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Influence of the antibiotic erythromycin on anaerobic treatment of a pharmaceutical wastewater.

A laboratory-scale anaerobic sequencing batch reactor was used to treat a model substrate mixture representing pharmaceutical wastewater at an organic loading rate of 2.9 g COD/(L d). After reaching stable operation the reactor was first exposed to low (1 mg/L) and, subsequently, to high (200 mg/L) concentrations of the antibiotic erythromycin. The addition of low levels of erythromycin resulted in a significant but limited reduction of biogas production by 5% and the higher level of erythromycin did not impact biogas production further, suggesting that a substantial fraction of the microbial populations in the ASBR were resistant to the antibiotic. Effluent soluble COD could not be accounted for in measured volatile fatty acids, perhaps suggesting the production of soluble microbial products. In batch tests evaluating the specific methanogenic activity, conversion of the model substrate mixture was only slightly affected by the presence of erythromycin. However, the conversion of butyric acid was inhibited when erythromycin was present. After 47 days of exposure to erythromycin, the conversion of butyric acid was inhibited to a lesser extent, suggesting the development of antibiotic resistance in the biomass. Exposure to antibiotics can affect specific substrate degradation pathways, leading to the accumulation of volatile fatty acids, soluble microbial products, and potentially to overall system instabilities.

Anti-Bacterial Agents↗

Relationships between pyruvate decarboxylation and branched-chain volatile acid synthesis in Ascaris mitochondria.

The rate of 14CO2 evolution from 1-[14C]pyruvate by intact Ascaris mitochondria was very slow, but increased with increasing concentrations of pyruvate. At all concentrations of pyruvate, in an aerobic environment, pyruvate decarboxylation was stimulated greatly by the addition of fumarate, malate, or succinate. However, under anaerobic conditions, only malate and fumarate stimulated pyruvate decarboxylation; succinate had no effect. This implies that the aerobic metabolism of succinate, presumably to other dicarboxylic acids, may be required for the stimulation. Incubation of sonicated mitochondria with pyruvate plus fumarate, under rate-limiting concentrations of NAD+, resulted in approximately equal quantities of pyruvate utilized and succinate formed, suggesting that pyruvate oxidation and fumarate reduction may be linked. Branched-chain, volatile fatty acids were not formed during incubations with either malate or succinate, or succinate plus acetate. However, incubations of intact Ascaris mitochondria with pyruvate plus succinate yielded 2-methylbutyrate and 2-methylvalerate, whereas incubations with pyruvate plus propionate yielded almost exclusively 2-methylvalerate. Oxygen dramatically inhibited the synthesis of the branched-chain acids from succinate plus pyruvate, attesting to the apparent anaerobic nature of Ascaris mitochondrial metabolism. Significantly, the addition of glucose plus ADP stimulated the formation of all volatile fatty acids. Therefore, the synthesis of branched-chain acids may be related directly to increased energy generation. Alternatively, they may function in the regulatory role of maintaining the mitochondrial redox balance.

Adenosine Diphosphate↗

Ranking the toxicity of fatty acids on Jurkat and Raji cells by flow cytometric analysis.

The fatty acids have an important role in the control of leukocyte metabolism and function. Higher concentrations of certain fatty acids, particularly polyunsaturated fatty acids (PUFAs) and volatile fatty acids, can cause cell death via apoptosis or, when concentrations are greater, necrosis. In this study, we determined the highest concentrations of various fatty acids that are non-toxic to two human leukemic cell lines, Jurkat (T-lymphocyte) and Raji (B-lymphocyte). Toxicity was evaluated by either loss of membrane integrity and/or DNA fragmentation using flow cytometric analysis. There were no remarkable differences for the toxicity of the fatty acids between B and T cell lines. The cytotoxicity of the fatty acids was related to the carbon chain length and number of double bonds: docosahexaenoic acid=eicosapentaenoic acid=arachidonic acid=gamma-linolenic acid=stearic acid=palmitic acid > linoleic acid=palmitoleic acid > vacenic acid=lauric acid > oleic acid > elaidic acid > capric acid > butyric acid > caprylic acid=caproic acid=propionic acid. The proportion of cells undergoing apoptosis or necrosis, induced by the fatty acids tested, remains to be investigated.

