most of the parameters and is preferrable from handling and spoilage point of view.

Beverages↗

[Effect of products of thermophilous methane fermentation on the fermentation of fruit must by Saccharomyces vini].

Experiments were carried out to study the effect of extracts from products of thermophilous methane fermentation at a dose of 0.7+2.0 ml/100 ml on the proliferation and fermentation activity of yeast Saccharomyces vini of the Yablochnaya-7 and Vishnevaya-33 race during their cultivation in the Hansen medium and in the apple and cranberry must with a normal and elevated content of sugar and acid. In some experiments the must was enriched in (NH4)2HPO4 at a dose of 0.3 g/l. Additions of small amounts of products of thermophilous methane fermentation accelerated fermentation of fruit musts with a normal sugar content and to a greater extent musts with an increased sugar content (27%). In the must enriched in (NH4)2HPO4 an almost complete (over 98%) fermentation of sugar developed for 27 days. In the must with an increased acidity (due to citric acid added to bring titrable acidity to 25 g/l) additions of the preparation also accerlerated the begining of the fermentation and increased its intensity.

Fruit↗

Pattern analysis techniques to process fermentation curves: application to discrimination of enological alcoholic fermentations.

In fermentation processes, kinetic curves are generally aimed at control purposes. However, these curves could also contain information about inherent features of the product (such as origin, quality, etc.). This article presents several pattern analysis techniques used to classify fermentation curves. An application to alcoholic fermentation is presented as an illustration: it aims at retrieving the origin of a must from its fermentation curve. The fermentation kinetics of five vineyard musts, harvested over 9 years on the same parcels, were recorded. From these curves two sets of variables were generated: The first (p(1)) gathers all the kinetic curve points. The second (p(2)) contains a restrained number of variables, generated by the expert knowledge of the enologist. The set p(2) was processed by two very different techniques: a linear one (factorial discriminant analysis) and a nonlinear one (artificial neural networks). The set p(1) was processed by a new chemometric technique, the discriminant partial least-squares regression. For all the sets and the techniques used the selection of variables was studied. The interest in the latter is largely demonstrated both by theoretical and practical discussions. The discrimination results (up to 94% of good predictions) enhance the interest of the on-line measurements and their use in such pattern analysis tools.

Algorithms↗

Fermentation behaviour and volatile compound production by agave and grape must yeasts in high sugar Agave tequilana and grape must fermentations.

Few studies have been performed on the characterization of yeasts involved in the production of agave distilled beverages and their individual fermentation properties. In this study, a comparison and evaluation of yeasts of different origins in the tequila and wine industries were carried out for technological traits. Fermentations were carried out in high (300 g l(-1)) and low (30 g l(-1)) sugar concentrations of Agave tequilana juice, in musts obtained from Fiano (white) and Aglianico (red) grapes and in YPD medium (with 270 g l(-1) of glucose added) as a control. Grape yeasts exhibited a reduced performance in high-sugar agave fermentation, while both agave and grape yeasts showed similar fermentation behaviour in grape musts. Production levels of volatile compounds by grape and agave yeasts differed in both fermentations.

Agave↗

[Fermentation process during the ensiling of green forage low in nitrate. 2. Fermentation process after supplementation of nitrate, nitrite, lactic acid bacteria and formic acid].

The effect of adding nitrate and nitrite (0.05% and 0.01% N in DM) of two inoculants or formic acid on the ensiling of orchardgrass and a grass-legumes-mixture, both low in nitrate, was proved in two experiments during ensiling starting in an early stage of fermentation silages without additives contained butyric acid, with increasing amounts up to 180 day of storage period. Silages with added nitrate or nitrite (0.1% N in DM) contained no butyric acid despite of a delay in the formation of lactic acid Nitrite was more effective as nitrate (both 0.05% N in DM) to suppress butyric acid fermentation. However, both additives, nitrite and nitrate, increased the lactic acid fermentation. Interpreting this facts it was hypothesized that the clostridia was inhibited by nitrate and nitrite. Addition of inoculants improved the formation of lactic acid, and decreased pH-value in comparison to control silages. The formation of butyric acid was limited, but not suppressed. The effect of inoculants differed in the intensity of fermentation and the formation of by-products of lactic acid fermentation. The effect of formic acid was not sure.

Animal Nutritional Physiological Phenomena↗

Starch and fiber fractions in selected food and feed ingredients affect their small intestinal digestibility and fermentability and their large bowel fermentability in vitro in a canine model.

The digestion of legumes, cereal grains, cereal and potato flours and grain-based foods in dogs was studied using two in vitro model systems. The first simulated the stomach and small intestine through the additions of acid and enzymes and large bowel fermentation through use of fecal inocula from dogs, and the second simulated small intestinal fermentation using canine ileal chyme as the bacterial source. All substrates were analyzed for total dietary fiber (TDF) including insoluble and soluble components, and starch fractions: rapidly digestible starch, slowly digestible starch, resistant starch (RS) and total starch. Legumes had high TDF and RS concentrations (mean 36.5 and 24.7%, respectively), resulting in lower ileal digestible starch and total digestible starch concentrations (mean 21 and 31%, respectively). Seventy-four percent of the TS in the cereal grains group was rapidly digestible starch plus slowly digestible starch compared with the flour group, where the corresponding value was 95%. This related to the processing of cereals to flours, in which TDF and RS concentrations were reduced markedly. This increased ileal digestible starch concentrations in the flour group (65%) versus the cereal grains group (60%). Ileal digestion of starch in grain-based food products like macaroni and spaghetti was high (96 and 92%, expressed as a percentage of TS, respectively). Fermentation of substrates with ileal microflora was influenced by substrate chemical composition, with the flour group exhibiting the highest organic matter disappearance values. The legume group had a high total short-chain fatty acid concentration (7.8 mmol/g organic matter fermented), perhaps as a result of fermentation of TDF as well as starch components. A database such as this one provides information about utilization of foods and feeds in the dog and potentially in humans.

Animals↗

Preservation of hatchery waste by lactic acid fermentation. 1. Laboratory scale fermentation.

In Experiment 1, two types of hatchery wastes, including cockerel chicks (CC), and shell waste (SW) blended with CC (60:40 CC:SW), were ground and mixed with a by-product carbohydrate (5, 10, and 15% wt/wt) with or without a bacterial culture and fermented for a period of 21 d. Cockerel chicks fermented with 15% carbohydrate and the culture had a pleasant smell and little H2S production. Elevated H2S concentrations were recorded for CC:SW samples at all carbohydrate levels when the culture was not added. No NH3 was detected from any treatments during fermentation. The addition of culture to the CC and CC:SW by-products resulted in pH values lower (P < 0.05) than those without culture on Day 21, and the 15% carbohydrate treatment significantly reduced pH beyond the 5% carbohydrate. Final proximate composition of CC and CC:SW samples with culture were not significantly different from those without culture added. In Experiment 2, carbohydrate was added at 10.0, 13.3, 16.7, and 20.0% wt/wt to CC and CC:SW in the presence of the bacterial culture. Shell waste alone was fermented with 15, 20, and 25% carbohydrate and the culture. Moisture level in this experiment was adjusted to approximately 70% for all treatments. The lowest pH for the CC and CC:SW treatments was observed at the 16.7% carbohydrate level. Shell waste pH was better maintained at the 20 and 25% carbohydrate levels. After fermentation for 21 d CC, CC:SW and SW treatments from Experiments 1 and 2 contained negligible Escherichia coli, and no Salmonella were detected.

Analysis of Variance↗