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B A Dehority

Publications and source records attributed to B A Dehority.

At least 19 recordsLinked to original sources
PubMed2003

Technical note: Specific PCR amplification of protozoal 18S rDNA sequences from DNA extracted from ruminal samples of cows.

A protozoa-specific primer (P-SSU-342f) was designed and paired with a eukarya-specific primer to amplify a 1,360-bp fragment of DNA encoding protozoal small subunit (SSU) ribosomal RNA from ruminal fluid of cows fed a mixed forage:grain diet or alfalfa hay. Sequencing of clones showed that P-SSU-342f is specific to ruminal protozoa and, with slight modifications, the primer will be useful for ecological studies of ruminal protozoa.

Animals↗
PubMed2001

Effect of feeding frequency on bacterial and fungal concentrations, pH, and other parameters in the rumen.

Three sheep were fed a pelleted high-roughage diet either once, 6, or 24 times per day in a 3 x 3 Latin square trial. During each 21-d period, 14 d were allowed for adaptation followed by a 7-d collection period, in which samples for microbial counts were taken on d 1 and 5 and several rumen parameters were measured on d 2 and 6. Bacterial concentrations were not different between feeding frequencies on the first sampling day but were higher (P < 0.05) on the second sampling day when the sheep were fed 24 times a day. Fungal concentrations were not different among feeding frequencies on either sampling day. No effects of feeding frequency were observed for the concentration of cellulolytic bacteria or fungi. On d 2, ruminal volume was larger (P < 0.05) with six feedings than with one feeding and fluid volume turnover was greater (P < 0.05) when sheep were fed 24 times per day. Rumen pH values were higher (P < 0.01) on both d 1 and 5 when the sheep were fed once a day and the percentage of rumen dry matter was highest (P < 0.02) with 24 feedings. These findings would suggest that if the same amount of a given diet is fed daily, the number of feedings does not markedly affect microbial concentrations, rumen volume, or liquid turnover time.

Adaptation, Physiological↗
PubMed2001

In vitro growth and starch digestion by Entodinium exiguum as influenced by the presence or absence of live bacteria.

In a preliminary study, the addition of antibiotics was shown to reduce bacterial concentrations in Entodinium exiguum cultures by more than 99% in 4 h, whereas the protozoal population was apparently unaffected. Using this procedure, the growth and amylolytic capability of Entodinium exiguum, in the presence or absence of live bacteria, was studied in vitro. Treatments for Trial 1 were protozoa plus antibiotics (PA), PA plus autoclaved bacteria (PAB), protozoa plus living bacteria (PLB), and only bacteria (BAC). Autoclaved or non-autoclaved cornstarch was used as an energy source. Treatment main effects were as follows: higher concentration of E. exiguum in PLB than in PA or PAB at 24 and 48 h (P < 0.01); PA and PAB were not different (P > 0.05); concentrations of E. exiguum higher in autoclaved cornstarch at 12 h (P < 0.05) but lower than in non-autoclaved cornstarch at 24 and 48 h (P < 0.01); and starch digestion in PLB was higher than in PA and PAB at all time periods, but only greater than BAC up to 24 h (P < 0.01). In Trial 2, only treatments PA, PLB, and BAC were tested. Rice starch and cornstarch were used as substrates. With rice starch, growth was higher in PLB than in PA at 24 and 48 h (P < 0.05). Starch digestion started earlier in PLB with rice starch (P < 0.05) but was complete for both substrates after 24 h. Up to 12 h (autoclaved cornstarch and rice starch) and 24 h (non-autoclaved cornstarch and cornstarch), the sum of digestion by bacteria and protozoa did not equal the extent of digestion in PLB, suggesting some kind of synergism. Total extent of digestion with protozoa was similar between the two sources; however, bacteria digested rice starch faster and to a greater extent than cornstarch. Approximate lag times with rice starch, autoclaved cornstarch, and non-autoclaved cornstarch were 6, 3, and 12 h for bacteria and < 6, 3, and 9 h for protozoa, respectively. Rate of digestion for non-autoclaved cornstarch was similar for bacteria and protozoa, whereas the rate of bacterial digestion was much faster with the other two substrates (autoclaved cornstarch and rice starch).

