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Effect of galactan on selected microbial populations and pH and volatile fatty acids in the ileum of the weanling pig.

Studies were conducted to determine the effect of galactan on the colonization of E. coli and lactobacilli and ileal pH and volatile fatty acid production in the digestive tract of the weanling pig. In each of two replicate trials, eight 21-d-old nursing pigs were cannulated in the terminal ileum. After a 7-d recovery period, the pigs were weaned and randomly assigned to two test diets: 1) a corn-soybean meal-based control diet and 2) a similar diet containing 1% galactan. On d 1 after weaning, all pigs were orally subjected to K88+ E. coli (2 x 10(9) colony forming units). Ileal digesta samples were collected on d 0, 2, 4, 6, 8, and 10 after weaning and assayed for total E. coli, K88+ E. coli, lactobacilli, pH, and VFA. At the end of the trials, the pigs were killed and digesta samples were collected from the stomach, duodenum, cecum, and colon. Assays similar to those performed on the digesta samples collected from ileal cannulas were performed. Pigs fed 1% galactan had lower (P < .10) ileal pH, lower (P < .05) total E. coli on d 6 and 8, and lower (P < .05) K88+ E. coli concentrations in the ileum than pigs fed the control diet. There were no differences in ADG or gain:feed ratio between diets. The VFA concentrations were not different in the ileum between diets. The VFA were higher (P < .10) on d 0 than on any other day of the study. Acetate and isobutyrate concentrations were lower (P < .10) in the cecum in pigs fed 1% galactan.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of volatile fatty acid infusion on development of the rumen epithelium in neonatal sheep.

The purpose of this study was to determine whether the continuous intraruminal infusion of calculated physiological concentrations of volatile fatty acids (VFA) stimulated the metabolic development of the neonatal rumen. Eight 1-wk-old lambs were assigned to one of three treatments: saline infusion (three lambs), VFA infusion (three lambs), or no infusion (two lambs). Rumen catheters were surgically implanted into lambs in the infusion groups. The amount of VFA infused, beginning at 2 wk of age, increased weekly in equal increments of 12.5% of the estimated net energy requirement until, at 6 wk of age, lambs received 50% of their estimated net energy requirement from the infused VFA. All lambs consumed milk replacer for ad libitum intake and had free access to water. The lambs that were infused with VFA tended to have longer rumen papillae. There were no differences in width or number of papillae per square centimeter across treatments. Rumen epithelial cells isolated from lambs that were infused with VFA tended to oxidize less glucose and produce more acetoacetate than did cells from lambs that were infused with saline or from uninfused lambs. beta-Hydroxybutyrate production by isolated rumen epithelial cells and concentrations of blood glucose, acetoacetate, and beta-hydroxybutyrate were not different among the three treatments. Thus, infusion of physiological concentrations of VFA appears to stimulate some aspects of rumen metabolic development.

Acetoacetates↗

Proportions of volatile fatty acids in relation to the chemical composition of feeds based on grass silage.

This experiment was designed to quantify the relationships between feed chemistry and the proportions of rumen volatile fatty acids (VFA) across a wide range of feedstuffs. In an experiment using 11 rumencannulated sheep, 16 test feeds were fed at three different inclusion rates in rations that were based on grass silage. The 17 periods of the experiment were each 14 d long. Eight rumen samples were taken every 24 h on d 13 and 14 of each period from which the mean daily proportions of VFA were derived. The effects of an increase in the proportion of test feed in the total feed on proportions of VFA were significant. The observed proportions of VFA were related to the chemical composition of the total feed by principal component regression. The inclusion of the amount of feed offered and the ratio of test feed to total feed in these regressions did not improve their precision; these terms were not significant. The significant terms in the regressions were crude protein, starch, sugar, and cellulose (calculated by difference). The R2 values achieved for the regressions between acetate, propionate, and butyrate (molar proportions) and feed composition were 77.5, 68.0, and 87.3%, respectively. These regressions provided an apparently robust basis for predicting molar proportions of VFA from feed chemistry in feeds based on grass silage.

