Effects of heat-stress-reducing systems on blood constituents, milk production and milk quality of Holstein and Jersey cows and heifers on pasture.

The objective of this study was to evaluate the influence of different heat-stress-reducing systems, i.e., sprinkler + artificial shade, shower + artificial shade, and artificial shade, on serum mineral, hormonal, hematological, and metabolite profiles, on milk production, and milk composition in lactating cows and pubertal heifers of Holstein and Jersey breeds. For this purpose, 12 animals were used: 3 Holstein cows with an average (mean ± SD) body weight of 600 ± 30 kg, 53 ± 11 months of age, and milk yield of 27 ± 3.5 kg/day; 3 Jersey cows with an average body weight of 370 ± 11 kg, 40 ± 6 months of age, and milk production of 11 ± 1.5 kg/day; 3 Holstein heifers (325 ± 25 kg and 16 ± 0.6 months of age); and 3 Jersey heifers (250 ± 25 kg and 13 ± 0.6 months of age). Animals were used in a replicated 3 × 3 Latin square design with a 3 × 2 × 2 factorial arrangement of treatments that included three treatments (sprinkler + artificial shade, shower + artificial shade, and artificial shade), two breeds (Holstein, Jersey), and two physiological stages (lactating cows, heifers). The experimental treatments influenced (P < 0.05) the concentrations of triiodothyronine, with the shower and shade systems showing greater and similar concentrations (99.5 and 96.3 µg/dL, respectively) when compared with sprinkler treatment (89.2 µg/dL). There was an effect (P < 0.05) of breed on the concentrations of Na + , K + , hemoglobin, hematocrit and mean corpuscular volume levels with the Holsteins having lower levels of Na + , K + , hemoglobin, hematocrit and mean corpuscular volume (101.1, 4.0 ng/mL, 11.2 g/dL, 24.7%, and 42.3 µm3, respectively) than the Jerseys (106.5 and 4.3 ng/mL, 12.4 g/dL, 27.7%, and 46.3 µm3, respectively. Total cholesterol and high-density lipoproteins were influenced by physiological stage (P < 0.05). Concentrations of cholesterol and high-density lipoproteins were higher for cows (94.1, and 56.9 mg/dL, respectively) than for heifers (56.9 and 42,9 mf/dL, respectively). Milk production and fat content were affected (P < 0.05) by breed (P < 0.05), with Holstein cows producing more milk (23.9 kg/day) than Jersey cows (12.0 kg/day), but Jersey cows had higher fat concentration (4.6%) than Holstein cows (3.0%). Therefore, the three different thermal-stress-reducing-systems tested were able to maintain the serum biomarkers within normal physiological ranges. However, the most appropriate thermal-stress-reducing-systems would be a sprinkler systema because it uses less water compared with the shower system.

Assessment of the Effects of Commercial or Locally Engineered Biochars Produced from Different Biomass Sources and Differing in Their Physical and Chemical Properties on Rumen Fermentation and Methane Production In Vitro.

In recent years, interest in using biochar as feed additives to mitigate enteric methane (CH4) emissions from ruminants has increased. It has been suggested that the mitigating potential of biochar is influenced by its physical (e.g., porosity-related) and chemical (e.g., redox-potential-related) properties. Thus, the aim of this in vitro study was to evaluate the effects of commercial or locally engineered biochars, produced from different biomass sources and differing in their physical and chemical characteristics, on rumen fermentation and CH4 production. For this purpose, a 24 h batch culture of ruminal fluid incubations was conducted in a complete randomized block design (repeated three times) that included a negative control (no additive), a positive control (monensin, 10 mg/mL), and four commercial and three locally engineered biochars, each evaluated at 1%, 2%, or 5% of the substrate's (i.e., the total mixed ration) dry matter. The evaluated biochars greatly differ in their chemical (i.e., moisture, ash, pH, redox potential, volatiles, carbon, fixed carbon, hydrogen, and sulfur) and physical (i.e., fine particles < 250 µm, bulk density, true density, porosity, electrical conductivity, specific surface area, and absorbed CO2) properties. Despite these differences and compared with the negative control, none of the biochars evaluated (regardless of the inclusion rate) influenced gas and CH4 production, volatile fatty acid characteristics (total concentration and profile), or ammonia-nitrogen (NH3-N) concentrations. As expected, monensin (i.e., the positive control) decreased (p < 0.05) CH4 production mainly because of a decreased (p < 0.05) acetate-to-propionate ratio. The results of this study reveal that despite the large differences in the physical and chemical properties of the biochars evaluated, their inclusion at different rates in vitro failed to modify rumen fermentation and decrease CH4 production. Based on these in vitro findings, it was concluded that biochar does not represent a viable strategy for mitigating enteric CH4 emissions.