Animals↗
PubMed2000

Antibiosis between ruminal bacteria and ruminal fungi.

Cellulose digestion, bacterial numbers, and fungal numbers were monitored over time in vitro by using a purified cellulose medium with and without antibiotics (penicillin and streptomycin). All fermentations were inoculated with a 1:10 dilution of whole rumen contents (WRC). Without antibiotics, cellulose digestion was higher (P < 0.01) at 24, 30, 48, and 72 h; fungi had almost disappeared by 24 h, while bacterial concentrations increased over 100-fold in 24 h and then decreased gradually up to 72 h. In those fermentations with added antibiotics, fungal concentrations increased 4-fold by 30 h and up to 42-fold at 72 h; bacterial concentrations were markedly reduced by 24 h and remained low through 72 h. Similar results were obtained with ground alfalfa as a substrate. In further studies, the in vitro fermentation of purified cellulose without antibiotics was stopped after 18 to 20 h, and the microbial population was killed by autoclaving. Antibiotics were added to half of the tubes, and all tubes were reinoculated with WRC. After 72 h, extensive cellulose digestion had occurred in those tubes without antibiotics, as compared to very low cellulose digestion with added antibiotics. The extent of this inhibition was found to increase in proportion to the length of the initial fermentation period, suggesting the production of a heat-stable inhibitory factor or factors. The inhibitory activity was present in rumen fluid, could be extracted from lyophilized rumen fluid (LRF) with water, and was stable in response to proteolytic enzymes. In addition, the water-extracted residue of LRF was found to contain growth factor activity for rumen fungi in vitro.

Animals↗
PubMed2000

Feasibility of using lignin isolated from forages by solubilization in acetyl bromide as a standard for lignin analyses.

Lignin concentration can be measured in plants by the acetyl bromide-soluble lignin spectrophotometric method; however, as with any spectrophotometric method, a reliable standard is needed. In the present experiments, lignin was extracted from each of the forages under study with the acetyl bromide reagent. The lignin isolated with acetyl bromide (LIAB) was then used as the reference standard in the acetyl bromide-soluble lignin (ABSL) analysis, which was compared with the acid detergent lignin (ADL) and potassium permanganate lignin (PerL) lignin analyses. Two maturity stages of each of the following forages were analyzed: Medicago sativa, Cynodon dactylon var. Coastal, Panicum maximum var. Centenário and var. Colonião, Cynodon plectostachyus, Pennisetum purpureum, Setaria nandi, and Avena sativa. In addition, one wood sample, Eucalyptus sp., was analyzed. In general, ABSL values were highest (P < 0.001), followed by PerL and ADL, which also differed from each other (P < 0.001). Correlations with in vitro dry matter digestibility of samples were highest with the ABSL method. Absorption spectra of LIAB, either from plants of different maturity stages or from different vegetable species, suggested the presence of differences among some of the lignins.

Acetates↗
PubMed1999

Feasibility of using total purines as a marker for ruminal bacteria.

A procedure for measuring total purine content of mixed ruminal bacteria was adapted for use in the determination of purines in pure cultures of ruminal bacteria. Recovery of adenine and guanine, alone or in mixture, was quite variable. The problem was traced to solubility of the silver salt of adenine in the acid wash solution. When the precipitating solution was used as the wash, recovery of the purines was over 97%. Recovery of a 1:1 mixture of adenine and guanine added to yeast RNA was 100.6+/-3.2%. Purine, protein, and bacterial concentrations were determined for 10 pure cultures of ruminal bacteria: Butyrivibrio fibrisolvens, D16f, H10b, and H17c; Fibrobacter succinogenes B21a; Lachnospira multiparus D25e; Lactobacillus lactis ARD26e; Prevotella ruminicola H15a; Ruminococcus albus 7; Ruminococcus flavefaciens B34b; and Streptococcus bovis ARD5d. The CV for the most-probable-number (MPN) assay (bacterial concentrations), purine analysis, and protein analysis were 55.86, 5.25 and 6.52%, respectively. Considerable variation was found among bacterial species and strains when purine and protein concentrations were compared as the amount per individual cell. More consistent values were obtained when these components were expressed on a dry matter basis. Purine:protein ratios for the 10 pure cultures ranged from .023 to .1299, with a mean value of .0883. For samples of mixed bacteria separated from ruminal fluid, this ratio was found to average .0306, which is approximately one-third of the value for the pure cultures. The value determined for the mixed bacterial sample is similar to previously reported values. Based on the ratio obtained with the pure cultures, the microbial protein flow out of the rumen has probably been overestimated in most previous reports. Limited studies suggest that the samples of mixed ruminal bacteria used as a standard are probably contaminated with feed particles containing protein, which results in lower purine:protein ratios.

