Comparison of microbial fermentation of high- and low-forage diets in Rusitec, single-flow continuous-culture fermenters and sheep rumen

Eight Rusitec and eight single-flow continuous-culture fermenters (SFCCF) were used to compare the ruminal fermentation of two diets composed of alfalfa hay and concentrate in proportions of 80 : 20 (F80) and 20 : 80 (F20). Results were validated with those obtained previously in sheep fed the same diets. Rusitec fermenters were fed once daily and SFCCF twice, but liquid dilution rates were similar in both types of fermenters. Mean values of pH over the 12 h postfeeding were higher (P < 0.001) in Rusitec than in SFCCF, with diet F80 showing higher values (P < 0.001) in both types of fermenters. Concentrations of total volatile fatty acids (VFA) were higher (P < 0.001) in SFCCF than in Rusitec, and in both systems were higher (P = 0.002) for diet F20 than for diet F80. There were significant differences between systems in the proportions of the main VFA, and a fermentation system × diet interaction (P < 0.001) was detected for all VFA with the exception of valerate. No differences (P = 0.145) between the two types of fermenters were detected in dry matter (DM) digestibility, but NDF, microbial N flow and its efficiency were higher (P = 0.001) in SFCCF compared to Rusitec. Whereas pH values and VFA concentrations remained fairly stable through the day in both in vitro systems, pH dropped and VFA increased shortly after feeding in sheep rumen reaching the minimum and maximal values, respectively, about 4 h after feeding. Both in vitro systems detected differences between diets similar to those found in sheep for liquid dilution rate, pH values, DM digestibility, microbial N flow and growth efficiency. In contrast, acetate/propionate ratios were lower for diet F20 than for F80 in sheep rumen (2.73 and 3.97) and SFCCF (3.07 and 4.80), but were higher for diet F20 compared to F80 (4.29 and 3.40) in Rusitec, with values considered to be unphysiological for high-concentrate diets. In vivo NDF digestibility was affected (P = 0.017) by diet, but no differences between diets (P > 0.05) were found in any in vitro system. A more precise control of pH in both types of fermenters and a reduction of concentrate retention time in Rusitec could probably improve the simulation of in vivo fermentation.     

Effect of plant oils and organic acids on rumen fermentation in vitro

We determined the effect of plant oils (rapeseed, sunflower, linseed) and organic acids (aspartic and malic) on the fermentation of diet consisting of hay, barley and sugar beet molasses. Rumen fluid was collected from two sheep (Slovak Merino) fed with the same diet twice daily. Mixed rumen microorganisms were incubated in fermentation fluid, which contained rumen fluid and Mc Dougall's buffer. All supplemented diets significantly increased pH, molar proportion of propionate, and numerically decreased methane production. Lactate production was also decreased significantly (except with malate). Incorporation of plant oils into aspartate- and malate-treated incubations negated the decrease of butyrate, lactate and the increase of pH and ammonia with malate treatment, as well asin vitro dry matter digestibility and pH with aspartate treatment. The effect of combined additives on methane production and molar proportion of propionate was lower compared with additives supplemented separately. Combination of additives had no additive effect on rumen fermentation. All additives decreased total protozoan counts in rumen fluid.     

Comparative studies on the degradation and mean retention time of solid and liquid phases in the forestomachs of dromedaries and sheep fed on low-quality roughages from Tunisia

The digestion of the acid detergent fiber (ADF) fraction of vetch-oat hay was studied in two dromedaries and three sheep, all rumen-fistulated and fed vetch-oat hay with a concentrate. Dromedaries and sheep consumed similar amounts of feed dry matter (DM) per kilogram of metabolic weight, but dromedaries drank less water than sheep. There were no differences in the volatile fatty acid (VFA) and ammonia concentrations in the rumen between dromedaries and sheep, but pH was higher in the dromedaries than in the sheep (P < 0.05). The mixture of VFA contained more propionate and butyrate and less acetate in dromedaries than in sheep (P < 0.05). The protozoal concentration was higher in the rumen of sheep than of dromedaries. This result was consistent with the higher N---NH₃, concentration in sheep. Entodinium was the most abundant species in both dromedaries and sheep. Specific rates of rumen liquid (PEG) and particle (chromium-mordanted hay) outflow were higher and lower, respectively, in dromedaries than in sheep. Also, dromedaries had higher in sacco, in vitro, and fecal digestibilities of vetch-oat hay dry matter (DM) and ADF than sheep. In sacco, the potential degradable fraction was higher, but the rate of degradation was not. The greater utilization of ADF in the rumen by dromedaries is discussed in relation to the higher cellulolytic activity of the rumen microorganisms, the longer retention time of feed particles and/or the greater buffering capacity of the rumen contents of the dromedary against fermentation acids.     

