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PubMed1998

Age effect on in vitro fermentation pattern and methane production in the caeca of chickens.

Age-dependent changes of the caecal fermentation pattern were studied in female chickens using in vitro batch incubation technique. Chickens were sequentially killed at the age of 1, 2, 3 and 4 months, their caecal contents added to a broth with starch and incubated at 39 degrees C for 20 h. Net productions of short-chain fatty acids (SCFA), succinate, ethanol, lactate, methane, hydrogen and ammonia were determined. Methanogenesis was absent in caeca of 1-month-old chickens. Production of methane started in the second month and doubled in the third month of age. The start of methanogenesis was accompanied by changes of the fermentation stoichiometry. The production of succinate ceased and that of ethanol decreased to less than one tenth. There were no major changes of the caecal fermentation pattern in the fourth month of age. The ammonia production increased in the second month, indicating increased deamination activity. No major shifts in SCFA molar composition dependent on age were found. Calculated hydrogen recoveries suggest a decrease of reductive acetogenesis until 3 months of age. It can thus be concluded that age and the onset of methane production affect the fermentation pattern in the caeca of chickens.

Aging↗
PubMed2000

Relationships between transit time in man and in vitro fermentation of dietary fiber by fecal bacteria.

OBJECTIVE: To assess the effects of drug-induced changes in mean transit time (MTT) on the activity of human fecal flora in vitro. METHODS: The activity of fecal flora was estimated by the ability of a fecal inoculum to ferment a substrate (beet fiber) in vitro in a batch system for 24 h. The inoculum was collected from 8 healthy volunteers studied during three 3-week randomized periods, who received a controlled diet alone (control period) or the same diet with either cisapride or loperamide. Cisapride and loperamide were adjusted in order to halve and double MTT measured during the control period. At the end of each period, the percentage disappearance of the initial added substrate and the concentration and the profile of short-chain fatty acids (SCFAs), were determined. RESULTS: In the control period, the pH of the inoculum and SCFA concentration were inversely related to MTT (P=0.0001). Individual SCFA production was also significantly related to MTT (P<0.01). Cisapride-reduced transit time was associated with a significant rise in the concentrations of total SCFAs (P<0.05), propionic and butyric acids (P<0.05) and the percentage substrate disappearance (P<0.05). Inverse relations were observed during the loperamide period. Moreover, MTT was inversely related to the percentage substrate disappearance (P<0.001), SCFA production (P<0.001) and butyrate production (P<0.0005). CONCLUSION: Changes in MTT alter bacterial activity and modify the bacterial pathways affecting the proportion of individual SCFAs. European Journal of Clinical Nutrition (2000) 54, 603-609

Adult↗
PubMed2000

In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria.

Anaerobic batch culture fermenters were used for a preliminary screening of the in vitro utilization by human gut microflora of dextran and novel oligodextrans (I, II and III) produced in the University of Reading (UK). Glucose and fructooligosaccharides (FOS) were used as reference carbohydrates. As expected, FOS acted as a good prebiotic in that it selectively increased numbers of bifidobacteria in the early stages of the fermentation. Dextran and oligodextrans each resulted in an enrichment of bifidobacteria in the batch cultures, with high levels of persistence up to 48 h. They also produced elevated levels of butyrate ranging from 5 to 14.85 mmol/l. To more effectively simulate conditions that prevail in different regions of the large intestine, a three-stage continuous culture cascade system was used to study further the fermentation of dextran, a low-molecular-mass oligodextran (IV) and maltodextrin. Oligodextran IV was shown to be the best substrate for bifidobacteria and lactobacilli with steady-state populations of bifidobacteria and lactobacilli being higher in all three vessels of the gut model than the respective populations resulting from dextran and maltodextrin. A maximum difference of 1.9 log was observed in vessel 1 for both bifidobacteria and lactobacilli in the case of dextran fermentation, with 1.4 log and 0.8 log in vessel 3 were the maximum differences for bifidobacteria and lactobacilli when maltodextrin was used as the carbohydrate source. Moreover, dextran and oligodextran appeared to stimulate butyrate production, with a maximum production up to 25.39 mmol/l in vessel 3 when fermenting dextran, followed by 21.70 mmol/l in the case of oligodextran IV and only 12.64 mmol/l in the case of maltodextrin.

