Microbiological evaluation of the intraruminal in sacculus digestion technique

The influence of nature of the feed sample, feeding frequency and pore size on the influx of bacteria and protozoa into synthetic fiber bags suspended in the rumens of sheep fed different diets was studied. Counts of total culturable bacteria in bags with a pore size of 10 microns were less than 30% of the ruminal counts for animals that were fed the lucerne hay and high-roughage diets. The maximum count (62 and 82% of the ruminal count) for these specific diets was obtained by using bags with a pore size of 53 microns. Protozoal counts in bags with pore sizes of 30 and 53 microns were equal to or higher than the ruminal counts for the lucerne hay and high-roughage diets but less than half of the ruminal count for the low-roughage diet. An interaction between incubation time, feeding frequency of the host animals, and the microbial populations developing inside the bags was also demonstrated. The results clearly show that the microbial population inside the bag differed from that of the surrounding ruminal ingesta and that caution must be taken in interpreting results on feed evaluation and especially on rates of degradation when using the in sacculus technique. Factors influencing the influx of bacteria and protozoa into bags with different pore sizes and containing a variety of substrates are discussed together with suggestions for the use of this technique.     

Rumen Microbial Ecology in Mule Deer

Mule deer rumen microbial populations from animals in the natural habitat in Utah and from captive deer fed various rations were studied. The microorganisms were characterized on the basis of morphology and Gram reaction. Rumen samples contained 13 identifiable types of bacteria and one genus of ciliate protozoa (Entodinium). Highest rumen bacterial populations were produced on rations containing barley. No differences in proportions of ruminal bacteria in the various morphological groups could be detected when animals were fed either natural browse plants or alfalfa hay. The total numbers of bacteria were similar for animals feeding on controlled diets of browse or hay and those in the natural habitat. Numbers of some bacterial types were directly related to ciliate protozoal numbers, whereas others were inversely related. Highest rumen ciliate protozoal populations were observed on rations containing barley. No differences in protozoal populations were noted between diets containing only browse or hay. Seasonal variations were noted in ciliate protozoal numbers from deer feeding in the natural habitat. The total number of ciliate protozoa decreased in the fall and winter and remained low until spring. There were indications that salt in the deer diet favorably affected rumen ciliate protozoa. Rather than revealing direct deer management applications, this study serves to stimulate and illuminate new approaches to research in range and wildlife nutrition.     

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Moderation of ruminal fermentation by ciliated protozoa in cattle fed a high-grain diet.

The objective of this study was to assess the influence of ciliated protozoa on ruminal fermentation in cattle fed high-grain diets. Six ruminally cannulated steers fed a corn-based grain diet (85% concentrate plus 15% alfalfa hay) at 12-h intervals were assigned randomly to two groups, ciliate free and faunated, in a crossover design. Defaunation was by ruminal emptying, omasal flushing, and treatment with sodium sulfosuccinate. Two to 3 weeks after defaunation, the ruminal contents of all steers were sampled before the morning feeding (0 h) and at 1, 2, 4, 6, 8, and 12 h after feeding to measure pH, analyze fermentation products, and monitor counts of ciliated protozoa and lactic acid-producing and -fermenting bacterial groups. Total numbers of ciliated protozoa in the faunated steers averaged 4.3 x 10(5)/g, and the protozoa consisted of nine genera. Ciliate-free steers had lower (P less than 0.01) ruminal pHs (pH 5.97) than faunated cattle (pH 6.45); however, the treatment-time interaction was not significant. Ruminal lactate and ammonia concentrations were similar in both groups. The total volatile fatty acid concentration was higher (P less than 0.05) in the ciliate-free steers than in the faunated steers and exhibited a treatment-time interaction (P less than 0.05). The acetate-to-propionate ratio was higher (P less than 0.05) in the faunated group than in the ciliate-free group and showed a treatment-time interaction (P less than 0.05). Total anaerobic bacterial counts were about fourfold higher in the ciliate-free group than in the faunated group.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Microflora and Concentrations of Volatile Fatty Acids in The Rumen of Cattle Fed on Single Component Rations