Cereals level and source effects on rumen fermentation,colostrum and milk properties,and blood metabolites in periparturient ewes

Optimal type and dietary inclusion rates of cereal grains for periparturient sheep are unknown. The objective was to determine effects of feeding diets with high (H) v. low (L) levels of ground corn grain (CN) v. combined ground wheat and barley grains (WB) on intake, rumen fermentation, colostrum and milk properties, and blood metabolites of periparturient sheep. Twenty Afshari×Merino ewes were used in a completely randomized design study from 24 days prepartum through 21 days postpartum. Ewes were kept indoors in individual boxes and received once daily at 0900 h total mixed rations. Treatments were mixed rations containing either (1) H or (2) L concentrate based on either (1) 100% CN or (2) 50 : 50 ratio of ground wheat : ground barley grains in a 2×2 factorial arrangement. Each treatment group had five ewes including two twin-lamb ewes and three single-lamb ewes. Postpartal dry matter intake (DMI) increased by feeding high CN v. high and low WB, while high v. low CN improved postpartum DMI. The DMI during lambing tended to increase with the high v. low WB. Feeding CN v. WB, and feeding both CN and WB at L v. H level increased fecal pH. Postpartal rumen pH was lower with the high v. low WB (5.7 v. 6.2). Rumen concentrations of propionate were lower and of acetate were higher with L v. H grain levels. Increased dietary grain reduced urine pH for WB (7.24 v. 7.83) but not for CN (7.63 v. 7.52) prepartum. Colostrum properties, postpartal urine pH, lamb weight at birth and 21 days of age, and placental weight and expulsion time were unaffected. Milk yield increased and milk fat yield tended to increase by H v. L grain diets. Plasma glucose was increased by feeding high v. low WB, whereas CN v. WB tended to reduce peripartal plasma non-esterified fatty acids (NEFA) and increased insulin to NEFA ratio. In conclusion, more cereal grains can be included in periparturient sheep diets and CN instead of WB may be fed to periparturient sheep to improve energy status. Findings suggest opportunities to optimize periparturient ewe physiology and performance through feeding certain cereals and avoiding high levels of WB.     

The effects of barley, unmolassed sugar-beet pulp and molasses supplements on organic matter, nitrogen and fibre digestion in the rumen of cattle given a silage diet

In a 4 × 4 Latin-square experiment with a 2 × 2 factorial arrangement of treatments, 4 cattle fitted with a rumen and duodenal cannula were given four grass-containing diets [480 g kg⁻¹ of the total dry matter (DM) intake] and barley (BU), barley + molasses (2:1) (BM), sugar-beet pulp (SU) or sugar-beet pulp + molasses (SM). Duodenal flow was estimated using Cr-mordanted straw and CoEDTA as markers, and microbial nitrogen entering the small intestine using purine bases of nucleic acids.

Molasses-containing diets had a higher (P < 0.01) organic matter (OM) digestibility. The proportion of digestible OM apparently disappearing in the rumen averaged 0.72 and was not significantly affected by the diet. When cattle received molasses, the quantity of microbial N entering the small intestine was higher (P < 0.05) and there was a trend towards a higher efficiency of microbial N synthesis (28.8 vs. 25.6 g N kg⁻¹ OM apparently digested in the rumen). When S diets were consumed, total non-ammonia N flow at the duodenum exceeded N intake by 7.0 g day⁻¹ and when B diets were consumed, it was 0.7 g day⁻¹ less than N intake. Feed N degradability in the rumen and apparent N digestibility of S diets were lower (P < 0.05; P < 0.001) than those of B diets.

