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[Acidity of the rumen liquid of calves after intra-ruminal administration of solutions for oral rehydration].

UNLABELLED: Investigations on the acidity of the rumen liquid after intraruminal administration of oral rehydration solutions (ORS) were carried out in 7 young calves. After voluntary intake of about one liter of milk replacer, three times a day, 1 liter of rehydration solution (A, B, C or D) was administered intraruminally by a stomach tube. Prior to every meal rumen fluid samples were aspirated for the determination of the pH, the concentrations of lactic acid and volatile fatty acids. Number of experiments: solution A: n = 2; B: n = 4; C: n = 2; D: n = 5. In 3 experiments, the pH of the rumen fluid was measured continuously for 4 hours using a special pH probe introduced intraruminally. After intraruminal administration of the oral rehydration solutions the pH values of the rumen fluid samples decreased from initial values of about 6.5 to values of about 5.0 or even lower in all calves. Simultaneously, the concentrations of the acids, mainly lactic acid, increased markedly, with some differences depending on the respective solution. The course of the pH measured continuously by means of the pH probe corresponded to the results obtained from the intermittent rumen fluid samples. CONCLUSIONS: The results confirm, that force-feeding of young calves with ORS as used in these experiments can induce ruminal acidosis and, therefore, should be avoided as far as possible.

Animals↗

[The effect of Rumensin on the metabolic profile of rumen fluid in feedlot cattle].

The effects were studied of sodium monensinate dosed 125 mg on the metabolic profile of rumen fluid. The experiment was conducted under current farming conditions in a pavilion cattle fattening house. The experiment, which lasted 367 days, comprised 985 bulls of the Bohemian Spotted breed. The feed ration was based on silage with an additive of dried poultry litter and concentrates. The additive was administered in concentrate mixture, 125 mg per head/day. After the start of the experiment want of appetite to sodium monensinate was observed. The animals took approximately four weeks to adapt completely to the additive. The following characteristics were investigated to study the metabolic profile of rumen fluid: actual acidity, total titration acidity, ammonia, total nitrogen, lactic acid, total volatile fatty acids, per cent acetic acid, per cent propionic acid, molar ratio acetic acid: propionic acid, per cent iso- and n-butyric acid, absolute number of infusoria and energy net yield of volatile fatty acids. Increased levels of the clinico-biochemical parameters of the metabolic profile of rumen fluid were found in lactic acid, propionic acid and energy net yield of volatile fatty acids. The per cent of propionic acid increased at some samplings by up to 116%. A drop was recorded in total nitrogen, per cent acetic acid, per cent butyric acid, molar ratio C2 : C3 and total number of infusoria. The decrease in the per cent of acetic acid ranged around 16% and the drop in butyric acid level amounted at some samplings up to 78%. No response to the additive was observed in the other characteristics of the metabolic profile of rumen fluid.

Acids↗

Rapid diagnosis of anaerobic empyema by direct gas-liquid chromatography of pleural fluid.

Sixty samples of pleural fluid from 52 patients were subjected to direct gas-liquid chromatographic studies, and results were correlated with findings from microbiologic cultures. Fourteen patients had anaerobic empyema, 22 had aerobic infections, and 16 had sterile pleural effusions. Multiple volatile fatty acids or succinic acid or both were found as markers of anaerobic infection in all but one instance. Aerobic infections and sterile pleural fluids were characterized by the absence of multiple volatile fatty acids or succinic acid. Infection with Bacteroides, when present, was characterized by a major product of succinic acid. One patient infected with Peptococcus magnus (which does not produce fatty acids or succinic acid) could not be diagnosed by gas-liquid chromatograms. Two patients without anaerobic bacteria in the pleural effusions but who had infections associated with Bacteroides fragilis outside the pleural space demonstrated succinic acid in the pleural fluid. With these exceptions, the presence of volatile fatty acids or succinic acid in pleural fluid was considered characteristic of anaerobic empyema. Direct gas-liquid chromatographic study of pleural fluids is, therefore, recommended as a routine procedure for rapid diagnosis of anaerobic empyema.

