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Cyclic changes in the osmolality and electrolyte composition in the gastrointestinal tract of the rock hyrax.

The rock hyrax has been shown to have a very unusual and complex digestive tract. The gastrointestinal tract is comparable to that of the simple and complex stomach of mammals as well as to that of birds. Determinations of osmolality and electrolytes have been made in different sections of the gut of the hyrax. However, with the exception of the elevated potassium levels observed in the cranial stomach, the hyrax poses no unusual osmotic or electrolyte concentrations when compared to man or other mammals. The greater cation concentration and hypertonicity of the cranial stomach was largely accounted for by the elevated potassium load in this gut segment. Sodium and potassium comprised the major cations present throughout the gastrointestinal tract. Chloride was the major anion observed in the foregut; however, the electroneutrality of the mid and hindgut of the hyrax was primarily maintained by the presence of large concentrations of volatile fatty acids produced in these portions of the gut. Volatile fatty acid also accounted for the high anion concentration and hypertonicity of the cranial stomach.

Animals↗

Influence of adjunct cultures on volatile free fatty acids in reduced-fat Edam cheeses.

The effects of the adjunct cultures Lactococcus lactis ssp. diacetylactis, Brevibacterium linens BL2, Lactobacillus helveticus LH212, and Lactobacillus reuteri ATCC 23272 on volatile free fatty acid production in reduced-fat Edam cheese were studied. Lipase activity evaluation using p-nitrophenyl fatty acid ester substrates indicated that L. lactis ssp. diacetylactis showed the highest activity among the 4 adjunct cultures. Full-fat and 33% reduced-fat control cheeses (no adjunct) were made along with 5 treatments of reduced-fat cheeses, which included individual, and a mixture of the adjunct cultures. Volatile free fatty acids of cheeses were analyzed using static headspace analysis with 4-bromofluorobenzene as an internal standard. Changes in volatile free fatty acid concentrations were found in headspace gas of cheeses after 3-and 6-mo ripening. Acetic acid was the most abundant acid detected throughout ripening. Full-fat cheese had the highest relative amount of propionic acid among the cheeses. Certain adjunct cultures had a definite role in lipolysis at particular times. Reduced-fat cheese with L. lactis ssp. diacetylactis at 3-mo showed the highest levels of butyric, isovaleric, n-valeric, iso-caproic, and n-caproic acid. Reduced-fat cheese with Lactobacillus reuteri at 6 mo produced the highest relative concentration of isocaproic, n-caproic, and heptanoic, and the highest relative concentration of total acids.

Acetic Acid↗

[Characterization of species of the genus Clostridium by gas chromatographic analysis of the fatty acids produced during metabolism].

Volatile and non volatile fatty acids deriving from the bacterial metabolism of different sugars were determined by gas-chromatography to better characterize Clostridium tertium and Clostridium ramosum. The data obtained were also used for numerical taxonomic analysis and dendrograms were elaborated to study the taxonomic relationships between the two species.

Carbohydrate Metabolism↗

Broiler chick responses to anorectic agents: 1. Dietary acetic and propionic acids and the digestive system.

The effects of various levels of acetic or propionic acid supplementation on feed intake and the gastrointestinal tract were studied in female broiler chicks. Voluntary feed intake and body weight gain decreased with the inclusion of the acids. Results indicate that acetic and propionic acids exert a similar anorectic effect on chicks fed diets differing in energy content. The inhibitory effect of these acids on voluntary feed intake could not be attributed to alterations in feed utilization, to differences in pancreatic alpha-amylase activity, or to differences in the acid-base balance. The increase in cecal capacity and volatile fatty acid (VFA) content is probably due to increased utilization of nonstarch polysaccharides at that site, the latter compensating for feed inhibition. This reflects a slight adaptive response of the cecum to anorectic agents.

Acetates↗

Biophysical model for organic acid excretion in Ascaris suum.