Rumen (P < 0.05) and total (P < 0.001) digestibility of neutral detergent fibre (NDF) and acid detergent fibre (ADF) was higher when S diets were given. The proportion of digestible fibre disappearing in the rumen was not affected by the diet. The rate and extent of silage and hay DM degradation were not significantly affected by the diet. However, dietary inclusion of molasses decreased (P < 0.05) the lag time of both hay and silage DM degradation.

The rumen dilution rate of liquid averaged 0.097 and that of particles, 0.049; neither was significantly different for either B and S diets or U and M diets. Duodenal liquid flow was higher (P < 0.05) for M diets.

Average rumen pH was not affected by the diet, but the molasses diets increased (P < 0.05) the range in rumen pH. The BM diet was associated with higher (P < 0.01) rumen ammonia concentration than the other diets. Low rumen ammonia concentrations (< 2 mM) were observed for long periods between feeds. The molar proportion of butyrate was higher on B diets and there was a trend towards a higher proportion of acetate and propionate on S diets. Molasses tended to increase the molar proportion of propionate and butyrate.     

The redox potential, rH and pH values in the gastrointestinal tract of small ruminants

Oxidation-reduction potentials (Eh), pH and rH in the gastrointestinal tract were measured in six goats and two sheep fed on ground barley and hay, with or without the addition of urea. Each ration was supplied for three weeks. The animals were slaughtered after morning feeding, the contents of relevant parts of the gastrointestinal tract were sampled, pH and Eh values were measured and rH values calculated. The range of the oxidation-reduction potential was rather extensive, from -300 to +186 mV. The variability of rH values was smaller, being between 4.6 and 12.9, except for three values. The following linear relationship holds for findings of Eh and pH: Eh (mV) = 292-69.9 pH with a correlation coefficient of r = 0.84. In those parts of the gastrointestinal tract, where the fermentation process occurs (the rumen, caecum and colon), the Eh and rH values are lower than those in the abomasum and duodenum. Urea addition has no effect on the oxidation-reduction equilibrium.     

Grain source and marginal changes in forage particle size modulate digestive processes and nutrient intake of dairy cows

This study investigated the effects of, and interactions between, dietary grain source and marginal changes in alfalfa hay (AH) particle size (PS) on digestive processes of dairy cows. A total of eight Holstein dairy cows (175 days in milk) were allocated in a replicated 4 × 4 Latin square design with four 21-day periods. The experiment was a 2 × 2 factorial arrangement with two levels of theoretical PS of AH (fine = 15 mm or long = 30 mm) each combined with two different sources of cereal grains (barley grain alone or barley plus corn grain in a 50 : 50 ratio). Results showed that cows consuming diets supplemented with corn had greater dry matter and nutrient intakes (P < 0.01), independent of forage PS. In addition, the apparent digestibility of fiber fractions was greater for diets supplemented with corn (P = 0.01). The feeding of barley grain-based diets was associated with greater apparent digestibility of non-fiber carbohydrates, and this variable was even greater when long AH was fed (P = 0.04). Moreover, the feeding of long AH resulted in longer time spent eating (P = 0.03) and higher pH (P < 0.01), as well as a tendency for higher acetate-to-propionate ratio in the rumen fluid (P = 0.06) at 3 h post feeding. In conclusion, the results indicated that the marginal increase of PS of AH may prolong eating time and improve rumen fermentation, particularly in diets based on barley grain. Partial substitution of barley grain by corn can improve feed intake and fiber digestibility in mid-lactation dairy cows.     

Repeated acidosis challenges and live yeast supplementation shape rumen microbiota and fermentations and modulate inflammatory status in sheep