The composition of the rumen microflora and the volatile fatty acids were examined in cattle free-grazing on grass or stall-fed on hay, grass pellets, oats or dried beet pulp with molasses. Total and viable counts of anaerobic bacteria were highest on the grass feeding, but viable counts as a percentage of total counts were highest when oats or beet pulp with molasses were fed. Counts of cellulolytic bacteria were lowest on these latter 2 diets, and highest on grass or grass pellet diets. Studies of the anaerobic flora showed that the composition in animals fed on grass pellets resembled more that found in animals free-grazing on grass than in those fed on hay. Counts of aerotolerant bacteria were only a small percentage of the total count, but were highest on the hay diet. On this latter diet and on grass-feeding the streptococci (identified as Streptococcus bovis) were predominant, but contrary to expectation, streptococci were found only in small numbers on the oats diet, where coryneform rods were the major type present. Although a period of 4–6 weeks was allowed for the animals to adapt to the feeds, the 2 periods of feeding on oats and dried beet pulp with molasses markedly affected the composition of the rumen flora in the subsequent periods of feeding grass pellets and hay. Ruinen volatile fatty acid analysis showed a propionogenic effect of oats and the highest percentage of butyric acid when beet pulp with molasses was fed. The expected propionogenic effect of grass pellets was not observed.     

Rumen microbial changes in cattle fed diets with or without salinomycin

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total and selective enumeration of anaerobic bacteria. VFA analysis, and enumeration of protozoa were collected during the 80:20 alfalfa hay-to-grain diet before salinomycin feeding, and during the 80:20, 50:50, and 20:80 hay-to-grain diets with salinomycin. At each sampling period, rumen samples were collected at 3 h after feeding on three consecutive days. Salinomycin feeding had no effect on rumen pH and total VFA concentration. The acetate-to-propionate ratio was significantly lower in salinomycin-fed steers than in the control. The molar proportion of butyrate increased in both control and salinomycin-fed steers. Total anaerobic bacterial counts were lower in salinomycin-fed steers than in the control steers after 8 weeks of salinomycin feeding. Salinomycin-resistant bacteria increased from 7.6 to 15.6% in salinomycin-fed steers but remained unchanged in control steers. Salinomycin had no effect on cellulolytic and lactate-utilizing bacteria, but the proportion of amylolytic bacteria was higher in salinomycin-fed steers than in control steers. The total number of protozoa decreased initially in salinomycin-fed steers. The initial reduction was due to reduced numbers of Entodinium species. Holotrichs were unaffected by salinomycin feeding.     

Rumen microbial changes in cattle fed diets with or without salinomycin.

Four rumen-fistulated steers, randomly assigned to two groups (control and salinomycin fed) were used to monitor the changes in rumen microbial populations and volatile fatty acids (VFA) concentrations associated with feeding salinomycin (0.22 mg X kg-1 X day-1). Steers were adapted to an alfalfa hay and grain (80:20) diet before supplementing the diet with salinomycin, and then the diet was changed to 50:50 and 20:80 ratios of alfalfa hay to grain at 2-week intervals. Rumen samples for total and selective enumeration of anaerobic bacteria. VFA analysis, and enumeration of protozoa were collected during the 80:20 alfalfa hay-to-grain diet before salinomycin feeding, and during the 80:20, 50:50, and 20:80 hay-to-grain diets with salinomycin. At each sampling period, rumen samples were collected at 3 h after feeding on three consecutive days. Salinomycin feeding had no effect on rumen pH and total VFA concentration. The acetate-to-propionate ratio was significantly lower in salinomycin-fed steers than in the control. The molar proportion of butyrate increased in both control and salinomycin-fed steers. Total anaerobic bacterial counts were lower in salinomycin-fed steers than in the control steers after 8 weeks of salinomycin feeding. Salinomycin-resistant bacteria increased from 7.6 to 15.6% in salinomycin-fed steers but remained unchanged in control steers. Salinomycin had no effect on cellulolytic and lactate-utilizing bacteria, but the proportion of amylolytic bacteria was higher in salinomycin-fed steers than in control steers. The total number of protozoa decreased initially in salinomycin-fed steers. The initial reduction was due to reduced numbers of Entodinium species. Holotrichs were unaffected by salinomycin feeding.     