To develop a model for the mechanisms of organic acid excretion in nematodes, we measured the concentrations of volatile fatty acids (VFAs), pH and electrical potentials across hypodermal and muscle membranes and across the composite body wall (consisting of hypodermis, muscle and cuticle) of Ascaris suum using standard chromatographic and microelectrode recording techniques. In incubates containing one parasite in 20 ml modified Ascaris Ringer's solution, the level of combined VFAs excreted into the medium increased linearly for about 18 h, then plateaued at a concentration of 4.2 mM; the medium acidified rapidly to a plateau at about pH 5.0 within 4-6 h. Following 24 h incubations, the concentrations of VFAs in the hypodermis, muscle, and pseudocoelomic compartments were 62.4 +/- 8.1, 62.3 +/- 7.8 and 74.4 +/- 3.2 mM, respectively. The pseudocoelomic fluid was more acidic (pH 6.52 +/- 0.06) than the hypodermis (pH 6.78 +/- 0.03) or muscle (pH 6.77 +/- 0.03). These data and the electrical potentials across hypodermal (-57.9 +/- 6.3 mV) and muscle (-30.3 +/- 0.8 mV) membranes were used to determine the equilibrium concentrations for protonated (HVFA) and anionic (VFA-) forms of the acids across these membranes and across the cuticle. Under these conditions, little transmembrane or transmural excretion of HVFAs is expected to occur in A. suum. However, a 16-27 mV driving force for VFA- excretion exists across hypodermal and muscle membranes, and a larger driving force is predicted to exist for these anions across the cuticle. This driving force could provide potential energy for VFA- excretion through anion channels which exist in muscle and hypodermal membranes of this parasite, or for facilitated transport systems.

Animals↗

The IWA Anaerobic Digestion Model No 1 (ADM1).

The IWA Anaerobic Digestion Modelling Task Group was established in 1997 at the 8th World Congress on Anaerobic Digestion (Sendai, Japan) with the goal of developing a generalised anaerobic digestion model. The structured model includes multiple steps describing biochemical as well as physicochemical processes. The biochemical steps include disintegration from homogeneous particulates to carbohydrates, proteins and lipids; extracellular hydrolysis of these particulate substrates to sugars, amino acids, and long chain fatty acids (LCFA), respectively; acidogenesis from sugars and amino acids to volatile fatty acids (VFAs) and hydrogen; acetogenesis of LCFA and VFAs to acetate; and separate methanogenesis steps from acetate and hydrogen/CO2. The physico-chemical equations describe ion association and dissociation, and gas-liquid transfer. Implemented as a differential and algebraic equation (DAE) set, there are 26 dynamic state concentration variables, and 8 implicit algebraic variables per reactor vessel or element. Implemented as differential equations (DE) only, there are 32 dynamic concentration state variables.

Bioreactors↗

[Diagnosis of anaerobic infection of the pleural fluid using gas-liquid chromatography].

BACKGROUND: An assessment of the efficacy of the detection of volatile and non-volatile fatty acids that are produced by microorganisms aimed at the possible diagnosis of pleural empyema. METHODS: 106 pleural fluids were examined by gas-liquid chromatography in contrast to conventional aerobic and anaerobic cultures. The sample previously prepared (1 microliter) was injected in an Hewlett-Packard 5890 chromatograph, that was equipped with a fused silica capillary column and a flame ionization detector. RESULTS: Growth of aerobic or anaerobic microorganisms was obtained in 13 (12.3%) of the samples, and volatile or non-volatile fatty acids were detected in all of them (sensitivity = 1). No growth was obtained in the 93 samples, but volatile or non-volatile fatty acids were detected in 64 (68.8%) of them (specificity = 0.31). A positive predictive value of 0.17 and a negative predictive value of 1 were obtained. Growth of anaerobic microorganisms was obtained in 9 samples (8.5%) and volatile fatty acids were detected in all of them (sensitivity = 1). Anaerobic microorganisms did not grow in 97 samples, however volatile fatty acids were detected in 2 (2.1%) of them. A positive predictive value of 0.82 and a negative of 1 were obtained. CONCLUSIONS: Referring to pleural fluid samples, the detection of volatile and non-volatile fatty acids by gas-liquid chromatography is a highly sensitive method, that may be used to quickly reject negative samples in about 30 mins. Furthermore, when anaerobic microorganisms exist, the high specificity of gas-liquid chromatography will make it possible to confirm their presence in the sample, mainly if there is a clinical suspect of anaerobic infection. Thus, the utilization of this sensitive method may also change the therapeutic behaviour.

