The effect of decanoic,dodecanoic and tetradecanoic acids on silage fermentation

The ability of decanoic (C₁₀), dodecanoic (C₁₂) and tetradecanoic (C₁₄) acids to restrict the fermentation of ryegrass and lucerne was investigated with laboratory silos. With ryegrass, addition of the C₁₂ acid alone at 1 kg t^?1 of fresh crop had no effect on extent or pattern of fermentation compared to that of the untreated crop, though at 2 kg t^?1 the acid appeared to reduce the content of butyric acid but not ammonia-N in the silage. Formic acid, added alone or with C₁₀, C₁₂ and C₁₄ acids restricted extent of fermentation and butyric acid production, but in this experiment addition of these acids had relatively little effect on the content of ammonia-N in the silages. With lucerne, extent of fermentation was similar for all treatments, though the untreated silage and those which received C₁₂ or formic acid alone at 1 kg t^?1 or 1 kg t^?1 of C₁₄ acid with 1 kg t^?1 of formic acid contained butyric acid in excess of 3% of the dry matter (DM). It is concluded that acids in the range C₁₂-C₁₄ are not effective in restricting silage fermentation and that their effectiveness is not increased by reduction in pH produced by the addition of formic acid. Microbiological assessments largely confirmed these findings.     

The use of fatty acids as grass silage additives

The effects of a range of fatty acids (C₁-C₁₈) on silage fermentation in laboratory silos have been examined. When acids were applied to ryegrass in equimolar concentrations (50 mM) in the C₁-C₆ range, only the C₆ acid (hexanoic) had a marked effect in inhibiting lactate production. When hexanoic acid was examined in a second experiment over the concentration range 9 mM to 50 mM, concentrations below 50 mM did not prevent acid production, and at the lower levels production of lactate was stimulated. When higher fatty acids hexadecanoic (16:0), octadecanoic (18:0) and octadecenoic (18:1) were added at similar concentrations (8 to 9 mM) and octadecadienoic (18:2) added at concentrations ranging from 8 mM to 29 mM to ryegrass samples, lactic acid fermentation was not inhibited.     

Determination of ethanol,volatile fatty acids,lactic and succinic acids in fermentation liquids by gas chromatography

A rapid and simple quantitative method was developed to determine, by gas chromatography, the concentrations in fermentation liquids of ethanol, the C₂-C₆ volatile fatty acids, and lactic and succinic acids. Aqueous samples were acidified with 250μlml^?1 metaphosphoric acid (5:1 ratio), centrifuged, and injected directly on to a column containing a porous aromatic polymer (Chromosorb 101) maintained at 200°C in a gas chromatograph fitted with a flame ionisation detector. It was unnecessary to purify samples further before injection, although distillation and ion-exchange methods were examined. Derivatisation of lactic and succinic acids before injection was not necessary, but the lowest level of detection of these two relatively non-volatile acids was about four times greater than that for the volatile fatty acids. The method described was suitable for the analysis of rumen fluid, methane digester fluid, silage extracts and other anaerobic fermentation fluids. The relative retention times are given for 23 organic acids and six other fermentation end-products.     

Studies of the effects on the fermentation pattern in the rumen of the addition of various sources and levels of the lower volatile fatty acids

The short term effects on rumen fermentation of the direct addition to the rumen of 1·5 M and 3 M of acetate, propionate and butyrate, given as the Ca²⁺, Na⁺, and NH⁺₄ salts and as the free acid were measured in a mature cow. Longer term effects of the oral addition of I·5 M of calcium and ammonium acetates were also measured in growing heifers. For all volatile fatty acids (VFA) each rate of addition consistently increased the level of VFA in rumen liquor, but these fell gradually over about 4 h. Total volatile fatty acids (TVFA) were lower when free acids were given than when their salts were used. There was little effect on rumen pH. Calcium acetate was unpalatable when offered orally, and it tended to depress TVFA levels. Mean effects of acetate salts given orally on molar percentage of acetate in rumen liquor were smaller than those found by direct addition.     