Apoptosis↗

[In vitro simulation of rabbit cecal fermentation in a semi- continuous flow fermentor. I. Role of food substrate pretreatment].

A Rusitec semi-continuous flow fermentor was used to study the influence of enzyme pretreatment of food substrates on the fermentation profile over a 2-week period following inoculation with rabbit caecal contents. Three types of substrate were examined: 1) homogenized commercial rabbit feed; 2) the solid remains of this feed after digestion with alpha-amylase for 24 h; and 3) substrate 2 digested for 4 h with pepsin (double enzyme treatment). One of a pair of nylon pouches containing 15 g substrate was replaced each day, thus producing a uniform 48-h fermentation. Fermentation of the untreated feed (1) for 5-6 days produced a fermentation profile quite different from that obtained in vivo in the rabbit caecum: propionic acid accounted for over 35% of total volatile fatty acid (VFA), and butyric acid for about 15%. Amylase digestion (2) gave a stable ferment profile closer to the in vivo profile, except that propionic and butyric acids were similar at 15% of total VFA. Digestion with both amylase and pepsin (3) produced a stable fermentation profile very close to the in vivo profile: C2 > 60%, C3 < 11% and 17% < C4 < 21%. The rate at which membrane constituents (acid detergent fibre, ADF) were lost in 48 h was similar to the digestibility coefficient measured in vivo by others for the same basic feed. Lastly, there was a high percentage (about 5%) of volatile C5 fatty acids; this could be due to the discontinuous fermentor input of one pouch per 24 h. Thus, feed pretreated with both amylase and pepsin simulates, in vitro, rabbit caecal fermentation in a semi-continuous Rusitec type fermentor.

Animal Feed↗

Gas chromatographic assessment of alcoholyzed fatty acids from yeasts: a new chemotaxonomic method.

An alternative chemotaxonomic method to methanolysis was developed for gas chromatographic assessment of fatty acids in whole yeast cells. Clinical and reference strains of the medically important yeasts Candida albicans, Torulopsis glabrata, and Saccharomyces cerevisiae were cultured for 48 h at 26 degrees C. Cellular lysis and transesterification were then performed with ethanol, propanol, butanol, or methanol. The relative recovery rates for cellular fatty acids, including the volatile acids C10:0 and C12:0, were similar after alcoholysis with ethanol, propanol, or butanol, while methanolysis gave lower recoveries of volatile fatty acids. Thus, after ethanolysis, the recovery of C10:0 acid (0.1, 1, and 10%) from a defined matrix (lyophilized Actinobacillus actinomycetemcomitans cells) varied from 97 to 102%, while the recovery of C10:0 after methanolysis varied from 49 to 75%. This indicated that with the frequently used methanolysis technique, there is a considerable loss of volatile fatty acids. These acids may be used as marker molecules for taxonomic differentiation between yeasts.

1-Propanol↗

Fecal numbers of bifidobacteria are higher in pigs fed Bifidobacterium longum with a high amylose cornstarch than with a low amylose cornstarch.