Adult↗

Effect of primary sludge fermentation products on mass balance for biological treatment.

Laboratory batch experiments were conducted at 20 degrees C to investigate the potential of primary sludge fermentation for the generation of readily biodegradable substrate and to evaluate the effect of fermentation products on mass balance for organic carbon, nitrogen and phosphorus, emphasizing COD fractionation. Fermentation converted between 18 to 30% of the initial volatile suspended solids in the sludge into soluble biodegradable COD. The volatile fatty fraction of the soluble COD was approximately 85% after the fermentation process. The average volatile fatty acid composition in fermentation involved 50% acetic acid, 33% propionic acid, 9% butyric acid and 8% valeric acid, indicating that the most important volatile fatty acid obtained during the biological fermentation process was acetate with more than half of total VFA concentration, which is one of the most important carbon sources for denitrification and biological nutrient removal processes. The recoverable fraction of the fermented sludge supernatant could potentially increase the readily biodegradable COD content of the primary effluent by 5%, together with a potential increase of the soluble nitrogen and phosphorus content by 2%.

Fatty Acids, Volatile↗

Rumen fermentation in vitro as influenced by long chain fatty acids.

Responses of rumen microbes to fatty acids were evaluated by production of total volatile fatty acid and ratio of acetate to propionate. Fermentations were under carbon dioxide for 20 h in 50-ml Erlenmeyer flasks in a Dubnoff metabolic shaking incubator. Flasks contained 20 ml medium, 1 ml reducing solution, 750 mg substrate (450 mg hay plus 300 mg grain), and varying amounts of long-chain fatty acids supplied as free acids, as calcium salts, or as triglycerides. They were inoculated with 5 ml rumen fluid obtained from a cow fed 3.6 kg grass hay, 2.3 kg grain, and .2 kg tallow daily. Volatile fatty acid production was decreased by long-chain fatty acids that contained less than 18 carbon atoms and by unsaturated long-chain fatty acids with 18 carbon atoms. Lauric acid decreased volatile fatty acid production by 69% and induced unusual acetate/propionate ratio (40:1). Stearic acid, however, did not affect volatile fatty acid production or acetate/propionate ratio. Within two series of long chain fatty acids (myristic, palmitic, stearic, arachidic; stearic, oleic, linolenic), melting point accounted for 93 to 95% of the variation of volatile fatty acid production and acetate/propionate. As calcium salts, long chain fatty acids caused small changes of fermentation. Our data support the proposition that hard fats and calcium salts of long-chain fatty acids do not interfere with ruminal fermentation.

Acetates↗

Metabolism of liquid whey fed to sheep.

Digestion was studied with three groups of four sheep each fed alfalfa hay. Treatments were whey only, whey plus water, and water only. Digestibilities of dry matter for the three rations were 71.3, 69.7, and 62.9%. Digestibilities of whey solids calculated by difference was 86.9% for whey only and 81.6% for whey plus water. Twelve sheep were in three trials to determine the intake of whey, water, and hay consumption. They were assigned to whey only, whey plus water, and water only. Hay, whey, and water consumption was 1.89, 8.73, and 0; 1.88, 7.62, and 1.21; and 2.05, 0, and 6.07 kg per animal daily. Three mature fistulated wether sheep were fed 6 liters of liquid whey daily. Three mature fistulated wether sheep uere fed 6 liters of liquid whey daily, and three similar sheep were fed water only. Rumen liquor samples were taken for analysis of volatile fatty acids at 0, 3, and 6 h after whey feeding on each of three alternate days. Alfalfa hay (1.2 kg/animal daily) was fed after the 6-h sampling. Molar percentage of acetic and valeric acids were lower and for butyric acid was higher for sheep fed whey. Propionic acid, acetate:propionate ratio, and total concentration were not affected significantly by whey feeding. Acetic acid decreased from 0 to 3 h, then partly returned to the prefeeding level at 6 h. The reverse was true for butyric acid. Valeric acid and total concentration of volatile fatty acids decreased from 0 to 6 h. Effects of day were unexplained.