Effect of Metabolizable Protein Supply on Milk Performance, Ruminal Fermentation, Apparent Total-Tract Digestibility, Energy and Nitrogen Utilization, and Enteric Methane Production of Ayrshire and Holstein Cows.

In North America, the nutrient requirements of dairy cattle are predicted using the Cornell Net Carbohydrate and Protein System (CNCPS) or the National Research Council (NRC). As Holstein is the most predominant dairy cattle breed, these models were developed based on the phenotypic, physiological, and genetic characteristics of this breed. However, these models may not be appropriate to predict the nutrient requirements of other breeds, such as Ayrshire, that are phenotypically and genetically different from Holstein. The objective of this study was to evaluate the effects of increasing the metabolizable protein (MP) supply using CNCPS on milk performance, ruminal fermentation, apparent total-tract digestibility, energy and N utilization, and enteric methane production in Ayrshire vs. Holstein lactating dairy cows. Eighteen (nine Ayrshire; nine Holstein) lactating cows were used in a replicated 3 × 3 Latin square design (35-d periods) and fed diets formulated to meet 85%, 100%, or 115% of MP daily requirement. Except for milk production, no breed × MP supply interaction was observed for the response variables. Dry matter intake (DMI) and the yields of energy-corrected milk (ECM), fat, and protein were less (p < 0.01) in Ayrshire vs. Holstein cows. However, feed efficiency and N use efficiency for milk production did not differ between the two breeds, averaging 1.75 kg ECM/kg DMI and 33.7 g milk N/100 g N intake, respectively. Methane yield and intensity and urinary N also did not differ between the two breeds, averaging 18.8 g CH4 /kg DMI, 10.8 g CH4 /kg ECM, and 27.6 g N/100 g N intake, respectively. Yields of ECM and milk protein increased (p ≤ 0.01) with increasing MP supply from 85% to 100% but no or small increases occurred when MP supply increased from 100 to 115%. Feed efficiency increased linearly with an increasing MP supply. Nitrogen use efficiency (g N milk/100g N intake) decreased linearly (by up to 5.4 percentage units, (p < 0.01) whereas urinary N excretion (g/d or g/100 g N intake) increased linearly (p < 0.01) with an increasing MP supply. Methane yield and emission intensity were not affected by MP supply. This study shows that feed efficiency, N use efficiency, CH4 (yield and intensity), and urinary N losses did not differ between Ayrshire and Holstein cows. Energy-corrected milk yield and feed efficiency increased, but N use efficiency decreased and urinary N losses increased with increasing dietary MP supply regardless of breed. Ayrshire and Holstein breeds responded similarly to increasing MP levels in the diet.

Effect of whole oilseeds in the diet on bacterial diversity in the solid fraction of the ruminal content of steers.

Our hypothesis was that different whole oilseeds included in the diet for steers confined could alter the diversity of rumen bacteria compared to a diet without oilseeds or an exclusively forage diet. It was aimed to evaluate the effects of oilseeds inclusion in the diet on bacterial diversity in the solid fraction of the ruminal content of steers, by gene sequences of the conserved 16S rDNA region. Six crossbred steers castrated males, fitted with ruminal cannula were used in a 6 × 6 Latin square design, using 21-day period. At the start of the experiment, the live weight of the animals averaged 416 ± 9.7 kg (mean ± SD). A total of 2,180,562 16S rDNA sequences were generated for the Bacteria domain by MiSeq sequencing. The bacterial diversity was composed of 24 bacterial phyla, with the most abundant being Firmicutes, Bacteroidetes, and Proteobacteria. Other phyla with less diversity were also identified including Eurychaeota, Tenericutes, SR1 Absconditalbacteria, Synergistetes, Actinobacteria, Saccharibacteria, Elusimicrobia, Cyanobacteria, Verrucomicrobia, Fusobacteria, Lentisphaerae. The similarity in the bacterial community averaged 50% for all the experimental diets. Steers-fed corn silage exhibited a great diversity of bacteria of the Firmicutes phylum. The steers-fed oilseeds in the diet had a great diversity of bacteria from the phylum Bacteroidetes and Proteobacteria. The inclusion of whole oilseeds in the steer diets can alter the rumen bacteria population by up to 50% of total diversity.