Acetates↗

Volatile fatty acids and the inhibition of Escherichia coli growth by rumen fluid.

Concentrations of volatile fatty acids (VFA) normally found in bovine rumen fluid inhibited growth of Escherichia coli in Antibiotic Medium 3. Acetic, propionic, and butyric acids each produced growth inhibition which was markedly pH-dependent. Little inhibition was observed at pH 7.0, and inhibition increased with decreasing pH. A combination of 60 mumoles of acetate, 20 mumoles of propionate, and 15 mumoles of butyrate per ml gave 96, 69, and 2% inhibition at pH 6.0, 6.5, and 7.0, respectively. Rumen fluid (50%) gave 89 and 48% inhibition at pH 6.0 and 6.5, respectively, and growth stimulation (22%) at pH 7.0. Rumen fluid inhibitory activity was heat-stable, was not precipitated by 63% ethyl alcohol, and was lost by dialysis and by treatment with anion-exchange resins but not with cation-exchange resins. These results are consistent with the idea that VFA are the inhibitory substances in rumen fluid. Previous results which indicated that rumen fluid VFA did not inhibit E. coli growth were due to lack of careful control of the final pH of the growth medium. The E. coli strain used does not grow in rumen fluid alone at pH 7.0.

Acetates↗

[Gas-liquid chromatography of non-volatile fatty acid produced by "Yersinia enterocolitica" (author's transl)].

The metabolic products (non-volatile fatty acids) of 24 strains belonging to four Yersinia enterocolitica groups (typical, sucrose negative, rhamnose positive, melibiose and rhamnose positive) were examined by gas liquid chromatography. All strains produced lactic, pyruvic and succinic acids. This technique did not allow any classification for taxonomic purpose. The evolution of lactic and succinic acid production was monitored for 72 h in shaked and non-shaked cultures. The concentration of both acids increased in the first 24 h, then decreased and increased again after 38 h. Shaked culture produced a lesser amount of lactic acid than did non-shaked culture. Only traces of succinic acid were detected in shaked culture. The quantities of metabolic products of the four strains grown in five media were significantly different. These observations suggest that the composition of the medium, the bacterial growth, inoculum size, incubation technique and the incubation time are important factors in metabolic production by aeroanerobic bacteria. Only constant results in the face of such varying factors could be of taxonomic value.

Chromatography, Gas↗

Effect of monensin sodium on the performance and proportions of rumen volatile fatty acids of Friesian bulls.

The effect of including monensin sodium at a dose rate of 40 mg/kg in the diet of Friesian bulls (mean initial liveweight 158 kg) fed a barley-based diet ad libitum was assessed in terms of growth rate, feed intake, feed and energy conversion efficiency and the concentration and molar proportions of ruminal volatile fatty acids over a 157-day period. Mean daily feed dry matter intake for the treated group (5.4 kg) was 24 per cent lower than the control group (7.1 kg). The treated group grew at a significantly slower rate than the control animals although feed conversion efficiency was improved by the treatment. A comparison with the results of other workers suggested that the dose rate used had been too high. There were no significant differences in the molar proportions of volatile fatty acids in rumen fluid as a result of monensin usage. While many workers have shown increases in the molar proportions of propionic acid the fact that the control group in this experiment had a very high value (48 per cent) may explain the lack of response.

Administration, Oral↗

Optimum design and operation of primary sludge fermentation schemes for volatile fatty acids production.