Ruminal microbial populations and fermentation characteristics in bison and cattle fed high- and low-quality forage

Ruminal microbial populations and fermentation products were compared between two ruminally cannulated bison (375 kg) and two ruminally cannulated Hereford steers (567 kg) on alfalfa or prairie hay diets. Differential media were used to enumerate carbohydrate-specific bacterial subgroups. Voluntary dry matter intake was higher (P=0.006) for cattle than for bison fed alfalfa, but prairie hay intake was not different (P=0.16) between the two species. Volatile fatty acid concentrations, pH, and ruminal ammonia were similar between bison and cattle on both diets. Total anaerobic bacteria and xylanolytic bacterial counts were higher (P<0.02) in bison than in cattle fed alfalfa. However, with the prairie hay diet, no differences in bacterial counts on any medium were observed between ruminant species. Both bison and cattle possessed a mixed A-B protozoan population with nearly identical protozoan numbers and distribution of genera. The similarities between bison and cattle consuming either high-or low-quality forage suggest that any differences in putative forage digestibility between the species are not due to differences in microbial counts.     

Protozoa involved in butyric rather than lactic fermentative pattern during latent acidosis in sheep

We used six ruminally cannulated Texel wethers to study the relative role of protozoa and lactate-metabolizing bacteria in ruminal fermentative patterns during an induced latent acidosis. The sheep were fed an alfalfa hay diet (H) and latent acidosis was induced, following a short transition period of one week, with a grain-rich acidotic diet (W, 60% wheat + 40% alfalfa hay). Ruminal pH, ruminal volatile fatty acids (VFA), lactate and NH3 concentrations, protozoa and lactate-utilizing bacterial counts, the relative proportions of three main bacteria implicated in lactate metabolism (a lactate-producing species, Streptococcus bovis, and two lactate-utilizing species, Selenomonas ruminantium, and Megasphaera elsdenii) using specific 16S-rRNA-targeting oligonucleotide probes, and lactate dehydrogenase (LDH) activity were determined for both diets. The pH parameters (mean, minimum, maximum, time and area under pH 6.0 and 5.5) measured with the W diet were indicative of a latent (i.e., subacute and maintained) acidosis. However, a butyric rather than lactic latent acidosis was observed in this study. Total ruminal lactate concentration remained at low levels with the acidotic diet (< 4 mmol x L(-1)), but changes were observed in VFA composition, which was oriented towards butyrate at the expense of acetate (P < 0.05), while propionate remained constant. In agreement with the low ruminal lactate concentration, no changes in the proportion of S. bovis 16S-rRNA were observed. The lactate-metabolizing bacterial population also remained fairly constant in number, proportion and activity. The increase in butyrate concentration was accompanied by a proliferation of entodiniomorphs (P < 0.01). These results suggest that the protozoa limited lactate accumulation and possibly also the decrease in pH during latent acidosis. Experiments with defaunated and faunated sheep could provide further evidence of the role of protozoa in the development of rumen latent acidosis.     

A Note on the Flora and Fauna in the Rumen of Steers Fed a Feedlot Bloat-provoking Ration and the Effect of Penicillin

