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.     

Studies on the biotin flow at the duodenum of dairy cows fed diets with different concentrate levels and types of forages

Biotin is involved in many vital metabolic pathways and must be provided for an efficient fermentation in the rumen, as well as for the intermediary metabolism of the host animal. Factors influencing ruminal biotin metabolism and output are widely unknown at present. Therefore, dairy cows fitted with permanent cannulas in the dorsal rumen and in the proximal duodenum were fed differently composed diets, and the biotin flow at the proximal duodenum was measured. The diets (on DM basis) consisted of 8.9 kg grass hay (Diet 1), 8.9 kg corn silage plus 2.0 kg concentrate (Diet 2), or 7.3 and 7.4 kg grass silage plus 10.0kg concentrate (Diets 3 and 4). The concentrate in Diets 3 and 4 contained 87% wheat and corn grain, respectively. The cows were pre-fed the rations for 21 days. Thereafter duodenal digesta was sampled every two h for 5 days. Cr2O3 served as a flow marker and the microbial proportion of total nitrogen at the duodenum was estimated by near infrared spectroscopy (NIRS). The duodenal flow of biotin was not related to biotin intake, but to the amount of fermented organic matter (FOM) and the amount of microbial protein (Biotin [mg/d] = 0.518 kg FOM - 0.300; r=0.85 and biotin [mg/d] = 0.012 x g microbial protein + 1.478; r = 0.84), irrespective of the composition of the diet fed. Mean daily biotin flow was 0.48 +/- 0.11 mg/kg FOM without any systematic effect of diet composition. The ruminal biotin balance, calculated as the difference between biotin flow at the duodenum and biotin intake, was positive (1.4 - 2.0 mg/d) in cows fed the mixed roughage/concentrate diets and negative (-0.71 mg/d) when the pure hay diet was fed.     

Studies on the biotin flow at the duodenum of dairy cows fed diets with different concentrate levels and types of forages

Abstract

Biotin is involved in many vital metabolic pathways and must be provided for an efficient fermentation in the rumen, as well as for the intermediary metabolism of the host animal. Factors influencing ruminal biotin metabolism and output are widely unknown at present. Therefore, dairy cows fitted with permanent cannulas in the dorsal rumen and in the proximal duodenum were fed differently composed diets, and the biotin flow at the proximal duodenum was measured. The diets (on DM basis) consisted of 8.9 kg grass hay (Diet 1), 8.9 kg corn silage plus 2.0 kg concentrate (Diet 2), or 7.3 and 7.4 kg grass silage plus 10.0 kg concentrate (Diets 3 and 4). The concentrate in Diets 3 and 4 contained 87% wheat and corn grain, respectively. The cows were pre-fed the rations for 21 days. Thereafter duodenal digesta was sampled every two h for 5 days. Cr₂O₃ served as a flow marker and the microbial proportion of total nitrogen at the duodenum was estimated by near infrared spectroscopy (NIRS). The duodenal flow of biotin was not related to biotin intake, but to the amount of fermented organic matter (FOM) and the amount of microbial protein (Biotin [mg/d] = 0.518 · kg FOM?0.300; r = 0.85 and biotin [mg/d] = 0.012 · g microbial protein + 1.478; r = 0.84), irrespective of the composition of the diet fed. Mean daily biotin flow was 0.48 ± 0.11 mg/kg FOM without any systematic effect of diet composition. The ruminal biotin balance, calculated as the difference between biotin flow at the duodenum and biotin intake, was positive (1.4 – 2.0 mg/d) in cows fed the mixed roughage/concentrate diets and negative (?0.71 mg/d) when the pure hay diet was fed.     

An investigation of the relationship between fecal and rumen bacterial concentrations in sheep