This study aimed to investigate the impact of repeated acidosis challenges (ACs) and the effect of live yeast supplementation (Saccharomyces cerevisiae I-1077, SC) on rumen fermentation, microbial ecosystem and inflammatory response. The experimental design involved two groups (SC, n=6; Control, n=6) of rumen fistulated wethers that were successively exposed to three ACs of 5 days each, preceded and followed by resting periods (RPs) of 23 days. AC diets consisted of 60% wheat-based concentrate and 40% hay, whereas RPs diets consisted of 20% concentrate and 80% hay. ACs induced changes in rumen fermentative parameters (pH, lactate and volatile fatty-acid concentrations and proportions) as well as in microbiota composition and diversity. The first challenge drove the fermentation pattern towards propionate. During successive challenges, rumen pH measures worsened in the control group and the fermentation profile was characterised by a higher butyrate proportion and changes in the microbiota. The first AC induced a strong release of rumen histamine and lipopolysaccharide that triggered the increase of acute-phase proteins in the plasma. This inflammatory status was maintained during all AC repetitions. Our study suggests that the response of sheep to an acidosis diet is greatly influenced by the feeding history of individuals. In live yeast-supplemented animals, the first AC was as drastic as in control sheep. However, during subsequent challenges, yeast supplementation contributed to stabilise fermentative parameters, promoted protozoal numbers and decreased lactate producing bacteria. At the systemic level, yeast helped normalising the inflammatory status of the animals.     

Deuterium oxide dilution accurately predicts water intake in sheep and goats

The aim of this study was to test whether the deuterium oxide dilution technique accurately predicts water intake in sheep and goats. Two other issues were also studied: (i) a comparison of water intake in sheep and goats and (ii) an assessment of whether observations of drinking behaviour can accurately measure the water intake. In this study, eight dry Boer goats and eight dry German Black Head Mutton ewes were kept under controlled stable conditions. Animals had access to hay and water ad libitum. Diurnal drinking behaviour was recorded by video. Individual daily water intake was measured and estimated for 2 weeks by re-weighing water buckets and from water kinetics using the deuterium oxide dilution technique, respectively. In addition, dry matter intakes were directly measured and were significantly higher in sheep than in goats. The average daily water consumption by drinking differed significantly between the two species, with higher intakes in sheep than in goats. Total body water expressed as a percentage of body mass did not differ between species. Measurement methods of total water intake (TWI) using deuterium oxide dilution and re-weighing water buckets did not differ significantly in both species (P = 0.926). Results obtained for measured and estimated TWI confirm that the isotope dilution technique gives reliable results for estimates of water intake in sheep and goats. The higher amount of water intake in sheep was also reflected by their drinking behaviour. Sheep spent approximately 0.3% per 24-h drinking, while Boer goats spent only 0.1%. However, measured and estimated TWIs were only moderately correlated to the daily time spent drinking. The lower water intake found in Boer goats confirms a superior water management capacity compared with Black Head Mutton sheep even under temperate conditions.     

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.     

Milk production is unaffected by replacing barley or sodium hydroxide wheat with maize cob silage in rations for dairy cows

Starch is an important energy-providing nutrient for dairy cows that is most commonly provided from cereal grains. However, ruminal fermentation of large amounts of easily degradable starch leads to excessive production and accumulation of volatile fatty acids (VFA). VFA not only play a vital role in the energy metabolism of dairy cows but are also the main cause of ruminal acidosis and depressed feed intake. The aim of the present study was to compare maize cob silage (MCS) as an energy supplement in rations for dairy cows with highly rumen-digestible rolled barley and with sodium hydroxide wheat (SHW), which has a higher proportion of by-pass starch than barley. Two studies were carried out: (1) a production study on 45 Danish Holstein cows and (2) an intensive study to determine digestibilities, rumen fermentation patterns and methane emission using three rumen-cannulated Danish Holstein cows. Both studies were organised as a 3×3 Latin square with three experimental periods and three different mixed rations. The rations consisted of grass-clover silage and maize silage (~60% of dry matter (DM)), rapeseed cake, soybean meal, sugar beet pulp and one of three different cereals as a major energy supplement: MCS, SHW or rolled barley (~25% of DM). When MCS replaced barley or SHW as an energy supplement in the mixed rations, it resulted in a lower dry matter intake; however, the apparent total tract digestibilities of DM, organic matter, NDF, starch and protein were not different between treatments. The energy-corrected milk yield was unaffected by treatment. The fat content of the milk on the MCS ration was not different from the SHW ration, whereas it was higher on the barley ration. The protein content of the milk decreased when MCS was used in the ration compared with barley and SHW. From ruminal VFA patterns and pH measures, it appeared that MCS possessed roughage qualities with respect to rumen environment, while at the same time being sufficiently energy rich to replace barley and SHW as a major energy supplement for milk production. The environmental impact, expressed as methane emissions, was not different when comparing MCS, SHW and barley.     