Bifidobacterium↗
PubMed1980

In vitro fermentation of feces from normal and chronically diarrheal horses.

Feces from 13 healthy horses and 8 horses with chronic diarrhea were subjected to an in vetro fermentation procedure that had been developed for rumen fluid. Fermentations were conducted over 6 hours in a closed system, with and without an essential amino acid (EAA) mixture being added to the basic starch-buffer medium. The addition of EAA caused no significant difference in results of fermentation of feces from healthy horses. For diarrheic animals, there was a significant (P less than 0.01) increase in gas and total volatile fatty acids production whether EAA were present or not, and alpha-amino nitrogen was utilized in significantly (P less than 0.01) greater amounts only if EAA was present. Fermentations were repeated on feces from five of the eight diarrheal horses after they had been treated with oral iodochlorhydroxyquin for 1 week, and had shown desirable clinical response. A significant difference was not shown between pre- and posttreatment fermentations, except for decreased butyrate production. The results are consistent with the hypothesis that chronic equine diarrhea is primarily a colonic disease and indicates that colonic maldigestion may, in part, be responsible for excess fecal water.

Amino Acids, Essential↗
PubMed2004

In vitro fermentation characteristics of a mixture of Raftilose and guar gum by human faecal bacteria.

BACKGROUND: The therapeutic effects of indigestible carbohydrates in the human colon are well known. Most fermentation studies have examined only single carbohydrates. Considering the idiosyncratic actions of individual carbohydrates and the dose required, it is unlikely that any one carbohydrate will provide ideal physiological actions without having some undesirable effects. Little is known of how mixing non-digestible carbohydrates affects their fermentation. This necessitates fermentation studies using different carbohydrates in mixtures. AIM OF THE STUDY: The aim of this study was to test the effect of mixing Raftilose (R) and guar gum (G) on short chain fatty acid (SCFA) production in in vitro cultures of human faecal bacteria. METHOD: The fermentation of the individual carbohydrates (10 mg/ml) was compared with that of a 50:50 mixture in anaerobic in vitro cultures of human faeces. RESULTS: Cultures of R/G mixtures produced significantly more n-butyrate than 100 mg G alone at 8 and 24 hours (p < 0.02). There was no significant difference in the production of n-butyrate between cultures of 100 mg R and the R/G mixture at 8 and 24 hours. R (100 mg) produced a propionic/butyric (p/b) acid ratio of 1.18 compared with 3.88 for 100 mg G, whereas R/G mixture produced a p/b ratio of 2.01. CONCLUSION: The fermentation of R/G mixture was different compared with 100 mg of R or G alone. There was no loss of n-butyrate in the culture containing R/G (50 mg of each) mixture compared with the 100 mg R culture.

Adult↗
PubMed1998

The effect of in vitro fermentation on specific gravity and sedimentation measurements of forage particles.

Dry matter degradability (DMD), gas production (GP), functional specific gravity (FSG), volume of gas associated (GA), water-holding capacity (WHC), and sedimentation measurements of orchard-grass (OG) and alfalfa (AA) hays (ground through a 8-mm screen) were studied before and after in vitro incubation with ruminal fluid for 2, 4, 8, 24, 48, and 72 h. The DMD was higher for AA than for OG (P < .001), but GP did not differ. The FSG of unfermented OG and AA was .59 and .73, respectively (P < .01). During fermentation, the FSG of OG increased more than did that of AA (from .93 to 1.39 for OG and from .97 to 1.27 for AA after 2 and 72 h, respectively), and GA decreased more rapidly (from .94 to -.04 mL/g DM and from .74 to .15 mL/g DM, respectively). The DMD was positively correlated with FSG (r = .83; P < .001) and, therefore, negatively with GA (r = -.72; P < .01). The WHC increased similarly in the two forages with fermentation time. Unfermented and fermented samples were incubated in sedimentation columns filled with distilled water for 19, 37, 75, 150, and 300 s. After 300 s of sedimentation time, the unfermented AA and OG samples tended to float (91.1 and 72.7% of DM, respectively). In contrast, fermented samples tended to sediment (90.7 and 90.9% of DM, respectively). There were only small effects of forage species and fermentation time on sedimentation tendency. Correlations between sedimentation measurements and DMD and FSG were not significant, with the only exception of DM recovered in the lower section of sedimentation columns after 75 s, which was particularly correlated with DMD (P < .01) and FSG (P < .05). The results suggest that degradation rate of fibrous particles is related to changes in FSG and GA and, therefore, could influence ruminal transit. However, FSG was unable to predict accurately the sedimentation behavior of samples.

