Effects of Bacillus subtilis natto on milk production,rumen fermentation and ruminal microbiome of dairy cows

Two experiments were conducted to evaluate the effects of Bacillus subtilis natto, which was initially isolated from fermented soybeans on milk production, rumen fermentation and ruminal microbiome in dairy cows. In Experiment 1, 36 early lactation Chinese Holstein dairy cows (56 ± 23 days in milk) were randomly assigned to three groups: Control, cows were fed total mixed ration (TMR); BSNLOW, TMR plus 0.5 × 10¹¹ colony-forming units (cfu) of B. subtilis natto/cow per day; and BSNHIGH, TMR plus 1.0 × 10¹¹ cfu of B. subtilis natto/cow per day. During the 70-day treatment period, daily milk production and daily milk composition were determined in individual cows. The results showed that supplementing dairy cows with 0.5 × 10¹¹ and 1.0 × 10¹¹ cfu of B. subtilis natto linearly increased (P < 0.01) milk production (25.2 and 26.4 kg/day v. 23.0 kg/day), 4% fat-corrected milk (27.3 and 28.1 kg/day v. 24.2 kg/day), energy-corrected milk (27.3 and 28.2 kg/day v. 24.2 kg/day), as well as milk fat (1.01 and 1.03 kg/day v. 0.88 kg/day), protein (0.77 and 0.82 kg/day v. 0.69 kg/day) and lactose yield (1.16 and 1.22 kg/day v. 1.06 kg/day) but decreased milk somatic cell counts (SCC) by 3.4% to 5.5% (P < 0.01) in BSNLOW and BSNHIGH treatments compared with Control. In Experiment 2, four rumen-cannulated dairy cows were fed the basal diet from 1 to 7 days (pre-trial period) and rumen samples were collected on days 6 and 7; the same cows then were fed 1.0 × 10¹¹ cfu/day B. subtilis natto from days 8 to 21 (trial period) and rumen samples were collected on days 20 and 21. B. subtilis natto was discontinued from days 22 to 28 (post-trial period) and rumen samples were collected on days 27 and 28. Compared with the pre- and post-periods, ruminal pH decreased by 2.7% to 3.0% during the trial period (P < 0.01), whereas ammonia nitrogen (NH₃-N), total volatile fatty acids and molar proportion of propionate (P < 0.01) and valerate (P < 0.05) increased. Molar proportion of acetate decreased and the acetate to propionate ratio was lower (P < 0.01) during the trial period. However, no differences for 24-h in sacco dry matter digestibility were detected among different periods (treatments) though NDF digestibility was reduced in the trial and post-trial periods (P < 0.01). Compared with pre-trial period, total ruminal bacteria, proteolytic and amylolytic bacteria in rumen enumerated by culture methods increased by 15.0%, 16.2% and 11.7%, respectively (P < 0.01) but protozoa decreased to 5.35 log₁₀ cfu/ml (P < 0.01) during the trial period. These results demonstrate that B. subtilis natto improves milk production and milk components yield, decreases SCC and promotes the growth of total ruminal bacteria, proteolytic and amylolytic bacteria, which indicate that B. subtilis natto has potential to be applied as a probiotic for dairy cows.     

Effect of rumen fill on intake of fresh perennial ryegrass in young and mature dairy cows grazing or zero-grazing fresh perennial ryegrass

Rumen fill may be a strong intake constraint for dairy cows fed on pasture, even though pasture is highly digestible in the grasslands of temperate climates. This constraint may also depend on the cows' maturity. Moreover, indoor feeding of fresh herbage may not always be a good model for the study of intake regulation at grazing. To test these hypotheses, four mature (6.3 ± 0.72 year old) and four young (3.8 ± 0.20 year old) dairy cows were offered fresh perennial ryegrass indoors or at grazing. The impact of rumen fill on intake was evaluated by addition of rumen inert bulk (RIB; coconut fiber, 15 l) compared to a control. The experimental design was a double 4 × 4 Latin square with four 14-day periods and a 2 × 2 factorial arrangement of two feeding methods (indoor feeding v. grazing), combined with the addition, or not, of RIB (RIB v. control), repeated for four mature and four young cows. Digestibility of offered herbage was 0.81. The average ytterbium measured dry matter intake (Yb DMI) was 19.0 and 15.5 kg/day for mature and young cows respectively (P = 0.019). The effect of RIB on predicted Yb DMI interacted with feeding method and cow age (P = 0.043). The presence of RIB decreased Yb DMI by 4.4 kg/day in mature cows at grazing and by 3.4 kg/day in young cows indoors, whereas it did not affect the Yb DMI of mature cows indoors or grazing young cows. Both grazing and young age constituted a clear constraint on the feeding behavior of the cows. Grazing cows had fewer ingestion and rumination sequences, which were longer and less evenly distributed throughout the day and night. Young cows had lower intake rates that were less adaptable to the feeding method and the presence of RIB. Mature cows clearly decreased their daily intake rate at grazing compared to indoor feeding, and with RIB compared to control, whereas the intake rate of young cows did not vary. These results indicate that rumen fill can represent a constraint on intake in grazing cows, even when highly digestible perennial ryegrass is offered. The study also shows that the impact of RIB on intake is highly dependent upon other constraints applied to the chewing behavior, which in this experiment were methods of offering herbage and cow age.     