With no acceptable method for collecting fresh rumen fluid from zoo ruminants, it was proposed that fecal bacterial concentrations may be correlated with rumen bacteria. If so, fecal bacterial concentrations could be used to study both the effects of diet on rumen bacteria as well as rumen abnormalities. Total and cellulolytic bacterial concentrations were determined in whole rumen contents and feces of sheep using a most‐probable‐number (MPN) assay. In a Latin square design, four crossbred ewes were fed diets of 100% long or chopped orchardgrass hay (OH) and 60% ground or whole shelled corn plus 40% chopped OH. In a second trial, the sheep were fed a pelleted complete feed at varying levels of intake i.e., control at 2.0% of body weight and at 1.8, 1.6, and 1.2% of body weight. Higher total rumen bacterial concentrations (P<0.01) were found on the high concentrate diets as compared with the high forage diets. Grinding the corn also increased total bacterial concentrations (P<0.05). Fecal concentrations of total bacteria were higher (P<0.01) with the high concentrate diets. Chopping the forage decreased the concentration of fecal cellulolytic bacteria (P<0.05) but had no effect on their concentration in the rumen. An inverse linear relationship (P<0.01) was observed between total bacterial concentrations in the feces and diet intake. Although relationships were observed between the rumen and feces for total and cellulolytic bacterial concentrations, they were dependent on diet, particle size, and level of intake. Thus, fecal bacterial concentrations cannot be used to reliably predict rumen bacterial concentrations. Zoo Biol 27:100–108, 2008. © 2008 Wiley‐Liss, Inc.     

Rumen Fungi and Forage Fiber Degradation

The role of anaerobic rumen fungi in in vitro forage fiber degradation was determined in a two forage × two inoculum source × five treatment factorial design. Forages used as substrates for rumen microorganisms were Coastal bermuda grass and alfalfa; inoculum sources were rumen fluid samples from a steer fed Coastal bermuda grass hay or alfalfa hay; treatments were whole rumen fluid (WRF), WRF plus streptomycin (0.2 mg/ml of rumen fluid) and penicillin (1.25 mg/ml of fluid), WRF plus cycloheximide (0.5 mg/ml of fluid), WRF plus streptomycin, penicillin, and cycloheximide, and McDougall buffer. Populations of fungi as shown by sporangial development were greater on bermuda grass leaves than on alfalfa leaflets regardless of inoculum source. However, endogenous fungal populations were greater from the alfalfa hay inoculum. Cycloheximide inhibited the fungi, whereas streptomycin and penicillin, which inhibit bacterial populations, resulted in an increase in numbers of sporangia in the alfalfa inoculum, suggesting an interaction between bacteria and fungi. Bacteria (i.e., WRF plus cycloheximide) were equal to the total population in degrading dry matter, neutral-detergent fiber (NDF), acid-detergent fiber (ADF), and cellulose for both inocula and both forages. Degradation of dry matter, NDF, ADF, and cellulose by anaerobic fungi (i.e., WRF plus streptomycin and penicillin) was less than that due to the total population or bacteria alone. However, NDF, ADF, and cellulose digestion was 1.3, 2.4, and 7.9 percentage units higher, respectively, for bermuda grass substrate with the alfalfa versus bermuda grass inoculum, suggesting a slight benefit by rumen fungi. No substantial loss of lignin (72% H₂SO₄ method) occurred due to fungal degradation. The most active fiber-digesting population in the rumen was the bacteria, even when streptomycin and penicillin treatment resulted in an increase in rumen fungi over untreated WRF. The development of large numbers of sporangia on fiber may not indicate a substantial role as digesters of forage.     

Removal of anaerobic fungi from the rumen of sheep by chemical treatment and the effect on feed consumption and in vivo fibre digestion

The population of anaerobic fungi in the rumen of sheep was reduced by the addition of tetronasin (an ionophore antibiotic) to a herbage diet. Fungi were reduced to undetectable levels (< 1 fungal zoospore per ml rumen fluid) by the combined addition of tetronasin and cycloheximide (a protein synthesis inhibitor) and the absence of fungi was maintained with low levels of tetronasin. Sheep with fungi present in the rumen ate 40% more of a straw-based diet (with a fibre digestibility in vivo of 51%) than they ate when without fungi (47% fibre digestibility). Counts of total viable bacteria, cellulolytic bacteria and ciliate protozoa in the rumen were not significantly different when anaerobic fungi were either present or absent.     