Bacterial species in the ruminal content of steers fed oilseeds in the diet.

The aim of this study was to evaluate bacterial species and diversity of methanogenic Archaea in the solid fraction of the ruminal content, through the gene sequences of the conserved 16S rDNA region, in response to the following diets: canola, cottonseed, sunflower, soybean, corn silage, and control diet. Six rumen-fistulated crossbred steers, with body weight (BW) of 416.33 ± 93.30 kg, were distributed in a 6 × 6 Latin square design. Regardless of the diet provided, amylolytic, proteolytic, and lactic bacteria were identified in the rumen fluid. Cellulolytic bacteria were predominant for all diets, reaching 47.75% of operational taxonomic units (OTUs) in animals fed with the cottonseed diet. Amylolytic bacteria reach 62.51% of OTU in animal fed sunflower diet, while proteolytic bacteria correspond to 65.96% of OTU in this same diet. Also, Megasphaera elsdenii bacterium was identified for all diets, with a greater percentage of OTU in steers fed the cottonseed diet. The diversity analysis of the species identified the methanogenic Archaea Methanobrevibacter ruminantium in all diets. We conclude that the control and corn silage diets have the most similar bacterial flora; diets with oilseeds had 47.5% similarity in rumen flora bacteria species. Animals fed with soybean showed a reduced number of methanogenic Archaea in the rumen content, which could be an alternative feed for cattle due to their low potential for energy losses with the production of methane.

Invited review: Current enteric methane mitigation options.

Ruminant livestock are an important source of anthropogenic methane (CH4). Decreasing the emissions of enteric CH4 from ruminant production is strategic to limit the global temperature increase to 1.5°C by 2050. Research in the area of enteric CH4 mitigation has grown exponentially in the last 2 decades, with various strategies for enteric CH4 abatement being investigated: production intensification, dietary manipulation (including supplementation and processing of concentrates and lipids, and management of forage and pastures), rumen manipulation (supplementation of ionophores, 3-nitrooxypropanol, macroalgae, alternative electron acceptors, and phytochemicals), and selection of low-CH4-producing animals. Other enteric CH4 mitigation strategies are at earlier stages of research but rapidly developing. Herein, we discuss and analyze the current status of available enteric CH4 mitigation strategies with an emphasis on opportunities and barriers to their implementation in confined and partial grazing production systems, and in extensive and fully grazing production systems. For each enteric CH4 mitigation strategy, we discuss its effectiveness to decrease total CH4 emissions and emissions on a per animal product basis, safety issues, impacts on the emissions of other greenhouse gases, as well as other economic, regulatory, and societal aspects that are key to implementation. Most research has been conducted with confined animals, and considerably more research is needed to develop, adapt, and evaluate antimethanogenic strategies for grazing systems. In general, few options are currently available for extensive production systems without feed supplementation. Continuous research and development are needed to develop enteric CH4 mitigation strategies that are locally applicable. Information is needed to calculate carbon footprints of interventions on a regional basis to evaluate the impact of mitigation strategies on net greenhouse gas emissions. Economically affordable enteric CH4 mitigation solutions are urgently needed. Successful implementation of safe and effective antimethanogenic strategies will also require delivery mechanisms and adequate technical support for producers, as well as consumer involvement and acceptance. The most appropriate metrics should be used in quantifying the overall climate outcomes associated with mitigation of enteric CH4 emissions. A holistic approach is required, and buy-in is needed at all levels of the supply chain.

Influence of Different Heat-Stress-Reducing Systems on Physiological and Behavioral Responses and Social Dominance of Holstein and Jersey Cows and Heifers on Pasture.

High ambient temperatures and relative humidity affect the behavior and physiology of the animal. This study investigated the influence of different heat-stress-reducing systems on the physiological, behavioral, and preferential responses of Holstein and Jersey cows and heifers on pasture. Experimental treatments were: (1) three heat-stress-reducing systems (sprinklers + artificial shade; showers + artificial shade; and artificial shade); (2) two breeds (Holstein and Jersey); and (3) two physiological stages (lactating cows and pubertal heifers). Physiological and behavioral responses to treatments were measured every 30 min on collection days. The frequency and duration of the use of the systems were recorded continuously 24 h/day for 3 days in each period. The air temperature and the relative humidity were 26 ± 4.1 °C and 74 ± 11.3%, respectively. The experimental treatments affected (p = 0.0354) standing idle, grazing behavior (p = 0.0435), and the frequency and duration of use of the systems by the animals (p < 0.0001). For all treatments, the respiratory rate and the coat surface temperature were highly and significantly correlated (p < 0.05) with the temperature and humidity index. In conclusion, under ambient conditions, dairy cows preferred using sprinklers or showers over artificial shade. These systems were more efficient at reducing the heat load and led to better behavioral and physiological responses.

