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.

Two-Stage Rumen Cannulation Technique in Dairy Cows.

OBJECTIVE: To describe a 2-stage rumen cannulation technique for dairy cows. STUDY DESIGN: Case series. ANIMALS: 172 dairy cows from 2 research institutions. METHODS: The 2-stage rumen cannulation technique first exteriorized a rumen segment within a wooden clamp, fixing the clamp to the skin with 6 mattress sutures. After 1 week, the necrotic rumen segment was removed, leaving a rumen fistula in which a 7.5 cm cannula was inserted. This was replaced by a 10 cm cannula a further 1 week later. The surgery took an average of 30 minutes. At least 1 assistant is required for the technique. RESULTS: The overall complication frequency was 7/172 (4%). One cow and 1 heifer aborted less than 10 days after surgery. Two late-pregnant heifers died from peritonitis after insertion of the 7.5 cm cannula because of incomplete adhesion of the rumen to the abdominal wall. The exteriorized rumen segment slipped back in the abdomen in 3 cows but was successfully re-clamped prior to insertion of the 7.5 cm cannula. CONCLUSION: A high success rate was achieved with this 2-stage cannulation technique. Postoperative complications were attributed to delayed adhesion of the rumen, perhaps because of stress-related factors (e.g., transport, mixing with other animals, transition period).

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 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.

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.

Intake and digestibility of fatty acids in late-lactating dairy cows fed flaxseed hulls supplemented with monensin.

Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids but there is little information on digestibility of its nutrients by dairy cows. Four rumen-cannulated multiparous Holstein cows averaging 665 ± 21 kg of body weight and 190 ± 5 d in milk at the beginning of the experiment were assigned to a 4 × 4 Latin square design with four 28-d experimental periods to determine the effects of feeding monensin and flaxseed hulls on total tract apparent digestibility of nutrients and fatty acids. The four treatments were: (1) diet CO: control with neither flaxseed hulls nor monensin added; (2) diet FH containing 19·8 g flaxseed hulls/100 g dry matter (DM); (3) diet MO with 16 mg monensin/kg DM; (4) diet HM containing 19·8 g flaxseed hulls/100 g DM and 16 mg monensin/kg DM. Diets provided similar amounts of protein and net energy of lactation. Digestibility of crude protein was higher for diets containing flaxseed hulls and for diets supplemented with monensin. Flaxseed hulls supplementation decreased digestibility of acid and neutral detergent fibre. Significantly higher digestibility of ether extract and individual fatty acids was observed for treatments with flaxseed hulls compared with treatments without flaxseed hulls. A combination of flaxseed hulls and monensin did not result in better fatty acid digestibility than when feeding only flaxseed hulls.

Digestion, milk production and milk fatty acid profile of dairy cows fed flax hulls and infused with flax oil in the abomasum.

Flax hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on digestion of flax hull based diets and nutritive value of flax hull for dairy production. Flax oil is rich in α-linolenic acid (LNA) and rumen bypass of flax oil contributes to increase n-3 FA proportions in milk. Therefore, the main objective of the experiment was to determine the effects of abomasal infusion of increasing amounts of flax oil on apparent digestibility, dry matter (DM) intake, milk production, milk composition, and milk FA profile with emphasis on the proportion of LNA when cows were supplemented or not with another source of LNA such as flax hull. Six multiparous Holstein cows averaging 650±36 kg body weight and 95±20 d in milk were assigned to a 6×6 Latin square design (21-d experimental periods) with a 2×3 factorial arrangement of treatments. Treatments were: 1) control, neither flax hull nor flax oil (CON), 2) diet containing (DM basis) 15·9% flaxseed hull (FHU); 3) CON with abomasal infusion of 250 g/d flax oil; 4) CON with abomasal infusion of 500 g/d flax oil; 5) FHU with abomasal infusion of 250 g/d flax oil; 6) FHU with abomasal infusion of 500 g/d flax oil. Infusion of flax oil in the abomasum resulted in a more pronounce decrease in DM intake for cows fed the CON diets than for those fed the FHU diets. Abomasal infusion of flax oil had little effect on digestibility and FHU supplementation increased digestibility of DM and crude protein. Milk yield was not changed by abomasal infusion of flax oil where it was decreased with FHU supplementation. Cows fed FHU had higher proportions of 18:0, cis9-18:1, trans dienes, trans monoenes and total trans in milk fat than those fed CON. Proportion of LNA was similar in milk fat of cows infused with 250 and 500 g/d flax oil in the abomasum. Independently of the basal diet, abomasal infusion of flax oil resulted in the lowest n-6:n-3 FA ratio in milk fat, suggesting that the most important factor for modification of milk FA profile was the amount of n-3 FA bypassing the rumen and not the amount of flax hull fed to dairy cows. Moreover, these data suggest that there is no advantage to supply more than 250 g/d of flax oil in the abomasum to increase the proportion of LNA in milk fat.