This paper presents a model-knowledge based algorithm for optimising the primary sludge fermentation process design and operation. This is a recently used method to obtain the volatile fatty acids (VFA), needed to improve biological nutrient removal processes, directly from the raw wastewater. The proposed algorithm consists in a heuristic reasoning algorithm based on the expert knowledge of the process. Only effluent VFA and the sludge blanket height (SBH) have to be set as design criteria, and the optimisation algorithm obtains the minimum return sludge and waste sludge flow rates which fulfil those design criteria. A pilot plant fed with municipal raw wastewater was operated in order to obtain experimental results supporting the developed algorithm groundwork. The experimental results indicate that when SBH was increased, higher solids retention time was obtained in the settler and VFA production increased. Higher recirculation flow-rates resulted in higher VFA production too. Finally, the developed algorithm has been tested by simulating different design conditions with very good results. It has been able to find the optimal operation conditions in all cases on which preset design conditions could be achieved. Furthermore, this is a general algorithm that can be applied to any fermentation-elutriation scheme with or without fermentation reactor.

Algorithms↗

Influence of supplemental endoglucanase or xylanase on volatile fatty acid production from ruminant feed by ruminal in vitro cultures.

This study employed two commercial enzyme preparations to examine the effects of endoglucanase, xylanase or their combination on in vitro volatile fatty acid (VFA) production by ruminal microbial populations. Batch ruminal cultures were established with one of various feedstuffs or with a fescue hay-based diet and ruminal fluid from a heifer fed a 40% forage:60% concentrate diet. Addition of xylanase at 135 xylanase units (XU) per ml increased total VFA production from the fescue hay-based diet (44.3 vs. 57.2 mM, p < 0.05) without changing the acetate to propionate (A:P) ratio. Addition of endoglucanase at 2, 3, 4, and 5 carboxymethyl cellulase units (CMCU) per ml increased total VFA production from the fescue hay-based diet on average by 36% (p < 0.05). Addition of 3, 4 and 5 CMCU/ml also decreased (p < 0.05) the A:P ratio. The combined addition of xylanase (135 XU/ml) and endoglucanase (5 CMCU/ml) increased total VFA production from the fescue hay-based diet (40.9 vs. 61.5 mM, p < 0.05) and reduced the A:P ratio (3.4 vs. 1.5, p < 0.05). The effects of endoglucanase and xylanase supplementation on in vitro VFA production varied across the various substrates used. However, endoglucanase supplementation consistently reduced the A:P ratio with all substrates tested. The effects of the enzyme combination were generally greater than either enzyme alone. We conclude that endoglucanase and xylanase activities differ in their ability to affect ruminal VFA production, and endoglucanase but not xylanase, may improve fermentation efficiency by reducing the A:P ratio.

Animal Feed↗

The effect of volatile fatty acids in acidified fermented piggery effluent on shiga-toxigenic and non-toxic resident strains of Escherichia coli.

AIMS: To examine the effects of acidified acidogenically fermented piggery effluent containing Volatile Fatty Acids (VFA) on shiga-toxigenic and resident strains of Escherichia coli (E. coli) as part of the development of a waste treatment process. METHODS AND RESULTS: Four shiga-toxigenic E. coli strains (O157:H7, 091.H-, 0111.H-, and 0123.H-) and four non-toxic resident enzootic strains were all killed by 3 h treatment with fermented piggery effluent liquor (153 mmol l(-1) total VFA) at pH 4.3. The shiga-toxigenic strains showed greater sensitivity after 1 h of treatment. The fermented liquor at pH 6.8 was not inhibitory. CONCLUSIONS: The shiga-toxigenic strains were no more resistant to the toxic effects of VFA than the non-toxic strains tested. SIGNIFICANCE AND IMPACT OF THE STUDY: Shiga-toxigenic strains and resident enzootic non-toxigenic strains are equally susceptible to inactivation by this waste treatment process and by acidified VFA in general.

Animals↗

Influence of methionine hydroxy analog and DL-methionine on rumen protozoa and volatile fatty acids.