Rubber seed oil and flaxseed oil supplementation alter digestion,ruminal fermentation and rumen fatty acid profile of dairy cows

Rubber seed oil (RO) that is rich in polyunsaturated fatty acids (FA) can improve milk production and milk FA profiles of dairy cows; however, the responses of digestion and ruminal fermentation to RO supplementation in vivo are still unknown. This experiment was conducted to investigate the effect of RO and flaxseed oil (FO) supplementation on nutrients digestibility, rumen fermentation parameters and rumen FA profile of dairy cows. Forty-eight mid-lactation Holstein dairy cows were randomly assigned to one of four treatments for 8 weeks, including basal diet (CON) or the basal dietary supplemented with 4% RO, 4% FO or 2% RO plus 2% FO on a DM basis. Compared with CON, dietary oil supplementation improved the total tract apparent digestibility of DM, neutral detergent fibre and ether extracts ( P < 0.05). Oil treatment groups had no effects on ruminal digesta pH value, ammonia N and microbial crude protein ( P > 0.05), whereas oil groups significantly changed the volatile fatty acid (VFA) profile by increasing the proportion of propionate whilst decreasing total VFA concentration, the proportion of acetate and the ratio of acetate to propionate ( P < 0.05). However, there were no differences in VFA proportions between the three oil groups (P > 0.05). In addition, dietary oil supplementation increased the total unsaturated FA proportion in the rumen by enhancing the proportion of trans-11 C18:1 vaccenic acid (VA), cis-9, trans-11 conjugated linoleic acid (CLA) and α-linolenic acid (ALA) ( P < 0.05). These results indicate that dietary supplementation with RO and FO could improve nutrients digestibility, ruminal fermentation and ruminal FA profile by enhancing the VA, cis-9, trans-11 CLA and ALA composition of lactating dairy cows. These findings provide a theoretical basis for the application of RO in livestock production.     

Effect of partially replacing a barley‐based concentrate with flaxseed‐based products on the rumen bacterial population of lactating Holstein dairy cows

Aims

The effects of partial replacement of a barley‐based concentrate with flaxseed‐based products on the rumen bacterial population of lactating Holstein dairy cows were evaluated.

Methods and Results

Treatments fed were CONT, a normal diet that included barley silage, alfalfa hay and a barley‐based concentrate that contained no flaxseed or faba beans; FLAX, inclusion of a nonextruded flaxseed‐based product containing 55·0% flaxseed, 37·8% field peas and 6·9% alfalfa; EXT, similar to FLAX, but the product was extruded and EXTT, similar to FLAX, but product was extruded and field peas were replaced by high‐tannin faba beans. The rumen bacterial population was evaluated by utilizing 16S rRNA gene sequencing. Most abundant phyla, families and genera were unaffected. However, some taxa were affected; for example, unsaturated fatty acid content was negatively correlated with Clostridiaceae, and tannin content was negatively correlated with BS11 and Paraprevotellaceae.

Conclusions

Predominant rumen bacterial taxa were not affected, but the abundance of some taxa found in lower proportions shifted, possibly due to sensitivity to unsaturated fatty acids or tannins.

Significance and Impact of the Study

Flaxseed‐based products were effective for partially replacing barley‐based concentrate in rations of lactating dairy cows. No negative effects of these products were observed on the abundance of predominant rumen bacterial taxa, with only minor shifts in less abundant bacteria.     