Effect of flaxseed lignans added to milk or fed to cows on oxidative degradation of dairy beverages enriched with polyunsaturated fatty acids.

Nutritional value is a priority in new product development. Using vegetable or marine oils, rich in polyunsaturated fatty acids, in dairy beverage formulations is an option to provide the consumers with healthier products. However, these formulations are prone to oxidation, which is responsible for rapid flavour degradation and the development of potentially toxic reaction products during storage. Flaxseed lignans, secoisolariciresinol diglucoside (SDG), and its mammalian metabolites have antioxidant activity and could be used in beverage formulations to prevent oxidation. Commercially available SDG extract was added to the formulation of dairy beverages enriched with flaxseed oil. As an alternative approach, dairy beverages were produced from milk naturally rich in SDG metabolites obtained through the alteration of cow diet. Resistance to oxidation was determined from the kinetics of hexanal and propanal production during heat and light exposure treatments. Increasing SDG concentration in dairy beverage slightly reduced redox potential but had no effect on oxygen consumption during oxidation treatments. The presence of SDG in dairy beverage significantly improved resistance to heat- and light-induced oxidation. However, purified enterolactone, a mammalian metabolite from SDG, prevented oxidation at much lower concentrations. The use of milk from dairy cow fed flaxseed meal did not improve resistance to oxidation in dairy beverage. Enterolactone concentration in milk was increased by the experimental diet but it remained too low to observe any significant effect on dairy beverage oxidation.

Ruminal fermentation characteristics and fatty acid profile of ruminal fluid and milk of dairy cows fed flaxseed hulls supplemented with monensin.

Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on its value for dairy production. Monensin supplementation is known to modify biohydrogenation of FA by rumen microbes. Therefore, the main objective of the experiment was to determine the effect of feeding a combination of monensin and flaxseed hulls on ruminal fermentation characteristics and FA profile of ruminal fluid and milk. Four ruminally fistulated multiparous Holstein cows averaging 665 ± 21 kg body weight and 190 ± 5 d in milk were assigned to a 4×4 Latin square design (28-d experimental periods) with a 2×2 factorial arrangement of treatments. Treatments were: 1) control, neither flaxseed hulls nor monensin; 2) diet containing (dry matter basis) 19·8% flaxseed hulls; 3) diet with monensin (16 mg/kg dry matter); 4) diet containing 19·8% (dry matter basis) flaxseed hulls and 16 mg monensin/kg. Flaxseed hull supplementation decreased the acetate to propionate ratio in ruminal fluid and monensin had no effect. Concentrations of trans-18:1 isomers (trans9,trans11,trans13/14+6/8) and cis9,12,15-18:3 in ruminal fluid and milk fat were higher and those of cis9,12-18:2 in milk fat tended (P=0·07) to be higher for cows supplemented with flaxseed hulls than for cows fed no flaxseed hulls. Monensin had little effect on milk fatty acid profile. A combination of flaxseed hulls and monensin did not result in better milk fatty acid profile than when feeding only flaxseed hulls.

The interaction of monensin and flaxseed hulls on ruminal and milk concentration of the mammalian lignan enterolactone in late-lactating dairy cows.