Three lactating Holstein cows with rumen cannulae were used in a 3 X 3 Latin square arrangement of treatments to study effects of methionine source on ciliated protozoa numbers and volatile fatty acid concentrations in the rumen. Cows were fed total mixed diets twice daily of 60% grain mix, 21% corn silage, and 19% alfalfa hay (dry basis). Methionine hydroxy analog, DL-methione, and sodium sulfate were added on an equal sulfur basis. Experimental periods were 14 d with four rumen samples taken daily on d 13 and 14. Total ciliated protozoa were higher when cows were fed DL-methionine than when fed methionine hydroxy analog or sodium sulfate. Ruminal concentrations of butyric acid and isobutyric acid were higher for the DL-methionine treatment than for analog or sodium sulfate treatments. Isovaleric acid concentrations were also higher in fluid from cows fed methionine compared with cows fed the analog.

Animals↗

Characterization of microbial populations and volatile fatty acid concentrations in the jejunum, ileum, and cecum of pigs weaned at 17 vs 24 days of age.

In a series of five 17-d replicate trials, a total of 54 cannulated and 12 noncannulated pigs were used to determine the effects of weaning age (17 d or 24 d) on pH, dry matter percentage, aerobic and anaerobic microflora, lactate, and volatile fatty acid (VFA) concentrations in the jejunum, ileum, and cecum of weanling pigs. At -14 d of age, cannulated pigs were surgically fitted with T-cannulas in the jejunum (n = 20), ileum (n = 18), or cecum (n = 16). Upon weaning, cannulated pigs were individually caged in an environmentally controlled room with ad libitum access to a phase starter diet and water. Noncannulated pigs were killed at weaning and samples were collected from the jejunum, ileum, and cecum. Digesta and fecal swabs from cannulated pigs were collected twice weekly. The pH of cecal contents was lower (P < 0.05) and dry matter percentage was greater (P < 0.05) than those ofjejunal or ileal contents. Pigs weaned at 24 d of age had increased (P < 0.05) E. coli populations 3 d postweaning compared to preweaning populations, regardless of site of collection, whereas this increase was not observed in pigs weaned at 17 d of age. Unweaned pigs maintained higher (P < 0.05) lactobacilli populations compared to weaned pigs; however, populations declined (P < 0.05) in both groups by 3 d postweaning, with pigs weaned at 24 d of age having lactobacilli populations greater than pigs weaned at 17 d of age. Fecal populations of E. coli and lactobacilli declined (P < 0.05), whereas fecal bifidobacteria populations increased (P < 0.05) postweaning, regardless of weaning age. Concentrations of total fecal anaerobes declined (P < 0.05) in pigs weaned at 17 d of age but were maintained in pigs weaned at 24 d of age. Volatile fatty acid concentrations were greater (P < 0.05) in the cecum than in the jejunum or ileum, and acetic acid concentrations decreased (P < 0.05) postweaning regardless of weaning age. A tendency for L+ lactate concentrations to be greater (P < 0.07) in the ileum and jejunum vs the cecum was observed. Results indicate that weaning and weaning age have significant effects on microbial populations and VFA concentrations.

Age Factors↗

Detection of alcohols and volatile fatty acids by head-space gas chromatography in identification of anaerobic bacteria.

A head-space gas chromatographic technique for the analysis of volatile bacterial metabolites is described. Bacteroides fragilis, Clostridium perfringens, and Propionibacterium acnes, cultured in a glucose-containing peptone yeast extract medium, were studied. The head-space technique was compared with the injection of the complete liquid culture medium, and solvent extracts thereof, into the gas chromatograph. Volatile fatty acids could be detected by all three methods, whereas alcohols produced by C. perfringens and P. acnes were detectable only in the head-space chromatograms. Both FFAP and Porapak Q were used as gas chromatography stationary phase. Porapak Q was found more suitable than FFAP for the separation of alcohols. The head-space technique requires a minimum of preparation before the analysis and is well suited to automation.

Alcohols↗

Effects of solids concentration, pH and carbon addition on the production rate and composition of volatile fatty acids in prefermenters using primary sewage sludge.