A study was made of the predominant culturable bacteria and ciliate protozoa present in the rumen of two steers that were regularly bloating on a pelleted ratio containing 22% alfalfa meal, 16% soybean oil meal, 61% barley, and 1% common salt. The ruminal microorganisms in the two animals differed as indicated by a high total culture count of bacteria, an almost complete absence of ciliate protozoa, a low pH, and a difference in the proportions of presumptively identified predominant bacterial groups in one animal (steer 26) as compared with the other (steer 32). The first exposure of the animals to procaine penicillin (75 or 150 mg per day on 2 successive days) resulted in an abnormal ruminal flora 31 hr after the first treatment as indicated by drastic drops in total and cellulolytic bacterial counts and a change in the proportions of predominant bacterial groups. The animals refused feed for 32 to 48 hr after the first treatment. After feed consumption resumed, further treatment with 75 mg penicillin on 4 successive days did not appear to greatly alter the flora and did not result in feed refusal in animal 32. In animal 26, amounts of penicillin progressing from 50 to 200 mg per day did not result in feed refusals and observations on rumen ingesta samples during this period indicated a decrease in total bacterial count, a great increase in numbers of ciliate protozoa, a higher pH, and a change in the proportions of predominant bacterial groups so that the ruminal picture was much more similar to that of animal 32 than to its own during the pre-penicillin period. Bloat was not relieved except during the period of feed refusal.

The results indicate that the ruminal flora rapidly adapts to penicillin and that bloat of the feedlot type can occur in animals with widely differing numbers and kinds of bacteria and protozoa. Feedlot bloat does not appear to be correlated with the occurrence or numbers of any of the individual predominant groups of bacteria cultured.

     

Adenosine Triphosphate and Deoxyribonucleic Acid in the Alimentary Tract of Cattle Fed Different Nitrogen Sources

Total microbial cell counts and the content of deoxyribonucleic acid (DNA) and adenosine triphosphate (ATP) were determined in ruminal and abomasal digesta and in faeces from heifers fed a basal diet of barley and straw supplemented with urea, casein, soybean-protein concentrate or feather meal. The concentration of ATP in the rumen content varied independently of the DNA content and total cell count, but dependently upon the nitrogen sources, the highest ATP/DNA ratio being obtained with casein. The ATP/DNA ratio in faeces was only one-tenth of that found in the digesta of the rumen and abomasum, indicating either an extremely low level of activity of the microbial cells or more probably a very large number of dead organisms. It is suggested that DNA and ATP might be useful indices of the microbial status in terms of biomass and metabolic activity. The total cell count should still be included in routine studies to determine the proportion of protozoa to bacteria.     

Effects of monolaurin on ruminal methanogens and selected bacterial species from cattle, as determined with the rumen simulation technique

Before being able to implement effective ruminal methane mitigation strategies via feed supplementation, the assessment of side effects on ruminal fermentation and rumen microbial populations is indispensable. In this respect we investigated the effects of monolaurin, a methane-mitigating lipid, on methanogens and important carbohydrate-degrading bacteria present in ruminal fluid of dairy cattle in continuous culture employing the rumen simulation technique. In six experimental runs, each lasting for 10 days, four diets with different carbohydrate composition, based on hay, maize, wheat and a maize-wheat mixture, either remained non-supplemented or were supplemented with monolaurin and incubated in a ruminal-fluid buffer mixture. Incubation liquid samples from days 6 to 10 of incubation were analyzed with relative quantitative polymerase chain reaction (qPCR) of 16S rRNA genes to assess monolaurin-induced shifts in specific rumen microbial populations in relation to the corresponding non-supplemented diets. Monolaurin completely inhibited Fibrobacter succinogenes in all diets while the response of the other cellulolytic bacteria varied in dependence of the diet. Megasphaera elsdenii remained unaffected by monolaurin in the two diets containing maize, but was slightly stimulated by monolaurin with the wheat and largely with the hay diet. The supply of monolaurin suppressed Methanomicrobiales below the detection limit with all diets, whereas relative 16S rRNA gene copy numbers of Methanobacteriales increased by 7-fold with monolaurin in case of the hay diet. Total Archaea were decreased by up to over 90%, but this was significant only for the wheat containing diets. Thus, monolaurin exerted variable effects mediated by unknown mechanisms on important ruminal microbes involved in carbohydrate degradation, along with its suppression of methane formation. The applicability of monolaurin for methane mitigation in ruminants thus depends on the extent to which adverse effects on carbohydrate-degrading bacteria actually impair the supply of digested carbohydrates to the animal.     