Four ruminally fistulated multiparous Holstein cows were assigned to a 4x4 Latin square design with a 2x2 factorial arrangement of treatments to study the effects of dietary supplementation of monensin and flaxseed hulls on ruminal and milk concentration of the mammalian lignan enterolactone (EL) and ruminal and faecal activity of beta-glucuronidase. The hypothesis was that monensin supplementation has no effect on the incorporation of EL into milk when cows are fed flaxseed hulls. Treatments were: 1) control, neither flaxseed hulls nor monensin (CO); 2) diet containing (dry matter basis) 20% flaxseed hulls (FH); 3) diet with monensin (16 mg/kg of dry matter; MO); 4) diet containing 20% (dry matter basis) flaxseed hulls and 16 mg/kg monensin (HM). Intake of dry matter was higher for CO and MO than for FH and HM and monensin had no effect. Milk production decreased in cows fed flaxseed hulls while monensin had no effect. Production of 4% fat-corrected milk and concentrations of milk fat, lactose, urea N, and total solids were similar among treatments. Although there was a decrease in ruminal activity of beta-glucuronidase when feeding flaxseed hulls, the metabolism of plant into mammalian lignans may be increased as shown by enhanced concentration of EL in the rumen and milk. Supplementation with flaxseed hulls then may contribute to favourably change milk composition for better human health by enhancing mammalian lignan EL concentration.

Weekly excretion of the mammalian lignan enterolactone in milk of dairy cows fed flaxseed meal.

Flaxseed meal (FM) is rich in the plant lignan secoisolariciresinol diglucoside (SDG) which is converted to the mammalian lignans enterodiol and enterolactone (EL) by ruminal microbiota. Feeding FM to dairy cows increases linearly EL concentration in milk but enterodiol is not detected. The objectives of the study were to determine the length of time to obtain peak EL concentration in the milk of dairy cows fed 20% FM and the length of time to return to EL baseline level in milk when cows are switched from high to low intake of flax SDG. A total of 12 multiparous lactating Holstein cows were assigned randomly to one of two feeding regimens: the control (CO) diet was fed for 6 weeks or the FM diet was fed from week 0 to 3 inclusive and then cows were switched to the control diet from week 3 to 6 inclusive. Milk samples were taken weekly for EL analysis. There was a significant interaction between feeding regimen and week for milk concentration of EL as a result of higher concentration of EL from week 1 to 3 for cows on the FM regimen compared with those on the CO regimen. Concentrations of milk EL on the FM regimen maintained uniform high levels from week 1 to 3 and they decreased significantly from week 3 to 4 when the CO diet was reintroduced in week 3. This study suggests that the conversion of SDG to the mammalian lignan EL and the transfer of EL to the mammary gland are well established after one week of feeding 20% FM in the diet of dairy cows and that milk concentration of EL returns to baseline level after one week of FM deprivation.

Ruminal metabolism of flaxseed ( Linum usitatissimum) lignans to the mammalian lignan enterolactone and its concentration in ruminal fluid, plasma, urine and milk of dairy cows.

Secoisolariciresinol diglucoside is the main flax (Linum usitatissimum) lignan that is converted to the mammalian lignans enterodiol (ED) and enterolactone (EL) by gastrointestinal microbiota. The objectives of the present study were to investigate the role of ruminal microbiota and the effects of flax oil on in vivo metabolism of flax lignans and concentration of EL in biological fluids. Four rumen-cannulated dairy cows were used in a 4 x 4 Latin square design. There were four periods of 21 d each and four treatments utilising flax hulls (1800 g/d) and oil (400 g/d) supplements. The treatments were: (1) oil and hulls administered in the rumen and abomasal infusion of water; (2) oil and hulls administered in the abomasum; (3) oil infused in the abomasum and hulls placed in the rumen; (4) oil placed in the rumen and hulls administered in the abomasum. Samples were collected during the last week of each period and subjected to chemical analysis. The site of supplementation of oil and hulls had no effect on ruminal EL concentration. Supplementing flax oil in the rumen and the abomasum led to similar EL concentrations in urine, plasma and milk. Concentrations of EL were higher in the urine, plasma and milk of cows supplemented with hulls in the rumen than in those placed with hulls in the abomasum. The present study demonstrated that ruminal microbiota play an important role in the metabolism of flax lignans.