Effects of fibre concentration of diets consisting of hay and slowly degradable concentrate on ruminal fermentation and digesta particle size in mid-lactation dairy cows

The objective of this study was to investigate the intestinal microbiota of growing kittens fed moderate- or high-protein diets using DNA-based qualitative and quantitative techniques. Kittens were weaned to a high-protein (HP; n = 7) or moderate-protein (MP; n = 10) diet at 8 weeks of age. Fresh faecal samples were collected at 8, 12, and 16 weeks of age. DNA was extracted and quantitative PCR used to quantify Bifidobacterium, Lactobacillus, Clostridium perfringens, and Escherichia coli concentrations. Denaturing gradient gel electrophoresis was performed to create a dendrogram and unrooted trees using Bionumerics 5.0 to identify similarity due to litter, age, or diet. Kittens fed HP had lower (p = 0.02) Bifidobacteria and Lactobacillus counts than MP-fed kittens. E. coli was lower (p = 0.02) in HP-fed kittens and tended to be affected by age (p = 0.09). Kittens were clustered by litter at 8 weeks of age, and then clustered by diet at 12 and 16 weeks of age. Our data suggest that faecal microbiota of growing kittens change after weaning and that dietary protein concentration affects E. coli, Bifidobacterium, and Lactobacillus populations. The relevance of these data in terms of intestinal health and disease remain to be determined and justifies further study.     

Effect of offering dairy cows diets differing in phosphorus concentration over four successive lactations: 2. Health, fertility, bone phosphorus reserves and nutrient utilisation

This experiment examined the long-term effects of offering diets containing low levels of dietary phosphorus (P) on dairy cow health, fertility and bone composition, and the effect of dietary P level on nutrient utilisation. One hundred winter-calving Holstein-Friesian dairy cows were offered diets containing either 'high' or 'low' levels of dietary P over a 4-year period. Rations offered during the winter included grass silage, maize silage (70 : 30 dry matter (DM) basis, approximately) and concentrates (10.0 to 12.0 kg/cow per day). During the summer periods in years 1 and 2, half of the cows grazed both day and night, while the remaining cows grazed by day, and were housed by night and offered grass silage. During years 3 and 4, all cows grazed both day and night during the summer period. Concentrate feed levels during the summer periods were 3.0 to 4.0 kg/cow per day. Different dietary P levels were achieved by offering concentrates containing either high or low P levels during the winter period (approximately 7.0 or 4.4 g P/kg DM, respectively) and during the summer period (approximately 6.8 or 3.6 g P/kg DM, respectively). Total ration P levels averaged 4.9 and 3.6 g P/kg DM for the high and low P winter diets, respectively, and 4.2 and 3.6 g P/kg DM for the high and low P summer diets, respectively. A total of 95, 70, 50 and 22 cows completed each of lactations from 1 to 4, respectively. Neither the incidence of lameness or mastitis, or milk somatic cell count, were affected by dietary P level (P > 0.05), while none of the fertility parameters recorded in any of lactations from 1 to 4 was affected by the dietary P level (P > 0.05). Dietary P level had no effect on the specific gravity, ash or calcium content of rib cortical bone cores (n = 78 cows), while the P content of cortical bone (g/kg fresh, g/kg DM and mg/ml fresh bone) was lower with cows offered low P diets (P < 0.05). Dietary P level had no significant effect on the digestibility of either the DM, nitrogen, energy or acid detergent fibre fraction of the diet (P > 0.05), while faecal P excretions were reduced by a mean of 27 g/cow per day with cows offered the low P diets during the winter period. The results of this study indicate that dietary P levels can be reduced to proportionately 0.8 (approximately) of current UK feeding standards (Agricultural and Food Research Council, 1991), with no detrimental effect on dairy cow health or fertility, while having only minor effects on bone composition.     

Effects of folic acid supplementation to rations differing in the concentrate to roughage ratio on ruminal fermentation,nutrient flow at the duodenum,and on serum and milk variables of dairy cows