Canola Meal versus Soybean Meal as Protein Supplements in the Diets of Lactating Dairy Cows Affects the Greenhouse Gas Intensity of Milk.

Soybean meal (SBM) and canola meal (CM) are protein supplements used in lactating dairy cow diets and, recently, an enteric methane-mitigating effect (i.e., lower Ym value) was reported for CM. Before recommending CM as a greenhouse gas (GHG) mitigation strategy, it is necessary to examine the net impact on total GHG emissions from milk production. The objective was to determine whether using CM rather than SBM in lactating dairy cow diets decreases GHG per kilogram of fat and protein corrected milk (FPCM), and whether the decrease depends upon where the meals are produced. Cradle to farm-gate life cycle assessments were conducted for a simulated dairy farm in eastern (Quebec) and western (Alberta) Canada. Scenarios examined the source of protein meal, location where meals were produced, and the methane-mitigating effect of CM. The Holos model was used to estimate GHG emissions from animals, manure, crop production, imported feeds, and energy use. GHG intensities (CO2e/kg FPCM) were 0.85-1.02 in the east and 1.07-1.11 in the west for the various scenarios, with enteric methane comprising 34 to 40% of total emissions. CM produced in western Canada with a low up-stream emission factor and low Ym value reduced CO2e/kg FPCM by 3% (western farm) to 6.6% (eastern farm) compared with SBM. We conclude that using CM rather than SBM in the diet of lactating dairy cows can be a GHG mitigation strategy depending upon where it is produced and whether it decreases enteric methane emissions.

Orange juice industry by-product silage can increase fat and protein in Holstein cow's milk.

We aimed to evaluate the effect of replacing corn silage by orange peel silage on nutrient intake, ruminal parameters and milk production of multiparous lactating Holstein cows. Eight fistulated Holstein cows averaging 587.5 ± 39.6 kg and 111 ± 22 d in milking were randomly assigned to a double 4 × 4 Latin square design carried out two times to determine the effects of feeding with orange peel silage (OPS) in substitution of whole plant corn silage (WPCS). The treatments were a control diet with WPCS only or diets with OPS replacing WPCS in the total mixed diet (250, 500, or 750 g/kg DM). All cows were fed the same 750 : 250 g/kg roughage : concentrate ratio. The DM intake and milk production were reduced with the OPS inclusion, with decreases in consumption of neutral detergent fibre and increased consumption of non-fibrous carbohydrates. Diets with 250 and 500 g/kg OPS showed similar milk production and protein content in milk to the standard WCPS diet, whilst 750 g/kg orange peel silage as roughage increased fat and protein contents significantly. The orange peel silage as a substitute for corn silage for feeding dairy cows did not show adverse changes in the rumen environment and showed promising results in the increase of fat in milk of Holstein cows.

Transcriptome adaptation of the bovine mammary gland to diets rich in unsaturated fatty acids shows greater impact of linseed oil over safflower oil on gene expression and metabolic pathways.