Twelve young male pigs consumed a purified diet containing wheat bran as fiber source. Starch provided 50% of total daily energy either as a low amylose cornstarch or as a high amylose (amylomaize) starch. The pigs were given a supplement of a freeze-dried probiotic organism (Bifidobacterium longum CSCC 1941). A block crossover design was used so that at any one time two groups of three pigs consumed either the high or low amylose cornstarch without probiotic and a further two groups of three pigs consumed either high or low amylose cornstarch with probiotic. Neither food intake nor body weight gain was affected by diet. Fecal output was higher when pigs were fed the high amylose cornstarch, but moisture content was unaffected. Fecal concentrations and excretion of total volatile fatty acids were higher when pigs were fed the high amylose cornstarch. Concentrations of acetate were unaffected by dietary starch, but those of propionate and butyrate were higher when the high amylose cornstarch was consumed. Fecal excretion of all three acids was higher during high amylose cornstarch feeding. Bifidobacteria were detected in the feces only when pigs were fed Bifidobacterium longum. Fecal bifidobacteria counts (expressed per gram of wet feces) and their daily fecal excretion were higher when pigs were fed high amylose cornstarch. Feeding the probiotic did not alter fecal starch or volatile fatty acids. None of the variables studied was affected by the order of feeding of starch or probiotic. The data show that a high amylose starch acts as a prebiotic in promoting the fecal excretion of probiotic organisms.

Amylose↗

Nutritional risk factors in the etiology of left displaced abomasum in dairy cows: a review.

The transition period occurring 2 wk prepartum through 2 to 4 wk postpartum is the major risk period in the etiology of left displaced abomasum. The prepartum depression of intake and the slow postpartum increase in intake are risk factors causing lower ruminal fill, reduced forage to concentrate ratio, and increased incidence of other postpartum disorders. Uncomplicated ketosis, retained placenta, metritis, and hypocalcemia at parturition are risk factors for left displaced abomasum. Excessive amounts of concentrate during the prepartum period increase the risk of left displaced abomasum, which may occur from the lower ruminal fill caused by greater prepartum intake depression and reduced forage to concentrate ratio, decreased ruminal motility from lower ruminal fill and higher volatile fatty acid concentration, and decreased abomasal motility and emptying from higher concentrations of volatile fatty acids. Effects of volatile fatty acids on motility may be exacerbated by low ruminal absorption of volatile fatty acids during the transition period. Minimal intake of concentrate during the prepartum period may increase the risk of left displaced abomasum through failure to increase the absorptive capacity of the ruminal papillae and failure of the microbial population of the rumen to adapt prior to the intake of high energy postpartum diets. Increased risk of left displaced abomasum in cows that are hypocalcemic at parturition may be due to decreased ruminal and abomasal motility.

Abomasum↗

Production response of lactating cows fed dried versus wet brewers' grain in diets with similar dry matter content.

Twenty-four Holstein-Friesian dairy cows (20 intact and 4 fitted with rumen cannula) during early lactation (56 +/- 25.3 d in milk) were assigned to two treatments to determine intake and production responses to feeding dried and wet brewers' grain. There were two cows fitted with a rumen cannula in each treatment. Cows were fed a total mixed ration twice daily containing either dried or wet brewers' grain at 15% of the dietary dry matter (DM). The diet contained 47% forage and 53% concentrate. The experimental design was a replicated 2 x 2 Latin square with two periods of 5 wk each. First 2 wk in each period were considered as adaptation to diets and data from the last 3 wk were used for treatment comparisons. Dried and wet brewers' diets contained 68.0 and 66.5% DM, respectively. Feeding brewers' grain dry or wet to dairy cows had no influence on feed intake (25.6 vs. 25.1 kg/d), fat corrected milk yield (40.1 vs. 40.7 kg/d), milk composition and feed consumption. The pH, ammonia, total volatile fatty acids and molar ratios of volatile fatty acids in the rumen fluid were not different between treatments. Fatty acid composition of milk fat from cows fed diets containing dry or wet brewers' grain was identical, except C18:2 and C18:3 fatty acids were lower in milk fat from cows fed wet brewers' grain compared with dried brewers' grain. The results from the present study suggest that the performance of cows fed either dried or wet brewers' grain at 15% of dietary DM was similar when diets had the same DM. The average price for dried and wet brewers' grain in the United States from July 2001 to June 2002 was dollars 145.3 and dollars 96.9/metric tonne DM, respectively. Using wet instead of dried brewers' grain will save dollars 49/metric tonne minus the difference in storage costs. Wet brewers' grain can be fed to dairy cows in areas that are close to the brewery and provides nutritive value similar to the dried brewers' grain.