Effect of wheat hay particle size and replacement of wheat hay with wheat silage on rumen pH,rumination and digestibility in ruminally cannulated non-lactating cows

This study examined the effects on intake, diurnal rumen pH changes, rumination and digestibility of feeding ruminally cannulated non-lactating cows in a Latin square design (four cows×four periods) with four total mixed rations (TMRs) typical for lactating cows. TMRs were based on: long wheat hay or short wheat hay, wheat silage or wheat silage+1.5% NaHCO₃ buffer, as the sole roughage source (30% of TMR dry matter (DM)). The level of physically effective NDF remaining above the 8 mm screen (peNDF) was similar in the long hay and silage-based TMRs (9.45% to 9.64% of DM) and lower in the short hay TMR (7.47% of DM). The four TMRs were offered individually at 95% of ad libitum intake to avoid orts within 24 h. Cows fed long hay consumed less DM than the short hay and silage groups (9.6 v. 10.5 and 10.8 kg/day, respectively) and sorted against large hay particles at 12 h post-feeding. Under the limitations of this study (non-lactating cows fed at restricted intake) short hay TMR prevented sorting within 12 h post-feeding, encouraged rumination per kg peNDF ingested, and had higher average rumen pH (6.24), whereas preventing sub acute ruminal acidosis (SARA, defined as pH<5.8 for at least 5 h/day). In contrast, the long hay and silage-based groups were under SARA. In vitro methane production of rumen fluid was higher in the hay-fed cows than in their silage-fed counterparts, and in all treatments lower at 1 h pre-feeding than at 6 h post-feeding. In vivo DM and NDF digestibility were similar for the short hay and silage TMRs, and higher than those of the long hay TMR. Under the conditions of this study, addition of 1.5% buffer to the wheat silage TMR had no effect on intake, rumen pH, creation of SARA and digestibility.     

Detection and monitoring of anaerobic rumen fungi using an ARISA method

Aim: To develop an automated ribosomal intergenic spacer region analysis (ARISA) method for the detection of anaerobic rumen fungi and also to demonstrate utility of the technique to monitor colonization and persistence of fungi, and diet‐induced changes in community structure. Methods and Results: The method could discriminate between three genera of anaerobic rumen fungal isolates, representing Orpinomyces, Piromyces and Neocallimastix species. Changes in anaerobic fungal composition were observed between animals fed a high‐fibre diet compared with a grain‐based diet. ARISA analysis of rumen samples from animals on grain showed a decrease in fungal diversity with a dominance of Orpinomyces and Piromyces spp. Clustering analysis of ARISA profile patterns grouped animals based on diet. A single strain of Orpinomyces was dosed into a cow and was detectable within the rumen fungal population for several weeks afterwards. Conclusions: The ARISA technique was capable of discriminating between pure cultures at the genus level. Diet composition has a significant influence on the diversity of anaerobic fungi in the rumen and the method can be used to monitor introduced strains. Significance and Impact of the Study: Through the use of ARISA analysis, a better understanding of the effect of diets on rumen anaerobic fungi populations is provided.     

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.     

Effects of increasing amounts of extruded linseed in the diet on apparent ruminal synthesis of some B vitamins in dairy cows

Many studies have shown that metabolic efficiency of ruminants can be significantly decreased when B-vitamin supply is insufficient. Under the present state of knowledge, the amounts of B vitamins available for intestinal absorption cannot be predicted based on diet composition. Therefore, in an attempt to increase our understanding of the effects of dietary factors, on B-vitamin supply for dairy cows, the effects of increasing amounts of extruded linseed in diets based on hay (permanent grassland hay, H; Experiment 1) or corn silage (CS; Experiment 2) on apparent ruminal synthesis (ARS) of thiamin, riboflavin, niacin, vitamin B₆, folates and vitamin B₁₂ were evaluated. In each experiment, four lactating Holstein cows fitted with cannulas in the rumen and the proximal duodenum were used in a 4 × 4 Latin square design. In both experiments, the dietary treatments consisted of an increasing supply of extruded linseed representing 0%, 5%, 10% or 15% of diet DM. The forage : concentrate ratios were 50 : 50 and 60 : 40 for Experiments 1 and 2, respectively. Duodenal flow was determined using YbCl₃ as a marker. The ARS of each B vitamin was calculated as duodenal flow – daily intake. In both experiments, treatments did not affect thiamin, riboflavin, niacin and vitamin B₁₂ duodenal flow or ARS. Increasing the dietary concentration of extruded linseed decreased folate intake in Experiment 1 and vitamin B₆ intake in Experiment 2 but resulted in a greater duodenal flow of vitamin B₆ and folates regardless of the forage used in basal diet. Greater dietary linseed concentrations decreased vitamin B₆ apparent degradation in the rumen in CS-based diet only and increased folate ARS in both H- and CS-based diets. Increasing linseed concentration of isonitrogenous and isoenergetic diets increased vitamin B₆ and folate supply to dairy cows, both with H- and CS-based diets.     