Animals↗
PubMed1998

Effect of ruminal cellulolytic bacterial concentrations on in situ digestion of forage cellulose.

To evaluate the effects of ruminal cellulolytic bacterial concentrations on in vivo cellulose digestion, varying percentages of flaked soybean hulls were substituted for orchardgrass hay in high-forage diets fed to sheep. In two experiments, total and cellulolytic ruminal bacterial concentrations were not affected by diet. No differences were found for in situ digestion of forage cellulose in the first experiment; however, in Exp. 2, ruminal pH and in situ cellulose digestion were lower (P<.01) with a 40% soybean hull diet. In Exp. 3 with four sheep, two diets were compared, one containing 19.6% cellulose from alfalfa meal and the other 64.3% purified wood cellulose. Ruminal pH was lower (P<.02), 9 and 24 h after feeding, for the high-cellulose diet. Total bacterial concentrations did not change with diet; however, the concentration of cellulolytic bacteria increased (P<.05) when the higher cellulose diet was fed. In situ cellulose digestion was not different between diets. In Exp. 4, 3% sodium bicarbonate was added to the high-cellulose diet, and it was fed twice a day. No differences were observed in pH between diets (P>.42). However, the concentration of total ruminal bacteria increased (P<.06), the concentration of cellulolytic bacteria increased (P<.03), and the percentage of cellulolytic bacteria increased (P<.04) when the buffered high-cellulose diet was fed. In situ digestion of alfalfa cellulose at 30 h was not different between diets (P>.60). These data indicate that the concentration of cellulolytic bacteria is not the limiting factor in the digestion of cellulose in the rumen.

Animal Feed↗
PubMed1998

Generation times of Epidinium caudatum and Entodinium caudatum, determined in vitro by transferring at various time intervals.

Generation times were determined in vitro with a pure culture of Epidinium caudatum and a mixed culture of Epidinium caudatum and Entodinium caudatum. Measurement of logarithmic growth from a small inoculum for Epidinium caudatum alone, or in coculture, resulted in generation times of 30.8 and 19.5 h, respectively. Epidinium concentrations, either alone or in coculture, were maintained when cultures were transferred every 12 h; however, concentrations decreased rapidly with transfers at 4, 6, or 8 h. For Entodinium caudatum, a generation time of 16.3 h was obtained from measurement of logarithmic growth. Based on sequential transfer data at varying time intervals, Epidinium caudatum and Entodinium caudatum seem to be capable of doubling in approximately 12 to 13 h. These values are markedly less than those previously reported and help explain the ability of these protozoa to maintain themselves in the rumen.

Animals↗
PubMed1996

Effect of prolonged high-concentrate feeding on ruminal protozoa concentrations.

Five ruminally cannulated steers, with ad libitum access to feed, were gradually adapted from an all-forage diet to a 75% concentrate diet over a 6-wk period. Three animals were then randomly assigned to an all-concentrate diet (87% whole corn) and the other two were fed a 90% concentrate plus 10% forage diet. These diets were fed for 17 wk and then reversed between groups for 11 additional weeks. Over the last 22 wk, addition of 10% forage to the all-concentrate diet had no effect on the concentration of total protozoa; however, Isotricha and Epidinium percentages increased (P < .05). Although concentrations varied markedly, protozoa persisted throughout the entire period of high-concentrate feeding (both diets) in three of the animals. In contrast, the other two animals were defaunated most of the time, except for the sporadic appearance of Entodinium species for 1- or 2-wk intervals. Average pH in these latter two animals was lower (P < .05) during the entire 28-wk high-concentrate feeding period. Because these two animals had a lower ruminal pH than the other three, even when fed all forage, it seems that the ruminal environment varies between individual animals. Thus, the maintenance of a protozoal population in animals fed high-concentrate diets may be related to physiological conditions such as rate and extent of salivary production, rate of fluid and particulate matter passage within each animal, and so on.