Ammonia↗

[Adaptation of rumen fermentation to monensin].

Adaptation of rumen fermentation to monensin feeding has been studied with rumen-fistulated sheep receiving a daily dose of 30 mg of monensin for a period of 21 d followed by a 28 d period during which 60 mg doses were administered. The ration consisted of 300 g of hay and 300 g of concentrates, fed at 9.00 h and 16.00 h. Monensin was placed in the rumen as an aqueous suspension, just prior to the morning feeding. Monensin infusion was preceded and followed by a period during which no monensin was infused. The following rumen fermentation parameters were determined: methane production, pH, volatile fatty acids (VFA) molar proportions, total volatile fatty acid concentration, lactate and ammonia concentrations and in sacco degradability of hay. Rumen gas expelled through the fistula was collected for 6 h per day and analysed. Total VFA concentration, molar proportions of individual VFA, pH, lactate and ammonia concentration were determined on rumen contents, sampled just prior the administration of monensin and 2 and 6 h later. In vitro incubations of 3 h were carried out with rumen fluid, sampled 1 h after feeding. In vivo and in vitro methane production was decreased by monensin feeding. The molar proportion of propionate in the rumen was increased, while acetate and butyrate percentages were lowered. The total VFA and ammonia concentrations were also decreased by monensin, but pH values were increased. In vitro production of propionate was stimulated by monensin administration and methanogenesis decreased. The organic matter in sacco degradability was not affected, probably because of the time difference between the introduction of bags and monensin in the rumen. These modifications of rumen fermentation persisted as long as monensin was given, indicating that in this experiment, there was no adaptation to the ionophore.

Ammonia↗

[Porcine pepsin on some chemical and microbiological indices of cheeses produced with its participation].

The authors report data about the contents of non-protein nitrogen, bound and soluble nitrogen, free amino acids, monocarbonyl compounds and volatile fatty acids as well as about the sum of dinitrophenylhydrazine derivatives of carbonyl compounds and about the microflora. The greatest differences between experimental and control cheeses (produced with rennin) in the contents of the above-mentioned constituents were observed within the first 30 days. On day 60 of experimentation, these differences had smoothed down. It is concluded that a I:I mixture of porcine pepsin and rennin may be used for the production of small medium hard cheeses without impairing their quality of nutritive value.

Amino Acids↗

[Variations in the pattern of ruminal volatile free fatty acids, acetoacetate and glucose in the blood plasma of lactating cows after a period of fasting and renewed feeding].

3 lactating cows were fasted for 4 days and then again received food. The trial was performed to investigate the effect of fasting on the concentration and molar proportion of volatile fatty acids in the rumen, on the concentration of free fatty acids and glucose in blood plasma and on the acetoacetate concentration in the blood. Fasting produced a considerable decrease in the total concentration of volatile fatty acids. Processes of ruminal fermentation were characterized, during the fasting period, by a continuous increase in the molar proportions of acetic acid, isobutyric acid and isovaleric acid whereas a significant decline observed to occur in the molar proportions of propionic acid and butyric acid. The reciprocal behaviour of acetate and propionate under conditions of fasting lead to a linear increase in the C2 to C3 ratio from 4.4 : 1 to 9,6 : 1. With renewed food supply all fermentation data returned to normal levels. The concentrations of blood glucose declined after 24 hrs of fasting, remained at the same level for the following 2 days and rose again to normal values on the 4th day of fasting. The concentration of acetoacetate decreased significantly on the first day of fasting to values that were 4 times as high as the initial value. The concentration of free fatty acids in blood plasma increased tremendously to a value 40 times as high as that observed at the beginning of the fasting period. It proved to be a good indicator for the energy requirements of the organism.

Acetoacetates↗

Effect of added caffeic acid and tyrosol on the fatty acid and volatile profiles of camellia oil following heating.