BACKGROUND: Nutritional strategies can decrease saturated fatty acids (SFAs) and increase health beneficial fatty acids (FAs) in bovine milk. The pathways/genes involved in these processes are not properly defined. Next-generation RNA-sequencing was used to investigate the bovine mammary gland transcriptome following supplemental feeding with 5% linseed oil (LSO) or 5% safflower oil (SFO). Holstein cows in mid-lactation were fed a control diet for 28 days (control period) followed by supplementation with 5% LSO (12 cows) or 5% SFO (12 cows) for 28 days (treatment period). Milk and mammary gland biopsies were sampled on days-14 (control period), +7 and +28 (treatment period). Milk was used to measure fat(FP)/protein(PP) percentages and individual FAs while RNA was subjected to sequencing. RESULTS: Milk FP was decreased by 30.38% (LSO) or 32.42% (SFO) while PP was unaffected (LSO) or increased (SFO). Several beneficial FAs were increased by LSO (C18:1n11t, CLA:10t12c, CLA:9c11t, C20:3n3, C20:5n3, C22:5n3) and SFO (C18:1n11t, CLA:10t12c, C20:1c11, C20:2, C20:3n3) while several SFAs (C4:0, C6:0, C8:0, C14:0, C16:0, C17:0, C24:0) were decreased by both treatments (P < 0.05). 1006 (460 up- and 546 down-regulated) and 199 (127 up- and 72 down-regulated) genes were significantly differentially regulated (DE) by LSO and SFO, respectively. Top regulated genes (≥ 2 fold change) by both treatments (FBP2, UCP2, TIEG2, ANGPTL4, ALDH1L2) are potential candidate genes for milk fat traits. Involvement of SCP2, PDK4, NQO1, F2RL1, DBI, CPT1A, CNTFR, CALB1, ACADVL, SPTLC3, PIK3CG, PIGZ, ADORA2B, TRIB3, HPGD, IGFBP2 and TXN in FA/lipid metabolism in dairy cows is being reported for the first time. Functional analysis indicated similar and different top enriched functions for DE genes. DE genes were predicted to significantly decrease synthesis of FA/lipid by both treatments and FA metabolism by LSO. Top canonical pathways associated with DE genes of both treatments might be involved in lipid/cholesterol metabolism. CONCLUSION: This study shows that rich α-linolenic acid LSO has a greater impact on mammary gland transcriptome by affecting more genes, pathways and processes as compared to SFO, rich in linoleic acid. Our study suggest that decrease in milk SFAs was due to down-regulation of genes in the FA/lipid synthesis and lipid metabolism pathways while increase in PUFAs was due to increased availability of ruminal biohydrogenation metabolites that were up taken and incorporated into milk or used as substrate for the synthesis of PUFAs.

Deep sequencing shows microRNA involvement in bovine mammary gland adaptation to diets supplemented with linseed oil or safflower oil.

BACKGROUND: Bovine milk fat composition is responsive to dietary manipulation providing an avenue to modify the content of fatty acids and especially some specific unsaturated fatty acid (USFA) isomers of benefit to human health. MicroRNAs (miRNAs) regulate gene expression but their specific roles in bovine mammary gland lipogenesis are unclear. The objective of this study was to determine the expression pattern of miRNAs following mammary gland adaptation to dietary supplementation with 5 % linseed or safflower oil using next generation RNA-sequencing. METHODS: Twenty-four Canadian Holstein dairy cows (twelve per treatment) in mid lactation were fed a control diet (total mixed ration of corn:grass silages) for 28 days followed by a treatment period (control diet supplemented with 5 % linseed or safflower oil) of 28 days. Milk samples were collected weekly for fat and individual fatty acid determination. RNA from mammary gland biopsies harvested on day-14 (control period) and on days +7 and +28 (treatment period) from six randomly selected cows per treatment was subjected to small RNA sequencing. RESULTS: Milk fat percentage decreased significantly (P < 0.001) during treatment with the two diets as compared to the control period. The individual saturated fatty acids C4:0, C6:0, C8:0, C14:0 and C16:0 decreased significantly (P < 0.05) while five USFAs (C14:1, C18:1n11t, C20:3n3, C20:5n3 and CLA:t10c12) increased remarkably (P < 0.05) in response to both treatments. Analysis of 361 million sequence reads generated 321 known bovine miRNAs and 176 novel miRNAs. The expression of fourteen and twenty-two miRNAs was affected (P < 0.05) by linseed and safflower oil treatments, respectively. Seven miRNAs including six up-regulated (bta-miR-199c, miR-199a-3p, miR-98, miR-378, miR-148b and miR-21-5p) and one down-regulated (bta-miR-200a) were found to be regulated (P < 0.05) by both treatments, and thus considered core differentially expressed (DE) miRNAs. The gene targets of core DE miRNAs have functions related to gene expression and general cellular metabolism (P < 0.05) and are enriched in four pathways of lipid metabolism (3-phosphoinositide biosynthesis, 3-phosphoinositide degradation, D-myo-inisitol-5-phosphate metabolism and the superpathway of inositol phosphate compounds). CONCLUSION: Our results suggest that DE miRNAs in this study might be important regulators of bovine mammary lipogenesis and metabolism. The novel miRNAs identified in this study will further enrich the bovine miRNome repertoire and contribute to understanding mammary gland biology.

mRNA Expression of lipogenic enzymes in mammary tissue and fatty acid profile in milk of dairy cows fed flax hulls and infused with flax oil in the abomasum.