Animal Feed↗

Fermentation of [14C]lactose in broiler chicks by cecal anaerobes.

Volatile fatty acids and lactic acid were products of [14C]lactose metabolism in 10-day-old broiler chicks. The [14C]lactose and unlabeled lactose were given to chicks in combination with cultures of cecal anaerobes (direct fed microbials) from adult broiler chickens. Aliquots of these cecal anaerobes had been used previously to reduce Salmonella typhimurium colonization in broiler chicks. In the current study, chicks were treated with anaerobes and anaerobes plus lactose on the day of hatch. Chicks given lactose plus anaerobes also received unlabeled lactose in the drinking water. Chicks in all treatment groups received [14C]lactose on Day 10 posthatch. After 5 h, most of the radiolabel was detected in the foregut and intestine. Significantly lower concentrations of [14C]lactic acid were detected in the serum and foregut of chicks given anaerobes and anaerobes plus lactose than in the similar samples from control chicks. Additionally, significantly less [14C]lactic acid was detected in the feces of chicks treated with anaerobes plus lactose than in feces from control chicks and chicks receiving only anaerobes. Significant differences in the total content of volatile fatty acids and lactic acid were detected in the serum, liver, intestine, and ceca of chicks treated with anaerobes or anaerobes plus lactose. These data indicate that cecal flora utilize lactose to produce volatile fatty acids and lactic acid.

Animals↗

Chemical changes in bovine colostrum preserved with formalin or by fermentation.

Fresh bovine colstrum from second and third milkings postpartum was inoculated with either a yogurt culture or Streptococcus lactis or preserved with .1% formalin in two separate experiments. There was a rapid decrease in pH of the fermented samples which corresponded to decreased lactose concentrations and increased lactate and titratable acidity. Titratable acidity in the fermented colostrums increased further after 10 days from the production of volatile fatty acids, but pH did not decrease. The pH of the yogurt fermentations in Experiment 2 decreased to 3.8, and the continued increase in titratable acidity was from lactic acid rather than volatile fatty acids. The lowest pH of all other fermentations was 4.1. The extent of metabolism of the formalin-preserved colostrum was much less than for the colostrum fermented by either culture.

Animals↗

[The effect of the early feeding of solid and bulk foods on the dynamic development of the digestive processes in calves ina large-scale breeding facility].

Digestive activities were studied in test calves (n = 12) in relation to age and feed intake. The calves were isolated after birth from adult cows. Since the 14th day of age, milk replacers were fed with concentrate feed mixture TG and alfalfa hay, i.e. the calves were in the period of plant feeding. The investigation lasted from the end of milk feeding (50th day) to the age of six months. Overall health condition was studied clinically. The characteristics of digestive activities studied in rumen fluid at weekly--monthly intervals were pH, ammonia content, total content of volatile fatty acids, content of particular volatile fatty acids, incidence and number of infusoria. At the age of 50 days, actual pH of rumen fluid was on the average 6.07 and it ranged from 5.84 to 6.76 in the next period. Ammonia content was the highest at the age of 50 days (21.49 mmol/l rumen fluid), then it dropped to 9.83 mmol/l rumen fluid at the age of 180 days. The average total concentration of volatile fatty acids (C2-C5) made 117.00 mmol/l during the first examination, it increased to 132.98 mmol/l at the age of 84 days, and it dropped to 94.16 mmol/l rumen fluid at the age of six months. A similar tendency was found in the volatile fatty acids. The infusoria were found out only at the age of 120 days--their numbers were 155 000 per ml; at the age of 180 days their numbers rose to 368 000 per ml rumen fluid. No disorders of health condition were recorded during the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Effects of dietary non-digestible oligosaccharides on microbial characteristics of ileal chyme and faeces in weaner pigs.