Camellia oil is widely used in some parts of the world partly because of its high oxidative stability. The effect of heating a refined camellia oil for 1 h at 120 degrees C or 2 h at 170 degrees C with exogenous antioxidant, namely, caffeic acid and tyrosol, was studied. Parameters used to assess the effect of heating were peroxide and K values, volatile formation, and fatty acid profile. Of these, volatile formation was the most sensitive index of change as seen in the number of volatiles and the total area count of volatiles in gas chromatograms. Hexanal was generally the dominant volatile in treated and untreated samples with a concentration of 2.13 and 5.34 mg kg(-1) in untreated oils heated at 120 and 170 degrees C, respectively. The hexanal content was significantly reduced in heated oils to which tyrosol and/or caffeic acid had been added. Using volatile formation as an index of oxidation, tyrosol was the more effective antioxidant of these compounds. This is contradictory to generally accepted antioxidant structure-activity relationships. Changes in fatty acid profiles after heating for up to 24 h at 180 degrees C were not significant.

Aldehydes↗

Ruminal digestion and fermentation of high-producing dairy cows with three different feeding systems combining pasture and total mixed rations.

Six multiparous Holstein cows fitted with rumen cannulas were used to study the effect of three feeding systems combining pasture and total mixed rations (TMR) on ruminal digestion in a 21-wk repeated measures experiment. The three treatments were: 1) pasture plus concentrate (PC), 2) pasture plus partial TMR (pTMR), and 3) TMR (nonpasture). Ruminal NH3-N concentration was lower on both the pTMR and TMR treatments (10.2 +/- 0.5 mg/dL) than on the PC treatment (19.9 +/- 0.5 mg/dL). Ruminal pH was not affected by treatments and averaged 5.87. Neither total volatile fatty acid concentration (137.5 mmol/L) nor individual volatile fatty acid proportions (63.1,20.6, and 12.0 mol/ 100 mol for acetate, propionate, and butyrate, respectively) differed among treatments. The pTMR treatment reduced the total potentially degradable fraction of dry matter (85.5 vs. 82.3%) and the potentially digestible fraction of neutral detergent fiber (82.1 vs. 74.9%) of pasture compared to the PC treatment. Ruminal NH3-N losses were reduced when combining pasture and TMR; however this combination decreased the ruminal digestion of pasture, indicating the presence of associative effects in the rumen.

Ammonia↗

Utilization and excretion of a new sweetener, fructooligosaccharide (Neosugar), in rats.

In order to study the digestibility of the fructooligosaccharide "Neosugar," [U-14C]Neosugar or [U-14C]sucrose was orally administered to germfree, conventional and antibiotic-treated rats and the radioactivities of expired 14CO2, urine and feces were determined 24 h later. More than 50% of the Neosugar was expired as CO2 in conventional rats. This was the same as for sucrose, but the time course was delayed by about 2 h. In germfree rats, no 14CO2 was released for the first 8 h, and 14CO2 released after 8 h probably reflected bacterial colonization of the gut. The radioactivity of the urine was about 3-4% in all groups, but that of the feces from germfree rats was about eight times higher than the level in conventional rats. When [U-14C]Neosugar was anaerobically incubated with the cecal contents of conventional rats, more than 10% of the added Neosugar was metabolized to CO2, about 66% to volatile fatty acids and about 7% to microbes. More than 58% of 1-14C-volatile fatty acids such as acetic acid, propionic acid or butyric acid injected directly into the cecum of conventional rats was excreted as CO2 within 24 h. These results indicate that Neosugar given orally to rats is metabolized mainly to volatile fatty acids and CO2 by intestinal microorganisms, and the volatile fatty acids produced are absorbed and further converted to CO2 in the body. Thus, the data indicate that Neosugar is partially utilized as an energy source.

Acetates↗

Effect of white-rot basidiomycetes-treated wheat straw on rumen fermentation in artificial rumen.