Animal Feed↗
PubMed1998

The use of cumulative gas and volatile fatty acid production to predict in vitro fermentation kinetics of Italian ryegrass leaf cell walls and contents at various time intervals.

Differences between the fermentation characteristics of cell contents (CC) and protease-treated cell walls (CW) of young leaves of Italian ryegrass (Lolium multiflorum Lam.) cultivar Multimo (tetraploid), were studied in vitro. Gas and volatile fatty acid (VFA) production rates were measured at regular intervals, as was the degradability of organic matter (OM) of CW. The measured VFA were used to predict the gas production and fermentable OM using stoichiometric calculations. For CW the volume and kinetics of measured gas production were the same as those predicted from the VFA formed. In contrast, the measured gas production for CC was consistently less than predicted, indicating that the stoichiometric equations were not valid for rapidly fermenting substrates. For both CC and CW, the relative rate of acetic acid production levelled off more slowly than for other VFA, resulting in an increasing gas yield (in ml/g fermentable OM) after 12 (CW)-24 (CC) h incubation. Consequently, the fermentation of OM was not linearly related to gas production kinetics. For CW, the kinetics of decline of degradable OM and fermentable OM were the same, after correction for a constant 'lost fraction' of degradable OM of 205 g/kg OM. This work indicates the value of detailed studies of fermentation processes to evaluate herbage quality. In particular, the role of CC and the difference between degradation and fermentation require further attention.

Cell Wall↗
PubMed2005

A novel galactooligosaccharide mixture increases the bifidobacterial population numbers in a continuous in vitro fermentation system and in the proximal colonic contents of pigs in vivo.

Prebiotics are nondigestible food ingredients that encourage proliferation of selected groups of the colonic microflora, thereby altering the composition toward a more beneficial community. In the present study, the prebiotic potential of a novel galactooligosaccharide (GOS) mixture, produced by the activity of galactosyltransferases from Bifidobacterium bifidum 41171 on lactose, was assessed in vitro and in a parallel continuous randomized pig trial. In situ fluorescent hybridization with 16S rRNA-targeted probes was used to investigate changes in total bacteria, bifidobacteria, lactobacilli, bacteroides, and Clostridium histolyticum group in response to supplementing the novel GOS mixture. In a 3-stage continuous culture system, the bifidobacterial numbers for the first 2 vessels, which represented the proximal and traverse colon, increased (P < 0.05) after the addition of the oligosaccharide mixture. In addition, the oligosaccharide mixture strongly inhibited the attachment of enterohepatic Escherichia coli (P < 0.01) and Salmonella enterica serotype Typhimurium (P < 0.01) to HT29 cells. Addition of the novel mixture at 4% (wt:wt) to a commercial diet increased the density of bifidobacteria (P < 0.001) and the acetate concentration (P < 0.001), and decreased the pH (P < 0.001) compared with the control diet and the control diet supplemented with inulin, suggesting a great prebiotic potential for the novel oligosaccharide mixture.

Animals↗
PubMed2005

Effect of insulin on in vitro fermentation activity of microrganism community of rumen ciliate Entodinium caudatum culture.