The present study was undertaken to determine the effects of dietary folic acid (FOL) supplementation on ruminal fermentation, duodenal nutrient flow, serum and milk variables, and on B-vitamin concentration in serum. The study was divided into two experiments: in Exp. 1 the forage to concentrate (F:C) ratio of the diet (DM basis) was 34:66 (high concentrate, HC), while in Exp. 2 the F:C ratio was 66:34 (high forage, HF). In addition, the cows received 0 or 1 g FOL/d. In Exp. 1, two German Holstein cows equipped with cannulas in the dorsal sac of the rumen and in the proximal duodenum were dry and five were lactating (186 ± 144 days in milk); in Exp. 2 four cows were dry and four were lactating (165 ± 57 days in milk). In cows fed the HC diet, FOL supplementation decreased the ruminally-fermented organic matter. Thus, less energy was available for ruminal microorganisms, which resulted in a reduced microbial crude protein flow at the duodenum. Feeding the HF diet, FOL supplementation only increased the apparent ruminal digestibility of acid detergent fibre (ADF). With the HF diet, FOL had no influence on the serum levels of glucose, non-esterified fatty acids, beta-hydroxybutyrate, urea, thiamine, riboflavin, pyridoxal-5′-phosphate, pyridoxic acid, pyridoxal, pyridoxine, pyridoxamine, pantothenic acid, nicotinamide or nicotinic acid, whereas supplementing FOL to the HC diet lowered the serum glucose and riboflavin levels. In both experiments, the supplementation of FOL had no effects on milk composition. Folic acid supplementation to both diets increased the concentrations of serum 5-methyl-tetrahydrofolate. However, no beneficial effects to dairy cows were obvious. Therefore, to achieve certain results, studies with a higher number of non-fistulated cows would be necessary.     

Essential oils for dairy calves: effects on performance,scours, rumen fermentation and intestinal fauna

The first cause of death of dairy calves is often diarrhea which is mainly caused by pathogenic bacteria, which can result in excessive use of antibiotics. However, facing the increase concern by the industry and consumers, the use of antibiotics not only to control pathogens, but also to manipulate growth, has become a challenge. Alternative additives, such essential oils, have the potential to decrease antibiotic use, without reducing performance or increasing mortality of dairy calves. The objective of this study was to evaluate the use of a commercial blend of essential oils, incorporated into the calf starter and/or milk replacer to monitor the effect on overall calf performance, fecal scores and rumen fermentation parameters. A total of 30 Holstein calves received 6 l/day of a liquid diet, consisting of a commercial milk replacer containing 20% CP : 15% fat (EE). Calves had free choice access to water and calf starter. Weaning occurred at week 8, and calves were followed until the 10^th week of age. Calves were assigned to one of the three treatment groups in a randomized block design. Treatments: (1) control without essential oils supplementation (C); (2) essential oils blend in the milk replacer at 400 mg/kg (MR) and (3) essential oils blend in the milk replacer (200 mg/kg) and starter feed (200 mg/kg) (MRS). From the 2^nd week, calves were weighed and body measurements were taken, while concentrate intake and fecal scores were monitored daily. Blood samples were drawn weekly for determination of glucose and β-hydroxybutyrate. Fecal samples were collected weekly and analyzed for lactic acid bacteria and Enterobacteria; and ruminal fluid for determination of pH, short chain fatty acids, ammonia-N and counts of amylolytic and cellulolytic bacteria, and protozoa. Performance, fecal scores and intestines microorganisms were not affected by the essential oils supplementation. Ruminal and blood parameters were also not affected, with the exception the rumen ammonia-N concentration, with higher values when essential oils were supplemented in a combination of milk replacer and starter feed. Most of the evaluated parameters were affected by age of calves, mainly as a response to the increase in concentrate intake as animals’ aged. Essential oils are promising substitutes for antibiotics. However, the dose and routes of administration deserve further studies, allowing a better animal performance and health to be achieved.     

Effect of changes in diet energy density on feed intake, milk yield and metabolic parameters in dairy cows in early lactation

The purpose of this experiment was to investigate how early lactating cows adjust their metabolism and production to acute, but moderate changes in the energy density of the diet. Sixty dairy cows were randomly assigned to one of four treatments: two change-over groups (HNH and NHN) and two control groups (HHH and NNN), where H and N refer to a high and normal energy density in the total mixed ration (TMR), respectively. The experimental period covered the first 9 weeks post calving, which was split up in three 3-week periods. Thus, cows assigned to HNH or NHN shifted TMR in weeks 4 and 7 after calving while cows assigned to HHH or NNN were fed the same TMR for all 9 weeks. Results from cows on treatment HNH were compared with group HHH while cows on treatment NHN were compared with group NNN. When the diet changed from N to H and H to N, cows increased and decreased their dry-matter intake (DMI), respectively compared with control groups. Cows adjusted milk yield accordingly to changes in DMI, although not always significantly. Energy-corrected milk yield was not significantly affected by any of the changes in the energy density of the diet but generally showed same tendencies as milk yield. Non-esterified fatty acids (NEFA), beta-hydroxybutyrate in blood and milk and triacylglycerol and glycogen content in the liver were not significantly affected by changes in the energy density of the diet, except from NEFA at one change. Glucose increased more when the diet changed from N to H and increased less when the diet changed from H to N, compared with control groups, although not always significantly. Collectively, these results suggest that cows adjust their DMI and partly milk yield according to the energy density of the diet and therefore only limited effects were observed in physiological parameters.     