Bacteria isolated from the duodenum, ileum, and cecum of young chicks.

Facultatively anaerobic and strictly anaerobic bacteria colonizing the intestinal tracts of 14-day-old chicks fed a corn-based diet were enumerated, isolated, and identified. Colony counts from anaerobic roll tubes (rumen fluid medium) or aerobic plates (brain heart infusion agar) recovered from homogenates of the duodenum, upper and lower ileum, and cecum varied appreciably among samples from individual birds. Anaerobic and aerobic counts from the duodenum and ileum were similar. Anaerobic counts were highest from the cecum (0.7 X 10(11) to 1.6 X 10(11)/g of dry tissue) and exceeded aerobic plate counts by a factor of at least 10(2). Facultatively anaerobic groups (Streptococcus, Staphylococcus, Lactobacillus, and Escherichia coli) comprised the predominant flora of the duodenum and ileum, although large numbers of anaerobes (9 to 39% of the small intestine isolates), represented by species of Eubacterium, Propionibacterium, Clostridium, Gemmiger, and Fusobacterium, were also recovered. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium, Clostridium Gemmiger, Fusobacterium, and Bacteriodes) made up nearly the entire microbial population of the cecum. Scanning electron microscopy of the intestinal epithelia of chicks revealed populations of microbes on the duodenal, ileal, and cecal mucosal surfaces.     

Effect of substituting wet corn gluten feed and corn stover for alfalfa hay in total mixed ration silage on lactation performance in dairy cows

Wet corn gluten feed (WCGF) is a high moisture feed containing rapidly digestible, non-forage fiber and protein. The objective of this study was to investigate the effect of substituting WCGF and corn stover for alfalfa hay in total mixed ration (TMR) silage on lactation performance and nitrogen balance in dairy cows. Nine multiparous Holstein dairy cows (BW = 532 ± 28.9 kg and day in milk = 136 ± 5.6 d; mean ± SD) were used in a replicated 3 × 3 Latin square design with 21-d periods (14 d of diet adaption and 7 d of sample collection). Groups were balanced for parity, day in milk, and milk production and consumed one of three treatment diets during each period. The treatment diets were fed as TMR and contained similar concentrate mixtures and corn silage but different proportions of roughage and WCGF. The three treatments were: (1) 0% WCGF, 0% corn stover, and 22.1% alfalfa hay (0% WCGF); (2) 6.9% WCGF, 3.4% corn stover, and 11.8% alfalfa hay (7% WCGF); and (3) 13.3% WCGF, 4.9% corn stover, and 3.9% alfalfa hay (13.3% WCGF). Compared to the 0% WCGF diet, the cows fed the 7% and 13.3% WCGF diets had a higher milk yield and concentration of milk fat, protein, lactose, and total solids. Effective degradability of DM was higher in the cows fed the 7% and 13.3% WCGF diets than it was with the 0% WCGF diet. Cows fed the 13.3% WCGF had a higher CP effective degradability and a lower rumen undegraded protein than cows fed the 0% WCGF diet. The concentration of ruminal volatile fatty acids and ammonia-N was higher in cows fed the 7% and 13.3% WCGF diets than cows fed the 0% WCGF diet. The fecal N was lower in cows fed the 7% and 13.3% WCGF diets than it was in cows fed the 0% WCGF diet. Milk N secretion and milk N as a percent of N intake were higher in cows fed the 13.3% WCGF diet than cows fed the 0% and 7% WCGF diets. In conclusion, it appears that feeding a TMR silage containing WCGF and corn stover in combination, replacing a portion of alfalfa hay, may improve lactation performance and nitrogen utilization for lactating dairy cows.     

Effects of common forage phenolic acids on Escherichia coli O157:H7 viability in bovine feces