Increasing evidence is emerging that the performance of enhanced biological phosphorus removal (EBPR) systems relies on not only the total amount but also the composition of volatile fatty acids (VFAs). Domestic wastewater often contains limited amounts of VFAs with acetic acid typically being the dominating species. Consequently, prefermenters are often employed to generate additional VFAs to meet the demand for carbon by EBPR and/or denitrification processes. Limited knowledge is currently available on the effects of operational conditions on the production rate and composition of VFAs in prefermenters. In this study, a series of controlled batch experiments were conducted with sludge from a full-scale prefermenter to determine the impact of solids concentration, pH and addition of molasses on prefermentation processes. It was found that an increase in solids concentration enhanced total VFA production with an increased propionic acid fraction. The optimal pH for prefermentation was in the range of 6-7 with significant productivity loss when pH was below 5.5. Molasses addition significantly increased the production of VFAs particularly the propionic acid. However, the fermentation rate was likely limited by the biological activity of the sludge rather than by the amount of molasses added.

Acetic Acid↗

Comparison of techniques to determine the clearance of ruminal volatile fatty acids.

The objective of this experiment was to compare measurements of fractional clearance rates obtained by using an unlabeled valerate-CoEDTA technique with measurements obtained by using a (13)C-labeled volatile fatty acids (VFA) technique. The exponential decay rate of the (13)C/(12)C ratio after pulse-dosing (13)C-acetate, (13)C-propionate, or (13)C-butyrate into the rumen was compared with the decay rate of rumen valerate concentration following a simultaneous pulse dose. The unlabeled valerate, CoEDTA, and each labeled VFA, one at a time, were concurrently mixed with the evacuated ruminal content of 6 lactating cows in two 3 x 3 Latin squares. The clearance of VFA by passage to the omasum was assumed to be equivalent to the decay in ruminal Co concentration and was around 50% of the total clearance. Acetate, propionate, and butyrate had similar fractional clearance rates (31.2, 33.4, 30.4%/h, respectively), but propionate had a higher absorption rate (19.2%/h) than butyrate (14.2%/h). Linear regression determination coefficients using the valerate clearance rate as an estimator for acetate, propionate, and butyrate rumen clearance were 0.51, 0.56, and 0.99, respectively. In a second experiment, the (13)C-valerate fractional clearance rate estimate (33.7%/h) was similar to the estimate obtained with unlabeled valerate (35.0%/h) by the valerate-Co technique. No (13)C enrichment of rumen microbes was noted 4 h after the intraruminal infusion of (13)C-valerate. Fractional VFA absorption rate estimates obtained in both techniques were similar, although both were lower than estimates reported in the literature by other methods.

Absorption↗

Effects of intraruminal infusions of mineral salts on volatile fatty acid production in steers fed high-grain and high-roughage diets.

Four rumen fistulated Holstein steers were used to assess the effects of intraruminal salt infusions on various rumen characteristics including volatile fatty acid production rates. In the first experiment, the basal diet consisted of 75% concentrate and 25% corn silage (high-grain diet). The experimental design was a 4 X 4 Latin square in which the four treatments were: 1) intraruminal infusion of 8 liters of water (control) or 8 liters of water plus 2) 288 g sodium bicarbonate, 3) 200 g sodium chloride, or 4) 600 g sodium chloride. In the second experiment, procedures were the same except the basal diet consisted of 64% alfalfa hay and 18% each of corn silage and concentrate (high-roughage diet). When the high-grain basal diet was fed, intraruminal infusions of mineral salts increased dilution rate of rumen fluid. Total fluid flow from the rumen also was increased by salt infusions with sodium bicarbonate and the most sodium chloride from (600 g/day) exerting greatest effects for both dietary regimens. Salt infusions reduced the molar percentage of rumen propionate and increased the molar percentage of acetate when the high-grain diet was fed. These same treatments were without effect on molar percentages of rumen acids when the high-roughage diet was fed. The change in the molar percentages of acetate and propionate on the high-grain ration was solely from reduction in propionate production. The lower production of propionate, from salt infusions, may have occurred because of the washout of readily fermentable materials. Dry matter disappearance from feeds placed in dacron bags and suspended in the rumen was unaffected by infusion of mineral salts.