Sulphate recycling and metabolism in sheep and cattle.

Merino wethers and Brahman x Shorthorn steers, offered lucerne or spear grass hay, were used to study the movements of sulphate through pools in plasma and ruminal liquor. The irreversible loss of sulphate from ruminal liquor was 60 and 76% of sulphur ingested for both species fed lucerne and spear grass respectively. The irreversible loss of sulphate from the plasma averaged 67 and 56% of sulphur ingested for animals fed lucerne and spear grass respectively. Daily recycling of sulphate to the rumen of sheep was 98 mg sulphur on the lucerne diet and 3.9 mg sulphur on the spear grass diet. Sulphate recycling in cattle fed lucerne was 533 mg sulphur; in cattle fed spear grass the value was 234 mg sulphur. Over 6 days following an intravenous injection of [35S]sulphate into sheep and cattle fed lucerne, 5-10% of the dose was excreted in the faeces and c. 10% was retained. Corresponding values for animals fed spear grass were 23-31% in faeces and 40-51% of the dose retained. After intraruminal injections of [35S]sulphate, animals fed lucerne excreted 15-18% of the dose in the faeces and retained 25-30% of the dose over 6 days. Values for animals fed spear grass were 22-26% in faeces and 62-70% retained. It was concluded that sulphate recycling to the rumen is a limiting factor in microbial synthesis for sheep fed low-quality roughage, and that secretion of endogenous sulphur into the postruminal tract of ruminants is of importance in the metabolism of sulphate.     

Effect of the tropical forage calliandra on microbial protein synthesis and ecology in the rumen

AIMS: To determine the effect of condensed tannins in Calliandra calothyrsus (calliandra) on rumen microbial function. METHODS AND RESULTS: Microbial populations, ruminal protein synthesis and fermentation end-products were measured in sheep fed roughage hay supplemented with calliandra (30%), with and without inclusions of polyethylene glycol (PEG) to counteract the effect of tannin. Molecular and conventional enumeration techniques were used to quantify rumen bacteria, fungi and protozoa, and protein synthesis was predicted from estimates of urinary purine excretion. The total number of cellulolytic bacteria, including populations of Fibrobacter succinogenes and Ruminococcus spp., was significantly lower in sheep supplemented with calliandra and these populations increased when animals were treated with PEG. By contrast, protozoa and fungi and the microbial group containing Bacteroides-Porphyromonas-Prevotella bacteria appeared to be less affected. The efficiency of microbial protein synthesis in the rumen was not altered significantly. CONCLUSION: Calliandra caused significant shifts in rumen microbial populations without changing the efficiency of protein synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: The effect of calliandra tannins on rumen digestion may result more from complexing with nutrients than direct inhibition of micro-organisms.     

Fermentation of plant cell walls by ruminal bacteria, protozoa and fungi and their interaction with fibre particle size

The objective of the experiment was to evaluate the contribution of various ruminal microbial groups to the fermentation of cell walls of corn stover with different particle sizes based on ruminal gas production in vitro. Physical, chemical, and antibiotical methods were used to differentiate groups of bacteria, protozoa and fungi in rumen fluid, offering following rumen microbial groups: whole rumen fluid (WRF), bacterial (B), protozoal (P), fungal (F), bacterial plus protozoal (B + P), bacterial plus fungal (B + F), protozoal plus fungal (P + F), and negative control (CON). Cell walls from corn stover were ground and ball milled to produce two different particle sizes. The results showed that digestion of the cell walls was undertaken by the interaction among ruminal bacteria, protozoa and fungi, and such co-actions seemed to fail alternation by one of three microbial groups or any combinations. However, B + P group showed a significant contribution to the degradation of milled cell walls, and B + F group revealed a great synergy effect on the ground cell walls degradation. Particle size of cell walls also had a considerable influence on their fermentation extent instead of the fermentative patterns by various rumen microbial groups.     