In the present study, the effect of flax hulls with or without flax oil bypassing the rumen on the expression of lipogenic genes in the mammary tissue of dairy cows was investigated. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four periods of 21 d each and four treatments: control diet with no flax hulls (CONT); diet with 9·88 % flax hulls in the DM (HULL); control diet with 500 g flax oil/d infused in the abomasum (COFO); diet with 9·88 % flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). A higher mRNA abundance of sterol regulatory element binding transcription factor, fatty acid (FA) synthase, lipoprotein lipase (LPL), PPARγ1, stearoyl-CoA desaturase (SCD) and acetyl-coenzyme A carboxylase-α was observed in cows fed HULL than in those fed CONT, and HUFO had the opposite effect. Compared with CONT, COFO and HUFO lowered the mRNA abundance of SCD, which may explain the lower proportions of MUFA in milk fat with flax oil infusion. The mRNA abundance of LPL in mammary tissue and proportions of long-chain FA in milk fat were higher in cows fed COFO than in those fed CONT. The highest proportions of trans FA were observed when cows were fed HULL. The present study demonstrates that flax hulls with or without flax oil infusion in the abomasum can affect the expression of lipogenic genes in the mammary tissue of dairy cows, which may contribute to the improvement of milk FA profile.

Effects of Adding Corn Dried Distiller Grains with Solubles (DDGS) to the Dairy Cow Diet and Effects of Bedding in Dairy Cow Slurry on Fugitive Methane Emissions.

The specific objectives of this experiment were to investigate the effects of adding 10% or 30% corn dried distillers grains with solubles (DDGS) to the dairy cow diet and the effects of bedding type (wood shavings, straw or peat moss) in dairy slurry on fugitive CH4 emissions. The addition of DDGS10 to the dairy cow diet significantly increased (29%) the daily amount of fat excreted in slurry compared to the control diet. The inclusion of DDGS30 in the diet increased the daily amounts of excreted DM, volatile solids (VS), fat, neutral detergent fiber (NDF), acid detergent fiber (ADF) and hemicellulose by 18%, 18%, 70%, 30%, 15% and 53%, respectively, compared to the control diet. During the storage experiment, daily fugitive CH4 emissions showed a significant increase of 15% (p < 0.05) for the slurry resulting from the corn DDGS30 diet. The addition of wood shavings and straw did not have a significant effect on daily fugitive CH4 emissions relative to the control diet, whereas the addition of peat moss caused a significant increase of 27% (p < 0.05) in fugitive CH4 emissions.

Effect of Corn Dried Distiller Grains with Solubles (DDGS) in Dairy Cow Diets on Manure Bioenergy Production Potential.

The main objective of this study was to obtain scientifically sound data on the bioenergy potential of dairy manures from cows fed different levels of corn dried distillers grains with solubles (DDGS). Three diets differing in corn DDGS content were formulated: 0% corn DDGS (DDGS0; control diet), 10% corn DDGS (DDGS10) and 30% corn DDGS (DDGS30). Bioenergy production was determined in psychrophilic (25 ± 1 °C) sequencing batch reactors (SBRs) fed 3 g COD L(-1)·day(-1) during a two-week feeding period followed by a two-week react period. Compared to the control diet, adding DDGS10 and DDGS30 to the dairy cow diet increased the daily amount of fat excreted in slurry by 29% and 70%, respectively. The addition of DDGS30 increased the cows' daily production of fresh feces and slurry by 15% and 11%, respectively. Furthermore, the incorporation of DDGS30 in the diet increased the daily amounts of dry matter (DM), volatile solids (VS), neutral detergent fiber (NDF), acid detergent fiber (ADF) and hemicellulose by 18%, 18%, 30%, 15% and 53%, respectively, compared to the control diet. While the addition of DDGS did not significantly affect the specific CH4 production per kg VS compared to the control diet, DDGS30 increased the per cow daily CH4 production by 14% compared to the control diet.

Mammary gene expression and activity of antioxidant enzymes and oxidative indicators in the blood, milk, mammary tissue and ruminal fluid of dairy cows fed flax meal.