Effect of forage particle length on rumen fermentation,sorting and chewing activity of late-lactation and non-lactating dairy cows

The objective of this study was to determine the effects of varying forage particle length on chewing activity, sorting behavior, rumen pH and rumen fill in late lactation and dry dairy cattle, fed rations with similar physically effective NDF but different mean particle length. Treatments consisted of three diets differing only in geometric mean length of forage: hay (5.40, 8.96 and 77.90 mm, for short (S), medium (M) and long (L) diets, respectively) for Experiment 1 (E1), and straw (10.16, 24.68 and 80.37 mm) for S, M and L diets, respectively, for Experiment 2 (E2). Hay or straw comprised the sole source of forage (50% and 75% of ration dry matter (DM) for E1 and E2, respectively). Both experiments used three rumen cannulated Holstein dairy cows, in late lactation for E1 and dry in E2, with 3 × 3 Latin square designs with 14 day periods. In E1, DM intake (DMI; 18.3 ± 2.1 kg/day; mean ± s.e.), pH (6.4 ± 0.1), time spent eating (280 ± 22.5 min/day), time spent ruminating (487 ± 17 min/day), and total time spent chewing (767 ± 34 min/day) were not different, whereas eating minutes per kilogram of DMI and NDF intake (NDFI) tended to increase linearly as forage length increased. Rumen digesta volume (l; 113.3 S, 117.8 M and 114.4 L ± 17.1) had a quadratic response, and rumen digesta weight tended to respond quadratically; however, differences were numerically small. In E2, DMI (8.3 ± 1.3 kg/day), pH (6.7 ± 0.1), time spent eating (236 ± 23.5 min/day), time spent ruminating (468 ± 45.2 min/day), total time spent chewing (704 ± 67.7 min/day) and minutes per kilogram NDFI were not different, whereas minutes per kilogram of DMI had a trend for a quadratic effect. Rumen digesta volume (111 ± 18.8 l) and weight (103 ± 17.4 kg) were not different. In both experiments, cows sorted against longer particles as determined by a particle length selection index; this behavior increased linearly as particle length increased. Greater forage particle length increased sorting behavior, but had no effect on rumen fermentation or chewing behavior.     

Rumen chemical and bacterial changes during stepwise adaptation to a high-concentrate diet in goats

The correlation between rumen chemical and bacterial changes was investigated during a four periodical stepwise adaptation to a high-concentrate diet (concentrate level at 0%, 30%, 50% and 70% for diet I to IV, respectively) in goats. The results showed that ruminal pH decreased from 6.7 to 5.5 after switching from diet I to II, and was maintained at about 5.5 on diet III. Denaturing gradient gel electrophoresis results showed that the rumen bacterial community was relatively stable during the initial three feeding periods, except for the appearance of three bands when diet changed from I to II, suggesting that an appropriate concentrate level can promote the proliferation of some bacteria. After 12 days of feeding diet III, total volatile fatty acid (VFA) concentration and butyrate proportion decreased. At days 2 and 3 of feeding diet IV, ruminal pH declined sharply to 5.3 and 4.7, respectively, and total VFA concentration decreased further while lactic acid concentration increased markedly, suggesting a relation between lactic acid accumulation and ruminal pH decline. At the same time, many bacteria disappeared, including most fibrolytic-related bacteria while Streptococcus bovis and Prevotella-like species dominated. Interestingly, Succinivibrio dextrinosolvens-like species maintained throughout the experiment, suggesting its tolerance to low pH. In conclusion, rumen bacterial community was relatively stable feeding 0% to 50% concentrate diets, and it was observed that appropriate concentrate levels in the diet could increase the diversity of rumen bacteria. However, concentrate-rich diets caused lactic acid accumulation and low ruminal pH that caused the disappearance of most fibrolytic-related bacteria sensitive to low pH while S. bovis and genus Prevotella persisted.     