Influence of diet and monensin on development of anaerobic fungi in the rumen, duodenum, cecum, and feces of cows

Three cows with fistulated rumens, duodenums, and ceca were fed five different diets: lucerne hay, lucerne hay plus whey (40:60), lucerne hay plus beets (50:50), corn silage plus monensin (40 ppm [40 g/kg] of dry matter intake), and lucerne hay plus monensin (80 ppm of dry matter intake). The fungal population was observed in the rumen, duodenum, cecum, and rectum and varied with diet; it was most abundant with lucerne hay alone and with corn silage plus monensin. The proportion of particles colonized by fungi in the duodenum, the cecum, and feces was measured by microscopic observation and varied from 5 to 50%, depending on the diet. The further sporangia attached to the plant particles were from the rumen, the more likely they were to be devoid of spores. Results confirmed the influence of diet on the development of the ruminal fungal population and showed that monensin does not eliminate these microorganisms. They also confirmed the presence of anaerobic fungi in the ruminant intestine. It is likely that anaerobic fungi leave the rumen attached to plant particles. However, large colonies of nonrhizoidal-type fungi were observed in cecum samples and in feces; at these sites, environmental conditions are perhaps more favorable for this type of fungus than they are in the rumen.     

How important is milk for near-weaned red kangaroos (<Emphasis Type="Italic">Macropus rufus</Emphasis>) fed different forages?

Red kangaroos (Macropus rufus) are large (>20 kg) herbivorous marsupials common to the arid and semi-arid regions of inland Australia, where drought is frequent. Young-at-foot (YAF) red kangaroos are the age/size class usually most affected by drought. Kangaroos at this YAF stage are making the transition from a milk-based diet to one of herbivory and an inability to adequately digest high-fibre feeds may contribute to their high mortalities during drought. We examined the role of milk in the nutrition of YAF red kangaroos fed forages of different fibre content and evaluated it as an extra energy and/or nitrogen source. Milk intake had little impact on the digestion of herbage by YAF red kangaroos fed low-fibre chopped lucerne (alfalfa) hay. Organic matter (OM) intake was 210+/-20 g day(-1) and 228+/-22 g day(-1), respectively, by YAF fed lucerne and lucerne with milk. Apparent digestibility of lucerne OM was ca. 55%, regardless of milk intake. Fed lucerne, with and without milk, YAF sustained growth rates of ca. 45 g day(-1). Conversely, even with a milk supplement, YAF red kangaroos ingested only 90+/-11 g day(-1) of high-fibre chopped oaten hay, of which they digested only ca. 36%. Despite milk intake, YAF fed chopped oaten hay lost between 0 and 75 g body mass day(-1) and were in negative nitrogen balance (-0.40+/-0.11 g N day(-1)). On all diets nitrogen loss was primarily as endogenous nitrogen (urinary and faecal) rather than as dietary nitrogen. Endogenous nitrogen losses were elevated in YAF fed chopped oaten hay, primarily as non-dietary faecal nitrogen. Overall, when high-quality feed was available, YAF were not markedly dependent on milk. However, YAF fed poor-quality chopped oaten hay would require up to 540 ml day(-1) of late-stage kangaroo milk to attain intakes of energy and nitrogen, and hence growth rates, comparable with those YAF fed lucerne.     

Influence of diet and monensin on development of anaerobic fungi in the rumen, duodenum, cecum, and feces of cows.