Effect of offering dairy cows diets differing in phosphorus concentration over four successive lactations: 1. Food intake, milk production, tissue changes and blood metabolites

The loss of phosphates from dairy farms contributes to the eutrophication of waterways. Whilst reducing the phosphorus (P) content of dairy cow diets has the potential to help reduce phosphate losses, diets containing inadequate dietary P may have a negative effect on cow health and performance. To address this issue, 100 winter-calving Holstein-Friesian dairy cows were offered diets containing either 'high' or 'low' levels of dietary P. The experiment was conducted over a 4-year period, with 80 primiparous cows commencing the study in year 1, while a further 20 primiparous cows commenced the study in year 2. Rations offered during the winter comprised grass silage, maize silage (70 : 30 dry matter (DM) basis, approximately) and concentrates (10.0 to 12.0 kg/cow per day). During the summer periods in years 1 and 2, half of the cows grazed both day and night, while the remaining cows grazed by day, and were housed by night and offered grass silage. During years 3 and 4, all cows grazed both day and night during the summer period. Concentrate feed levels during the summer periods were 3.0 to 4.0 kg/cow per day. Different dietary P levels were achieved by offering concentrates containing either high or low P levels during the winter period (approximately 7.0 or 4.4 g P/kg DM respectively), and during the summer period (approximately 6.8 or 3.6 g P/kg DM, respectively). Total ration P levels averaged 4.9 and 3.6 g P/kg DM for the 'high' and 'low' P winter diets respectively, and 4.2 and 3.6 g P/kg DM for the 'high' and 'low' P summer diets respectively. A total of 95, 70, 50 and 22 cows completed each of lactations 1 to 4 respectively. Dietary P level had no significant effect on food intake, milk output or milk composition (P > 0.05). Plasma P concentrations were significantly lower with cows offered the 'low' P diet in each of lactations 1 to 4 (P < 0.05). In each of lactations 3 and 4, cows offered the 'low' P diet tended to have lower condition scores and live weights than those offered the 'high' P diet. The results of this experiment highlight that the P content of dairy cow diets can be substantially reduced with no detrimental effect on dairy cow performance.     

Microbial biodiversity of the liquid fraction of rumen content from lactating cows

Host and dietary interactions with the rumen microbiome can affect the efficacy of supplements, and their effect on the composition of the bacterial population is still unknown. A 16S rRNA metagenomic approach and Next-Generation Sequencing (NGS) technology were used to investigate the bacterial microbiome composition in the liquid fraction of the rumen content collected via stomach tubing. To investigate biodiversity, samples were taken from three groups of four lactating dairy cows given a supplement of either 50 g of potato protein (Ctrl group), or 50 g of lyophilized Saccharomyces cerevisiae (LY group) or 50 g of dried S. cerevisiae (DY group) in a potato protein support. Rumen samples were collected after 15 days of dietary treatments and milk production was similar between the three groups. Taxonomic distribution analysis revealed a prevalence of the Firmicutes phylum in all cows (79.76%) and a significantly (P<0.05) higher presence of the genus Bacillus in the DY group. Volatile fatty-acid concentration was not significantly different between groups, possibly because of relatively high inter-animal variability or limited effect of the treatments or both, and the correlation analysis with bacterial taxa showed significant associations, in particular between many Firmicutes genera and butyrate. Limited differences were observed between dietary treatments, but the lack of microbiome data before yeast administration does not allow to draw firm conclusions on the effect of dietary treatments.     