Effects of expander-treating a barley-based concentrate on ruminal fermentation, bacterial N synthesis, escape of dietary N, and performance of dairy cows

Three primiparous dairy cows in early lactation with cannulas in rumen, duodenum and ileum were used in a 3×3 Latin square design to study effects of expander treatment of a barley-based concentrate. The concentrate was either pelleted at 75–80°C or expander treated at 125–130°C prior to pelleting. The diets consisted of 6.7 kg DM of grass silage and 10 kg DM of (1) 100% pelleted, (2) 50% pelleted and 50% expanded or (3) 100% expanded concentrate. The diets were offered as a mixed ration in four equal meals daily. Ruminal fermentation, bacterial N synthesis, duodenal, ileal and faecal flow of nutrients, and animal performance were monitored. Expander treatment numerically increased ruminal digestion of starch, which explained the observed increase in ruminal VFA concentration and the lowered ruminal pH (P<0.05). The proportion of butyrate in rumen liquid increased, whereas the proportion of propionate decreased in the expanded compared to the pelleted treatment (P<0.05). Expander treatment tended to increase rumen volume and rumen NDF pool size. Ruminal digestion of NDF was numerically lower in the expanded than in the pelleted treatment. No differences in bacterial N synthesis or efficiency of synthesis were observed among treatments. Expander treatment numerically increased the duodenal flow of non-ammonia N (NAN) and amino acid N (AAN), and seemed to increase the flow of non-ammonia non-bacterial N (NANBN) to the duodenum to a similar extent as was indicated by nylon bag studies. Milk production and milk fat and protein content were increased by the expander treatment (P<0.05), indicating that expander treatment increased the supply of nutrients for milk production.     

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.     

Effect of rumen ciliates on the digestive utilization of various carbohydrate-rich diets and on the end-products formed in the rumen. II. Utilization of inulin, saccharose and lactose

Three diets rich in inulin, saccharose and lactose, respectively, were given to 10 rumen-fistulated sheep. Two animals were defaunated, two were inoculated with either Polyplastron multivesibulatum or Entodinium sp., and two others were inoculated with both. The latter two were bred in conventional conditions. All animals ingested the same amounts of carbohydrates in the three diets (21-22 g/kg P0.75/day). Dietary nitrogen content was similar (table 1). The ciliate population was improved with the inulin diet (fig. 1; table 2). With a mixed population, the Entodinium improved with the inulin diet (fig 1; table 2). With a mixed population, the Entodinium sp. genus was always predominant. Holotrich protozoa (mainly Isotricha) in the rumen of the conventional sheep represented 15 to 30% of the total ciliate biomass, indicating that they were able to metabolize these soluble sugars. We also observed that P. multivesiculatum can ferment cellulose and all the soluble carbohydrates proposed in these diets. However, Entodinium sp. development occurred mainly in the presence of the sugard produced during carbohydrate hydrolysis by other ciliates or bacteria. The highest organic matter digestibility, noted in faunated animals (table 3) was confirmed by the VFA concentration in the rumen (table 4). This could be explained either by an activation of bacterial metabolism due to predation or by the direct effect of ciliates on fermentations, or both. Modifications in the VFA composition varied with ciliate inoculation, showing that ciliate metabolism may vary with the nature of the energy in the diet or that the observed results depended on various opposite effects in which the intensity of each component was influenced by the diet. In general, the acetic acid molar proportion increased and propionic acid decreased when there was a considerable Entodinium sp. population. The effect on butyric acid was low with these diets. Higher ammonia and lactic acid concentrations were observed in the rumen of faunated than defaunated sheep, irrespective of the ciliate inoculum.     