Determination of volatile fatty acids C1–C6, and lactic acid in silage juice

A method for the determination of volatile fatty acids, C₁–C₆, and lactic acid in silage juice is described. The procedure is simple and capable of application to large numbers of samples. It involves separation of the free acids by ion exchange, formation of their benzyl esters by the addition of benzyl bromide to a solution of the tetrabutylammonium salts in acetone and ultimate determination by gas liquid chromatography.     

Rumen volatile fatty acids and milk composition from cows fed hay, haylage, or urea-treated corn silage.

Alfalfa-brome hay, haylage, .5% urea-treated corn silage, or .5% urea plus 1% dried whey-treated corn silage was fed as the only forage to one of four groups of 10 lactating cows per group for a lactation trial of 10 wk. Rumen samples were collected via stomach tube 3 to 4 h after the morning feeding. The pH of the rumen samples from cows fed hay was higher than for cows fed haylage, urea-treated corn silage, and urea-whey corn silage, 6.69 versus 6.36, 6.40, and 6.50. Total volatile fatty acids and propionate were highest from cows fed urea-whey corn silage and were higher on all three fermented forages than cows fed hay. Acetate/propionate ratio was highest from cows fed hay and lowest from cows fed corn silages. Butyrate was highest from cows fed haylage or hay. Milk protein composition was not affected by ration although nonprotein nitrogen of milk was highest from cows fed the urea-treated corn silages. Oleic acid and total unsaturated fatty acids were lowest in milk fat from cows fed hay while palmitic acid was highest from cows fed hay and haylage. These results suggest that type of forage fed may cause small changes in rumen fermentation and in milk composition. The importance of these changes is unknown but may affect properties of dairy products produced from this milk.     

Inoculant effects on alfalfa silage: in vitro gas and volatile fatty acid production

Alfalfa silages from 2 similar trials were analyzed for in vitro ruminal gas production. In both trials, there were 15 treatments: alfalfa treated at ensiling with 1 of 14 lactic acid bacterial inoculants or untreated alfalfa. First-cut (477 g of dry matter/kg) and second-cut (393 g of dry matter/kg) alfalfa were ensiled in glass jars for a minimum of 35 d at room temperature (∼22°C). At opening, a portion of each silage was wet-ground with a mixer. Each silage was then assessed for in vitro ruminal gas production in 3 replicate runs with the wet-ground silage, 1 on the fresh silage and 2 on frozen and thawed silage. In vitro gas production was measured in 160-mL sealed serum vials incubated at 39°C. One gram of silage was incubated with 17.1 mL of nutrient solution, 0.9 mL of reducing solution, and 12 mL of ruminal inoculum (1:2 vol.vol mixture of rumen fluid and buffer). Gas production was measured manually by using a pressure gauge at 3, 6, 9, 24, 48, and 96 h. At 96 h, the rumen fluid was analyzed for pH and volatile fatty acids. In the 2 trials, the untreated control silage produced either numerically the highest or one of the highest levels of gas production per unit of dry matter incubated. In first-cut silage, 9 of the inoculant treatments at 9 h and 4 treatments at 96 h had reduced gas production compared with the control. In second-cut silage, 10 inoculant treatments at both 9 and 96 h had reduced gas production compared with the control. Furthermore, in first-cut silage, the fraction of total gas production at 3, 6, and 9 h was numerically the highest for the control, and only 4 treatments were not significantly lower than the control at 9 h. In second-cut silage, 2 of 14 inoculated treatments produced faster fractional rates of gas production than the control, but most inoculated treatments had numerically slower fractional rates (4 significant) in the first 9 h. The in vitro fermented wet-ground control silages had one of the highest acetate:propionate ratios in both trials, significantly higher than 12 and 8 of the inoculated treatments in first- and second-cut silage, respectively. The response in acetate:propionate ratio in both cuts was similar, even though the control silage was highest in lactic acid in one trial and lowest in the other. Overall, inoculation of crops at ensiling appears to affect in vitro ruminal fermentation of wet-ground silages, even in the absence of large effects during silage fermentation.     