Fructooligosaccharides (FOS) and transgalactooligosaccharides (TOS), which are non-digestible oligosaccharides (NDO), were included at 10 and 40 g/kg in an NDO--free control diet at the expense of purified cellulose. Each of the 5 diets was fed to 4 weaner pigs and microbial characteristics of their ileal chyme and faeces were assessed. The NDO-pigs had lower ileal pH than the control pigs. Dietary NDO did not affect the ileal volatile fatty acid concentration, though FOS-pigs had a higher concentration of lactic acid and relatively more iso-valeric acid and less acetic acid than TOS-pigs. The NDO-pigs had lower ileal aerobic bacterial counts than the control pigs, whilst the FOS-pigs had a larger ileal anaerobic bacterial counts than the TOS-pigs. The NDO-pigs had an higher faecal pH and their faecal volatile fatty acid pool contained relatively more iso-butyric acid and iso-valeric acid than the control pigs. The TOS-pigs tended to have higher faecal anaerobic bacterial counts and had a smaller concentration of faecal volatile fatty acid than the FOS-pigs. We concluded that whilst effects at the faecal level may have been partly due to a reduced intake of cellulose, dietary NDO can exert precaecal prebiotic effects in weaner pigs.

Animals↗

Influence of ensiling temperature, simulated rainfall, and delayed sealing on fermentation characteristics and aerobic stability of corn silage.

The aim of this study was to determine how delayed silo sealing, high ensiling temperatures, and rainfall at harvest affect the fermentation and aerobic stability of corn silage. One-half of each of 4 replicated, 6 x 1.5 m plots of a corn hybrid was harvested at 35% dry matter (Dry), and each of the other halves was harvested after they were sprinkled with sufficient water to simulate 4 mm of rainfall (Wet). Six representative (2 kg) subsamples were taken from the Wet and Dry forage piles and ensiled immediately (Prompt). Three hours later, 6 additional representative (2 kg) samples were taken from each pile and ensiled (Delay). Half of the bags from each moisture x sealing time treatment combination were stored for 82 d in a 40 degrees C incubator (Hot) and the other half were stored in a 20 degrees C air-conditioned room (Cool). A 2 (moisture treatments) x 2 (sealing times) x 2 (ensiling temperatures) factorial design with 3 replicates per treatment was used for the study. Wetting the corn silage increased concentrations of NH(3)-N, ethanol, and acetic acid. Ensiling at 40 instead of 20 degrees C increased pH, in vitro digestibility, and concentrations of NH(3)-N, residual water-soluble carbohydrates and acid detergent insoluble crude protein. The higher ensiling temperature also reduced concentrations of neutral and acid detergent fiber and lactic and acetic acid. Delayed sealing reduced concentrations of NH(3)-N and total volatile fatty acids. Wetting, high temperature ensiling, and delayed sealing each reduced yeast counts slightly, and marginally (8 h) increased aerobic stability. Hot-Wet-Delay silages were more stable than other silages but had the lowest lactic to acetic acid ratio, and total volatile fatty acid concentration. This study indicates that the fermentation of corn silage is adversely affected by wet conditions at harvest and high ensiling temperatures, whereas delayed silo sealing for 3 h caused no adverse effects.

Acetic Acid↗

Effects of cellobiose and monensin on in vitro fermentation of organic acids by mixed ruminal bacteria.