Ruminant animals are carriers of Escherichia coli O157:H7, and the transmission of E. coli O157:H7 from cattle to the environment and to humans is a concern. It is unclear if diet can influence the survivability of E. coli O157:H7 in the gastrointestinal system or in feces in the environment. Feces from cattle fed bromegrass hay or corn silage diets were inoculated with E. coli O157:H7, and the survival of this pathogen was analyzed. When animals consumed bromegrass hay for <1 month, viable E. coli O157:H7 was not recovered after 28 days postinoculation, but when animals consumed the diet for >1 month, E. coli O157:H7 cells were recovered for >120 days. Viable E. coli O157:H7 cells in feces from animals fed corn silage were detected until day 45 and differed little with the time on the diet. To determine if forage phenolic acids affected the viability of E. coli O157:H7, feces from animals fed corn silage or cracked corn were amended with common forage phenolic acids. When 0.5% trans-cinnamic acid or 0.5% para-coumaric acid was added to feces from silage-fed animals, the E. coli O157:H7 death rate was increased significantly (17-fold and 23-fold, respectively) compared to that with no addition. In feces from animals fed cracked corn, E. coli O157:H7 death rates were increased significantly with the addition of 0.1% and 0.5% trans-cinnamic acid (7- and 13-fold), 0.1% and 0.5% p-coumaric acid (3- and 8-fold), and 0.5% ferulic acid (3-fold). These data suggest that phenolic acids common to forage plants can decrease viable counts of E. coli O157:H7 shed in feces.     

Comparative denaturing gradient gel electrophoresis analysis of fungal communities associated with whole plant corn silage.

Significant portions of grain produced for livestock consumption are convened into ensiled forage. Silage producers have long recognized the positive effects of using an inoculant to insure the proper transformation of forage into a palatable and digestible feedstuff. When silage is fed from a storage structure, exposure to air stimulates the growth of epiphytic aerobes that may result in the loss of up to 50% of the dry matter. Moreover, fungi have been found to be associated with ensiled forage, but their growth is normally suppressed by the anaerobic conditions. However, the introduction of oxygen results in a fungal bloom, and the fungi and the associated metabolites may result in lost productivity in the livestock consuming the contaminated forage. In this study, we report on the diversity of the fungal community associated with whole plant corn silage during the ensiling process, and the effect of two different bacterial inoculants as compared with the uninoculated natural epiphytic fermentation on the distribution of the fungi associated with the silage. The fungal community from duplicate mini-silo packages of the same treatment was analyzed by denaturing gradient gel electrophoresis and direct sequencing of the resulting operational taxonomic units. This method proved useful in analyzing the complex microbial communities associated with the forage in that it was possible to determine that one inoculant dramatically influenced the fungal community associated with whole plant corn silage.     

Rumen fungi: morphological types from Georgia cattle and the attack on forage cell walls

Fungal colonies developing in anaerobic media from zoospores in rumen fluid from cows eating Cynodon dactylon or Medicago sativa included types showing monocentric and polycentric growth. High energy supplements added to diets of Sorghum bicolor silage increased fungal numbers in the rumen, but increases were also affected by the history and predisposition of the animal. Mixed fungal types in rumen fluid and pure cultures of isolates showing monocentric and polycentric growth degraded and weakened lignocellulosic tissues and penetrated the cuticle of C. dactylon leaf blades. By weakening or degrading recalcitrant structures in forages, rumen fungi may alter physical parameters of plants that influence utilization of fibre by ruminants.     

Evaluation of the effects of dietary particle fractions on fermentation profile and concentration of microbiota in the rumen of dairy cows fed grass silage-based diets

The study evaluated the effects of three different theoretical particle lengths (TPL) of grass silage on the distribution of particle fractions of the diet and the resulting effects on fermentation profile and concentrations of protozoa and mixed bacterial mass in the rumen of three lactating Holstein cows fed total mixed rations (45% grass silage, 5% grass hay and 50% concentrate) ad libitum. Decreasing TPL of grass silage (long, medium, short) reduced particles retained on the 19-mm sieve of the Penn State Particle Separator, while particle fractions from 8 mm to 19 mm and smaller than 8 mm were increased. Different TPL did not affect pH and the concentration of volatile fatty acids in the rumen. However, lowering the TPL from long to medium increased significantly the bicarbonate concentration, acetate proportion and protozoal number in the rumen, whereas the proportion of bacterial protein in ruminal digesta and its amino acid concentration were significantly increased by the short TPL. For the current feeding conditions, it can be concluded that increasing the fraction of particles between 8 and 19 mm and probably even the fraction below 8 mm by decreasing TPL of grass silage do not adversely affect rumen conditions and can be beneficial in terms of optimising concentration and activity of ruminal microbiota in high-yielding dairy cows.     

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.     