Effects of niacin supplementation and dietary concentrate proportion on body temperature,ruminal pH and milk performance of primiparous dairy cows

The objective of this study was to investigate the effects of niacin and dietary concentrate proportion on body temperature, ruminal pH and milk production of dairy cows. In a 2 × 2 factorial design, 20 primiparous Holstein cows (179 ± 12 days in milk) were assigned to four dietary treatments aimed to receive either 0 or 24 g niacin and 30% (low) or 60% (high) concentrate with the rest being a partial mixed ration (PMR) composed of 60% corn and 40% grass silage (on dry matter basis). Ambient temperature and relative humidity were determined and combined by the calculation of temperature humidity index. Respiration rates, rectal, skin and subcutaneous temperatures were measured. Milk production and composition were determined. Ruminal pH and temperature were recorded at a frequency of 5 min using wireless devices for continuous intra-ruminal measurement (boluses). pH values were corrected for pH sensor drift. The climatic conditions varied considerably but temporarily indicated mild heat stress. Niacin did not affect skin, rectal and subcutaneous temperatures but tended to increase respiration rates. High concentrate reduced skin temperatures at rump, thigh and neck by 0.1–0.3°C. Due to the technical disturbances, not all bolus data could be subjected to statistical evaluation. However, both niacin and high concentrate influenced mean ruminal pH. High concentrate increased the time spent with a pH below 5.6 and ruminal temperatures (0.2–0.3°C). Niacin and high concentrate enhanced milk, protein and lactose yield but reduced milk fat and protein content. Milk fat yield was slightly reduced by high concentrate but increased due to niacin supplementation. In conclusion, niacin did not affect body temperature but stimulated milk performance. High concentrate partially influenced body temperatures and had beneficial effects on milk production.     

Effect of dietary Quebracho tannin extract on feed intake,digestibility, excretion of urinary purine derivatives and milk production in dairy cows

The aim of this study was to evaluate the effects of dietary Quebracho tannin extract (QTE) on feed intake, apparent total tract digestibility (ATTD), excretion of urinary purine derivatives (PD) and milk composition and yield in dairy cows. Fifty Holstein cows were divided into two groups. To reach a similar performance of both groups, cows were divided according to their milk yield, body weight, days in milk and number of lactations at the start of the experiment averaging 33.2 ± 8.2 kg/d, 637 ± 58 kg, 114 ± 73 d and 2.3 ± 1.6 lactations, respectively. The cows were fed a basal diet as total mixed ration containing on dry matter (DM) basis 34% grass silage, 32% maize silage and 34% concentrate feeds. Three dietary treatments were tested, the control (CON, basal diet without QTE), QTE₁₅ (basal diet with QTE at 15 g/kg DM) and QTE₃₀ (basal diet with QTE at 30 g/kg DM). Two treatments were arranged along six periods each 21 d (13 d adaptation phase and 8 d sampling phase). The ATTD of DM and organic matter were reduced only in Diet QTE₃₀, whereas both QTE treatments reduced ATTD of fibre and nitrogen (N), indicating that QTE impaired rumen fermentation. Nevertheless, feed intake was unaffected by QTE. In Diet CON, urinary N excretion accounted for 29.8% of N intake and decreased in treatments QTE₁₅ and QTE₃₀ to 27.5% and 17.9%, respectively. Daily faecal N excretion increased in treatments CON, QTE₁₅ and QTE₃₀ from 211 to 237 and 273 g/d, respectively, which amounted to 39.0%, 42.4% and 51.7% of the N intake, respectively. Hence, QTE shifted N excretion from urine to faeces, whereas the proportion of ingested N appearing in milk was not affected by QTE (average 30.7% of N intake). Daily PD excretion as indicator for microbial crude protein (CP) flow at the duodenum decreased in treatment QTE₃₀ compared with Diet CON from 413 to 280 mmol/d. The ratios of total PD to creatinine suggest that urinary PD excretion was already lower when feeding Diet QTE₁₅. While there was no effect of Diet QTE₁₅, treatment QTE₃₀ reduced milk yield, milk fat and protein. Both QTE treatments reduced milk urea concentration, which suggest that ruminal degradation of dietary CP was reduced. In summary, adding QTE at dosages of 15 and 30 g/kg DM to diets of lactating dairy cows to improve feed and protein use efficiency is not recommended.     

Effects of starch-rich or lipid-supplemented diets that induce milk fat depression on rumen biohydrogenation of fatty acids and methanogenesis in lactating dairy cows