This study evaluated three white-rot basidiomycetes for their potential to improve the ruminal degradation of wheat straw. Pleurotus ostreatus (PO), Pleurotus ostreatus-mutant (PO-M) and Trametes gibbosa (TG) were incubated on wheat straw for 30 days at 28 degrees C. Neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein and in vitro dry matter digestibility (IVDMD) were determined. The results demonstrated increasing crude protein and ash contents (%) in the fungi-treated straws. The IVDMD values were also increased. Compared to untreated wheat straw (UWS), the NDF and ADF contents were reduced in fungi-treated straw (TWS). Out of the three fractions-hemicellulose, cellulose and lignin-hemicellulose showed the largest proportionate loss and lignin the smallest in all three fungi-treated straws. TWS with Pleurotus ostreatus (TWS-PO), Pleurotus ostreatus-mutant (TWS-PO-M) and Trametes gibbosa (TWS-TG) together with barley (80:20%) were used as the experimental diets in the artificial rumen. UWS with barley (80:20%) served as the control diet. The results revealed significantly higher IVDMD values, NDF, ADF and cellulose digestibilities (%) with the experimental diets. The production of propionic acid decreased, n-butyric, n-valeric and isovaleric acids (mmol/day-1) increased and the volatile fatty acid (VFA) production expressed in mol VFAs.kg-1 digested dry matter decreased in experimental diets. The total microbial production also decreased at fermentation in the experimental diets.

Animals↗

Sites of organic acid production and absorption in gastrointestinal tract of the pig.

The relationship between diet, pH, and microbial digestion of carbohydrate was examined in 24 pigs fed either a conventional or a low-protein, high-cellulose experimental diet and sacrificed 2, 4, 8, or 12 h after a meal. In animals fed the control diet contents of the cranial half of the stomach demonstrated marked, cyclic fluctuations in pH and high concentrations of organic acids. Contents of the caudal (glandular) half were lower in both pH and organic acid concentration. Despite concentrations of volatile fatty acids (VFAs) as high as 250 meq/liter in the large intestine, the pH remained relatively neutral. The VFA levels remained relatively constant throughout the length of the colon. The VFA transport across isolated gastric and large intestinal mucosa also was examined. All four types of gastric mucosa absorbed and transported VFA at substantial rates. Mucosa of pig cecum and colon transported VFA at much greater rates than gastric mucosa and greater rates than previously determined in equine large intestinal mucosa or even bovine rumen epithelium. Comparison with results of earlier studies in the pony suggested that the higher concentration of VFA in the large intestinal contents of pigs was due to the more rapid rate of digesta passage rather than to less efficient absorption of fatty acids.

Animals↗

Broiler chick responses to anorectic agents: dietary acetic and propionic acids and the blood metabolites.

The effects of dietary supplementation with acetic or propionic acid, given at a level of 3% of diets containing 2,500 or 2,750 kcal of metabolizable energy per kg, on levels of blood glucose, and on lipid, protein, and energy metabolisms were studied in female broiler chicks. Voluntary feed and energy intakes, as well as body weight gain, decreased significantly with the inclusion of the acids in either diet, in decreasing order of effect: propionate > acetate > nonsupplemented controls. The relative weight of the abdominal adipose tissue decreased significantly with the inclusion of either acid, but was unaffected by dietary energy level. In contrast, the weight of the liver was higher in birds fed the low-energy diet, but was unaffected by the anorectic agents. Inclusion of the acids increased the concentration of volatile fatty acids in the cecum and blood plasma. The rate of clearance of plasma propionate was greater than that of acetate. Anorectic agents and dietary energy density exhibited a synergistic effect on feed intake. These agents altered blood metabolites involved in lipid and energy metabolisms, but not in that of protein. Plasma metabolites and enzymes, particularly beta-hydroxybutyrate and alkaline phosphatase, reflected the degree of feed restriction, supporting the hypothesis that the hypophagic effect of these acids is due to metabolic intervention.

3-Hydroxybutyric Acid↗