The objective of this study was to determine the effects of cellobiose and monensin on the in vitro fermentation of organic acids (L-aspartate, fumarate, and DL-malate) by mixed ruminal bacteria. Ruminal fluid was collected from a steer fed 36.7 kg of forage and 4.5 kg of concentrate supplement once per day. Ruminal fluid was centrifuged to sediment feed particles and protozoa, and the resulting supernatant, which contained bacteria, was added (33%, vol/vol) to anaerobic media (500 ml). Incubations (n = 2) were performed in batch culture at 39 degrees C and sampled at 0, 2, 4, 6, 8, 12, and 24 h. Organic acids were added to achieve a final concentration of 7.5 mM. Cellobiose was added to obtain a final concentration of 5 mM, and monensin dissolved in ethanol was included at concentrations of 0 or 5 ppm. Addition of cellobiose to organic acid fermentations increased the rate of organic acid utilization by the mixed bacterial population. Total concentrations of volatile fatty acids were increased by the addition of cellobiose to all fermentations. A lag period (< or = 8 h) occurred in fermentations that were treated with monensin before organic acids were utilized. Total concentrations of volatile fatty acids were increased, and the acetate to propionate ratio was decreased, by monensin treatment. When cellobiose and monensin were added together, propionate production and organic acid utilization were increased. Both cellobiose and monensin affected the in vitro fermentation of organic acids by mixed ruminal bacteria by providing a carbon and energy source and by influencing electron disposal.

Acetates↗

[Possible interesting dietetic effects of lactulose as a feed additive in pig feed].

In reared piglets (n = 10), fattening pigs (n = 20) and sows (n = 8) the effects of lactulose as feed additive on the quality and composition of chyme and faeces were tested to find out potential dietetic advantages of its use. The lactulose concentration in the diets varied between 27-29 g/kg dry matter in piglets and fattening pigs and 55-140 g/kg dry matter in the diet of sows. In piglets and fattening pigs the lactulose intake did not alter the dry matter content and pH in faeces; in spite of the higher concentration lactulose did not result in diarrhea in sows or adult minipigs, there was only a slight decrease in the dry matter content of faeces. In contrast to findings in human beings only traces of lactulose reached the hind gut, although higher lactulose concentrations were measured at the end of the small intestine (in piglets: 10-40 g, in fattening pigs 40-70 g lactulose per kg dry matter of chyme). The oral intake of lactulose did not increase the concentration of lactic acid or volatile fatty acids in the content of the small intestine and did not reduce the pH in the chyme, but in the contents of cecum and colon ascendens higher concentrations of volatile fatty acids were measured (in piglets: significantly higher values, in fattening pigs slightly increased concentrations). In piglets as well as in fattening pigs lactulose resulted in reduced concentrations of ammonia in the content of small and large intestine, further more the concentrations of lipopolysaccharides were increased in the colonal chyme, when pigs ingested the lactulose containing diet. Due to the fact that only small amounts of lactulose reached the hind gut there are doubts, whether lactulose has comparable dietetic effects in pigs as known from human beings (for example laxative effects, forced elimination of bacteria like salmonella). The results obtained here in pigs underline the necessity of experimental studies in the target species (including parameters of digestive physiology, chemical/microbial composition of the chyme) before substances are used and recommended for dietetic purposes in feeding practice. In spite of generally adopted similarity of the alimentary tract in pigs and human beings there are eventually some important differences within the intestine, its digestive capacity or in reactions on ingested substances leading to unexpected results.

Animal Feed↗

Factors influencing rumen microbial growth rates and yields: effect of amino acid additions to a purified diet with nitrogen from urea.

Effects of isonitrogenous urea and amino acid additions upon microbial growth in rumen contents from a cow fed a purified diet in which urea was the sole nitrogen source were studied. Incorporation of amino acids into microbial cells, volatile fatty acids, and carbon dioxide was estimated. Rates of microbial growth, volatile fatty acid production, and effects of amino acids upon microbial nitrogen yields were highest right after feeding and decreased with time after feeding. Microbial growth and amounts of amino acids incorporated into microbial cells, volatile fatty acids and carbon dioxide were related closely to quantity of starch remaining in the rumen. High amounts of starch increased microbial protein synthesis from carbon-14 labeled amino acids and reduced amounts of amino acid fermentation. Estimated microbial protein yields per day were 326.0, 444.4, 497.3, and 527.3 g when 0, 15, 30, and 45 mg amino acid nitrogen replaced urea nitrogen during incubation. Respective values for microbial cells per mole estimated adenosine triphosphate were 15.2, 19.2, 21.0, and 24.5. Microbial cell yields per kg carbohydrate digested were 139.0, 189.5, 212.0, and 224.8 g for 0, 15, 30, and 45 mg amino acid nitrogen. Addition of small amounts of amino acids to a diet containing urea as the sole nitrogen source improved considerably rumen microbial protein yields.