The influence of insulin (17.4 nmol l-1) on total gas and methane production, the concentration of total and individual fatty acids and dry matter degradability was investigated in the rumen ciliate culture of Entodinium caudatum. The experimental groups consisted of control group (without insulin) and two groups with insulin application--single shot and long-term application (over 30 days). Fermentation activity of each experimental group was observed on two subgroups: whole protozoan culture (protozoa plus bacteria) and bacterial fraction (bacteria without protozoa). Long-term application of insulin significantly increased methane production and DM degradability in the whole protozoan culture. Total VFA concentration was significantly increased by long-term as well as single-dose application of insulin (by 255% and 158%, respectively). The growth of the protozoa was not influenced by insulin treatments. It can be concluded that the fermentation activity of the community of the rumen ciliate Entodinium caudatum culture was marked stimulated by application of insulin.

Animals↗
PubMed2006

In vitro fermentation studies for selection and evaluation of Bacillus strains as starter cultures for the production of okpehe, a traditional African fermented condiment.

Selected Bacillus and Enterococcus strains, isolated from traditional okpehe fermentations, were studied for their suitability as starter cultures in laboratory-scale fermentations of Prosopis africana seeds for the production of okpehe, a traditional fermented vegetable product of Nigeria. The strains were selected on the basis of highest proteolytic activity, as determined with the APIZYM (BioMerieux) test. The choice of starter strains was narrowed to Bacillus subtilis strains BFE 5301 and BFE 5372. These were determined as the best starter combination because of rapid growth, high amylolytic and proteolytic activities, high levels of polyglutamic acid production by strain BFE 5372, as well as bacteriocin production by strain BFE 5301. Other mixed culture fermentations did not yield sensorically acceptable products. Although a monoculture fermentation, using only B. subtilis strain BFE 5372, produced okpehe with very good sensory characteristics, the growth of B. cereus could be detected after 48 h fermentation, indicating that this starter did not sufficiently contribute to product safety. Mixed culture fermentation with the combination of bacteriocin-producing starter B. subtilis BFE 5301 and the non-bacteriocin-producing B. subtilis BFE 5372, produced a product with good sensory characteristics, in which growth of B. cereus was delayed. The bacteriocin produced by B. subtilis strain BFE 5301 was identified as subtilisin, using subtilisin-specific primers and PCR amplification of the subtilisin gene. The bacteriocin was heat-stable at 100 degrees C for 10 min and exhibited highest activity at pH values lower or equal to pH 6.0. The bacteriocin was sensitive to the proteolytic enzymes trypsin and alpha-chymotrypsin at concentrations of 10 mg/ml.

Bacillus subtilis↗
PubMed1994

Effects of 3 chemical treatments on in vitro fermentation of rice straw by mixed rumen microbes in the presence or absence of anaerobic rumen fungi.

Rice straw (Rs) was treated by a sodium chlorite/acetic acid mixture (Sct), ammoniation (At) and alkaline hydrogen peroxide (Athp) methods. The objective was to compare their degradation and fermentation products in the presence or suppression of anaerobic fungi. Significant differences (P < 0.01) in degradation of straws were observed during all periods of incubation with Sct having the highest digestibilities during the 48-h and 72-h incubations. The degradation of Sct straw was highest in both of the incubations with whole rumen fluid (WRF) and WRF plus cycloheximide. In sacco digestion followed the order of Sct > At > Rs > Athp. Suppressing fungal activity with cycloheximide resulted in a decrease in dry matter degradation, with concomitant decrease in total volatile fatty-acid concentration. While the suppression of fungal activity with cycloheximide depressed acetate and butyrate production, it favored an increase in propionate production.

Acetates↗
PubMed2006

In vitro fermentation of sugar beet arabinan and arabino-oligosaccharides by the human gut microflora.

