Vitamin Requirements of Several Cellulolytic Rumen Bacteria

Abstract

Four strains of cellulolytic bacteria recently isolated from in vitro rumen fermentations were used in this study. Nine water-soluble vitamins were tested in singledeletion and single-addition plus biotin experiments, each with and without charcoalextracted casein hydrolysate. Bacteroides succinogenes A3C and B21a required only biotin under the above experimental conditions. Ruminococcus flavefaciens B34b showed an absolute requirement for biotin and was stimulated by p-aminobenzoic acid (PABA) in the single-deletion experiments. In the single-addition plus biotin experiments, PABA and, to a lesser extent, vitamin B12 appeared to be required for maximal growth. The presence or absence of casein hydrolysate did not affect the vitamin requirements for the aforementioned three strains. In the single-deletion experiments, R. flavefaciens Cla showed an absolute requirement for biotin and, when casein hydrolysate was omitted, for B12. When casein hydrolysate was present, no requirement for B12 could be observed. In the single-addition experiments where the basal medium contained biotin and casein hydrolysate or B12 , PABA was required for maximal growth; however, the single deletion of PABA caused only slight retardation of growth. Investigation of the B12 or casein hydrolysate requirement of Cla revealed that a mixture of purified amino acids simulating casein hydrolysate satisfied this requirement. Subsequent work indicated that this requirement could be satisfied by the amino acid methionine.

In situidentification of carboxymethyl cellulose-digesting bacteria in the rumen of cattle fed alfalfa or triticale

“…Filter paper was used to cultivate R. albus, R. flavefaciens, and F. succinogenes for CMCase staining. B. fibrisolvens (OR79) was cultured as described by Scott & Dehority (1965). Cells taken at the end of exponential growth phase were used.…”

Interspecies H2 transfer in cellulose degradation between fibrolytic bacteria and H2-utilizing microorganisms from the human colon

“…All microorganisms were grown under strict anaerobic conditions according to Hungate [8]. The medium used to grow cellulolytic and methanogenic strains was that described by Lowe et al [9] and modi¢ed in our laboratory by addition of 10 ml l 31 of vitamin solution [10], no addition of coenzyme M solution and replacement of trace elements and volatile fatty acid solutions according to Balch et al [10] and Scott and Dehority [11], respectively. The sole carbon source was either Whatman No.…”

A method for the selective enumeration and isolation of ruminal Lactobacillus and Streptococcus

“…and Lactobacillus spp., enabling the isolation and enumeration of these specific populations. (Scott and Dehority 1965), supplemented with 5% (v/v) clarified rumen fluid, 0·3% (w/v) glucose, 0·3% (w/v) cellobiose and 0·6% (w/v) starch, was used as total count medium and incubated in an anaerobic chamber with a CO 2 : H 2 (90% : 10%) atmosphere. The clarified rumen fluid was prepared by straining ruminal fluid through two layers of cheese cloth, autoclaving the resultant fluid and then centrifuging the fluid at 6000 g to remove particulate matter.…”