Amino Acids↗

DETERMINATION OF STEAM-VOLATILE ORGANIC ACIDS IN FERMENTATION MEDIA BY GAS-LIQUID CHROMATOGRAPHY.

Five gas chromatographic liquid phases (25% Carbowax 20 M plus 4% H(3)PO(4), 17.5% dioctyl sebacate plus 7.5% sebacic acid, 17.5% dioctyl sebacate plus 7.5% docosanoic acid, 5% Tween 80, and 20% LAC-296 [poly (diethylene glycol adipate)] plus 2% H(3)PO(4)) were studied with respect to their utility in the separation and quantitation of steam-volatile organic acids commonly produced in fermentation. Optimal operating conditions and column stability for routine analysis were established. An Aerograph Hy-Fi gas chromatograph was used for all work, except the studies with Tween 80 in which an Aerograph A-90-C was employed. Chromatographic traces are presented of volatile fatty acid analyses with each of the liquid phases. Complete separation of all isomers of the fatty acids from C(2) to C(5) was accomplished by the Carbowax 20 M plus H(3)PO(4), dioctyl sebacate plus sebacic acid, and dioctyl sebacate plus docosanoic acid columns. The latter two liquid phases were extremely unstable and proved to be unsatisfactory for analysis of aqueous samples. A column of Carbowax 20 M + H(3)PO(4) separated steam-volatile organic acids completely. The volatile fatty acid isomers were separated by 5% Tween 80 somewhat less completely, and the peak shapes were not as sharp and symmetrical as that desired for good quantitative work. LAC-296 (20%) plus 2% H(3)PO(4) proved to be the most satisfactory of the liquid phases for routine analysis of deproteinated in vitro rumen fermentation media. The column has been used for routine analysis of rumen fermentation fluid and in vitro rumen incubation fluid. All the organic acids from C(2) to C(5), except isobutyric, could be quantitated with this column. Stability of the column with the aqueous solutions was extremely good. The standard deviation of the analysis of each volatile acid component in a fermentation fluid was less than 0.5 molar per cent. The short-chain organic acids (C(2) to C(5)) were shown to be extremely stable in aqueous solution for as long as 6 months after preparation for gas chromatographic analysis by protein precipitation with metaphosphoric acid-H(2)SO(4) and refrigeration at 4 C in stoppered tubes.

Animals↗

Effects of citrus pulp in high urea rations for steers.

Effects of pelleted and conventional citrus pulp as a replacement for corn, with soybean meal added to keep protein comparable, were tested in rations with 5% urea and 33.33% sugarcane bagasse for fistulated steers. Thus, all rations were low in readily fermented carbohydrates other than those of corn or citrus pulp. Evaluation criteria were concentrations of urea in blood and of pH, ammonia, and volatile fatty acids of rumen fluid. Citrus pulp for diets 1, 2, 3, and 4 was 0, 19, 38, or 55%. Rumen fluid and blood were sampled 1 h before and 2, 4, 7, and 12 h after feed was placed directly into the rumen. No differences between pelleted and conventional pulp or among time trends were significant except that for both forms rumen ammonia was lower with the two highest percents of citrus pulp. Addition of citrus pulp at 0, 19, 38, or 55% of the ration reduced rumen pH (6.85, 6.65, 6.61, 6.51). Blood urea and rumen ammonia decreased in steers fed 19, 38, or 55% pulp; thus, the acetic to propionic ratio was higher. Butyric acid changed only in the time trend. Total volatile fatty acid concentrations were higher at 19, 38, and 55% than at 0% pulp. They were higher at 38 and 55 than at 19%.