The effects of flax meal (FM) on the activity of antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)) in the blood, mammary tissue and ruminal fluid, and oxidative stress indicators (thiobarbituric acid-reactive substances(TBARS) and 1,1-diphenyl-2-picrylhydrazyl-scavenging activity) in the milk, plasma and ruminal fluid of dairy cows were determined.The mRNA abundance of the antioxidant enzymes and oxidative stress-related genes was assessed in mammary tissue. A total of eight Holstein cows were used in a double 4 x 4 Latin square design. There were four treatments in the diet: control with no FM(CON) or 5% FM (5FM), 10% FM (10FM) and 15% FM (15FM). There was an interaction between treatment and time for plasma GPx and CAT activities. Cows supplemented with FM had a linear reduction in TBARS at 2 h after feeding, and there was no treatment effect at 0, 4 and 6 h after feeding. TBARS production decreased in the milk of cows fed the 5FM and 10FM diets. There was a linear increase in nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) mRNA abundance in mammary tissue with FM supplementation.A linear trend for increased mRNA abundance of the CAT gene was observed with higher concentrations of FM. The mRNA abundance of CAT, GPx1, GPx3, SOD1, SOD2, SOD3 and nuclear factor of k light polypeptide gene enhancer in B-cells (NFKB) genes was not affected by the treatment. These findings suggest that FM supplementation can improve the oxidative status of Holstein cows as suggested by decreased TBARS production in ruminal fluid 2 h post-feeding and increased NFE2L2/nuclear factor-E2-related factor 2 (Nrf2) mRNA abundance in mammary tissue.

Diet-induced alterations in total and metabolically active microbes within the rumen of dairy cows.

DNA-based techniques are widely used to study microbial populations; however, this approach is not specific to active microbes, because DNA may originate from inactive and/or dead cells. Using cDNA and DNA, respectively, we aimed to discriminate the active microbes from the total microbial community within the rumen of dairy cows fed diets with increasing proportions of corn silage (CS). Nine multiparous lactating Holstein cows fitted with ruminal cannulas were used in a replicated 3×3 Latin square (32-d period; 21-d adaptation) design to investigate diet-induced shifts in microbial populations by targeting the rDNA gene. Cows were fed a total mixed ration with the forage portion being either barley silage (0% CS), a 50∶50 mixture of barley silage and corn silage (50% CS), or corn silage (100% CS). No differences were found for total microbes analyzed by quantitative PCR, but changes were observed within the active ones. Feeding more CS to dairy cows was accompanied by an increase in Prevotella rRNA transcripts (P = 0.10) and a decrease in the protozoal rRNA transcripts (P<0.05). Although they were distributed differently among diets, 78% of the amplicons detected in DNA- and cDNA-based fingerprints were common to total and active bacterial communities. These may represent a bacterial core of abundant and active cells that drive the fermentation processes. In contrast, 10% of amplicons were specific to total bacteria and may represent inactive or dead cells, whereas 12% were only found within the active bacterial community and may constitute slow-growing bacteria with high metabolic activity. It appears that cDNA-based analysis is more discriminative to identify diet-induced shifts within the microbial community. This approach allows the detection of diet-induced changes in the microbial populations as well as particular bacterial amplicons that remained undetected using DNA-based methods.

Effects of abomasal infusion of flaxseed (Linum usitatissimum) oil on microbial ß-glucuronidase activity and concentration of the mammalian lignan enterolactone in ruminal fluid, plasma, urine and milk of dairy cows.

Ruminal microbiota plays an important role in the conversion of plant lignans into mammalian lignans. The main mammalian lignan present in the milk of dairy cows fed flax products is enterolactone (EL). The objectives of the present study were to investigate the effects of abomasal infusion of flax oil on the metabolism of flax lignans and concentrations of EL in biological fluids of dairy cows. A total of six rumen-cannulated dairy cows were assigned within a 2 × 3 factorial arrangement of six treatments utilising flax hulls (0 and 15·9 % of DM) and abomasal infusion of flax oil (0, 250 and 500 g/d). There were six periods of 21 d each. Samples were collected during the last 7 d of each period and subjected to chemical analysis. Flax hull supplementation increased concentrations of EL in ruminal fluid, plasma, urine and milk, while flax oil infusion had no effect. Post-feeding, ß-glucuronidase activity in the ruminal fluid of cows infused with 250 g flax oil was significantly lower for cows fed hulls than for those fed the control diet. The present study demonstrated that the presence of a rich source of n-3 fatty acids such as flax oil in the small intestine does not interfere with the absorption of the mammalian lignan EL and that lower ruminal ß-glucuronidase activity had no effect on the conversion of flax lignans into EL in the rumen of dairy cows.