Animals↗
PubMed1996

Effects of feed and water deprivation on ruminal characteristics and microbial population of newly weaned and feedlot-adapted calves.

Two experiments were conducted to determine the effects of length of feed and water deprivation on ruminal microbes and ruminal characteristics. In Exp. 1, treatments were as follows: 1) weaned, but not trucked (0 h), 2) weaned, trucked, and fasted for 48 h (48 h), and 3) weaned, trucked, and fasted for 72 h (72 h). On d 0 (arrival back at the feedlot after treatment), DMI, ruminal contents (P < .05) decreased as length of feed and water deprivation increased. However, on d 4, there were no longer any differences (P > .10) in DMI or weight of ruminal contents. On d 0, percentage of ruminal DM in the 48- and 72-h treatment groups was lower (P < .01) than in the 0-h treatment group, but there were no differences (P > .10) by d 4. After the steers used in Exp. 1 had been in the feedlot for 28 h, they were reallotted, and the three treatments (except for weaning) were repeated; the 0-h group remained at the feedlot. On d 0, DMI of the 48-h group was lower (P < .05) than that of the 0-h group; DMI of the 72-h group was intermediate. On d 0, the 0-h treatment group had a greater ruminal DM percentage than the 48- and 72-h treatment groups (P < .05); however, due to large within-treatment variation the total weight of ruminal contents did not differ (P > .10) among treatments. There were no decreases (P > .10) in cellulolytic or total bacterial concentration or in the ruminal numbers of cellulolytic or total bacteria due to length of feed and water deprivation on d 0 in either Exp. 1 or 2. In both trials, 48 and 72 h of feed and water deprivation decreased (P < .05) protozoal numbers on d 0 and 4 compared with the 0-h treatment group.

Aging↗
PubMed1996

Interactions between Fibrobacter succinogenes, Prevotella ruminicola, and Ruminococcus flavefaciens in the digestion of cellulose from forages.

The synergistic and inhibitory interactions observed between Fibrobacter succinogenes A3c, Prevotella ruminicola H2b, and Ruminococcus flavefaciens B34b in the digestion of forage cellulose were studied in detail. Orchardgrass and alfalfa hays, both at two maturity stages, were used as substrates. Sequential inoculation procedures were developed whereby a second inoculation was made after the initial fermentation was killed. Total cellulose digestion from sequential addition of the organisms was then compared to values obtained in simultaneous co-culture. When the noncellulolytic P. ruminicola was co-cultured with either of the two cellulolytic species (F. succinogenes or R. flavefaciens) forage cellulose digestion numerically increased over that of the cellulolytic species alone. In contrast, decreases from co-culture values were noted with sequential addition of the organisms. When F. succinogenes and R. flavefaciens were co-cultured, cellulose digestion was reduced compared to F. succinogenes alone. However, no such reduction was observed when the organisms were added sequentially. Further experiments indicated that this inhibitory activity is only produced when the organisms are co-cultured and is stable to autoclaving at 121 degrees C for 20 min. Inhibition of this type could be the result of bacterocin production by one of the organisms; however, most bacterocins are destroyed by autoclaving.

Animal Feed↗
PubMed1994

Degradation and utilization of forage hemicellulose by rumen bacteria, singly in coculture or added sequentially.