Effects of dehydrated lucerne and soya bean meal on milk production and composition,nutrient digestion,and methane and nitrogen losses in dairy cows receiving two different forages

Dehydrated lucerne is used as a protein source in dairy cow rations, but little is known about the effects of lucerne on greenhouse gas production by animals. Eight Holstein dairy cows (average weight: 582 kg) were used in a replicated 4×4 Latin square design. They received diets based on either maize silage (M) or grass silage (G) (45% of diet on dry matter (DM) basis), with either soya bean meal (15% of diet DM) completed with beet pulp (15% of diet DM) (SP) or dehydrated lucerne (L) (30% of diet DM) as protein sources; MSP, ML, GSP and GL diets were calculated to meet energy requirements for milk production by dairy cows and degradable protein for rumen microbes. Dry matter intake (DMI) did not differ among diets (18.0 kg/day DMI); milk production was higher with SP diets than with L diets (26.0 v. 24.1 kg/day), but milk production did not vary with forage type. Milk fatty-acid (FA) composition was modified by both forage and protein sources: L and G diets resulted in less saturated FA, less linoleic acid, more trans-monounsaturated FA, and more linolenic acid than SP and M diets, respectively. Enteric methane (CH₄) production, measured by the SF₆ tracer method, was higher for G diets than for M diets, but did not differ with protein source. The same effects were observed when CH₄ was expressed per kg milk. Minor effects of diets on rumen fermentation pattern were observed. Manure CH₄ emissions estimated from faecal organic matter were negatively related to diet digestibility and were thus higher for L than SP diets, and higher for M than G diets; the resulting difference in total CH₄ production was small. Owing to diet formulation constraints, N intake was higher for SP than for L diets; interaction between forage type and protein source was significant for N intake. The same statistical effects were found for N in milk. Faecal and urinary N losses were determined from total faeces and urine collection. Faecal N output was lower for M than for G diets but did not differ between protein sources. Urinary N output did not differ between forage types, but was lower for cows fed L diets than for cows fed SP diets, potentially resulting in lower ammonia emissions with L diets. Replacing soya bean meal plus beet pulp with dehydrated lucerne did not change CH₄ production, but resulted in more N in faeces and less N in urine.     

Effects of different sources of physically effective fiber on rumen microbial populations

Physically effective fiber is needed by dairy cattle to prevent ruminal acidosis. This study aimed to examine the effects of different sources of physically effective fiber on the populations of fibrolytic bacteria and methanogens. Five ruminally cannulated Holstein cows were each fed five diets differing in physically effective fiber sources over 15 weeks (21 days/period) in a Latin Square design: (1) 44.1% corn silage, (2) 34.0% corn silage plus 11.5% alfalfa hay, (3) 34.0% corn silage plus 5.1% wheat straw, (4) 36.1% corn silage plus 10.1% wheat straw, and (5) 34.0% corn silage plus 5.5% corn stover. The impact of the physically effective fiber sources on total bacteria and archaea were examined using denaturing gradient gel electrophoresis. Specific real-time PCR assays were used to quantify total bacteria, total archaea, the genus Butyrivibrio, Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens and three uncultured rumen bacteria that were identified from adhering ruminal fractions in a previous study. No significant differences were observed among the different sources of physical effective fiber with respect to the microbial populations quantified. Any of the physically effective fiber sources may be fed to dairy cattle without negative impact on the ruminal microbial community.     

Evaluation of the effects of dietary particle fractions on fermentation profile and concentration of microbiota in the rumen of dairy cows fed grass silage-based diets

The study evaluated the effects of three different theoretical particle lengths (TPL) of grass silage on the distribution of particle fractions of the diet and the resulting effects on fermentation profile and concentrations of protozoa and mixed bacterial mass in the rumen of three lactating Holstein cows fed total mixed rations (45% grass silage, 5% grass hay and 50% concentrate) ad libitum. Decreasing TPL of grass silage (long, medium, short) reduced particles retained on the 19-mm sieve of the Penn State Particle Separator, while particle fractions from 8 mm to 19 mm and smaller than 8 mm were increased. Different TPL did not affect pH and the concentration of volatile fatty acids in the rumen. However, lowering the TPL from long to medium increased significantly the bicarbonate concentration, acetate proportion and protozoal number in the rumen, whereas the proportion of bacterial protein in ruminal digesta and its amino acid concentration were significantly increased by the short TPL. For the current feeding conditions, it can be concluded that increasing the fraction of particles between 8 and 19 mm and probably even the fraction below 8 mm by decreasing TPL of grass silage do not adversely affect rumen conditions and can be beneficial in terms of optimising concentration and activity of ruminal microbiota in high-yielding dairy cows.