Bacteria, Aerobic↗

Effect of fats and fatty acid combinations on ruminal fermentation in semi-continuous in vitro cultures.

Four in vitro trials were conducted to determine how ruminal fermentation is affected by source of fat, level of fat, and combinations of fatty acids. Trials I and II examined how volatile fatty acids (VFA) were changed by three sources of fat (blended animal-vegetable fat, corn oil and tallow fatty acids) each added to a hay substrate at six levels (0, 2, 4, 6, 8 and 10%). Increasing blended fat caused no changes in VFA levels except to decrease butyric acid from 12.1 to 9.9% of the total VFA (P less than .05). Corn oil and tallow fatty acids both increased propionic acid, causing the ratio of acetic to propionic acids (A/P) to decrease (P less than .01). Trial III tested different ratios of oleic/stearic and linoleic/stearic acids to determine if certain combinations were better for fermentation. There was no evidence of synergism among fatty acids since increasing the ratio of unsaturates steadily reduced A/P. Trial IV was designed to determine how changes in VFA levels reflect changes in fiber digestibility of substrates containing added fat. Volatile fatty acids having significant regressions with fiber digestibility were acetic acid (r = .648), propionic acid (r = -.670), total VFA concentration (r = .742) and A/P (r = .831). Results are interpreted to show that blended animal-vegetable fats are less toxic in the rumen than equal levels of other lipids, and the beneficial effects of blended fat cannot be attributed to a unique combination of fatty acids acting synergistically.

Animals↗

Production responses of dairy cows fed diets supplemented with ammonium salts of iso C-4 and C-5 acids.

A three-university study examined effects of ammonium salts of volatile fatty acids on dairy cows for a full lactation cycle. Objectives were a) to determine the most efficacious amount and ratio of two components, ammonium salts of C-5 acids and ammonium isobutyrate, and b) to compare responses of this optimum blend with the control in a response surface design. This design tested six blends of the two components defined by the center point and the five points of an equilateral pentagon from which response surfaces would be computed and a control. Each treatment consisted of 23 cows for a total of 161 multiparous Holstein cows. All cows were fed corn silage for ad libitum consumption, 1.8 kg of urea-mineral-vitamin-premix plus supplement based on corn grain and corn gluten meal according to milk production. Response surfaces, based on milk yield, defined an optimum blend composed of 61 g ammonium salts of C-5 acids plus 28 g ammonium isobutyrate per cow per day equivalent to 120 g aqueous blend (74% solids). The optimum blend was similar to the center point blend. Cows receiving the center point blend peaked higher and produced more milk, 4% fat-corrected milk, milk protein, and total solids than control cows. For the first trimester of lactation, milk yields for control and center point blend were 28.6 and 31.8 kg. The 305-day milk, 4% fat-corrected milk yields, and dry matter intakes (kg) were: control 20.5, 19.7, 17.3; center point blend 23.2, 21.6, 17.5. Increased milk production was associated with improved feed utilization. No effect on health or reproduction was observed.

Animal Feed↗

Effects of monensin, pyromellitic diimide and 2-bromoethanesulfonic acid on rumen fermentation in vitro.

The effects of monensin, 2-bromoethanesulfonic acid (2-BES) and pyromellitic diimide (diimide) on gas and volatile fatty acid (VFA) production by the rumen microbiota were compared in mixed culture. Oat hay, a hay-concentrate mixture (48% hay, 43% concentrate) and a soluble carbohydrate mixture were used as substrates for microbial growth. The highest concentrations of diimide (10 ppm) and 2-BES (30 microM) decreased methanogenesis by 97 and 76%, respectively, while H2 accumulation was increased 30- and 20-fold, respectively. The effect of monensin on methanogenesis was less dramatic as 10 ppm decreased CH4 accumulation 16% and H2 did not accumulate. Diimide and 2-BES decreased the acetate:propionate ratio with the hay (P less than .05) and soluble carbohydrate mixture (P less than .025). The numbers of saccharolytic, cellulolytic and methanogenic bacteria from sheep fed a diet with diimide (60 ppm) did not differ significantly from sheep fed a control diet. Semicontinuous culture incubations indicated that the mixed rumen microbial population could adapt and degrade diimide after 24 h of incubation.