Generation of volatile fatty acids by axillary bacteria

It is generally accepted that short-chain (C(2)-C(5)) volatile fatty acids (VFAs) are among the causal molecules of axillary malodour. It is also widely acknowledged that malodour generation is attributable to the biotransformation of odourless natural secretions, into volatile odorous products, by axillary bacteria. However, little information is available on the biochemical origins of VFAs on axillary skin. In these studies, assay systems were developed to investigate the generation of VFAs from substrates readily available to the bacteria resident on axillary skin. Propionibacteria and staphylococci were shown to ferment glycerol and lactic acid to the short-chain (C(2)-C(3)) VFAs, acetic and propionic acid. Furthermore, staphylococci are capable of converting branched aliphatic amino acids, such as leucine, to highly odorous short-chain (C(4)-C(5)) methyl-branched VFAs, such as isovaleric acid, which are traditionally associated with the acidic note of axillary malodour. However, in vitro kinetic data indicates that these pathways contribute less to axillary VFA levels, than fatty acid biotransformations by a recently defined sub-group of the Corynebacterium genus, corynebacteria (A). The results of these studies provide new understanding on the biochemical origins of VFA-based axillary malodour which, in turn, should lead to the development of novel deodorant systems.     

Development of normal and branched chain volatile fatty acids during the fermentation process in the manufacture of fish sauce

The formation of volatile fatty acids (VFAs) in fish sauce was investigated. When fish were allowed to spoil prior to salting, very high concentrations of VFAs were produced. The increase was probably caused by microbial action. Addition of salt to spoiled fish seemed to inhibit microbial action, thus suppressing the formation of VFAs. The contents of short and long straight chain VFAs in aerobically fermented sauces were significantly higher than in anaerobically fermented sauces, suggesting that straight chain VFAs developed from fish fats. Certain branched VFAs were derived from the degradation of specific amino acids, such as isobutyric acid from valine and isovaleric acid from leucine. Alanine and isoleucine did not have a clear influence on the production of VFAs. © 2001 Society of Chemical Industry     

Development and origin of the volatile fatty acids in budu

The origin and mechanism of formation of the volatile fatty acids (VFA) present in budu were investigated. The acids did not appear to derive from the breakdown of the fish lipid. When the fish was allowed to spoil, a single bacterial species predominated, and as the fermentation progressed, the appropriate VFA were formed. Using (U^?14C)-protein hydrolysate it was shown that amino acids are the precursors of the n-butanoic and n-pentanoic acid and also contributed to the formation of other acids. The bacterium did not use glucose as a carbon source nor did any particular combination of unlabelled amino acids produce the fatty acids. The metabolic route by which the fatty acids are produced from the amino acids is not known. An experiment that allowed spoilage to occur, prior to salting, in the normal environment involved in the preparation of budu, showed that the VFA were produced.     

The effect of Lactobacillus buchneri and other additives on the fermentation and aerobic stability of barley silage