Assessment of the effect of condensed (acacia and quebracho) and hydrolysable (chestnut and valonea) tannins on rumen fermentation and methane production in vitro.

BACKGROUND: Tannins added to animal diets may have a positive effect on energy and protein utilisation in the rumen. The objective of this study was to examine the impact of different sources and concentrations (20, 50, 100, 150 and 200 g kg⁻¹ dry matter (DM)) of condensed (acacia and quebracho) and hydrolysable (chestnut and valonea) tannins on rumen microbial fermentation in vitro. The experiment also included a negative control with no tannins (control) and a positive control with monensin (10 mg L⁻¹). RESULTS: In vitro gas production and total volatile fatty acid (VFA) concentration decreased as tannin concentration increased. Addition of acacia, chestnut or valonea tannins at ≥ 50 g kg⁻¹ or quebracho tannins at ≥ 100 g kg⁻¹ resulted in a decrease (up to 40%) in methane (CH4) production compared with the control. Valonea tannins were the only tannin source that reduced (-11%) CH4 production at 50 g kg⁻¹ without affecting VFA concentration. Tannin treatments reduced ammonia (NH3) and branched-chain VFA concentrations, indicating a reduction in ruminal protein degradation. Monensin reduced CH4 production (-37%) and NH3 concentration (-20%) without affecting total VFA concentration. CONCLUSION: Supplying acacia, chestnut or valonea tannins at 50 g kg⁻¹ has the potential to reduce CH4 production and ruminal protein degradation with minimum detrimental effects on efficiency of ruminal fermentation.

Effects of cinnamon leaf, oregano and sweet orange essential oils on fermentation and aerobic stability of barley silage.

BACKGROUND: Silage additives are marketed with the primary aim of improving the fermentation and/or aerobic stability of silage. The objective of this study was to evaluate the impact of three different essential oils (EOs; cinnamon leaf (CIN), oregano (ORE) and sweet orange (SO)) on the fermentation characteristics and stability of barley silage. Chopped whole-plant barley (Hordem vulgare L.) forage was ensiled either untreated (0 mg kg⁻¹ dry matter (DM)) or treated with CIN, ORE or SO (37.5, 75 and 120 mg kg⁻¹ DM). RESULTS: Moulds were not detected in any treatments, including the control, after 7 days of air exposure. All EOs at a concentration of 120 mg kg⁻¹ silage DM decreased (P = 0.001) yeast populations in comparison with the control during air exposure. Net gas, methane and ammonia concentrations in vitro did not differ among treatments. Changes in volatile fatty acid concentrations were small, and in situ data showed no changes in DM and neutral detergent fibre digestion rates for CIN, ORE or SO at concentrations up to 120 mg kg⁻¹ DM. CONCLUSION: The findings from this study show that a concentration of 120 mg EO kg⁻¹ DM decreased yeast counts during aerobic stability tests. However, all EO treatments had minimal effects on data from in vitro and in situ incubations.

Mammary gene expression and activity of antioxidant enzymes and concentration of the mammalian lignan enterolactone in milk and plasma of dairy cows fed flax lignans and infused with flax oil in the abomasum.

The objectives of the study were to investigate the effects of dietary supplementation of flax hulls and/or flax oil on the activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX)) in plasma and the mammary gland and the relative mRNA abundance of antioxidant genes in the mammary gland of dairy cows. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four treatments: control with no flax hulls (CONT), 9·88% flax hulls in the DM (HULL), control with 500 g flax oil/d infused in the abomasum (COFO), 9·88% flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). Plasma GPX activity tended to decrease with flax oil supplementation. Cows fed HULL had higher levels of CAT, GPX1 and SOD1 mRNA in the mammary gland and lower mRNA abundance of GPX3, SOD2 and SOD3 compared with those fed CONT. Abundance of CAT, GPX1, GPX3, SOD2 and SOD3 mRNA was down-regulated in the mammary gland of cows fed HUFO compared to those fed CONT. The mRNA abundance of CAT, GPX1, GPX3 and SOD3 was lower in the mammary gland of cows fed COFO than in the mammary gland of cows fed CONT. The present study demonstrates that flax hulls contribute to increasing the abundance of some antioxidant genes, which can contribute to protecting against oxidative stress damage occurring in the mammary gland and other tissues of dairy cows.