Alkanesulfonates↗

Effects of pH and pH fluctuations on microbial fermentation and nutrient flow from a dual-flow continuous culture system.

Eight dual-flow continuous culture fermenters (1400 ml) were used in two consecutive periods to study the effects of pH and pH fluctuations on microbial fermentation and nutrient flow. Fermenters were maintained at 39 degrees C, with solid and liquid dilution rates of 5 and 10%/h, respectively, and fed continuously a 60% alfalfa hay and 40% concentrate diet (18.9% crude protein, 36.6% neutral detergent fiber, 17.6% acid detergent fiber). Treatments were high pH (constant at 6.4); low pH (constant at 5.7); cycles of 4 h at pH 6.4 and 4 h at pH 5.7; and pH constant at 6.4, except for two 30-min drops per day to pH 5.7, followed by a 3-h slow recovery to pH 6.4. The low pH (constant at 5.7) produced lower apparent dry matter, neutral detergent fiber and acid detergent fiber digestion, lower total and branch-chained volatile fatty acid concentrations, and lower acetate and higher propionate proportions than high pH (constant at 6.4). There were no differences in these estimates between constant high pH and the two treatments that alternated high pH and low pH. The constant low pH reduced protein degradation and increased nonammonia N and dietary N flow compared with constant high pH. The pH treatments had no effect on bacterial N flow or efficiency of microbial protein synthesis. Flow of essential amino acids was highest for constant low pH and lowest for constant high pH. Results indicate that constant low pH reduced fiber and protein digestion and increased the flow of total and some individual amino acids. However, the effects of transitory decreases of pH were either small or insignificant with the conditions tested in this study.

Amino Acids↗

Amino acid deamination by ruminal Megasphaera elsdenii strains.

When ruminal fluid from a cow fed timothy hay was serially diluted (10-fold increments into anaerobic broth containing 15 mg ml(-1) Trypticase), the low dilutions (< or =10(-6)) had optical densities greater than 2.0 and ammonia concentrations greater than 100 m M. The optical densities and ammonia concentrations of the 10(-8) and 10(-9) dilutions were very low, but large cocci were observed in the 10(-8) dilution. The large cocci were isolated and identified by 16S rDNA sequencing as Megasphaera elsdenii. The freshly isolated strain (JL1) grew well on Trypticase, but less than 4% of the amino acid nitrogen in Trypticase was converted to ammonia. Optical density and ammonia production were twice as great if Casamino acids were provided, and similar results were obtained with seven other strains (B159, AW106, YT91, LC1, T81, J1, and YZ70). Specific activities of deamination (based on Casamino acids) of the eight strains ranged from 100 (strain JL1) to 325 (strain B159) nmol mg protein(-1) min(-1). None of the strains could utilize branched-chain amino acids as an energy source for growth, but specific activities of branched-chain amino acid deamination ranged from 15 to 65 nmol mg protein(-1) min(-1). All eight of the M. elsdenii strains grew well in the presence of 5 micro M monensin, and only two of the strains were strongly inhibited by 20 micro M monensin. On the basis of these results, it appears that M. elsdenii is deficient in peptidase activity and can utilize only a few amino acids. Some M. elsdenii strains produced ammonia and branched-chain volatile fatty acids nearly as fast as obligate amino acid-fermenting ruminal bacteria, but the extent of this production was at least fourfold lower. Because all of the strains could tolerate 5 micro M monensin, it is unlikely that this feed additive would significantly inhibit M. elsdenii in vivo.

Amino Acids↗

Effect of fermented feed on shedding of Enterobacteriaceae by fattening pigs.

