Liquid Chromatographic Procedure for Fermentation Product Analysis in the Identification of Anaerobic Bacteria

Ehrlich, Garry G.; Goerlitz, Donald F.; Bourell, James H.; Eisen, Grant V.; Godsy, E. Michael

doi:10.1128/aem.42.5.878-885.1981

Cited by 114 publications

(34 citation statements)

References 7 publications

(6 reference statements)

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“…Samples were fixed with Hayem solution (mg mL −1 : HgCl 2 , 2.5; Na 2 SO 4 , 25; NaCl, 5.0) for bacterial counting and with 1:10 diluted formalin (40 w/v of water) for protozoal counting. Short‐chain fatty acid (SCFA) analyses in fermentation fluid were determined25 using a HPLC (LaChrom, L‐7000 series; Hitachi Ltd, Tokyo, Japan) to separate C 2 (acetate), C 3 (propionate), C 4 (butyrate), isoC 4 , C 5 (valerate) and isoC 5 .…”

Section: Methodsmentioning

confidence: 99%

Nutrient and energy content, in vitro ruminal fermentation characteristics and methanogenic potential of alpine forage plant species during early summer

Jayanegara

Marquardt

Kreuzer

et al. 2011

J. Sci. Food Agric.

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Section: Methodsmentioning

confidence: 99%

Nutrient and energy content, in vitro ruminal fermentation characteristics and methanogenic potential of alpine forage plant species during early summer

Jayanegara

Marquardt

Kreuzer

et al. 2011

J. Sci. Food Agric.

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“…After the frozen samples for VFA analysis were thawed, a 1-mL aliquot of the medium was centrifuged at approximately 2,000 × g for 5 min, and 360 mL of the supernatant was transferred to a microcentrifuge tube containing 40 mL of 50 mM H 2 SO 4 . After mixing and standing at room temperature for 10 min, the centrifugation was repeated and the supernatant was analyzed for VFA by the HPLC method of Ehrlich et al (1981) using an HPX-87H column (7.8 × 300 mm) at 30°C, isocratic elution with 5 mM H 2 SO 4 , and UV detection at 210 nm. A mixture of acetic, propionic, isobutyric, and butyric acids was included as a calibration standard in all analyses.…”

Section: Methodsmentioning

confidence: 99%

Effect of maturity on digestion kinetics of water-soluble and water-insoluble fractions of alfalfa and brome hay.

Stefanon

Pell

Schofield

1996

Journal of Animal Science

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Alfalfa and bromegrass, each harvested at five different stages of maturity, were separated into water-insoluble and -soluble fractions. The NDF concentrations ranged from 19 to 43% for alfalfa and from 42 to 58% for brome. The rates of digestion, by mixed ruminal microflora, of the unfractionated forage and of the water-insoluble and -soluble fractions were measured in vitro using pressure sensors to monitor gas production. Both forages showed the expected decline in fiber digestibility with increasing maturity. A dual-pool logistic model gave pool sizes, specific rates, and a single lag time for both the faster- and slower-digesting fractions. The main difference between alfalfa and brome was in the soluble pool. This pool produced approximately 40% of the total gas in alfalfa, 25% in brome. The specific digestion rates of the brome soluble pool were approximately 50% higher than those for alfalfa. Net VFA production showed a somewhat higher acetate: propionate ratio for brome (3.2) compared with alfalfa (2.2), but there was little change with increasing maturity within a given forage. Gas production curves for the unfractionated forage showed a 0 to 10% positive deviation from curves created by adding data from separate digestion of the insoluble and soluble forage fractions. Gas measurements offer a promising approach to the study of the water-soluble extracts of forages and the interaction of the soluble- and insoluble-fractions during fermentation.

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“…Other analytical methods. Soluble fermentation products were determined by high-pressure liquid chromatographic procedures (6). We used an Aminex ion-exclusion column (HPX-87H; Bio-Rad Laboratories, Richmond, Calif.) at 35°C and eluted with 0.013 N H2SO4 (0.55 ml/min; 77 kg of F per cm2).…”

Section: Methodsmentioning

confidence: 99%

Acetate production from hydrogen and [13C]carbon dioxide by the microflora of human feces

Lajoie

Bank

Miller

et al. 1988

Appl Environ Microbiol

130

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Fecal suspensions from humans were incubated with 13Co2 and H2. The suspensions were from subjects who harbored 108 and 1010 methanogens per g (dry weight) of feces, respectively, and from a subject who did not harbor methanogens. Quantitative nuclear magnetic resonance spectroscopy showed that acetate labeled in both the methyl and carboxyl groups was formed by suspensions from the subject without methanogens and the subject with the lower concentrations of methanogens. The amounts of labeled acetate formed were in agreement with the amounts expected based on measurements of H2 utilization. No labeled acetate was formed by suspensions from the subject with the higher concentrations of methanogens, and essentially all of the H2 used was accounted for by CH4 production. Suspensions from the subject with lower concentrations of methanogens produced both methane and acetate from H2 and CO2. The results indicate that reduction of CO2 to acetate may be a major pathway for microbial production of acetate in the human colon except when very high concentrations of methanogens (ca. 1010 per g [dry weight] of feces) are present. Double-labeled acetate was also formed from H2 and "3CO2 by fecal suspensions from nonmethanogenic and moderately methanogenic rats.

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Liquid Chromatographic Procedure for Fermentation Product Analysis in the Identification of Anaerobic Bacteria

Cited by 114 publications

References 7 publications

Nutrient and energy content, in vitro ruminal fermentation characteristics and methanogenic potential of alpine forage plant species during early summer

Nutrient and energy content, in vitro ruminal fermentation characteristics and methanogenic potential of alpine forage plant species during early summer

Effect of maturity on digestion kinetics of water-soluble and water-insoluble fractions of alfalfa and brome hay.

Acetate production from hydrogen and [13C]carbon dioxide by the microflora of human feces

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