Animals↗

[Feeding behavior, rumination, reticulo-ruminal fermentation and plasma volatile fatty acids, compared in goats and sheep; influence of the diet].

Two-year old Alpine she-goats (n = 3) and Texel ewes (n = 3) were compared as to eating behaviour, rumination and volatile fatty acids (VFA) in rumen and blood. The animals were fed once daily with two different proportions (20 and 80%) of barley and hay. Dry matter intake was fixed at 48 g D.M/kg P0.75 per day. In similar feeding and environmental conditions, eating behaviour and rumination of goats and sheep did not differ much: the goats tended to eat faster and there were more rumination periods in the sheep. Latency time, mean duration of rumination periods, daily rumination time and circadian pattern of rumination did not differ significantly between the two species (fig. 1). With both diets we observed a higher VFA concentration and a lower acetate/propionate ratio in the rumen of the ewes; however, rumen pH was lower only in those eating the 80% barley ration (fig. 2). Blood VFA in the jugular vein did not differ between sheep and goats (fig. 3). The proportion of cereals and hay in the diets affected rumen fermentation and rumination pattern in both species. With a higher concentrate/roughage ratio, rumen and plasma VFA increased, while the pH and acetate/propionate ratio in the lumen juice, the number of rumination periods and daily rumination time decreased. When the animals were fed the 80% barley ration, there was practically no rumination in the first 9 h after the single meal. During this time, rumen pH was minimal and VFA levels in the rumen and blood were maximal.

Animal Feed↗

[Effect of wood waste on the digestibility of glycides and level of volatile fatty acids in sheep].

Sheep were subject to an experiment concerning the effect of treated beech sawdust and thickened aqueous hydrolyzate of beech wood (xylocel) on the digestibility of glycides and on the level of volatile fatty acids (VFA) in the rumen content. The results prove that the use of treated beech sawdust as a replacer for ground barley straw favourably influenced the digestibility of monosaccharides, cellulose, lignin and total dry matter of the diet. Sawdust reduced the concentration of total VFA in the rumen content but the ratio of acetic acid to propionic acid (A:P) was lower and the energetic efficiency of the VFA produced was higher in the diet containing sawdust. The addition of urea to the diet with beech sawdust conditioned less effectively, as distinct from the case mentioned above, did not give a clear effect. Urea encouraged the digestibility of pentosanes, glucose and cellulose, but reduced the energetic efficiency of the VFA produced. Xylocel had no clear effect on the digestibility of sugars either. The higher A:P ratio and the lower energetic efficiency of the VFA produced testify to the fact that xylocel in combination with beet molasses was not a sufficient replacer of sugar-beet glycides.

Animal Feed↗

Volatile fatty acid impacts on nitrite oxidation and carbon dioxide fixation in activated sludge.

Batch test were performed to assess nitrite removal, nitrate formation, CO2 fixation, gaseous nitrogen production and microbial density in activated sludge exposed to volatile fatty acid (VFA) mixtures. Nitrite removal and nitrate formation were both affected by the presence of VFAs, but to different degrees. Nitrate formation rates were reduced to a greater extent (79%) than nitrite removal rates (36%) resulting in an apparent unbalanced nitrite oxidation reaction. Since the total bacterial density and the nitrite oxidizing bacteria (NOB, Nitrospira) concentration remained essentially constant under all test conditions, the reduction in rates was not due to heterotrophic uptake of nitrogen or to a decrease in the NOB population. In contrast to the nitrogen results, VFAs were not found to impact CO2 fixation efficiency. It appeared that nitrite oxidation occurred when VFAs were present since the oxidation of nitrite provides energy for CO2 fixation. However, nitrate produced from the oxidation of nitrite was reduced to gaseous nitrogen products. N2O gas was detected in the presence of VFAs which was a clear indication that VFAs stimulated an alternative pathway, such as aerobic denitrification, during biotransformation of nitrogen in activated sludge.

Bacteria↗