Epidemiological studies showed that the use of fermented feed could significantly reduce Salmonella prevalence in pigs compared to the use of normal feed. Experimental challenge experiments with Salmonella have however never been conducted to reveal the efficacy of fermented feed in reducing Salmonella shedding and/or reducing the number of Salmonella-positive pigs. A longitudinal study was conducted to measure the effect of fermented feed, in particular of its components lactic acid and Lactobacillus plantarum, on gastrointestinal bacterial ecology (Salmonella, Enterobacteriaceae, lactobacilli, volatile fatty acids (VFAs), pH). Seeder pigs were used as a mode for Salmonella transmission within a pig herd. Bacteriological measurements were performed in faeces of the pigs. The results showed that fermented feed affected/reduced the Enterobacteriaceae population in faeces of the pigs. No differences were found in the number of positive pigs infected or in the number of shedding with Salmonella serovar Typhimurium fed fermented feed and between the normal feed group. S. serovar Goldcoast could not establish an infection in the seeder pigs in the fermented feed group as well as in the normal feed group. The pH of the faeces in the fermented feed groups was significantly higher than the pH of the faeces of the normal feed groups. The role of the undissociated form of the faecal VFAs on the significantly lower Enterobacteriaceae number in faeces of the pigs of the fermented feed groups could not be demonstrated because of the significant higher pH in the faeces of the pigs fed fermented feed.

Animal Feed↗

Intestinal absorption of end products from digestion of carbohydrates and proteins in the pig.

The kinetics of appearance of various nutrients in the portal vein during the postprandial period was studied in conscious pigs by means of a technique based on measurement of the porto-arterial differences in nutrient concentrations simultaneously with that of the portal blood flow rate. The rate and level of appearance of sugars in the portal vein varied with the carbohydrate ingested. It was very rapid after intake of glucose and sucrose, slower after that of maize starch and very slow after that of lactose. The absorption of the latter became very rapid again if it was hydrolysed prior to its ingestion. During absorption, some sugars (fructose or galactose) released from the corresponding sucrose and lactose, respectively during digestion, were partly metabolized into glucose by the enterocyte. The rate of absorption of amino acids released in the digestive tract varied according to the origin of the food ingested, i.e. it was more rapid after intake of wheat or fish proteins than after that of barley. In the case of barley the absorption rate of amino acids differed from that of glucose of the starch. The profile of the amino acid mixtures appearing in the portal vein during absorption differed a little from the profiles of those present in the ingested proteins in the case of essential amino acids and differed much in the case of non essential amino acids. Some essential amino acids (histidine, aromatic amino acids) appeared more rapidly and others more slowly, (lysine, sulphur amino acids, arginine). Because of transaminations, only small amounts of glutamic acid occurred in the portal vein whereas the amounts of alanine as compared to those ingested, were very large. The hierarchy of amino acid absorption was the same whatever the protein studied (fish, wheat, barley). The appearance in the portal vein of alpha-amino nitrogen from enzyme hydrolysates perfused through the duodenum was more rapid than after perfusion of a mixture of free amino acids. During digestion of diets containing 6% cellulose, large amounts of volatile fatty acids (about 2200 mmol/24 h, i.e. 2500 kJ/24h) and lactic acid (60-85 g/24 h) appeared in the portal vein.

Amino Acids↗

Caecal microflora and fermentation pattern in exclusively milk-fed young rabbits.

Thirty-two SPF New Zealand White rabbits of both sexes were used in this experiment. They were sequentially slaughtered at 22, 29, 35 and 42 days of age. The rabbits were only milked by their mother and by foster mothers (no access to solid food) throughout the experiment. They exhibited a caecal fermentation pattern that turned towards a proteolytic metabolic activity (high levels in NH3 and in branched-chain fatty acids and valeric acid: 17 mmol.L-1 and 1 mmol.L-1, on average, respectively). The absence of caecal cellulolytic microflora and the low concentration of the total volatile fatty acids (tVFA) could be explained by the lack of substrate that would have been brought by a solid feed. From day 29 onwards, the low tVFA (12.5 mmol.L-1) and high NH3 concentrations (16.5 mmol.L-1) explained the high pH value (6.8 on average). This could be considered as a pathological value in weaned rabbits, but in our case no clinical signs of diarrhoea were observed. The evolution of the colibacilli flora according to age was similar to that usually described and thus was not correlated with pH, tVFA or cellulolytic flora. The evolution of the colibacilli population (from 10(7) bact/g on day 22 to 10(3) on day 42) seemed dependent on ontogenic factors rather than on the composition of the caecal media (pH, etc.).