Optimizing milk production efficiency implies diets allowing low methane (CH₄) emissions and high dairy performance. We hypothesize that nature of energy (starch v. lipids) and lipid supplement types (monounsaturated fatty acid (MUFA) v. polyunsaturated fatty acid (PUFA) mitigate CH₄ emissions and can induce low milk fat content via different pathways. The main objective of this experiment was to study the effects of starch-rich or lipid-supplemented diets that induce milk fat depression (MFD) on rumen biohydrogenation (RBH) of unsaturated fatty acids (FA) and enteric CH₄ emissions in dairy cows. Four multiparous lactating Holstein cows (days in milk=61±11 days) were used in a 4×4 Latin square design with four periods of 28 days. Four dietary treatments, three of which are likely to induce MFD, were based (dry matter basis) on 56% maize silage, 4% hay and 40% concentrates rich in: (1) saturated fatty acid (SFA) from Ca salts of palm oil (PALM); (2) starch from maize grain and wheat (MFD-Starch); (3) MUFA (cis-9 C18:1) from extruded rapeseeds (MFD-RS); and (4) PUFA (C18:2n-6) from extruded sunflower seeds (MFD-SF). Intake and milk production were measured daily. Milk composition and FA profile, CH₄ emissions and total-tract digestibility were measured simultaneously when animals were in open-circuit respiration chambers. Fermentation parameters were analysed from rumen fluid samples taken before feeding. Dry matter intake, milk production, fat and protein contents, and CH₄ emissions were similar among the four diets. We observed a higher milk SFA concentration with PALM and MFD-Starch, and lower milk MUFA and trans-10 C18:1 concentrations in comparison to MFD-RS and MFD-SF diets, while trans-11 C18:1 remained unchanged among diets. Milk total trans FA concentration was greater for MFD-SF than for PALM and MFD-Starch, with the value for MFD-RS being intermediate. Milk C18:3n-3 content was higher for MFD-RS than MFD-SF. The MFD seems more severe with MFD-SF and MFD-RS than PALM and MFD-Starch diets, because of a decrease in milk SFA concentration and a stronger shift from trans-11 C18:1 to trans-10 C18:1 in milk. The MFD-SF diet increased milk trans FA (+60%), trans-10 C18:1 (+31%), trans-10,cis-12 CLA (+27%) and PUFA (+36%) concentrations more than MFD-RS, which explains the numerically lowest milk fat yield and indicates that RBH pathways of PUFA differ between these two diets. Maize silage-based diets rich in starch or different unsaturated FA induced MFD with changes in milk FA profiles, but did not modify CH₄ emissions.     

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.     

Effects of protein sources on concentrations of hydrogen sulphide in the rumen headspace gas of dairy cows

Two Latin square design experiments investigated the relationship between hydrogen sulphide concentration in the rumen headspace gas of dairy cows and the early stages of protein degradation in the rumen. In Expt 1, three protein sources differing in rumen N (nitrogen) degradability (maize gluten feed (MGF); sunflower meal (SFM); and soyabean meal (SBM)) were used, whereas in Expt 2 four different batches of the same feed (MGF) differing in colour (CIE L*, a*, b* (CIELAB) scale) were used. After allowing the concentration of hydrogen sulphide in rumen gas to decline close to zero, a fixed amount of protein sources was offered to cows and the concentrations of hydrogen sulphide were recorded in rumen headspace gas at 30-min intervals. In Expt 1, the concentration of hydrogen sulphide showed considerable variation between protein sources, with MGF having the highest concentration followed by SFM and SBM resulting in very low concentrations. The N wash losses (zero time measurements with nylon bags) ranked the feeds in the same way, from MGF (highest; 61%) to SBM (lowest; 26%). There were marked differences in the degradation of cystine and methionine between protein sources, although the degradation of cystine was always higher than for methionine. MGF (Expt 2) led to increased concentrations of hydrogen sulphide, with peak concentrations achieved between 1 and 2 h after feeding. The concentrations of hydrogen sulphide were higher for MGF1, intermediate for MGF2 and lower for MGF3 and MGF4, agreeing with colour scale. Differences in the early stages of dietary sulphur degradation corresponded with differences in hydrogen sulphide concentrations in rumen gas. The results suggest that hydrogen sulphide concentrations in the rumen headspace gas could be useful to evaluate nutritional parameters not measured by the in sacco technique, contributing to a better understanding of the response of dairy cows to different protein supplements.     