In Vitro Lactate Metabolism by Ruminal Ingesta

Ruminal ingesta (300 ml) obtained from a fistulated cow fed alfalfa hay (H), 3.6 kg of grain mixture with corn silage fed ad libitum (S), 2.5:1 grain-alfalfa hay mixture (G), or a 2.5:1 grain-alfalfa hay mixture providing 545 g of sodium and calcium lactate daily (L) were incubated for 8 hr with nonpolymerized sodium lactate or 17% polymerized lactic acid neutralized to pH 6.7. Polymerization had no effect on the rate of lactate utilization. The initial rates of lactate metabolism for the H, G, S, and L ingesta were 0.72, 0.95, 1.8, and 3.4 meq per 100 ml of rumen fluid per hr, respectively. Lactate-2-(14)C was incubated for 4 hr with each type of ruminal ingesta. Of the label recovered in the volatile fatty acids (VFA), 74.1, 61.2, 49.3, and 38.9% was recovered in acetate, and 9.4, 19.8, 23.3, and 51.9% was recovered in propionate with H, G, S, and L ingesta, respectively. The balance of label was distributed between butyrate and valerate. The titratable VFA did not follow this pattern of production. With the hay ingesta, lactate metabolism resulted in a net loss of acetate and a large increase in butyrate. Little propionate was produced. The G, S, and L ingesta metabolized lactate to yield progressively more propionate and less butyrate. Evidence was gathered to suggest that acetate was the primary end product of lactate metabolism but that oxidation of lactate to pyruvate dictated the synthesis of butyrate from acetate to maintain an oxidation-reduction balance. It was noted that acetate and butyrate production from lactate was pH-dependent, with acetate production maximal at pH 7.4 and butyrate at 6.2. Propionate production was largely unaffected within this pH range.     

Indigestion in Young Calves. II. The Influence Of Ground Barley,Coarse and Fine Hay

Four experiments comprising 86 calves have been carried out in order to examine the influence of ground barley, coarse and fine hay, when raising young calves on a high lactose milk replacer causing diarrhoea. The frequency of diarrhoea decreased when feeding barley (P < 0.01) or fine hay (P < 0.001), but increased when feeding coarse hay (P<0.01). Barley increased (P < 0.01) live weight gain and carcass weight, reduced (P < 0.001) rumen pH, favoured gram-positive cocci and rods in the rumen, caused hyperkeratosis and gave increased (P < 0.01) empty reticulo-rumen weights, the latter being supposed to be enhanced by hyperkeratosis. The calves ate more (P < 0.001) fine than coarse hay, fine hay being superior (P < 0.01) in promoting growth of the rumen. All hay maintained a predominantly gram-negative rumen flora, but somewhat different from that on the milk replacer, and counteracted (P < 0.01) low pH in the rumen and apparently also hyperkeratosis due to barley. The results with barley indicated no relationship between diarrhoea and hyperkeratosis.     

Effects of the replacement of concentrate and fibre-rich hay by high-quality hay on chewing,rumination and nutrient digestibility in non-lactating Holstein cows

The aim of this study was to investigate the effects of high-quality hay with an elevated sugar content alone or with graded amounts of concentrate feed on chewing and ruminating activity, apparent total tract digestibility (ATTD) and ruminal pH at different time points after feeding in the free ruminal liquid (FRL) and the particle-associated ruminal liquid (PARL). Eight rumen cannulated non-lactating Holstein cows were arranged in a Latin square design in four experimental runs lasting 25 d each. The four diets tested were 60NQ (60% normal-quality hay + 40% concentrate), 60HQ (60% high-quality hay + 40% concentrate), 75HQ (75% high-quality hay + 25% concentrate) and 100HQ (100% high-quality hay). Normal and high-quality hays differed in sugar contents (11.3% vs. 18.7% in dry matter [DM]), neutral detergent fibre (NDF; 57.7% vs. 46.3% in DM), acid detergent fibre (ADF, 35.0% vs. 23.5% in DM) and crude protein (CP, 11.3% vs. 23.5% in DM). Data showed that ATTD of DM, CP, NDF and ADF was higher with the high-quality hay diets. Time spent eating was reduced with high-quality hay. However, time spent ruminating was longest in Group 100HQ. In all groups, ruminal pH of FRL and PARL decreased with time after the morning feeding. But 10 h later, pH of Group 100HQ was higher again compared with the other groups. Considering the average pH in FRL over all measured time points, cows in Groups 60NQ and 100HQ had higher pH values of 6.85 and 6.83, respectively. Regarding pH values in PARL, animals of Group 60NQ displayed the highest pH value (6.68), whereas the lowest value of 6.21 was found in Group 60HQ. Overall, results suggest that high-quality hay maintains the diet’s structural effectiveness by stimulating rumination and stabilising ruminal pH while greatly improving ATTD. However, the structural effectiveness of the high-quality hay gets impaired with increasing proportion of concentrate feed in the diet.