Ammonia↗

Production of PHA from starchy wastewater via organic acids.

Polyhydroxyalkanoate (PHA) was produced from a starchy wastewater in a two-step process of microbial acidogenesis and acid polymerization. The starchy organic waste was first digested in a thermophilic upflow anaerobic sludge blanket (UASB) reactor to form acetic (60-80%), propionic (10-30%) and butyric (5-40%) acids. The total volatile fatty acids reached 4000 mg l(-1) at a chemical oxygen demand (COD) loading rate of 25-35 g l(-1) day(-1). A carbon balance indicates that up to 43% of the organic carbon in the starchy waste went to the organic acids and the rest to biogas, volatile suspended solids and residual sludge accumulated in the reactor. The acid composition profile was affected by COD loading rate: a medium rate around 9 g l(-1) day(-1) gave a high propionic acid content (29% wt) and a high rate around 26 g l(-1) day(-1) led to a high butyric acid content (34% wt). The acids in the effluent solution after microfiltration were utilized and polymerized into PHA by bacterium Alcaligenes eutrophus in a second reactor. Fifty grams of PHA was produced from 100 g total organic carbon (TOC) utilized, a yield of 28% based on TOC, which is comparable with 55 g PHA per 100 g TOC of pure butyric and propionic acids used. PHA formation from individual acids was further investigated in a semi-batch reactor with three acid feeding rates. With a limited nitrogen source (80-100 mg NH(3) per liter), the active biomass of A. eutrophus, not including the accumulated PHA in cells, was maintained at a constant level (8-9 g l(-1)) while PHA content in the cell mass increased continuously in 45 h; 48% PHA with butyric acid and 53% PHA with propionic acid, respectively. Polyhydroxybutyrate was formed from butyric acid and poly(hydroxybutyrate-hydroxyvalerate) formed from propionic acid with 38% hydroxyvalerate.

Alcaligenes↗

Faecal steroids and colorectal cancer: the effect of lactulose on faecal bacterial metabolism in a continuous culture model of the large intestine.

The effect of lactulose on intestinal bacterial metabolism in two identical single-stage chemostats has been studied. The study was designed as a single stage crossover. Both cultures were inoculated and treated in an identical manner except that whilst one fermenter was subject to pH control the other was not and vice versa. Complex bacterial populations were maintained for 23 days and many of the metabolic functions carried out by the micro-organisms in vivo were demonstrated in vitro. The predominant organisms belonged to the genera Bacteroides, Bifidobacterium and Clostridium with abundant levels of anaerobic Gram-positive rods. The redox potential in each fermenter showed considerable variation with maximum values of below -300 eV being attained. An indication of the stability of the bacterial communities was the production of short-chain volatile fatty acids; during steady-state conditions the mean ratio of the major acids acetic, propionic and butyric being 3.90:0.69:1.00 and 3.65:0.76:1.00 in each fermenter, respectively. During steady-state conditions, 7 alpha-dehydroxylation of the primary bile acid chenodeoxycholic acid was maintained at a constant rate with lithocholic acid representing over 85% of total steroid. Addition of the soluble fibre lactulose to the cultures had a profound effect on many of the parameters tested. Without pH control the culture pH dropped to below 5.0 and this coincided with a substantial decrease in total anaerobes, especially Bacteroides spp., an increase in Lactobacillus spp. (concomitant with an increase in lactic acid), a decrease in the concentration of short-chain volatile fatty acids and a substantial decrease in 7 alpha-dehydroxylation of chenodeoxycholic acid. These results show that it is possible to maintain viable intestinal bacteria in vitro using a continuous culture chemostat and that the cultures respond to changing conditions as evinced by the inhibition of 7 alpha-dehydroxylation of chenodeoxycholic acid on addition of lactulose. This indicates that the model may be of further use in studying the modulation of secondary bile acid formation, a possible risk factor in colorectal carcinogenesis.

Amino Acids↗