Whole-plant barley (39.4% dry matter) was treated with various chemical and biological additives to assess their effects on silage fermentation and aerobic stability. Treatments were untreated forage, forage treated with several amounts of Lactobacillus buchneri and enzymes (L. buchneri at 1 x 10(5), 5 x 10(5), and 1 x 10(6) cfu/g of fresh forage), forage treated with an inoculant containing (Lactobacillus plantarum, Pediococcus pentosaceus, Propionibacterium freudenreichii, and enzymes), or forage treated with a buffered propionic acid-based additive (0.2% of fresh weight). Sixty-nine d after ensiling, silages treated with L. buchneri and enzymes had lower pH, but had higher concentrations of acetic and propionic acids and higher concentrations of ethanol when compared with untreated silage. Silage treated with the multistrain inoculant containing L. plantarum had lower pH and higher concentrations of lactic acid, but lower concentrations of ammonia-N, neutral detergent fiber, and acid detergent fiber than did untreated silage. The addition of the buffered propionic acid additive resulted in silage with higher concentrations of lactic and acetic acid compared with untreated silage. Numbers of yeasts in all silages were low at silo opening (less than 3.0 log cfu/g) and were numerically the lowest in silages treated with L. buchneri but only treatment with the intermediate and high level of L. buchneri improved the aerobic stability of silage. Because of the altered fermentation pattern, inoculation with L. buchneri, when applied at equal to or more than 5 x 10(5) cfu/g, and enzymes improved the aerobic stability of barley silage.     

The effect of inoculum level on the formation of higher alcohols,fatty acids and esters in the malt whisky fermentation

The levels of higher alcohols, fatty acids and esters formed in a small-scale whisky fermentation which had been inoculated with different amounts of yeast was investigated. The total level of higher alcohols increased with increasing inoculum levels. However, the relative levels of propanol, isobutanol, amyl alcohols and 2-phenyl ethanol were unaffected. The levels of octanoic, decanoic and dodecanoic acids and their ethyl esters were depressed at inoculum levels above 2 × 10⁷ cells ml^?1. Varying the inoculum levels did not have a consistent effect on the acetate esters of the higher alcohols. However, the highest values were obtained at inoculum levels 4 × 10⁷ cells ml^?1.     

Effects of feeding or infusing ammonium salts of volatile fatty acids on ruminal fermentation, plasma characteristics, and milk production of cows

Eight rumen-fistulated Holstein cows, averaging 77 d postpartum, were used in a replicated 4 X 4 Latin square design with 28-d periods to investigate the effect of ammonium salts of isobutyrate, 2-methylbutyrate, isovalerate, and n-valerate on animal performance and their possible mechanism(s) and site(s) of action. Each cow was fed ad libitum a complete mixed diet of 55% corn silage and 45% concentrate on a dry basis that was supplemented with 1.8 kg of premix daily. Treatments were 1) control, 2) ammonium salts of volatile fatty acids in premix, 3) ammonium salts of volatile fatty acids ruminally infused, or 4) ammonium salts of volatile fatty acids abomasally infused. Mean ruminal fluid pH and concentrations of ammonia and volatile fatty acids for treatment comparisons were not different. Plasma concentrations of isobutyrate, 2-methylbutyrate, and valerate differed among treatments, but there was no significant effect on dry matter intake, milk production, milk composition, or efficiency of feed utilization. Apparent nutrient digestibility; disappearance of dry matter, cellulose, and nitrogen from polyester bags suspended in the rumen; and plasma concentrations of glucose, free fatty acids, and growth hormone also were not significantly affected by treatment.     

Effects of volatile fatty acids on propionate metabolism and gluconeogenesis in caprine hepatocytes

Isolated caprine hepatocytes were incubated with fatty acids of various chain lengths. Short-chain fatty acids effects on rates of gluconeogenesis and oxidation from [2-14C]propionate were determined. Additions of glucose (2.5 mM) had no effect on hepatic [2-14C]propionate metabolism in the presence and absence of amino acids. A complete mixture of amino acids increased label incorporation from [2-14C]propionate into [14C]glucose by 22%. Butyrate inhibited [2-14C]propionate metabolism and increased the apparent Michaelis constant for [2-14C]propionate incorporation into [14C]glucose from 2.4 +/- 1.5 to 5.6 +/- .9 mM. Butyrate's effects on propionate were similar in the presence and absence of L-carnitine (1 mM). Isobutyrate, 2-methylbutyrate, and valerate (1.25 mM) had no effect on [14C]glucose production but decreased 14CO2 production to 57, 61, and 54% of the control [2-14C]propionate (1.25 mM). This inhibition on 14CO2 production was not competitive. Isovalerate had no effect on either [2-14C]propionate incorporation into glucose or CO2. An increase in ratio of [14C]glucose to 14CO2 from [2-14C]propionate demonstrated that short-chain fatty acids other than butyrate do not inhibit gluconeogenesis from propionate. In addition, fatty acids that generate a net synthesis of intracellular oxaloacetate may partition propionate carbons toward gluconeogenic rather than oxidative pathways in goat hepatocytes.     