Aging↗

Effect of nisin on two cultures of rumen ciliates.

The effect of nisin (in the form of Nisaplin) was determined using two species of rumen ciliate protozoa in vitro, on their co-culture bacterial population, and volatile fatty acid concentration. Nisaplin did not affect the in vitro growth of Entodinium caudatum at concentrations of 50-400 mg/L during short-term treatment (5 d). Long-term application (30 d) of Nisaplin (100 mg/L) significantly decreased growth of the Epidinium ecaudatum forma caudatum et ecaudatum but not growth of E. caudatum. Nisaplin moderately supported the growth of E. caudatum after omission of wheat gluten (source of amino acids for protozoan growth). An inhibition of Gram-positive facultative anaerobic bacterial population in the protozoan cultures (lactobacilli, enterococci, staphylococci and amylolytic streptococci) was observed during long-term Nisaplin treatment. The concentration of volatile fatty acids significantly increased during the long-term Nisaplin treatment of both cultures. The propionate concentration in the mixture of volatile fatty acids was nearly twice higher on the account of the decreased concentration (from 74 to 63%) of acetate.

Animals↗

Effect of acid speciation on solid waste liquefaction in an anaerobic acid digester.

Laboratory scale experiments were carried out to evaluate and determine the suitable working conditions for the enhancement of volatile fatty acids (VFAs) production with the stabilization of solid pineapple waste (peel) simultaneously when operating two-phase anaerobic digestion. It was found that acid production, enhanced when the digester was operated at neutral pH. VFAs, could be obtained at about 50 g/kg-waste within 14 days when operating at pH 6.5-7.5 compared to 33 g/kg-waste for digester operated at pH 5.0-6.0 for 32 days. The total volatile solid reduction of 42-48% was obtained with a 51-57% volume reduction at neutral pH. The effect of undissociated acids on the activity of acetogenic bacteria was also studied. The inhibitory level of undissociated acids for acidogenesis was found to be about 2300, 650, and 120 mg/L at pH 5, 6, and 7, respectively. The results revealed that at low pH, undissociated acids are inhibitory. It was found that for efficient operation of two-phase anaerobic digestion, both VFAs concentration and pH should be controlled.

Acids↗

The role of pH in mesophilic anaerobic sludge solubilization.

The effect of pH on anaerobic solubilization of synthetic sludge (dog food) and domestic primary sludge was investigated and compared. Anaerobic solubilization was carried out in Continuously Stirred Anaerobic Reactors at mesophilic temperature (35 degrees C) and pH was fixed at 6.5 (pH-controlled). The aim of pH control in the reactors consisting of synthetic and primary sludge, was the evaluation of retardation in hydrolysis/acidogenesis at low pH values. Since synthetic and primary sludge have different biodegradation characteristics, the results were compared. In both sludges, acetic acid was the main Volatile Fatty Acids (VFAs) produced. Volatile Suspended Solid (VSS) reduction was found as 67% in about 20 days in the anaerobic digestion of synthetic sludge. whereas for the same interval VSS reduction could only be achieved by 32% in primary sludge at 35 degrees C in the pH-controlled reactors. When both types of sludges were used as substrates, the reactors removed VSS with a corresponding production of VFAs and Soluble Chemical Oxygen Demand (SCOD). However, in the pH-controlled reactors production of VFAs and SCOD was ceased after 5 days in primary sludge whereas VFAs and SCOD production continued after 5 days in synthetic sludge which indicated that hydrolysis and fermentation in the anaerobic solubilization reactors were not complete and continued longer. On the other hand, in the pH-controlled reactor of primary sludge, methanogenic phase could operate after 5 days of operation as hydrolysis/acidogenesis stopped.

Bacteria, Anaerobic↗