Three cows with fistulated rumens, duodenums, and ceca were fed five different diets: lucerne hay, lucerne hay plus whey (40:60), lucerne hay plus beets (50:50), corn silage plus monensin (40 ppm [40 g/kg] of dry matter intake), and lucerne hay plus monensin (80 ppm of dry matter intake). The fungal population was observed in the rumen, duodenum, cecum, and rectum and varied with diet; it was most abundant with lucerne hay alone and with corn silage plus monensin. The proportion of particles colonized by fungi in the duodenum, the cecum, and feces was measured by microscopic observation and varied from 5 to 50%, depending on the diet. The further sporangia attached to the plant particles were from the rumen, the more likely they were to be devoid of spores. Results confirmed the influence of diet on the development of the ruminal fungal population and showed that monensin does not eliminate these microorganisms. They also confirmed the presence of anaerobic fungi in the ruminant intestine. It is likely that anaerobic fungi leave the rumen attached to plant particles. However, large colonies of nonrhizoidal-type fungi were observed in cecum samples and in feces; at these sites, environmental conditions are perhaps more favorable for this type of fungus than they are in the rumen.     

Effects of the forage-to-concentrate ratio of the diet on feeding behaviour in young Blond d'Aquitaine bulls

The activities of bulls, their feeding behaviour and their ruminal pH were examined at several stages during the finishing period, according to the forage-to-concentrate ratio of their diet. Twenty-four bulls of the Blond d'Aquitaine breed (initial body weight = 326 ± 21 kg) were assigned to six balanced pens with a space allowance of 9.4 m2 per bull during the finishing period. They were fed three different diets with achieved forage-to-concentrate ratios of (i) 8% straw and 92% concentrate, (ii) 44% hay and 56% concentrate and (iii) 57% maize silage and 43% concentrate. Bulls had ad libitum access to feed dispensed once daily. Offered and refusals were weighed on 5 consecutive days per week. The bulls were slaughtered at the common final live weight of 650 kg and the finishing period lasted 138, 181 and 155 days for straw-concentrate, hay-concentrate and maize silage-concentrate diets, respectively. The time budget was estimated four times by scan sampling with a 10-min interval. Feeding behaviour was appraised using data from electronic feeding gates. Ruminal pH was measured from a ruminal fluid sample collected by rumenocentesis. On average, the bulls spent 78% of the time lying or standing still, and 11% of the time eating. The forage-to-concentrate ratio of the diet influenced only those activities directly linked to feeding, i.e. eating and drinking. Bulls fed a high-concentrate diet spent less time eating than the other bulls (47 min v. >2 h) and took shorter meals (7 min v. 17 min). The bulls fed the straw-concentrate diet spread their meals over the entire day, whereas the others maintained two major peaks of eating activity, the main one in the morning after feed dispensing, the other one at the end of the diurnal period. Intake rate ranged widely between diets, from 58 g/min on average for the diets based on hay or maize silage up to 173 g/min for the high-concentrate diet. The concentrate-diet bulls also had a lower ruminal pH during the first 2 months of the finishing period. The dispersion of meals based on a high-acidosis-risk diet may be a way to limit the decrease in ruminal pH.     

Effect of the tropical forage calliandra on microbial protein synthesis and ecology in the rumen

C.S. MCSWEENEY, B. PALMER, R. BUNCH AND D.O. KRAUSE. 2001 .
Aims: To determine the effect of condensed tannins in Calliandra calothyrsus (calliandra) on rumen microbial function.
Methods and Results: Microbial populations, ruminal protein synthesis and fermentation end-products were measured in sheep fed roughage hay supplemented with calliandra (30%), with and without inclusions of polyethylene glycol (PEG) to counteract the effect of tannin. Molecular and conventional enumeration techniques were used to quantify rumen bacteria, fungi and protozoa, and protein synthesis was predicted from estimates of urinary purine excretion. The total number of cellulolytic bacteria, including populations of Fibrobacter succinogenes and Ruminococcus spp., was significantly lower in sheep supplemented with calliandra and these populations increased when animals were treated with PEG. By contrast, protozoa and fungi and the microbial group containing Bacteroides - Porphyromonas - Prevotella bacteria appeared to be less affected. The efficiency of microbial protein synthesis in the rumen was not altered significantly.
Conclusions: Calliandra caused significant shifts in rumen microbial populations without changing the efficiency of protein synthesis.
Significance and Impact of the Study: The effect of calliandra tannins on rumen digestion may result more from complexing with nutrients than direct inhibition of micro-organisms.