酸肉发酵中挥发性风味物质的变化及对品质的影响

该文以SPME-GC-MS和模糊数学感官评价研究长时间发酵对酸肉挥发性风味物质的影响及对感官品质的变化。结果显示,不同发酵时段酸肉挥发性风味物质种类和含量处于动态变化中,发酵90 d时挥发性物质总数较高并保持稳定,主要有酯类、醛类和酸类等。结合特征性风味成分变化、剖面分析及主成分分析显示,新鲜猪肉风味主要受醛类影响;发酵30 d时,酸肉呈青草味,主要受酯类和醇类影响;发酵30～90 d,主要受酮类和酸类影响,发酵肉香成为酸肉主体风味,同时有青草味及瓜果清香等味道辅助,构成了酸肉独特的风味特征。发酵150 d左右主要受碳氢化合物、酯类、醇类和其他类影响,呈橡胶、脂肪等刺激性气味。综上,酸肉以发酵方式保藏时间不宜超过90 d,以发酵60 d左右风味品质较好。该文从风味和感官品质角度分析为传统酸肉以发酵的方式保藏提供有关风味化学的研究数据。     

Effects of addition of malic or citric acids on fermentation quality and chemical characteristics of alfalfa silage

We studied the effects on alfalfa preservation and chemical composition of the addition of different levels of malic acid and citric acid at ensiling as well as the utilization efficiency of these 2 organic acids after fermentation. Alfalfa was harvested at early bloom stage. After wilting to a dry matter content of approximately 40%, the alfalfa was chopped into 1- to 2-cm pieces for ensiling. Four levels (0, 0.1, 0.5, and 1% of fresh weight) of malic acid or citric acid were applied to chopped alfalfa at ensiling with 4 replicates for each treatment, and the treated alfalfa forages were ensiled for 60 d in vacuum-sealed polyethylene bags (dimensions: 200 mm × 300 mm) packed with 200 to 230 g of fresh alfalfa per mini silo and an initial density of 0.534 g/cm³. The application of malic or citric acids at ensiling for 60 d led to lower silage pH than was observed in the control silage (0% of malic or citric acids). Application of the 2 organic acids led to higher lactic acid concentration in alfalfa silage than in the control silage except with the application rate of 1% of fresh weight. Silages treated with both organic acids had lower nonprotein nitrogen concentrations than the control silages, and the nonprotein nitrogen concentrations in ensiled forages decreased with the increase in malic or citric acid application rates. The application of the 2 organic acid additives led to lower saturated fatty acid proportions and higher polyunsaturated fatty acid proportions in ensiled alfalfa than in the control silage. The amount of malic and citric acids degraded during ensiling of alfalfa was 1.45 and 0.63 g, respectively. At the application rate of 0.5% of fresh weight, residues of malic acid and citric acid in alfalfa silage were 11.1 and 13.6 g/kg of dry matter. These results indicate that including malic or citric acids at the ensiling of alfalfa effectively improved silage fermentation quality, limited proteolysis, improved fatty acid composition of the ensiled forage, and could provide animals with additional feed additives proven to promote animal performance. However, when the application rate of both organic acids reached 1%, the concentration of lactic acid in silages decreased notably. Additionally, 0.5 and 1% application rates also increased the yeast count in ensiled alfalfa.