Effects of increasing diet fermentability on intake,digestion, rumen fermentation,blood metabolites and milk production of heat-stressed dairy cows

Heat stress is a major problem for dairy cows in hot climates, thus coping strategies are essential. This study evaluated the effects of increasing diet fermentability on intake, total tract digestibility, ruminal pH and volatile fatty acids (VFA) profile, blood metabolite profile and milk production and composition of lactating dairy cows managed under conditions of ambient heat stress. Nine multiparous cows (650 ± 56 kg BW; mean ± SD) averaging 102 ± 13 days in milk and producing 54 ± 6 kg/day were randomly assigned to a triplicate 3 × 3 Latin square. During each 21-day period, cows were offered one of three total mixed rations that varied in diet fermentability. The three levels of diet fermentability were achieved by increasing the proportion of pellets containing ground wheat and barley in the dietary DM from 11.7% (low), to 23.3% (moderate), and 35.0% (high) by replacing ground corn grain. Each period had 14 day of adaptation and 7 day of sampling. The ambient temperature–humidity index ( ≥ 72) indicated that the cows were in heat stress almost the entire duration of the study. Also, rectal temperature of cows was elevated at 39.2°C, another indication of heat stress. Increasing diet fermentability linearly decreased dry matter intake (22.8, 22.5, 21.8 kg/day for low, moderate and high, respectively; P ≤ 0.05) but increased non-fibre carbohydrate digestibility (P ≤ 0.05) and tended to increase digestibility of DM (P = 0.10) and crude protein (P = 0.06). As a result, the intake of digestible DM was not affected by the treatments. The production of 3.5% fat corrected milk (32.6, 33.7, and 31.5 kg/day) was quadratically (P ≤ 0.05) affected by diet fermentability with lower production for the high diet compared with the other two, which were similar. Rumen pH (ruminocentesis) and proportions of butyrate and isovalerate linearly decreased whereas propionate proportion linearly increased with increasing diet fermentability (P ≤ 0.05). The rumen concentration of NH₃-N (11.0, 9.0, and 8.7 mg/dL) and blood concentration of urea linearly decreased with increasing diet fermentability (P ≤ 0.05). The activity of alkaline phosphatase increased (65.1, 83.2, and 84.9 U/l) and concentration of malondialdehyde decreased (2.39, 1.90 and 1.87 µmol/l) linearly with increasing diet fermentability (P ≤ 0.05), which indicated possible attenuation of the effects of oxidative stress with increasing diet fermentability. Overall, a modest increase of diet fermentability improved nitrogen metabolism, milk protein production and oxidative stress of heat-stressed dairy cows, but a further increase in diet fermentability decreased milk yield.     

Magnesium absorption as influenced by the rumen passage kinetics in lactating dairy cows fed modified levels of fibre and protein

The potassium sensitive magnesium absorption through the rumen wall may be influenced by additional dietary properties, such as diet type, forage type or forage to concentrate ratio. These properties are likely associated to rumen passage kinetics modified by dietary fibre content. The study aimed to assess the effects of rumen passage kinetics on apparent Mg absorption and retention in lactating dairy cows fed modified levels of fibre. Six lactating Red-Holstein and Holstein cows, including four fitted with ruminal cannulas were randomly assigned to a 3 × 3 cross-over design. The experimental diets consisted of early harvested low NDF (341 g NDF/kg DM) and late harvested high NDF (572 g NDF/kg DM) grass silage (80% DM) and of concentrates (20% of DM). As the low-fibre diet was excessive in protein, a third high-fibre diet was formulated to be balanced in digestible protein with the low-fibre diet to avoid any eventual confounding effects of NDF and protein excess. All diets were formulated to contain iso-Ca, -P, -Mg, -K and -Na. Passage kinetics of solid and liquid phase of rumen digesta were evaluated using ruminal marker disappearance profiles. Cows fed the low-fibre diet had compared to the other diets, an up to 40% lower solid and 26% lower liquid phase volume of rumen digesta and a 10% numerically higher fractional rumen liquid passage rate. Rumen pH lost 0.6 units and Mg concentration in the rumen liquid phase tripled when cows were fed the low-fibre diet. Faecal Mg excretion was up to 14% higher in cows fed the low-fibre diet and Mg absorbability was 12% compared to up to 19% in other diets. Urinary Mg excretion in cows fed the low-fibre diet was half of the ones in the other treatments, but Mg retention was not affected. Dietary protein excess neither affected rumen passage kinetics nor Mg absorption and retention. Absorption of Mg was correlated with rumen liquid volume which both decreased with decreasing daily NDF intake (NDFi, 11.8 ± 2.4 l/kg NDFi). Consequently, daily Mg absorption decreased by 1.32 ± 0.28 g/kg decreasing NDFi. To conclude, in addition to the known antagonistic effect of dietary K, the present data indicate that Mg absorption was dependent from NDFi which modified rumen liquid volume, but was independent of dietary protein excess likely associated to low NDF herbages.