Procedures for sequential addition experiments were developed to study the mechanisms involved in the synergistic and inhibitory interactions observed in forage hemicellulose digestion by rumen bacterial cocultures. One organism was allowed to ferment a forage substrate, the culture tube was sterilized and then inoculated with a second organism. No differences were found in the extent of degradation or utilization between fermentations sterilized by oxidation or heat, and based on ease of handling, heat was used in all subsequent experiments. Studies were conducted with Fibrobacter succinogenes A3c, Ruminococcus flavefaciens B34b and Prevotella ruminicola H2b, singly and in all possible combinations. Results from the sequential addition studies substantiated earlier suggestions that the increase observed in hemicellulose utilization results from initial solubilization of the hemicellulose from the forage by the non-utilizer and subsequent utilization of subsequent utilization of this solubilized polysaccharide by the utilizing, but non-degrading organism.

Animal Feed↗
PubMed1994

Ruminal characteristics, microbial populations, and digestive capabilities of newly weaned, stressed calves.

Eight ruminally fistulated steers, 7 to 8 mo old, were used in a completely randomized, 2 x 2 factorial experiment to determine the effects of energy density and protein source in receiving diets on in situ DM, NDF, and N disappearance, concentrations of ruminal bacteria, protozoa, ammonia, and pH. Two energy densities (1.80 and 1.48 Mcal/kg of NEm) and two protein sources (spray-dried blood meal [SDBM] and soybean meal [SBM]) were compared. Fistulated steers were weaned, transported by truck, and held in a sale barn before their arrival at the feedlot. On d 0 (day of arrival at the feedlot), DMI was 62% of DMI on d 7 after arrival. Overall, feeding a high-energy diet resulted in lower (P < .01) in situ DM disappearance (DMD) of orchardgrass than feeding a low-energy diet at both 24 and 48 h. In situ 24-h DMD averaged 46.6% on d-3 and 41.6% on d 0, whereas 48-h in situ DMD on d -3 and 0 averaged 58.2 and 58.6%, respectively, indicating the ruminal microbial population was not inhibited in its ability to digest available substrate. Additionally, there were no differences (P > .10) in 48-h in situ NDF disappearance between d -3 and 0 (58.8 vs 57.8%), respectively. No differences (P > .10) occurred in the concentration of total bacteria, or cellulolytic bacteria, due to feed and water deprivation. Concentration of total protozoa was lower (P < .05) on d 0 than at any other time. Entodinium averaged 72.5% of genera before weaning, and more than 90% of genera found on all treatments by d 21. Diplodinium and Epidinium percentages tended to decline after weaning. Isotricha concentrations were low and Dasytricha were eliminated after d 7. In conclusion, the concentration of ruminal bacteria and the ability to digest available substrate were not decreased immediately after weaning, trucking, and 24 h of feed and water deprivation.

Ammonia↗
PubMed1994

The influence of restricted feeding on site and extent of digestion and flow of nitrogenous compounds to the duodenum in steers.

The effects of restricted feeding on site and extent of digestion, flow of N-containing compounds to the duodenum, and ruminal metabolism were determined. In Trial 1, corn silage-based diets were fed at one of three intakes to three ruminally and duodenally fistulated steers (420 +/- 4.4 kg BW) in a repeated 3 x 3 Latin square experiment. Feeding levels were 100% of ad libitum intake, 90% of ad libitum intake, and 80% of ad libitum intake. Site of digestion was not affected by restricted feeding; however, extent of DM, NDF, and apparent N digestion were increased (P < .03) 2.8, 4.1, and 6.2 percentage units for each 1-kg restriction in DM intake. Restricted feeding increased (P < .06) ruminal pH and ammonia concentrations, decreased (P < .05) ruminal liquid dilution rate, and had no major effects on bacterial concentrations. In Trial 2, all-concentrate diets based on whole shelled corn were fed at 100, 90, or 80% of ad libitum intake to six ruminally and duodenally fistulated steers (440 +/- 4.1 kg BW) in a replicated 3 x 3 Latin square experiment. Restricted feeding increased (P < .06) apparent ruminal DM digestion; however, true ruminal OM digestibility was not affected. Extent of apparent N digestion increased 3.0 percentage units for each 1-kg restriction in DMI. Ruminal pH was not affected by restricted feeding; however, ruminal ammonia concentrations increased (P < .06) and ruminal liquid dilution rate decreased (P < .04) with restricted feeding. In both Trials, total N flow to the duodenum was approximately 100% of N intake for the 80% of ad libitum intake. Flow of N exceeded N intake in a linear (P < .01) manner as DMI increased. Decreases in duodenal N flow when intake was restricted resulted from decreases (P < .01) in bacterial N flowing to the duodenum, and also in decreases (P < .01) in non-ammonia, non-bacterial N flow when whole shelled corn-based diets were fed. We concluded that restrictions in digestible energy intake are less than the DM restriction when corn silage-based diets are fed but are equal to the DM restriction when whole shelled corn-based diets are fed. Also, restricted feeding strategies reduce the flow of N to the duodenum from both bacterial and feed origins.