AIMS: To determine the fermentation profiles by human gut bacteria of arabino-oligosaccharides of varying degree of polymerization. MATERIALS AND METHODS: Sugar beet arabinan was hydrolyzed with a commercial pectinase and eight fractions, of varying molecular weight, were isolated by gel-filtration chromatography. Hydrolysis fractions, arabinose, arabinan and fructo-oligosaccharides were fermented anaerobically by gut bacteria. Total bacteria, bifidobacteria, bacteroides, lactobacilli and the Clostridium perfringens/histolyticum sub. grp. were enumerated using fluorescent in situ hybridization. RESULTS: Bifidobacteria were stimulated to different extents depending on molecular weight, i.e. maximum increase in bifidobacteria after 48 h was seen on the lower molecular weight fractions. Lactobacilli fluctuated depending on the initial inoculum levels. Bacteroides numbers varied according to fraction; arabinan, arabinose and higher oligosaccharides (degree of polymerization, dp > 8) resulted in significant increases at 24 h. Only carbohydrate mixtures with dp of 1-2 resulted in significant increases at 48 h (log 8.77 +/- 0.23). Clostridia decreased on all substrates. CONCLUSIONS: Arabino-oligosaccharides can be considered as potential prebiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: Arabinan is widely available as it is a component of sugar beet pulp, a co-product from the sugar beet industry. Generation of prebiotic functionality from arabinan would represent significant added value to a renewable resource.

Arabinose↗
PubMed2006

In vitro fermentation of breast milk oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri.

It has been proposed that human milk oligosaccharides (HMO) function as a prebiotic for bifidobacteria, yet this activity has not been adequately investigated. In this study, Bifidobacterium infantis was shown to ferment purified HMO as a sole carbon source, while another gut commensal, Lactobacillus gasseri, did not ferment HMO. Our results support the hypothesis that HMO selectively amplify bacterial populations in the infant intestine.

Bifidobacterium↗
PubMed1978

Maintenance of a certain rumen protozoal population in a continuous in vitro fermentation system.

A continuous culture system suitable for maintaining certain rumen protozoa was developed by modifying the procedure of Walter and Pilgrim (R.A. Weller and A.F. Pilgrim, Br. J. Nutr. 32:341-351, 1974) to include a dialyzing system. The concentration of ciliate protozoa, the pH value, and concentrations of volatile fatty acid and ammonia-N could be maintained within normal rumen limits for more than 15 days by appropriate choice of mechanical agitation and of the amount of substrate and physical form of the substrate bags. The average concentration of protozoa in the free fluid around the substrate was about 10% that in the fluid squeezed from solid digesta residues. More than 10(6) protozoa per ml was present in the fresh substrates only 2 h after supplying the substrate. These facts suggest that sequestration of the protozoa among particulate digesta is an important factor in maintaining the concentration of protozoa.

Animals↗
PubMed1994

Effects of beet pulp and animal by-products on milk yield and in vitro fermentation by rumen microorganisms.

Forty-six Holstein cows (30 primiparous) were assigned to one of four dietary treatments arranged as a 2 x 2 factorial experiment during wk 4 to 17 of lactation. Main effects were corn versus dried sugar beet pulp and soybean meal versus animal by-product meal (mixture of meat and bone meal, feather meal, and blood meal). Beet pulp replaced half of the corn at 15% of dietary DM. Diet DM (mean of four treatments) contained 18% alfalfa pellets, 17.4% alfalfa hay, 17.2% corn silage, and 47.1% concentrate. Milk yield did not differ among treatments (mean 32.0 kg/d). Dry matter intake, milk CP percentage, and milk CP yield decreased 5.6, 3.7 and 5.2%, respectively, but milk fat percentage increased 4.7% when beet pulp replaced corn. Animal by-products did not affect DMI or milk fat, but milk CP percentage decreased 3.0%. The same diets were evaluated in a continuous culture system. Fungal extract (Aspergillus oryzae), added as the third treatment, had little effect on fermentation. Digestion of DM, OM, NDF, and ADF were not affected by dietary treatments. Molar proportion of acetate was greater when corn was replaced by beet pulp. Although flow of NAN from fermenters increased by 3.2% with beet pulp and 3.1% with animal by-products, milk CP percentages decreased.

Animal Feed↗