Animal Feed↗
PubMed1993

Effects of diet and hindgut defaunation on diet digestibility and microbial concentrations in the cecum and colon of the horse.

The effects of diet and hindgut defaunation (removal of protozoa from the hindgut) on diet digestibility (Trial 1) and on total and cellulolytic bacterial and fungal concentrations in the cecum and colon (Trial 2) were investigated. A high-forage (HF) diet, 90% alfalfa hay-10% concentrate, or a higher-concentrate (HC) diet, 60% alfalfa hay-40% concentrate, was limit-fed. In Trial 1, defaunation resulted in a slight decrease in DM digestibility (P < .1) and had no effect on cellulose digestibility. Dry matter digestibility was higher (P < .001) with the HC diet; however, no differences were observed in cellulose digestion. For the faunated periods, protozoal concentrations were similar in the cecum and greater in the colon for both diets (P < .05). A diet x location interaction was observed for the genera Buetschlia and Blepharocorys. In Trial 2, defaunation had no effect on either total or cellulolytic bacterial concentrations in the cecum or colon. Total bacterial concentrations were higher (P < .06) in the colon when ponies were fed the HC diet. Defaunation did not affect total fungal concentrations in the cecum; however, fungal concentrations in the colon were slightly higher (P < .1) when the ponies were defaunated. Diet had no effect on total or cellulolytic fungal concentrations. Both total and cellulolytic fungal concentrations were approximately 10-fold higher in the colon than in the cecum (P < .01). Protozoa do not seem to play an essential role in the fermentation of feedstuffs in the equine hindgut.

Animal Feed↗
PubMed1992

In vitro and in vivo studies of factors affecting digestion of feeds in synthetic fiber bags.

Dry matter disappearance (DMD) and cellulose disappearance (CD) from alfalfa hay were measured simultaneously in synthetic fiber bags incubated in vitro. In general, DMD, CD, and pH were reduced (P less than .05) inside the bags compared with values in the fermentation flasks. Bags became fully expanded with gas soon after the fermentation started, which seemed to inhibit fluid movement into and out of the bag. By creating a vacuum in the flask, fluid exchange was enhanced, and DMD, CD, and pH increased (P less than .01) inside the bags. Sample size, type of fabric, and fabric pore size affected pH, DMD, and CD. Inoculum source (forage or concentrate diet) did not affect DMD in either bags or flasks. In contrast, DMD in bags incubated in vivo was affected (P less than .01) by animal diet. When bags were incubated in vivo inside a rigid plastic container, both pH and DMD were reduced (P less than .01) compared with bags suspended freely in the rumen. Direct physical action of ruminal contents on the bags apparently enhanced exchange of fluid. Bacterial concentrations in bags suspended freely in the rumen were greater than those in bags incubated inside containers. Synthetic fiber bags may be useful in comparing digestibility between diets; however, caution is needed in evaluating such data in absolute terms.

Animal Feed↗
PubMed1992

Relationship between rumen bacterial concentrations and total numbers.

Based on data from 33 sheep, bacterial concentrations per gram of rumen contents were correlated highly with total bacterial numbers (r = .99). Diets ranged from all forage to 90% concentrate. Similar data were obtained from cattle fed a corn silage diet (r = .95); however, the slope of the regression lines differed between species. Weight of rumen contents was correlated poorly with total bacterial numbers.

Animals↗